Olfaction-associated quality of life: Polish adaptation and validation of a Questionnaire of Olfactory Disorders (QOD-PL) in patients with chronic rhinosinusitis.

BACKGROUND: The focus on health-related quality of life (HRQOL) in medical research is becoming more and more intensive, with attention being paid to the patient's subjective feelings and assessment of one's health status. Smell disorders can significantly impact human life. The Questionnaire of Olfactory Disorders (QOD) is a self-monitoring questionnaire that provides subjective information about olfactory disorders. OBJECTIVES: This study aimed to check the reliability and validation of the Polish version of QOD (QOD-PL) for use in patients with olfactory impairment in Poland. MATERIAL AND METHODS: A total of 158 patients (76 females, mean age (Mage) 45.97 ±16.37 years), suffering from chronic rhinosinusitis (CRS), whose olfactory function was measured using the Sniffin' Sticks test (SST), were studied. All patients completed 3 validated questionnaires: Importance of Olfaction (IO), Sino-Nasal Outcome Test-22 (SNOT-22) and QOD-PL. RESULTS: Internal consistency and test-retest reliability of the entire QOD-PL scale were good (Cronbach's α = 0.88). The convergent validity of the QOD-PL and its subscales correlated with IO and SNOT-22. The life quality statements in the QOD-PL (QOD-PL-LQ) score, its negative statements, and the mean score for VAS scales were significantly and positively correlated with all symptoms measured with SNOT-22 and none of the IO scales. CONCLUSIONS: The QOD-PL is a reliable, valid and important tool for assessing HRQOL in patients with olfactory disorders. However, it is important to note that not all of its subscales can be considered and interpreted separately.

Telomere-to-telomere genome sequence of the model mould pathogen Aspergillus fumigatus.

The pathogenic fungus Aspergillus fumigatus is a major etiological agent of fungal invasive and chronic diseases affecting tens of millions of individuals worldwide. Draft genome sequences of two clinical isolates (Af293 and A1163) are commonly used as reference genomes for analyses of clinical and environmental strains. However, the reference sequences lack coverage of centromeres, an accurate sequence for ribosomal repeats, and a comprehensive annotation of chromosomal rearrangements such as translocations and inversions. Here, we used PacBio Single Molecule Real-Time (SMRT), Oxford Nanopore and Illumina HiSeq sequencing for de novo genome assembly and polishing of two laboratory reference strains of A. fumigatus, CEA10 (parental isolate of A1163) and its descendant A1160. We generated full length chromosome assemblies and a comprehensive telomere-to-telomere coverage for CEA10 and near complete assembly of A1160 including ribosomal repeats and the sequences of centromeres, which we discovered to be composed of long transposon elements. We envision these high-quality reference genomes will become fundamental resources to study A. fumigatus biology, pathogenicity and virulence, and to discover more effective treatments against diseases caused by this fungus.

Functional Genomic and Biochemical Analysis Reveals Pleiotropic Effect of Congo Red on Aspergillus fumigatus.

Inhibition of fungal growth by Congo red (CR) has been putatively associated with specific binding to ß-1,3-glucans, which blocks cell wall polysaccharide synthesis. In this study, we searched for transcription factors (TFs) that regulate the response to CR and interrogated their regulon. During the investigation of the susceptibility to CR of the TF mutant library, several CR-resistant and -hypersensitive mutants were discovered and further studied. Abnormal distorted swollen conidia called Quasimodo cells were seen in the presence of CR. Quasimodo cells in the resistant mutants were larger than the ones in the sensitive and parental strains; consequently, the conidia of the resistant mutants absorbed more CR than the germinating conidia of the sensitive or parental strains. Accordingly, this higher absorption rate by Quasimodo cells resulted in the removal of CR from the culture medium, allowing a subset of conidia to germinate and grow. In contrast, all resting conidia of the sensitive mutants and the parental strain were killed. This result indicated that the heterogeneity of the conidial population is essential to promote the survival of Aspergillus fumigatus in the presence of CR. Moreover, amorphous surface cell wall polysaccharides such as galactosaminogalactan control the influx of CR inside the cells and, accordingly, resistance to the drug. Finally, long-term incubation with CR led to the discovery of a new CR-induced growth effect, called drug-induced growth stimulation (DIGS), since the growth of one of them could be stimulated after recovery from CR stress.IMPORTANCE The compound Congo red (CR) has been historically used for coloring treatment and histological examination as well to inhibit the growth of yeast and filamentous fungi. It has been thought that CR binds to ß-1,3-glucans in the fungal cell wall, disrupting the organization of the cell wall structure. However, other processes have been implicated in affecting CR sensitivity. Here, we explore CR susceptibility through screening a library of genetic null mutants. We find several previously uncharacterized genetic regulators important for CR susceptibility. Through biochemical and molecular characterization, we find cell membrane permeability to be important. Additionally, we characterize a novel cell type, Quasimodo cells, that occurs upon CR exposure. These cells take up CR, allowing the growth of the remaining fungi. Finally, we find that priming with CR can enhance long-term growth in one mutant.

