Leaf Extract from European Olive (Olea europaea L.) Post-Transcriptionally Suppresses the Epithelial-Mesenchymal Transition and Sensitizes Gastric Cancer Cells to Chemotherapy.

The overall survival of patients with the advanced and recurrent gastric cancer (GC) remains unfavorable. In particular, this is due to cancer spreading and resistance to chemotherapy associated with the epithelial-mesenchymal transition (EMT) of tumor cells. EMT can be identified by the transcriptome profiling of GC for EMT markers. Indeed, analysis of the TCGA and GTEx databases (n = 408) and a cohort of GC patients (n = 43) revealed that expression of the CDH2 gene was significantly decreased in the tumors vs. non-tumor tissues and correlated with the overall survival of GC patients. Expression of the EMT-promoting transcription factors SNAIL and ZEB1 was significantly increased in GC. These data suggest that targeting the EMT might be an attractive therapeutic approach for patients with GC. Previously, we demonstrated a potent anti-cancer activity of the olive leaf extract (OLE). However, its effect on the EMT regulation in GC remained unknown. Here, we showed that OLE efficiently potentiated the inhibitory effect of the chemotherapeutic agents 5-fluorouracil (5-FU) and cisplatin (Cis) on the EMT and their pro-apoptotic activity, as was demonstrated by changes in the expression of the EMT markers (E- and N-cadherins, vimentin, claudin-1) in GC cells treated with the aforementioned chemotherapeutic agents in the presence of OLE. Thus, culturing GC cells with 5-FU + OLE or Cis + OLE attenuated the invasive properties of cancer cells. Importantly, upregulation of expression of the apoptotic markers (PARP cleaved form) and increase in the number of cells undergoing apoptosis (annexin V-positive) were observed for GC cells treated with a combination of OLE and 5-FU or Cis. Collectively, our data illustrate that OLE efficiently interferes with the EMT in GC cells and potentiates the pro-apoptotic activity of certain chemotherapeutic agents used for GC therapy.

Determining the reliability of diagnosis and treatment using artificial intelligence software with panoramic radiographs.

Purpose: The objective of this study was to evaluate the accuracy and effectiveness of an artificial intelligence (AI) program in identifying dental conditions using panoramic radiographs (PRs), as well as to assess the appropriateness of its treatment recommendations. Material and Methods: PRs from 100 patients (representing 4497 teeth) with known clinical examination findings were randomly selected from a university database. Three dentomaxillofacial radiologists and the Diagnocat AI software evaluated these PRs. The evaluations were focused on various dental conditions and treatments, including canal filling, caries, cast post and core, dental calculus, fillings, furcation lesions, implants, lack of interproximal tooth contact, open margins, overhangs, periapical lesions, periodontal bone loss, short fillings, voids in root fillings, overfillings, pontics, root fragments, impacted teeth, artificial crowns, missing teeth, and healthy teeth. Results: The AI demonstrated almost perfect agreement (exceeding 0.81) in most of the assessments when compared to the ground truth. The sensitivity was very high (above 0.8) for the evaluation of healthy teeth, artificial crowns, dental calculus, missing teeth, fillings, lack of interproximal contact, periodontal bone loss, and implants. However, the sensitivity was low for the assessment of caries, periapical lesions, pontic voids in the root canal, and overhangs. Conclusion: Despite the limitations of this study, the synthesized data suggest that AI-based decision support systems can serve as a valuable tool in detecting dental conditions, when used with PR for clinical dental applications.

Co-loading of Temozolomide with Oleuropein or rutin into polylactic acid core-shell nanofiber webs inhibit glioblastoma cell by controlled release.

