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Objectives/hypothesis: Unsedated transnasal tracheoscopy (TNT) has emerged as a technique in otolaryngology-head and neck surgery for an awake airway examination in the office setting. This study investigates the safety, procedural success rate, indications, and findings of TNTs performed over a 3-year period at an academic medical center., Study Design: Retrospective chart review., Methods: After institutional review board approval, billing records were reviewed for patients who underwent TNT from 2007-2009 in the University of Wisconsin-Madison Department of Otolaryngology-Head and Neck Surgery. Hospital charts for these patients were obtained, and data regarding patient demographics, complications, procedural success rate, indications, and findings were recorded., Results: Sixty-eight TNTs were performed on 44 patients over the last 3 years (25 males, 19 females; age range, 16-91 years). No complications were noted. Ninety-one percent of procedures were able to be completed. Indications for TNT were to: 1) detect airway stenosis or pathology, 2) evaluate the larynx and trachea prior to airway surgery, 3) monitor postoperative results of airway interventions, and 4) evaluate the airway prior to tracheotomy tube decannulation., Conclusions: TNT is a safe procedure that can be performed on the unsedated patient using only topical anesthesia and is an attractive alternative to rigid bronchoscopy. The procedural success rate was high, indicating good patient tolerance. The indications for TNT, including its use as a tool for surgical planning, have become better defined. TNT has become a standard tool in the management of patients with airway pathology in our practices., (Copyright © 2012 The American Laryngological, Rhinological, and Otological Society, Inc.)

Ectomycorrhizal (ECM) hyphal communities have not been well characterized. Furthermore, there have been few studies where the ECM hyphal community is compared to fungi detected as sporocarps or ECM-colonized root tips. We investigated fungi present as hyphae in a well-studied California Quercus-Pinus woodland. Hyphal species present were compared to those found as sporocarps and ECM root tips at the same site. Hyphae were extracted from root-restrictive nylon mesh in-growth bags buried in the soil near mature Quercus douglasii, Quercus wislizeni, and Pinus sabiniana. Taxa were identified using PCR, cloning, and DNA sequencing of internal transcribed spacer and 28s rDNA. Among the 33 species detected, rhizomorph-forming ECM fungi dominated the hyphal community, especially species of Thelephoraceae and Boletales. Most fungi in soils near Quercus spp. and P. sabiniana were ECM basidiomycetes, but we detected two ECM ascomycetes and three non-mycorrhizal fungi. Many ECM species present as hyphae were also previously detected at this site as sporocarps (18%) or on ECM root tips (58%). However, the hyphal community was mostly dominated by different taxa than either the sporocarp or ECM root communities.

The Internal Transcribed Spacer (ITS) regions of ribosomal DNA are widely used as markers for phylogenetic analyses and environmental sampling from a variety of organisms including fungi, plants, and animals. In theory, concerted evolution homogenizes multicopy genes so that little or no variation exists within populations or individuals. However, contrary to theory, ITS variation has been confirmed in populations and individuals from a diverse range of eukaryotes. The presence of intraspecific and intra-individual variation in multicopy genes has important implications for ecological and phylogenetic studies, yet relatively little is known about natural variation of these genes, particularly at the community level. In this study, we examined intraspecific and intra-sporocarp ITS variation by DNA sequencing from sporocarps and pooled roots from 68 species of ectomycorrhizal fungi collected at a single site in a Quercus woodland. We detected ITS variation in 27 species, roughly 40% of the taxa examined. Although intraspecific ITS variation was generally low (0.16-2.85%, mean = 0.74%), it was widespread within this fungal community. We detected ITS variation in both sporocarps and ectomycorrhizal roots, and variation was present within species of Ascomycota and Basidiomycota, two distantly related lineages within the Fungi. We discuss the implications of such widespread ITS variability with special reference to DNA-based environmental sampling from diverse fungal communities.

Quercus woodlands are key components of California's wild landscapes, yet little is known about ectomycorrhizal (EM) fungi in these ecosystems. We examined the EM community associated with Quercus douglasii using sporocarp surveys and by pooling EM roots and subjecting them to DNA extraction, polymerase chain reaction (PCR), cloning, restriction fragment length polymorphism (RFLP) screening and DNA sequencing. Ectomycorrhizal root symbionts were sampled four times in 2003-04. During this time, the below-ground community structure was relatively stable; we found no evidence of taxa adapted to winter or spring conditions and only one species varied widely in occurrence between years. The EM community from sporocarps and roots was diverse (161 species), rich in Ascomycota (46 species), and dominated by fungi with cryptic sporocarps. This included a large number of resupinate and hypogeous taxa, many of which were detected both above- and below-ground.

Genea and Genabea are considered ectomycorrhizal (EM) symbionts of higher plants, but because of their hypogeous habit, dark coloration and the small size of their ascomata, relatively little is known about these genera. Ascomata of six morphological species of Genea and one of Genabea were frequently collected at a single site in xeric Quercus woodlands of California's Sierra Nevada foothills. While most collections were easily referred to known species, those putatively identified as Genea harknessii and Genea arenaria were problematic. Genea harknessii collections appeared relatively homogenous based on morphology, but significant ITS variation revealed by rDNA sequencing suggested cryptic species diversity. Specimens of G. arenaria approximated the brief, original species description except for abundant clumps of septate setae formed at the apex of peridial warts. To verify the identity of this species we reexamined the holotype and analyzed morphology and ITS sequences of G. arenaria ascomata from a wide geographic range. To authenticate the EM status of Genea and Genabea with Quercus we collected healthy EM of Quercus douglasii and Quercus wislizenii and compared their ITS sequences to those from ascomata. We detected nine distinct ITS types of Genea and Genabea on roots. Two new species described here as Genea bihymeniata sp. nov. and Genea cazaresii sp. nov., were discovered during study of herbarium specimens. A phylogenetic analysis of 28 s rDNA from Genea and Genabea indicated three distinct lineages: Genea, Genabea and a third represented by Genea intermedia. For the latter we propose Gilkeya gen. nov. to accommodate the single known species, Gilkeya compacta comb. nov. A dichotomous key to all known Genea, Genabea and Gilkeya spp. from western North America is presented.