Comparison of the clinical differences between COVID-19, SARS, influenza, and the common cold: A systematic literature review.

BACKGROUND: This review focuses on the frequency of symptoms in COVID-19 in comparison to SARS, influenza and common cold. OBJECTIVES: To evaluate and compare the knowledge about the clinical features, symptoms and differences between patients with COVID-19, SARS, influenza, and common cold. The research can help ear, nose and throat specialists and other health practitioners manage patients during the COVID-19 pandemic. MATERIAL AND METHODS: The biomedical databases used in the study included PubMed and MEDLINE. Statistical analysis using the Z-score test assessed which symptoms were more characteristic of COVID-19 than other viral diseases. RESULTS: Among individuals with COVID-19, the most frequently reported symptoms were cough (70%), fever (45%), muscular pain (29%), and headache (21%), whereas sore throat (12%), and rhinorrhea (4%) were observed at lower rates. Fever was identified as most frequent in COVID-19 (74%), appearing at a higher rate in those cases than in influenza (68%) or the common cold (40%) (p < 0.05). In comparison to other viral diseases, sore throat was rarely reported in COVID-19 and SARS (12% and 18%, respectively) (p < 0.05). In influenza and common cold, a cough was identified in 93% and 80% of cases (p < 0.05). Headache, rhinorrhea, muscular pain, and sore throat were more common in influenza (91%, 91%, 94%, and 84%, respectively) and common cold (89%, 81%, 94%, and 84%, respectively) than in COVID-19 (21%, 4%, 29%, and 12%, respectively) and SARS (45%, 12%, 55%, and 18%, respectively) (p < 0.05). CONCLUSION: The results of the analysis show that a greater number of general symptoms should lead to a diagnosis of influenza or common cold rather than COVID-19.

Functional and transcriptional profiling of non-coding RNAs in yeast reveal context-dependent phenotypes and in trans effects on the protein regulatory network.

Non-coding RNAs (ncRNAs), including the more recently identified Stable Unannotated Transcripts (SUTs) and Cryptic Unstable Transcripts (CUTs), are increasingly being shown to play pivotal roles in the transcriptional and post-transcriptional regulation of genes in eukaryotes. Here, we carried out a large-scale screening of ncRNAs in Saccharomyces cerevisiae, and provide evidence for SUT and CUT function. Phenotypic data on 372 ncRNA deletion strains in 23 different growth conditions were collected, identifying ncRNAs responsible for significant cellular fitness changes. Transcriptome profiles were assembled for 18 haploid ncRNA deletion mutants and 2 essential ncRNA heterozygous deletants. Guided by the resulting RNA-seq data we analysed the genome-wide dysregulation of protein coding genes and non-coding transcripts. Novel functional ncRNAs, SUT125, SUT126, SUT035 and SUT532 that act in trans by modulating transcription factors were identified. Furthermore, we described the impact of SUTs and CUTs in modulating coding gene expression in response to different environmental conditions, regulating important biological process such as respiration (SUT125, SUT126, SUT035, SUT432), steroid biosynthesis (CUT494, SUT053, SUT468) or rRNA processing (SUT075 and snR30). Overall, these data capture and integrate the regulatory and phenotypic network of ncRNAs and protein-coding genes, providing genome-wide evidence of the impact of ncRNAs on cellular homeostasis.