Glioblastoma (GB) has susceptibility to post-surgical recurrence. Therefore, local treatment methods are required against recurrent GB cells in the post-surgical area. In this study, we developed a nanofiber-based local therapy against GB cells using Oleuropein (OL), and rutin and their combinations with Temozolomide (TMZ). The polylactic acid (PLA) core-shell nanofiber webs were encapsulated with OL (PLAOL), rutin (PLArutin), and TMZ (PLATMZ) by an electrospinning process. A SEM visualized the morphology and the total immersion method determined the release characteristics of PLA webs. Real-time cell tracking analysis for cell growth, dual Acridine Orange/Propidium Iodide staining for cell viability, a scratch wound healing assay for migration capacity, and a sphere formation assay for tumor spheroid aggressiveness were used. All polymeric nanofiber webs had core-shell structures with an average diameter between 133 ± 30.7-139 ± 20.5 nm. All PLA webs promoted apoptotic cell death, suppressed cell migration, and spheres growth (p < 0.0001). PLAOL and PLATMZ suppressed GB cell viability with a controlled release that increased over 120 h, while PLArutin caused rapid cell inhibition (p < 0.0001). Collectively, our findings suggest that core-shell nano-webs could be a novel and effective therapeutic tool for the controlled release of OL and TMZ against recurrent GB cells.

Association among Orthodontic Malocclusions, Paranasal Sinuses Anatomic Variations and Adenoid Vegetation in Children Using CBCT.

The aim of this study is to evaluate the relationship between orthodontic malocclusion, paranasal sinus (PS) variations, and adenoid vegetation in a group of pediatric patients with chronic rhinosinusitis. Clinical and radiographical data were retrospectively evaluated and 58 patients were diagnosed as having chronic sinus disease. Cone-beam computed tomography (CBCT) images were acquired with Newtom-3G. Anatomical variations of the PS were assessed on every section. Additionally, for cephalometric analysis, the images were imported into the InVivoDental software program. A total of 252 anatomical variations, which encompassed 19 different types, were detected in the current study. Concha bullosa was the most common anatomical variation, at 72.4%. Septum deviation was the second most common one, at 67.2%. The Class III group exhibited a significantly higher prevalence of concha bullosa and secondary middle turbinate than the other groups. While adenoid vegetation was most common in the Class III group, sinusitis and antral disease were most common in the Class II group. Overall, Class III subjects exhibited fewer PS variations. In conclusion, concha bullosa emerged as the most prevalent anatomical variation, with distinctive patterns observed across different malocclusion groups. Therefore, CBCT is useful, especially in pediatric patients, due to its low dose advantage.

Investigation of the Effectiveness of Oleuropein in a Three-Dimensional In Vitro Hepatocellular Tumor Sphere Model.

OBJECTIVES: This study was conducted to examine the dose-related effects over time of oleuropein on the proliferation and area of tumor spheroids in hepatocellular carcinoma cells. MATERIALS AND METHODS: We examined the possible effects of 100 to 500 µM dose concentrations of oleuropein on HepG2 cell proliferation using a real-time cell analyzer. A 3-dimensional hepatocellular carcinoma tumor spheroid model was established by seeding HepG2 cells at a density of 160 cells/well in custom 96-well microplates with low attachment surfaces and culturing for 3 days. Tumor spheres were treated with increasing oleuropein doses for 72 hours, and images were captured every 24 hours. The dose-dependent effects of oleuropein on tumor sphere size were analyzed by measuring the area of tumor spheres with ImageJ software. We conducted oleuropein viability and cytotoxicity analyses using calcein acetoxymethyl ester-based and propidium iodide-based staining in the tumor model. RESULTS: Oleuropein inhibited cell proliferation; as the dose concentration of oleuropein increased, so did its capacity to inhibit cell proliferation (P < .001). The size of untreated tumor spheres increased at 72 hours (P < .001). However, treatment with 100 to 500 µM oleuropein reduced tumor size by 63.56% to 88.06% compared with untreated cells at the end of 72 hours (P < .001). With increasing concentrations, oleuropein inhibited the viability of tumor spheres, eliminating necrotic death caused by tumor hypoxia. CONCLUSIONS: Overall, oleuropein reduced the size of tumors by inhibiting tumor proliferation and viability. In this context, oleuropein could be a candidate molecule for further extensive studies to reduce hepatocellular carcinoma tumors to meet Milan criteria for liver transplant.

