Morphological and molecular identification to secure cultivar maintenance and management of self-sterile Rubus arcticus.

BACKGROUND AND AIMS: Preservation of cultivar purity creates a particular challenge for plants that are self-incompatible, require insects for cross-pollination, and have easily germinating seeds and vigorously spreading rhizomes. As the fields must be planted with mixed populations, and a balance must be maintained between the cultivars to achieve effective pollination, methods for field monitoring of the relative density of different cultivars must be practical. Furthermore, a DNA-based method is needed for cultivar verification in the collections and outside of the growing season. The aim of this study was to develop both types of methods for Rubus arcticus (arctic bramble). METHODS: Morphological parameters were measured from six cultivars grown on three farms. Observations from the flowers and fruits included: petal and sepal number, flower diameter, arrangement of petals, size of calyx in relation to corolla, fruit weight, yield and soluble sugars. Observations from the leaves included: width and height of middle leaflet, shape of the base of terminal leaflet, shape of terminal leaflet, leaf margin serration and fingertip touch. The applicability of simple sequence repeat (SSR) or microsatellite DNA markers developed for red raspberry was tested on eight arctic bramble cultivars. KEY RESULTS AND CONCLUSIONS: Morphological and molecular identification methods were developed for R. arcticus. The best morphological characteristics were the length-to-width ratio of the middle leaflet and leaf margin serration. A particular characteristic, fingertip touch, was shown by electron microscopy to be related to the density and quality of the leaf hairs. Red raspberry SSR marker no. 126 proved to be applicable for differentiation of the eight arctic bramble cultivars tested. These identification methods are critical to secure the maintenance and management of R. arcticus. However, the challenges faced and approaches taken are equally applicable to other species with similar biology.

Spatial genetic structure of aquatic bryophytes in a connected lake system.

Using genetic markers, we investigated the genetic structure of three clonal aquatic moss species, Calliergon megalophyllum Mikut., Fontinalis antipyretica Hedw. and F. hypnoides Hartm. on two scales: among populations in a connected lake system (large-scale spatial genetic structure) and among individuals within populations (fine-scale spatial genetic structure). Mean genetic diversities per population were 0.138, 0.247 and 0.271, respectively, and total diversities equalled 0.223, 0.385 and 0.421, respectively. Relative differentiation levels (FST values of 0.173, 0.280 and 0.142, respectively) were significant but showed that there is a moderate amount of gene flow taking place within the lake system connected with narrow streams. Bayesian STRUCTURE analysis provided some indication that the direction of water flow influences population genetic structuring in the studied aquatic mosses. We propose that dispersal leading to gene flow in C. megalophyllum, F. antipyretica and F. hypnoides takes place both along water via connecting streams and by animal vectors, such as waterfowl. Nevertheless, the slight genetic structuring pattern along the direction of water flow suggests that dispersal of shoots or their fragments along water is a means of dispersal in these mosses. The absence of sexual reproduction and spores may have caused the observed spatial genetic structure within populations, including aggregations of similar genotypes (clones or closely related genotypes) at short distances in populations otherwise showing an isolation by distance effect. Regardless of the results pointing to the dominance of vegetative propagation, it is impossible to completely rule out the potential role of rare long-distance spore dispersal from areas where the species are fertile.

Permanent Genetic Resources added to Molecular Ecology Resources Database 1 December 2009-31 January 2010.

This article documents the addition of 220 microsatellite marker loci to the Molecular Ecology Resources Database. Loci were developed for the following species: Allanblackia floribunda, Amblyraja radiata, Bactrocera cucurbitae, Brachycaudus helichrysi, Calopogonium mucunoides, Dissodactylus primitivus, Elodea canadensis, Ephydatia fluviatilis, Galapaganus howdenae howdenae, Hoplostethus atlanticus, Ischnura elegans, Larimichthys polyactis, Opheodrys vernalis, Pelteobagrus fulvidraco, Phragmidium violaceum, Pistacia vera, and Thunnus thynnus. These loci were cross-tested on the following species: Allanblackia gabonensis, Allanblackia stanerana, Neoceratitis cyanescens, Dacus ciliatus, Dacus demmerezi, Bactrocera zonata, Ceratitis capitata, Ceratitis rosa, Ceratits catoirii, Dacus punctatifrons, Ephydatia mülleri, Spongilla lacustris, Geodia cydonium, Axinella sp., Ischnura graellsii, Ischnura ramburii, Ischnura pumilio, Pistacia integerrima and Pistacia terebinthus.

