Methodological Approaches Frame Insights into Endophyte Richness and Community Composition.

Isolating microbes is vital to study microbiomes, but insights into microbial diversity and ecology can be constrained by recalcitrant or unculturable strains. Culture-free methods (e.g., next-generation sequencing, NGS) have become popular in part because they detect greater richness than culturing alone. Both approaches are used widely to characterize microfungi within healthy leaves (foliar endophytes), but methodological differences among studies can constrain large-scale insights into endophyte ecology. We examined endophytes in a temperate plant community to quantify how certain methodological factors, such as the choice of cultivation media for culturing and storage period after leaf collection, affect inferences regarding endophyte communities; how such effects vary among plant taxa; and how complementary culturing and NGS can be when subsets of the same plant tissue are used for each. We found that endophyte richness and composition from culturing were consistent across five media types. Insights from culturing and NGS were largely robust to differences in storage period (1, 5, and 10 days). Although endophyte richness, composition, and taxonomic diversity identified via culturing vs. NGS differed markedly, both methods revealed host-structured communities. Studies differing only in cultivation media or storage period thus can be compared to estimate endophyte richness, composition, and turnover at scales larger than those of individual studies alone. Our data show that it is likely more important to sample more host species, rather than sampling fewer species more intensively, to quantify endophyte diversity in given locations, with the richest insights into endophyte ecology emerging when culturing and NGS are paired.

Probabilities of ossicular discontinuity in chronic suppurative otitis media using pure-tone audiometry.

OBJECTIVE: To analyze the likelihood ratios (LRs) and predictive values of preoperative air-bone gap (ABG) levels on the presence of gross ossicular discontinuity (OD) among chronic suppurative otitis media (CSOM) patients. SETTING: Tertiary hospitals. METHODS: Records of 276 patients with CSOM 7 to 75 years old undergoing their first tympanomastoidectomy were reviewed. Association of preoperative audiogram on the presence of OD was analyzed using logistic regression analysis and chi 2 tests. Frequency-specific ABG values were compared with the presence of OD. RESULTS: In CSOM without cholesteatoma, the ABG of 20 dB or less at 500 Hz (LR [-], 0.119; 95% confidence interval [CI], 0.016-0.867) and 30 dB or less at 1 kHz (LR [-], 0.276; 95% CI, 0.087-0.876) decreased probability of OD from 33 to 5.6% and 15.5%, respectively. Air-bone gap levels of greater than 30 dB at 2 kHz (LR [+], 2.8; 95% CI, 1.4-5.9) and greater than 40 dB at 4 kHz (LR [+], 2.2; 95% CI, 1.2-3.9) increased the probability of OD from 33 to 51 to 89%. In the presence of cholesteatoma, the chance of OD was 88%. The ability of ABG to alter probability of OD was not significant in the presence of cholesteatoma. CONCLUSION: Narrow ABG at lower frequencies suggested absence of OD. Wide ABG at higher frequencies suggested presence of OD. Simple tympanoplasty can be done to patients with a small chance of OD as assessed by pure-tone audiometry, whereas a mandatory exploration of the ossicular chain with possible reconstruction was suggested on subjects with a high chance of OD. The presence of cholesteatoma warrants ossicular chain exploration.

Comparative study of cochlear damage with three perimodiolar electrode designs.

OBJECTIVE: To describe intracochlear insertion trauma caused by three perimodiolar cochlear implant electrodes. STUDY DESIGN: Descriptive histological study of 15 human cadaver temporal bones. METHODS: Fifteen cadaver temporal bones underwent surface preparation and were implanted with one of the following perimodiolar electrode arrays: Combi 40+PM (MedEl Corporation), HiFocus II (Advanced Bionics Corporation), or Contour (Cochlear Corporation). A cryosectioning technique was used to study horizontal sections at 200 microm intervals with the electrode in place. Image-enhanced videofluoroscopy and computer-assisted morphometrics were used to assess the mechanism of insertion trauma and to determine electrode position within the modiolus. RESULTS: Histological examination revealed varying degrees of damage to the spiral ligament, basilar membrane, and osseous spiral lamina. Using a novel grading system for electrode trauma, there was no statistically significant difference among the three electrodes. A literature search of histological studies of a commonly used "standard" electrode showed damage equal to or greater than that seen in the current study. CONCLUSIONS: Insertion trauma caused by periomodiolar electrodes occurs to an acceptable degree. Refinement of electrodes based on mechanisms of trauma may be able to further reduce damage.

Modiolar proximity of three perimodiolar cochlear implant electrodes.

A new generation of cochlear implant electrodes has been designed to position the stimulating contacts close to the modiolus in order to reduce power consumption and increase stimulation selectivity. The purpose of this study was to assess electrode position in the cochlea for three recently designed electrodes. Fifteen cadaveric temporal bones were implanted with one of three perimodiolar electrode arrays: Nucleus Contour; Med-El Combi40+ PM (developmental version); and Clarion HiFocus II. Image-enhanced videofluoroscopy and computer morphometrics were used to assess stimulating contact position relative to the modiolus. The mean distance (+/- 1 standard deviation) to the modiolus for all electrode contacts was 0.33 (+/-0.24), 0.30 (+/-0.27) and 0.16 mm (+/-0.19) for the Contour, Combi40+ PM and HiFocus II arrays, respectively. In addition, dynamic videofluoroscopy was used to correlate device-specific insertion characteristics with contact-to-modiolus distance. All three devices were successful in terms of locating electrode contacts very close to the modiolar wall.