Evaluation of the influence of hyoid bone position, volume, and types on pharyngeal airway volume and cephalometric measurements.

OBJECTIVES: We aimed to explore the position, morphological, and morphometric properties of the hyoid bone (HB) and to investigate the effect of HB on the pharyngeal airway (PA) volume and cephalometric measurements. METHODS: A total of 305 patients with CT images were included in the study. DICOM images were transferred to the InVivoDental three-dimensional imaging software. The position of the HB was determined based on the cervical vertebra level and in volume render tab, the bone was classified into six types after all structures around the HB were removed. Also, final bone volume was recorded. In the same tab, the pharyngeal airway volume was divided and measured in three groups (nasopharynx-oropharynx-hypopharynx). The linear and angular measurements were performed on the 3D cephalometric analysis tab. RESULTS: HB was most commonly located in C3 vertebra level (80.3%). While B-type was found to be the most frequent (34%), V-type was the least frequent (8%). The volume of the HB was found to be significantly higher in male (3205 mm3) than female (2606 mm3) patients. Also, it was significantly higher in the C4 vertebra group. The vertical height of the face was positively correlated with the HB volume, C4 level position, and increased oro-nasopharyngeal airway volume. CONCLUSION: The volume of the HB is found to differ significantly between genders and can potentially serve as a valuable diagnostic tool for understanding respiratory disorders. Its morphometric features are associated with increased face height and airway volume; however, are not related with the skeletal malocclusion classes.

Influence of voxel size and filter type on detecting vertical root fracture using cone-beam computed tomography.

Vertical root fractures (VRFs) can start at any level of the root and progress longitudinally to the coronal attachment. This study aimed to investigate the effects of different exposure parameters used when obtaining CBCT scans in detecting simulated VRFs. Hence, 80 intact human mandibular single-rooted pre-molar teeth without root fractures were included in the study. No statistically significant difference was found between the filters in terms of VRF detection in the group with the roots with only root canal filling (Groups 1 and 5); however, 100 voxels were found to be more successful in terms of VRF detection than other voxel sizes. Results of this study suggest that using lower voxel sizes leads to an accurate diagnosis of vertical root fracture, in addition, our results revealed that using AR filters did not improve the diagnostic accuracy in detecting VRFs.

Automatic Segmentation of Teeth, Crown-Bridge Restorations, Dental Implants, Restorative Fillings, Dental Caries, Residual Roots, and Root Canal Fillings on Orthopantomographs: Convenience and Pitfalls.

BACKGROUND: The aim of our study is to provide successful automatic segmentation of various objects on orthopantomographs (OPGs). METHODS: 8138 OPGs obtained from the archives of the Department of Dentomaxillofacial Radiology were included. OPGs were converted into PNGs and transferred to the segmentation tool's database. All teeth, crown-bridge restorations, dental implants, composite-amalgam fillings, dental caries, residual roots, and root canal fillings were manually segmented by two experts with the manual drawing semantic segmentation technique. RESULTS: The intra-class correlation coefficient (ICC) for both inter- and intra-observers for manual segmentation was excellent (ICC > 0.75). The intra-observer ICC was found to be 0.994, while the inter-observer reliability was 0.989. No significant difference was detected amongst observers (p = 0.947). The calculated DSC and accuracy values across all OPGs were 0.85 and 0.95 for the tooth segmentation, 0.88 and 0.99 for dental caries, 0.87 and 0.99 for dental restorations, 0.93 and 0.99 for crown-bridge restorations, 0.94 and 0.99 for dental implants, 0.78 and 0.99 for root canal fillings, and 0.78 and 0.99 for residual roots, respectively. CONCLUSIONS: Thanks to faster and automated diagnoses on 2D as well as 3D dental images, dentists will have higher diagnosis rates in a shorter time even without excluding cases.

Olea europaea L. Leaf Extract Attenuates Temozolomide-Induced Senescence-Associated Secretion Phenotype in Glioblastoma.