Important research questions in allergy and related diseases: 3-chronic rhinosinusitis and nasal polyposis - a GALEN study.

Chronic rhinosinusitis is one of the most common health care challenges, with significant direct medical costs and severe impact on lower airway disease and general health outcomes. The diagnosis of chronic rhinosinusitis (CRS) currently is based on clinical signs, nasal endoscopy and CT scanning, and therapeutic recommendations are focussing on 2 classes of drugs, corticosteroids and antibiotics. A better understanding of the pathogenesis and the factors amplifying mucosal inflammation therefore seems to be crucial for the development of new diagnostic and therapeutic tools. In an effort to extend knowledge in this area, the WP 2.7.2 of the GA(2)LEN network of excellence currently collects data and samples of 1000 CRS patients and 250 control subjects. The main objective of this project is to characterize patients with upper airway disease on the basis of clinical parameters, infectious agents, inflammatory mechanisms and remodeling processes. This collaborative research will result in better knowledge on patient phenotypes, pathomechanisms, and subtypes in chronic rhinosinusitis. This review summarizes the state of the art on chronic rhinosinusitis and nasal polyposis in different aspects of the disease. It defines potential gaps in the current research, and points to future research perspectives and targets.

Excessive variation in Y chromosomal DNA in Rumex acetosa (Polygonaceae).

Rumex acetosa is one of the few angiosperms that possesses sex chromosomes. The same types of abundant repetitive sequences cover both heterochromatic Y chromosomes present in males. The aim of this study was to investigate genetic variation in paternally inherited Y chromosomal DNA and in maternally inherited cpDNA, and to find out whether the examined genomic regions are suited to a phylogeographic study in R. acetosa. DNA sequence polymorphisms present in the 850-bp heterochromatic segment on the Y chromosomes were compared to variation in the 409-bp long chloroplast section (trnL- trnF spacer) in R. acetosa originating from several European locations and from the Altai mountains in Russia. A great amount of genetic variation was detected within the Y chromosomal region while only four chloroplast genotypes were detected. Although the chloroplast haplotypes possessed some geographic pattern, no clear phylogeographic pattern was detected based on the variable Y chromosomes. The mean Y chromosomal nucleotide diversity among all samples equaled 6.6 %, and the mean proportion of polymorphic sites per individual equaled 8.2 % among SNP sites and 1.7 % among all sites investigated. The high number of substitutions detected in the Y chromosomal DNA shows that this heterochromatic sequence has a high mutation rate. The diversity pattern indicates that gene flow via pollen is extensive and it blurs any geographical pattern in the Y chromosomal variation. The high number of repeats and uncertainty concerning the extent of recombination between the two Y chromosomes impair the usability of the Y chromosomal segment for phylogeographic or population genetic studies.

In vivo relationship between collagenase-2 and interleukin-8 but not tumour necrosis factor-alpha in chronic rhinosinusitis with nasal polyposis.

