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Purpose To describe the minimum inhibitory concentration (MIC) of fungal isolates to natamycin and voriconazole, and to compare these MICs to previous ocular susceptibility studies. Design Experimental laboratory study using isolates from a randomized clinical trial. Methods The Mycotic Ulcer Treatment Trial I was a randomized, double-masked, multicenter trial comparing topical natamycin and voriconazole for fungal keratitis treatment. Susceptibility testing to natamycin and voriconazole were performed according to Clinical and Laboratory Standards Institute methods. The relationship between organism and MIC was assessed. A literature review was performed to compare results to previous ocular susceptibility studies. Results Of the 323 patients enrolled in the trial, MICs were available for 221 (68%). Fusarium (N=126) and Aspergillus species (N=52) were the most commonly isolated organisms. MICs to natamycin and voriconazole were significantly different across all genera (P<0.001). The MIC median (MIC50) and 90th percentile (MIC90) for natamycin were equal to or higher than voriconazole for all organisms, except Curvularia species. Compared to other organisms, Fusarium species isolates had the highest MICs to voriconazole and A. flavus isolates had the highest MICs to natamycin. Our results were similar to previous reports except the voriconazole MIC90 against Aspergillus species was 2-fold higher and the natamycin MIC90 against A. fumigatus was 4-fold higher in our study. Conclusion In this large susceptibility study, Fusarium isolates were least susceptible to voriconazole and A. flavus isolates were least susceptible to natamycin when compared to other filamentous fungi. In the future, susceptibility testing may help guide therapy if performed in a timely manner.

Mammalian spermatogenesis is a highly ordered process that occurs in mitotic, meiotic, and postmeiotic phases. The unique mechanisms responsible for this tightly regulated developmental process suggest the presence of an intrinsic genetic program composed of spermatogenic cell-specific genes. In this study, we analyzed the mouse round spermatid UniGene library currently containing 2124 gene-oriented transcript clusters, predicting that 467 of them are testis-specific genes, and systematically identified 28 novel genes with evident testis-specific expression by in silico and in vitro approaches. We analyzed these genes by Northern blot hybridization and cDNA cloning, demonstrating the presence of additional transcript sequences in five genes and multiple transcript isoforms in six genes. Genomic analysis revealed lack of human orthologues for 10 genes, implying a relationship between these genes and male reproduction unique to mouse. We found that all of the novel genes are expressed in developmentally regulated and stage-specific patterns, suggesting that they are primary regulators of male germ cell development. Using computational bioinformatics tools, we found that 20 gene products are potentially involved in various processes during spermatogenesis or fertilization. Taken together, we predict that over 20% of the genes from the round spermatid library are testis-specific, have discovered the 28 authentic, novel genes with probable spermatogenic cell-specific expression by the integrative approach, and provide new and thorough information about the novel genes by various in vitro and in silico analyses. Thus, the study establishes on a comprehensive scale a new basis for studies to uncover molecular mechanisms underlying the reproductive process.In sexual reproduction, diploid cells divide to form haploid cells, and the haploid cells from two individuals fertilize to form new diploid cells. This process, producing unpredictably dissimilar offspring, requires intricate and elaborate molecular and cellular events such as genetic recombination and the formation of gametes specialized for fertilization. Male germ cell development or spermatogenesis is a tightly regulated developmental process that occurs in successive mitotic, meiotic, and postmeiotic phases (1-3). The process occurs in the epithelial lining of seminiferous tubules, in testis. Spermatogonial stem cells, located around the outer region next to the basal lamina surrounding the seminiferous tubule, undergo mitosis, and some of them differentiate into later stage spermatogonia that gradually become primary spermatocytes. These cells continue through the first meiotic division to become secondary spermatocytes. Subsequently, the second meiotic division occurs in rapid succession to produce spermatids. These haploid spermatids are then remodeled into spermatozoa by spermiogenesis. The major events in spermiogenesis are chromatin condensation and morphological changes. This tightly regulated process accompanying meiotic progression and the drastic chang...

Purpose To assess the association between minimum inhibitory concentration (MIC) and clinical outcomes in a fungal keratitis clinical trial. Design Experimental study using data from a randomized comparative trial. Participants Of the 323 patients enrolled in the trial, we were able to obtain MIC values from 221 patients with monocular fungal keratitis. Methods The Mycotic Ulcer Treatment Trial I (MUTT I) was a randomized, double-masked clinical trial comparing clinical outcomes of monotherapy with topical natamycin versus voriconazole for the treatment of fungal keratitis. Speciation and determination of MIC to natamycin and voriconazole were performed according to Clinical and Laboratory Standards Institute guidelines. The relationship between MIC and clinical outcome was assessed. Main Outcome Measures The primary outcome was 3-month best spectacle-corrected visual acuity. Secondary outcomes included 3-month infiltrate/scar size, corneal perforation and/or therapeutic penetrating keratoplasty (TPK), and time to re-epithelialization. Results A 2-fold increase in MIC was associated with a larger 3-month infiltrate/scar size (0.21 mm, 95% confidence interval [CI] 0.10–0.31, P <0.001) and increased odds of perforation (odds ratio [OR] 1.32, 95% CI 1.04–1.69, P=0.02). No correlation was found between MIC and 3-month visual acuity. For natamycin-treated cases, an association was found between higher natamycin MIC with larger 3-month infiltrate/scar size (0.29 mm, 95% CI 0.15–0.43, P<0.001) and increased perforations (OR 2.41, 95% CI 1.46–3.97, P<0.001). Among voriconazole-treated cases, the voriconazole MIC did not correlate with any of the measured outcomes in the study. Conclusion Decreased susceptibility to natamycin was associated with increased infiltrate/scar size and increased odds of perforation. There was no association between susceptibility to voriconazole and outcome.