Objectives: The purpose of this study was to investigate the effect of Olea europaea L. leaf extract (OLE) on senescence and senescence-associated secretory phenotype (SASP) caused by temozolomide (TMZ) in glioblastoma (GB). Materials and Methods: A senescence ß-galactosidase assay and a colony formation assay were used to determine the effects of OLE, TMZ, and OLE + TMZ on the cellular senescence and aggressiveness of GB cell lines T98G and U87MG. mRNA expression levels of p53, a senescence factor, interleukin (IL)-6, matrix metalloproteinases (MMP)-9, and nuclear factor kappa B1 (NF-κB1) as SASP factors and Bcl-2 and Bax as senolytic markers were assessed using quantitative reverse transcription-real-time polymerase chain reaction. Cells were double-stained with acridine orange and propidium iodide to observe the cell morphology. Results: TMZ increased the senescence rate of GB cells (p<0.001). Besides, OLE + TMZ reduced the proportion of senescent cells (p<0.001) and their capability to form colonies compared to TMZ-only-treated cells. Additionally, OLE + TMZ co-treatment elevated the mRNA expression levels of MMP-9, IL-6, NF-κB1, p53, and the Bax/Bcl-2 ratio compared to TMZ-only treatment. Especially in U87MG cells, involvement of OLE in TMZ treatments increased more than six times in the Bax/Bcl-2 ratio compared to TMZ-only, which induced the apoptosis-like morphological features (p<0.0001). Conclusion: Collectively, our findings presented the inhibitory effect of OLE on TMZ-mediated SASP-factor production in GB and, accordingly, its potential contribution to elongate the time of recurrence.

Diabetes Mellitus-Mediated MALAT1 Expression Induces Glioblastoma Aggressiveness.

AIM: To describe the role of metastasis-associated lung adenocarcinoma transcript 1 (MALAT1) in glioblastoma (GB) progression in patients concurrently diagnosed with diabetes mellitus (DM). MATERIAL AND METHODS: Formalin-fixed paraffin-embedded (FFPE) tumor samples of 47 patients diagnosed with GB only and 13 patients diagnosed with GB and DM (GB-DM) were enrolled in this study. Data for p53 and Ki67 immunohistochemical staining of the tumors and blood HbA1c levels of patients with DM were retrospectively collected. MALAT1 expression was assessed using quantitative real-time polymerase chain reaction. RESULTS: The coexistence of GB and DM induced the nuclear expression of p53 and Ki67 compared with GB only. MALAT1 expression was higher in GB-DM tumors than in GB only tumors. The expression of MALAT1 and HbA1c levels were positively correlated. Additionally, MALAT1 was positively correlated with tumoral p53 and Ki67. The disease-free survival of patients with GB-DM with high MALAT1 expression was shorter than that of those diagnosed with GB only and with a lower MALAT1 expression. CONCLUSION: Our findings suggest that one of the mechanisms of the facilitating effect of DM on GB tumor aggressiveness is via MALAT1 expression.

Degenerative changes of the mandibular condyle in relation to the temporomandibular joint space, gender and age: A multicenter CBCT study.

BACKGROUND: Cone-beam computed tomography (CBCT) is used to provide multiplanar views of the temporomandibular joint (TMJ) bone components as well as TMJ pathologies without superposition, magnification or distortion. OBJECTIVES: The study aimed to analyze degenerative changes in the condylar surface, and their relationship with patient age and gender, and the TMJ space measurements by using CBCT images. MATERIAL AND METHODS: A total of 258 individuals were retrospectively analyzed. The degenerative bone changes of the condylar head were evaluated and classified on the right and left sides. The shortest distances from the anterior, superior and posterior parts of the condylar head to the glenoid fossa were measured to represent the TMJ space. Univariate and multivariate logistic regression analyses then evaluated the effect of age and gender on the presence of degenerative changes. RESULTS: Condylar flattening was most frequently observed (413 TMJs, 53.5%). However, the presence or absence of the change types did not differ according to the sides. The mean values of the TMJ space measurements on the right and left sides were narrower in the group with changes than in the group without changes. Nonetheless, no statistically significant difference in the TMJ space was found between the groups (p > 0.05). CONCLUSIONS: An increased risk of radiographically detectable degenerative alterations in left TMJs was detected for males and for increasing age. Degenerative changes in the condylar surface may affect the dimensions of the TMJ space.