Concurrent Squamous Cell Carcinoma and Leiomyosarcoma in the Larynx at Relapse 15 Years Post-radiation for Primary Laryngeal Cancer.

Malignant tumours of the larynx account for 50% of all head and neck neoplasms. Squamous cell carcinoma is the most common malignancy of the larynx and accounts for 95-98%. Leiomyosarcoma (LMS) originating in larynx is a very rare malignant tumour derived from the smooth muscle tissue. The authors present a case of a 75-year man with coexistent squamous cell carcinoma and leiomyosarcoma in the larynx 15 years after radiation therapy for promany laryngeal cancer. Two cases of the synchronous presence of these neoplasms have been reported so far in English literature. This paper presents the clinical picture, radiological and histopathological diagnoses, and treatment of this case. Key Words: Squamous cell carcinoma, Leiomyosarcoma, Larynx.

The negative cofactor 2 complex is a key regulator of drug resistance in Aspergillus fumigatus.

The frequency of antifungal resistance, particularly to the azole class of ergosterol biosynthetic inhibitors, is a growing global health problem. Survival rates for those infected with resistant isolates are exceptionally low. Beyond modification of the drug target, our understanding of the molecular basis of azole resistance in the fungal pathogen Aspergillus fumigatus is limited. We reasoned that clinically relevant antifungal resistance could derive from transcriptional rewiring, promoting drug resistance without concomitant reductions in pathogenicity. Here we report a genome-wide annotation of transcriptional regulators in A. fumigatus and construction of a library of 484 transcription factor null mutants. We identify 12 regulators that have a demonstrable role in itraconazole susceptibility and show that loss of the negative cofactor 2 complex leads to resistance, not only to the azoles but also the salvage therapeutics amphotericin B and terbinafine without significantly affecting pathogenicity.

Anatomical and Clinical Implications in Neocondyle Stability After a Condylectomy.

: A condylectomy of the mandibular condyle is considered to be the treatment of choice in most cases of condylar head hyperactivity. The aim of the procedure is to remove the growth center of the mandible which is responsible for the mandibular enlargement and asymmetry. This surgical procedure has an impact on the condyle shape and position, but the restoration of mandibular movement and a stable joint position (namely, the proper alignment of the newly shaped condylar head within the condyle fossa) should also be considered important surgical outcomes. In this article, the authors present their own experience in performing condylectomies with an arthroplasty procedure and a special forced suturing technique (FST) in terms of achieving early, accurate mandibular movement and maintaining a stable condyle position in early and late outcomes. MATERIALS AND METHODS: A modified high condylectomy with arthroplasty and FST results had been studied in anatomical, radiological, and clinical model. RESULTS: Early findings after FST are promising. A slight improvement in lateral jaw movement was noted after condylectomy with arthroplasty (P < 0.05) both in early and late follow-up. Incisal opening, mandibular protrusion, and lateral movement were sustained. A stable condyle position within the fossa was achieved in each case of condylectomy with arthroplasty (P < 0.05). CONCLUSIONS: The FST condylectomy and reattachment of the lateral pterygoid muscle in a new, wider position provided an improvement in lateral jaw movement as well as in incisal opening and mandibular protrusion in early follow-up examination compared to the presurgical values. It seems that the FST enabled a better new condylar head position in the glenoid fossa and improved early functional mandibular movement.

Fast and Reliable PCR Amplification from Aspergillus fumigatus Spore Suspension Without Traditional DNA Extraction.

Aspergillus fumigatus is an opportunistic human pathogenic mold. DNA extraction from this fungus is usually performed by mechanical perturbation of cells, as it possesses a rigid and complex cell wall. While this is not problematic for single isolates, it can be time consuming for large numbers of strains if using traditional DNA extraction procedures. Therefore, in this article we describe a fast and efficient thermal-shock method to release DNA from spores of A. fumigatus and other filamentous fungi without the need for complex extraction methods. This is especially important for high-throughput PCR analyses of mutants in 96- or 384-well formats in a very short period of time without any concern about sample cross-contamination. This method is currently being used to validate the protein-coding gene and non-coding RNA knockout libraries in A. fumigatus generated in our laboratory, and could be used in the future for diagnostics purposes. © 2019 The Authors.