BACKGROUND: The characteristic feature of chronic rhinosinusitis with nasal polyposis (CRSwNP) is eosinophilic inflammation of the sinus mucosa; a type of inflammation also seen in asthmatic airways. Similar histopathologic findings of airway remodelling are present in both diseases. Remodelling is tightly controlled by matrix metalloproteinases (MMP). Increase of collagenase-2 (MMP-8) expression in the bronchial epithelial cells has been described in asthmatic patients, but it has not been studied in CRSwNP. METHODS: The concentrations and degree of activation of MMP-8 were analysed by immunofluorometric assay and Western blotting, respectively, in sinus mucus samples from CRSwNP patients and in nasal lavages from healthy controls in relation to inductive cytokines interleukin-8 (IL-8) and tumour necrosis factor-alpha (TNF-alpha). RESULTS: Significantly elevated levels of MMP-8 and IL-8 but not TNF-alpha were found in CRSwNP patients relative to controls. In particular, the activation of mesenchymal-type MMP-8 but not polymorphonuclear-type MMP-8 was associated with elevated IL-8 levels. CONCLUSIONS: The IL-8 and MMP-8 seemingly form an inductive cytokine-proteinase cascade in CRSwNP pathogenesis. Together they provide a target for novel therapies and a diagnostic tool for monitoring CRSwNP treatment.

Does the triad of fungi, bacteria and exposure to moisture have an impact on chronic hyperplastic sinusitis?

UNLABELLED: Here we evaluated a possible relationship between chronic hyperplastic sinusitis (CHS) and moisture exposure and secondly a seasonal variation of fungal and bacterial findings in the healthy nose. In 28 CHS patients sinus mucus was collected during endoscopic sinus surgery. Samples from the nasal cavities of 19 healthy volunteers were collected by nasal lavage (NAL) in January and in September. Bacterial culture and fungal staining and culture were carried out. Histological samples from the sinus mucosa were obtained. Patients' medical history and environmental factors were enquired. Mold odor or moisture problems in the home or work environment were reported by 46% of the CHS patients. Patients who reported moisture exposure did not differ significantly from those who had not been exposed with regards to microbiological findings, tissue eosinophilia, and earlier operations. Cladosporium (16%) and Alternaria (11%) were found in NAL fluid collected in the autumn from the control subjects. No fungi were isolated from samples taken during the winter. An association between CHS or fungal sinusitis and moisture damage was not apparent in the present study. The fungal findings in the nasal cavity reflect the environmental exposure. This should be taken into account when NAL is used for microbiological studies. PRACTICAL IMPLICATIONS: Living in a moldy house or working in a similar environment may increase the risk of respiratory symptoms and infections. However, our results suggest that chronic hyperplastic sinusitis and fungal sinusitis are not associated with moisture exposure. The nose is a good collector of particles in the air. Especially samples taken by the nasal lavage method reflect the environmental exposure. This should be taken into account when this method is used for microbiological studies.

Usefulness of mean aortic valve gradient and left ventricular diastolic filling pattern for distinguishing symptomatic from asymptomatic patients.

Consecutive, symptomatic (n = 15) and asymptomatic (n = 25) men with aortic stenosis (valve area < 1.2 cm2) and no clinical evidence of myocardial ischemia underwent radionuclide angiography at rest and during supine bicycle ergometry. Ejection fraction, diastolic filling pattern and aortic valve area/gradient were measured on enrollment and when patients became symptomatic (n = 10) or underwent valve replacement (n = 22) during a 2-year follow-up period. Both groups had similar heart rate, blood pressure and ejection fractions, but mean aortic gradients were higher in symptomatic (53 +/- 4 mm Hg) than asymptomatic (37 +/- 2 mm Hg) subjects p < 0.01. Functional limitation evoked by exercise was prevalent even in the asymptomatic group but symptomatic patients exercised to lower work levels than asymptomatic subjects (184 +/- 27 and 307 +/- 32 kg.m/min, respectively, p = 0.02). Ejection fraction failed to increase with exercise in either group. Symptomatic subjects had supranormalization of early diastolic filling with shorter time to the peak filling rate than asymptomatic subjects (137 +/- 16 and 172 +/- 9 ms, respectively, p < 0.05) and a greater first 1/3 filling fraction. The 10 patients who became symptomatic during follow-up had higher first 1/3 filling fractions (53 +/- 7 and 42 +/- 5%, respectively) and mean gradients (41 +/- 4 and 33 +/- 2 mm Hg, respectively) than subjects who remained asymptomatic, p < 0.05. High mean aortic gradients, impaired exercise tolerance and enhanced early diastolic filling distinguish symptomatic from asymptomatic patients.