Olea europaea Leaf Phenolics Oleuropein, Hydroxytyrosol, Tyrosol, and Rutin Induce Apoptosis and Additionally Affect Temozolomide against Glioblastoma: In Particular, Oleuropein Inhibits Spheroid Growth by Attenuating Stem-like Cell Phenotype.

The effects of Olea europaea leaf extract (OLE) phenolics, including oleuropein (OL), hydroxytyrosol (HT), tyrosol (TYR), and rutin against glioblastoma (GB), independently and in combination with temozolomide (TMZ), were investigated in T98G and A172 cells. Cell growth was assessed by WST-1, real-time cell analysis, colony formation, and cell cycle distribution assays. A dual acridine orange propidium iodide (AO/PI) staining and annexin V assay determined cell viability. A sphere-forming assay, an intracellular oxidative stress assay, and the RNA expression of CD133 and OCT4 investigated the GB stem-like cell (GSC) phenotype. A scratch wound-healing assay evaluated migration capacity. OL was as effective as OLE in terms of apoptosis promotion (p < 0.001) and GSC inhibition (p < 0.001). HT inhibited cell viability, GSC phenotype, and migration rate (p < 0.001), but its anti-GB effect was less than the total effect of OLE alone. Rutin decreased reactive oxygen species production and inhibited colony formation and cell migration (p < 0.001). TYR demonstrated the least effect. The additive effects of OL, HT, TYR and rutin with TMZ were significant (p < 0.001). Our data suggest that OL may represent a novel therapeutic approach against GB cells, while HT and rutin show promise in increasing the efficacy of TMZ therapy.

A comparative study of paranasal sinus and nasal cavity anatomic variations between the Polish and Turkish Cypriot Population with CBCT.

BACKGROUND: Genetic and environmental factors especially climatic conditions are thought to influence the shape and size of the paranasal sinuses and anatomic variations may create both a diagnostic and therapeutic challenge. However, no study has been published about the climatic adaptation of the paranasal sinus region in different populations. This study aimed to compare the prevalence of anatomical variants in the paranasal sinus and nasal cavity using Cone-Beam Computed Tomography (CBCT) between Polish and Turkish Cypriot populations. METHODS: The material consisted of volumes acquired utilizing Galileos (Sirona, Germany) as well as Newtom 3G (QR Verona, Newtom, Italy) CBCT units. There were examined 356 Polish and 359 Turkish Cypriot patients in whom paranasal sinuses were included in the field of view. Paranasal sinus anatomic variations were assessed in both populations. RESULTS: In the Polish population, the most common anatomic variation was septum deviation followed by the Agger nasi cell and concha bullosa with a prevalence of 87.7%, 83.2%, and 54.8% respectively. For the Turkish Cypriot population, the most common anatomic variation was Agger nasi cell followed by concha bullosa and supraorbital ethmoid cells with a prevalence of 81.6%, 68%, and 57.8% respectively. Many anatomic variations were found to show substantial differences among both populations. Incidence rates of hyperpneumatization of the frontal sinus, septum pneumatization, supraorbital ethmoid cells, concha bullosa, uncinate bulla, and internal carotid artery protrusion into the sphenoid sinus were significantly higher in the Turkish Cypriot group, while the incidence of Haller cell, frontal sinus hypoplasia, maxillary sinus hypoplasia, ethmomaxillary sinus, sphenomaxillary plate, and septum deviation were significantly higher in Polish population. CONCLUSION: According to the Köppen-Geiger world climatic map, the climate is warmer and drier in Turkish Cypriote populations than in the Polish population. These climatic differences influence the paranasal sinus variations between the Turkish Cypriot and Polish populations that must be taken into account by rhinologic surgeons especially when performing frontal and sphenoid sinus surgery.