High-Throughput Gene Replacement in Aspergillus fumigatus.

Aspergillus fumigatus is a human pathogen and the principal etiologic agent of invasive and chronic aspergillosis leading to several hundreds of thousands of deaths every year. Very few antifungals are available to treat infections caused by A. fumigatus, and resistance is developing to those we have. Our understanding of the molecular mechanisms that drive pathogenicity and drug resistance have been hampered by the lack of large mutant collections, which limits our ability to perform functional genomics analysis. Here we present a high-throughput gene knockout method that combines a highly reproducible fusion PCR method to enable generation of gene replacement cassettes with a multiwell format transformation procedure. This process can be used to generate 96 null mutants within 5 days by a single person at a cost of less than £18 ($24) per mutant and is being employed in our laboratory to generate a barcoded genome-wide knockout library in A. fumigatus. © 2019 The Authors.

[A case of rare laryngeal neurogenic tumor].

Laryngeal schwannoma is a rare benign tumour of the larynx. Schwannomas derive from the Schwann cells. Magnetic resonance imaging (MRI) is the best imaging tool for suggesting the diagnosis. We report case of laryngeal schwannoma. The case was a 61-year-old man with a 1-year history of dysphonia and stridor. Laryngoscopy revealed a submucosal mass of the glottic area. A computed tomography scan of the larynx showed an 26(CC) x 18 (TR) x 24 (AP)mm expansile mass in the glottic area. Histopathological examination diagnosed a schwannoma of the larynx. External surgeries were successfully performed. Diagnosis and specific treatment are discussed.

Reliability of computed tomography scans in the diagnosis of chronic rhinosinusitis.

BACKGROUND: Paranasal computed tomography (CT) has become the investigation method of choice to confirm or exclude the diagnosis of chronic rhinosinusitis (CRS) on the basis of its ability to deliver objective data regarding the presence of inflamed mucosa or polyps. OBJECTIVES: The aim of the study was to assess the reliability of CT scan findings among untreated CRS patients without the presence of polyps in a nasal endoscopy. MATERIAL AND METHODS: Among patients with clinically demonstrated CRS considered for surgery, 93 subjects who had had 2 CT scans performed at different time points in the diagnostic process were enrolled into the study. Paranasal sinus involvement on both CT scans was scored using the Lund-Mackay (L-M) and modified Lund-Mackay scales. Both CT exams served to assess the extent of the potential endoscopic sinus surgery. RESULTS: The time interval between CT scans ranged from 31 to 1,162 days (mean: 338 days). The L-M scores from the 1st CT examination correlated statistically with the results of the 2nd CT (r = 0.86; p < 0.05). When compared to the 1st scan, the L-M score in the 2nd CT scan remained the same in 36 patients (39%), increased in 23 patients (25%) and decreased in 34 patients (36%). There was no statistically significant correlation between the change in the L-M scores and the time interval between CT examinations. CONCLUSIONS: The present study indicates that mucosal thickening within paranasal sinuses among untreated patients with CRS is stable over shortand middle-time intervals, regardless of the initial intensity of the disease. The time delay between the CT examination and qualification for surgery does not influence the decision regarding the performance of the operation. The results suggest the conclusion that repeating CT scans in symptomatic, untreated patients with CRS should be seriously considered.

Large-scale profiling of noncoding RNA function in yeast.