Blood-Based Biomarkers in Afp Normal/Stable Hepatocellular Carcinoma: Diagnostic and Prognostic Relevance of Mir-10b for Patients on Liver Transplant List.

BACKGROUND: As a diagnostic criteria of hepatocellular carcinoma (HCC), the exact threshold of alpha-fetoprotein (AFP) is controversial. In additional, not all HCC tumors are AFP positive or secrete elevated amounts of AFP into the serum. However, the diagnosis of HCC is quite important on the liver transplant list. Therefore, the purpose of this study was to investigate the expression of circulating micro RNAs (miRNAs) in AFP-stable HCC patients. Thus, we aimed to determine a diagnostic biomarker in these patients. METHODS: Sixteen miRNAs were evaluated using a real-time quantitative reverse transcription polymerase chain reaction system in AFP-stable HCC and AFP-trending HCC patients. RESULTS: In our study, 46.7% (n = 28) of the patients diagnosed with HCC had stable/normal AFP levels. We detected that high expression of miR-24, miR-10b and the low expression of miR-143 were independently and significantly associated with HCC in AFP-stable compared with AFP trending (P < .05). Additionally, we demonstrated that the overexpression of miR-10b was associated with poor disease-free survival in HCC (P = 0.001). CONCLUSIONS: Although more clinical validations are needed for the diagnosis of HCC, our current results indicate that the coexistence of high expression of miR-10b and miR-24 may help clinicians adjust in the diagnosis of HCC in patients who are on the liver transplant list but awaiting biopsy for the diagnosis of HCC.

A Fused Deep Learning Architecture for the Detection of the Relationship between the Mandibular Third Molar and the Mandibular Canal.

The study aimed to generate a fused deep learning algorithm that detects and classifies the relationship between the mandibular third molar and mandibular canal on orthopantomographs. Radiographs (n = 1880) were randomly selected from the hospital archive. Two dentomaxillofacial radiologists annotated the data via MATLAB and classified them into four groups according to the overlap of the root of the mandibular third molar and mandibular canal. Each radiograph was segmented using a U-Net-like architecture. The segmented images were classified by AlexNet. Accuracy, the weighted intersection over union score, the dice coefficient, specificity, sensitivity, and area under curve metrics were used to quantify the performance of the models. Also, three dental practitioners were asked to classify the same test data, their success rate was assessed using the Intraclass Correlation Coefficient. The segmentation network achieved a global accuracy of 0.99 and a weighted intersection over union score of 0.98, average dice score overall images was 0.91. The classification network achieved an accuracy of 0.80, per class sensitivity of 0.74, 0.83, 0.86, 0.67, per class specificity of 0.92, 0.95, 0.88, 0.96 and AUC score of 0.85. The most successful dental practitioner achieved a success rate of 0.79. The fused segmentation and classification networks produced encouraging results. The final model achieved almost the same classification performance as dental practitioners. Better diagnostic accuracy of the combined artificial intelligence tools may help to improve the prediction of the risk factors, especially for recognizing such anatomical variations.

Early-stage colon cancer with high MALAT1 expression is associated with the 5-Fluorouracil resistance and future metastasis.

BACKGROUND: This study aimed to investigate the role of long noncoding RNA (LncRNA) expression profiles to predict relapse and 5-FU response in patients with stage I/II colon cancer (CC). METHODS AND RESULTS: The expression level of 15 LncRNA was analyzed in stage I/II colon tumors of 126 CC patients. To confirm the findings in-vitro, 5FU-resistant HT29 cells were generated by subjecting HT-29 cells to the increasing concentrations of 5FU for 6 months. The 5FU resistance was observed in WST-1 and Annexin V analyses. The colony formation and wound healing assays were assessed to determine the metastatic properties of the cells. Expression levels of LncRNAs and mRNA of EMT-related genes were determined by RT-PCR. The role of LncRNA on metastasis and 5FU sensitivity were confirmed in pcDNA3.0-PTENP1 and si-MALAT1 expressed 5FU-resistant HT29 cell lineages. RESULTS: High MALAT1 (p = 0.0002) and low PTENP1 (p = 0.0044) expressions were significantly associated with 5-FU resistance and tumor relapse in stage I/II CC. The invasiveness and colony-forming characteristics of 5-FU-resistant cell lineages were higher as compared to the parent HT-29. Moreover, the expression of MALAT1 (p = 0.0009) was increased while the expression of PTENP1 (p = 0.0158) decreased in 5FU-resistant-HT-29 cells. Si-MALAT1 treatment increased cell sensitivity to 5FU, whereas it decreased invasive behaviors of 5 FU-resistant-HT-29 cells. CONCLUSION: MALAT1 may be a biomarker in predicting recurrence in early-stage CC. Our findings suggest that a cell-based therapy to target MALAT1 could be established for these patients to prevent metastasis and 5-FU resistance.

Investigation of the Defective Growth Pattern and Multidrug Resistance in a Clinical Isolate of Candida glabrata Using Whole-Genome Sequencing and Computational Biology Applications.

Candida glabrata is increasingly isolated from blood cultures, and multidrug-resistant isolates have important implications for therapy. This study describes a cholesterol-dependent clinical C. glabrata isolate (ML72254) that did not grow without blood (containing cholesterol) on routine mycological media and that showed azole and amphotericin B (AmB) resistance. Matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) and whole-genome sequencing (WGS) were used for species identification. A modified Etest method (Mueller-Hinton agar supplemented with 5% sheep blood) was used for antifungal susceptibility testing. WGS data were processed via the Galaxy platform, and the genomic variations of ML72254 were retrieved. A computational biology workflow utilizing web-based applications (PROVEAN, AlphaFold Colab, and Missense3D) was constructed to predict possible deleterious effects of these missense variations on protein functions. The predictive ability of this workflow was tested with previously reported missense variations in ergosterol synthesis genes of C. glabrata. ML72254 was identified as C. glabrata sensu stricto with MALDI-TOF, and WGS confirmed this identification. The MICs of fluconazole, voriconazole, and amphotericin B were >256, >32, and >32 µg/mL, respectively. A novel frameshift mutation in the ERG1 gene (Pro314fs) and many missense variations were detected in the ergosterol synthesis genes. None of the missense variations in the ML72254 ergosterol synthesis genes were deleterious, and the Pro314fs mutation was identified as the causative molecular change for a cholesterol-dependent and multidrug-resistant phenotype. This study verified that web-based computational biology solutions can be powerful tools for examining the possible impacts of missense mutations in C. glabrata. IMPORTANCE In this study, a cholesterol-dependent C. glabrata clinical isolate that confers azole and AmB resistance was investigated using artificial intelligence (AI) technologies and cloud computing applications. This is the first of the known cholesterol-dependent C. glabrata isolate to be found in Turkey. Cholesterol-dependent C. glabrata isolates are rarely isolated in clinical samples; they can easily be overlooked during routine laboratory procedures. Microbiologists therefore need to be alert when discrepancies occur between microscopic examination and growth on routine media. In addition, because these isolates confer antifungal resistance, patient management requires extra care.

Comparison of Intracranial and Extracranial Carotid Artery Calcifications between Obstructive Sleep Apnea Patients and Healthy Individuals: A Combined Cone-Beam Computed Tomography and Polysomnographic Study.