Noncoding RNAs (ncRNAs) are emerging as key regulators of cellular function. We have exploited the recently developed barcoded ncRNA gene deletion strain collections in the yeast Saccharomyces cerevisiae to investigate the numerous ncRNAs in yeast with no known function. The ncRNA deletion collection contains deletions of tRNAs, snoRNAs, snRNAs, stable unannotated transcripts (SUTs), cryptic unstable transcripts (CUTs) and other annotated ncRNAs encompassing 532 different individual ncRNA deletions. We have profiled the fitness of the diploid heterozygous ncRNA deletion strain collection in six conditions using batch and continuous liquid culture, as well as the haploid ncRNA deletion strain collections arrayed individually onto solid rich media. These analyses revealed many novel environmental-specific haplo-insufficient and haplo-proficient phenotypes providing key information on the importance of each specific ncRNA in every condition. Co-fitness analysis using fitness data from the heterozygous ncRNA deletion strain collection identified two ncRNA groups required for growth during heat stress and nutrient deprivation. The extensive fitness data for each ncRNA deletion strain has been compiled into an easy to navigate database called Yeast ncRNA Analysis (YNCA). By expanding the original ncRNA deletion strain collection we identified four novel essential ncRNAs; SUT527, SUT075, SUT367 and SUT259/691. We defined the effects of each new essential ncRNA on adjacent gene expression in the heterozygote background identifying both repression and induction of nearby genes. Additionally, we discovered a function for SUT527 in the expression, 3' end formation and localization of SEC4, an essential protein coding mRNA. Finally, using plasmid complementation we rescued the SUT075 lethal phenotype revealing that this ncRNA acts in trans. Overall, our findings provide important new insights into the function of ncRNAs.

History of genome editing in yeast.

For thousands of years humans have used the budding yeast Saccharomyces cerevisiae for the production of bread and alcohol; however, in the last 30-40 years our understanding of the yeast biology has dramatically increased, enabling us to modify its genome. Although S. cerevisiae has been the main focus of many research groups, other non-conventional yeasts have also been studied and exploited for biotechnological purposes. Our experiments and knowledge have evolved from recombination to high-throughput PCR-based transformations to highly accurate CRISPR methods in order to alter yeast traits for either research or industrial purposes. Since the release of the genome sequence of S. cerevisiae in 1996, the precise and targeted genome editing has increased significantly. In this 'Budding topic' we discuss the significant developments of genome editing in yeast, mainly focusing on Cre-loxP mediated recombination, delitto perfetto and CRISPR/Cas.

Corticosteroid treatment is associated with increased filamentous fungal burden in allergic fungal disease.

BACKGROUND: Allergic diseases caused by fungi are common. The best understood conditions are allergic bronchopulmonary aspergillosis and severe asthma with fungal sensitization. Our knowledge of the fungal microbiome (mycobiome) is limited to a few studies involving healthy individuals, asthmatics, and smokers. No study has yet examined the mycobiome in fungal lung disease. OBJECTIVES: The main aim of this study was to determine the mycobiome in lungs of individuals with well-characterized fungal disease. A secondary objective was to determine possible effects of treatment on the mycobiome. METHODS: After bronchoscopy, ribosomal internal transcribed spacer region 1 DNA was amplified and sequenced and fungal load determined by real-time PCR. Clinical and treatment variables were correlated with the main species identified. Bronchopulmonary aspergillosis (n = 16), severe asthma with fungal sensitization (n = 16), severe asthma not sensitized to fungi (n = 9), mild asthma patients (n = 7), and 10 healthy control subjects were studied. RESULTS: The mycobiome was highly varied with severe asthmatics carrying higher loads of fungus. Healthy individuals had low fungal loads, mostly poorly characterized Malasezziales. The most common fungus in asthmatics was Aspergillus fumigatus complex and this taxon accounted for the increased burden of fungus in the high-level samples. Corticosteroid treatment was significantly associated with increased fungal load (P < .01). CONCLUSIONS: The mycobiome is highly variable. Highest loads of fungus are observed in severe asthmatics and the most common fungus is Aspergillus fumigatus complex. Individuals receiving steroid therapy had significantly higher levels of Aspergillus and total fungus in their bronchoalveolar lavage.

A resource for functional profiling of noncoding RNA in the yeast Saccharomyces cerevisiae.

Eukaryotic genomes are extensively transcribed, generating many different RNAs with no known function. We have constructed 1502 molecular barcoded ncRNA gene deletion strains encompassing 443 ncRNAs in the yeast Saccharomyces cerevisiae as tools for ncRNA functional analysis. This resource includes deletions of small nuclear RNAs (snRNAs), transfer RNAs (tRNAs), small nucleolar RNAs (snoRNAs), and other annotated ncRNAs as well as the more recently identified stable unannotated transcripts (SUTs) and cryptic unstable transcripts (CUTs) whose functions are largely unknown. Specifically, deletions have been constructed for ncRNAs found in the intergenic regions, not overlapping genes or their promoters (i.e., at least 200 bp minimum distance from the closest gene start codon). The deletion strains carry molecular barcodes designed to be complementary with the protein gene deletion collection enabling parallel analysis experiments. These strains will be useful for the numerous genomic and molecular techniques that utilize deletion strains, including genome-wide phenotypic screens under different growth conditions, pooled chemogenomic screens with drugs or chemicals, synthetic genetic array analysis to uncover novel genetic interactions, and synthetic dosage lethality screens to analyze gene dosage. Overall, we created a valuable resource for the RNA community and for future ncRNA research.

Correction: Mitochondrial Complex I Is a Global Regulator of Secondary Metabolism, Virulence and Azole Sensitivity in Fungi.

[This corrects the article DOI: 10.1371/journal.pone.0158724.].

Investigation of sinonasal anatomy via low-dose multidetector CT examination in chronic rhinosinusitis patients with higher risk for perioperative complications.

The aim of the study was to compare visualisation of the surgically relevant anatomical structures via low- and standard-dose multidetector CT protocol in patients with chronic rhinosinusitis (CRS) and higher risk for perioperative complications (i.e. presence of bronchial asthma, history of sinus surgery and advanced nasal polyposis). 135 adult CRS patients were divided randomly into standard-dose (120 kVp, 100 mAs) or low-dose CT groups (120 kVp, 45 mAs). The detectability of the vital anatomical structures (anterior ethmoid artery, optic nerve, cribriform plate and lamina papyracea) was scored using a five-point scale (from excellent to unacceptable) by a radiologist and sinus surgeon. Polyp sizes were quantified endoscopically according to the Lildholdt's scale (LS). Olfactory function was tested with the "Sniffin' Sticks" test. On the low-dose CT images, detectability ranged from 2.42 (better than poor) for cribriform plate among anosmic cases to 4.11 (better than good) for lamina papyracea in cases without nasal polyps. Identification of lamina papyracea on low-dose scans was significantly worse in each group and the same was the case with cribriform plates in patients with advanced polyposis and anosmia. Cribriform plates were the most poorly identified (between poor and average) among all the structures on low-dose images. Identification of anterior ethmoid artery (AEA) with reduced dose was insignificantly worse than with standard-dose examination. The AEA was scored as an average-defined structure and was the second weakest visualised. In conclusion, preoperatively, low-dose protocols may not sufficiently visualise the surgically relevant anatomical structures in patients with CRS and bronchial asthma, advanced nasal polyps (LS > 2) and history of sinus surgery. Low mAs value enables comparable detectability of sinonasal landmarks with standard-dose protocols in patients without analysed risk factors. In the context of planned surgery, the current preferences of the tube should be carefully evaluated for different patient constitutions to minimise the risk of complications.

The usefulness of free dicom viewers in rhinologic practice.

Indroduction: The planning of the course of surgery in patients with paranasal sinus disorders requires the meticulous evaluation of computed tomography (CT) scans. However, ENT doctors still often receive unsatisfactory DICOM images, which require multiplanar reconstruction (MPR). THE AIM: The objective of the present study was to evaluate and test DICOM viewers with MPR function currently available on the Internet for free. MATERIAL AND METHODS: Detailed Internet searches and analysis of databases were carried out to find the maximum number of browsers. Available software was downloaded to assess, qualitatively and quantitatively, the parameters and functions useful in rhinologic practice. Additionally, a questionnaire was developed in order to ascertain the expectations and habits of ENT doctors in relation to DICOM browsers and CT scan evaluation. RESULTS AND CONCLUSIONS: Among the hundreds of browsers, 17 programs were chosen to be included in the final comparison. The outcomes of the survey proved that many free DICOM tools are effective alternatives to commercially available programs. However, the results do not enable us to predict precisely which tool will meet the needs of each physician. The survey among ENT doctors highlighted their limited awareness of software with MPR functions, in particular those that scored best in the test.