Purpose: This study aimed to compare the presence and grades of intra- and extracranial carotid artery calcifications between obstructive sleep apnea (OSA) and non-OSA patients. Methods: CBCT records of 190 patients (95 OSA patients and 95 non-OSA patients) were retrospectively collected and analyzed. Patient demographic data, including age and gender for both study groups and body mass index (BMI), and apnea-hypopnea index (AHI) for OSA patients were recorded. The presence of intra- and extracranial carotid artery calcifications and the number of calcifications were noted according to the grading scale. Results: There was a significant difference in carotid artery calcifications between OSA patients and healthy individuals. A total of 56.8% of the OSA patients showed at least one carotid artery calcification, whereas 13.8% of healthy individuals showed at least one carotid artery calcification (p < 0.05). For intracranial calcifications, OSA patients showed a significantly higher prevalence than healthy individuals (p < 0.05). The results showed that as the apnea-hypopnea index increases in OSA patients, the incidence of carotid artery calcification increases simultaneously. AHI > 30 patients showed the highest percentage of calcifications. Conclusion: In conclusion, OSA patients showed a higher prevalence of calcified carotid artery calcifications than healthy individuals. The results can be interpreted as the higher AHI, the more carotid artery calcification occurs. As these lesions can be a precursor of future strokes, 3D MDCT/CBCT images should evaluate meticulously not only extracranial but also intracranially, especially in OSA patients.

AI-based automatic segmentation of craniomaxillofacial anatomy from CBCT scans for automatic detection of pharyngeal airway evaluations in OSA patients.

This study aims to generate and also validate an automatic detection algorithm for pharyngeal airway on CBCT data using an AI software (Diagnocat) which will procure a measurement method. The second aim is to validate the newly developed artificial intelligence system in comparison to commercially available software for 3D CBCT evaluation. A Convolutional Neural Network-based machine learning algorithm was used for the segmentation of the pharyngeal airways in OSA and non-OSA patients. Radiologists used semi-automatic software to manually determine the airway and their measurements were compared with the AI. OSA patients were classified as minimal, mild, moderate, and severe groups, and the mean airway volumes of the groups were compared. The narrowest points of the airway (mm), the field of the airway (mm2), and volume of the airway (cc) of both OSA and non-OSA patients were also compared. There was no statistically significant difference between the manual technique and Diagnocat measurements in all groups (p > 0.05). Inter-class correlation coefficients were 0.954 for manual and automatic segmentation, 0.956 for Diagnocat and automatic segmentation, 0.972 for Diagnocat and manual segmentation. Although there was no statistically significant difference in total airway volume measurements between the manual measurements, automatic measurements, and DC measurements in non-OSA and OSA patients, we evaluated the output images to understand why the mean value for the total airway was higher in DC measurement. It was seen that the DC algorithm also measures the epiglottis volume and the posterior nasal aperture volume due to the low soft-tissue contrast in CBCT images and that leads to higher values in airway volume measurement.

Frequency of azole resistance in clinical and environmental strains of Aspergillus fumigatus in Turkey: a multicentre study.

OBJECTIVES: Aspergillus fumigatus causes several diseases in humans and azole resistance in A. fumigatus strains is an important issue. The aim of this multicentre epidemiological study was to investigate the prevalence of azole resistance in clinical and environmental A. fumigatus isolates in Turkey. METHODS: Twenty-one centres participated in this study from 1 May 2018 to 1 October 2019. One participant from each centre was asked to collect environmental and clinical A. fumigatus isolates. Azole resistance was screened for using EUCAST agar screening methodology (EUCAST E.DEF 10.1) and was confirmed by the EUCAST E.DEF 9.3 reference microdilution method. Isolates with a phenotypic resistance pattern were sequenced for the cyp51A gene and microsatellite genotyping was used to determine the genetic relationships between the resistant strains. RESULTS: In total, resistance was found in 1.3% of the strains that were isolated from environmental samples and 3.3% of the strains that were isolated from clinical samples. Mutations in the cyp51A gene were detected in 9 (47.4%) of the 19 azole-resistant isolates, all of which were found to be TR34/L98H mutations. Microsatellite genotyping clearly differentiated the strains with the TR34/L98H mutation in the cyp51A gene from the strains with no mutation in this gene. CONCLUSIONS: The rate of observed azole resistance of A. fumigatus isolates was low in this study, but the fact that more than half of the examined strains had the wild-type cyp51A gene supports the idea that other mechanisms of resistance are gradually increasing.