Towards solving the genetic diagnosis odyssey in Iranian patients with congenital anomalies.

Understanding the underlying causes of congenital anomalies (CAs) can be a complex diagnostic journey. We aimed to assess the efficiency of exome sequencing (ES) and chromosomal microarray analysis (CMA) in patients with CAs among a population with a high fraction of consanguineous marriage. Depending on the patient's symptoms and family history, karyotype/Quantitative Fluorescence- Polymerase Chain Reaction (QF-PCR) (n = 84), CMA (n = 81), ES (n = 79) or combined CMA and ES (n = 24) were performed on 168 probands (66 prenatal and 102 postnatal) with CAs. Twelve (14.28%) probands were diagnosed by karyotype/QF-PCR and seven (8.64%) others were diagnosed by CMA. ES findings were conclusive in 39 (49.36%) families, and 61.90% of them were novel variants. Also, 64.28% of these variants were identified in genes that follow recessive inheritance in CAs. The diagnostic rate (DR) of ES was significantly higher than that of CMA in children from consanguineous families (P = 0·0001). The highest DR by CMA was obtained in the non-consanguineous postnatal subgroup and by ES in the consanguineous prenatal subgroup. In a population that is highly consanguineous, our results suggest that ES may have a higher diagnostic yield than CMA and should be considered as the first-tier test in the evaluation of patients with congenital anomalies.

Comparison of the Impacts of Benson Relaxation Technique and Foot Reflexology Massage on Sleep Quality of Patients with Systolic Heart Failure: A Randomized Clinical Trial.

Background: Cardiovascular diseases can affect sleep quality. The use of non-pharmacological methods to improve the sleep quality of heart failure patients is essential. Therefore, this study compared the effects of the Benson relaxation technique and foot reflexology massage on sleep quality of those patients. Materials and Methods: In this clinical trial study, 93 patients with systolic heart failure referred to Javad Al-Aeme heart clinic in Torbat Heydarieh were selected with purposive sampling method and divided into the foot reflexology massage, Benson relaxation technique, and control groups using the balanced blocking randomization method. The Benson relaxation technique and the foot reflexology massage were performed for 20 and 30 minutes three days per week for four weeks, respectively. The control group only received routine care. Sleep quality was measured by the Pittsburgh sleep quality index and then compared among the groups before and after the intervention. The data were analyzed using ANOVA, Tukey's post hoc tests, paired t test, and Chi-square at the significant level of p < 0.05. Results: After interventions, the mean (SD) quality of sleep significantly increased in both intervention groups (Reflexology: pre-intervention 10.80 (3.40), post-intervention 6.60 (3.10), Benson relaxation: pre-intervention 15.50 (2.40) post-intervention 12 (2.60), compared to the control group (pre-intervention 10.50 (1.90) post-intervention 9.40(1.70) (p < 0.001). There was no significant difference between the two intervention groups (p = 0.53). Conclusions: The interventions improved the quality of sleep in patients with systolic heart failure. Therefore, these methods can be used as a suitable complementary treatment to improve the quality of sleep by nurses and midwives.

Persistent Molecular Disease in Adult Patients With AML Evaluated With Whole-Exome and Targeted Error-Corrected DNA Sequencing.

PURPOSE: Persistent molecular disease (PMD) after induction chemotherapy predicts relapse in AML. In this study, we used whole-exome sequencing (WES) and targeted error-corrected sequencing to assess the frequency and mutational patterns of PMD in 30 patients with AML. MATERIALS AND METHODS: The study cohort included 30 patients with adult AML younger than 65 years who were uniformly treated with standard induction chemotherapy. Tumor/normal WES was performed for all patients at presentation. PMD analysis was evaluated in bone marrow samples obtained during clinicopathologic remission using repeat WES and analysis of patient-specific mutations and error-corrected sequencing of 40 recurrently mutated AML genes (MyeloSeq). RESULTS: WES for patient-specific mutations detected PMD in 63% of patients (19/30) using a minimum variant allele fraction (VAF) of 2.5%. In comparison, MyeloSeq identified persistent mutations above 0.1% VAF in 77% of patients (23/30). PMD was usually present at relatively high levels (>2.5% VAFs), such that WES and MyeloSeq agreed for 73% of patients despite differences in detection limits. Mutations in DNMT3A, ASXL1, and TET2 (ie, DTA mutations) were persistent in 16 of 17 patients, but WES also detected non-DTA mutations in 14 of these patients, which for some patients distinguished residual AML cells from clonal hematopoiesis. Surprisingly, MyeloSeq detected additional variants not identified at presentation in 73% of patients that were consistent with new clonal cell populations after chemotherapy. CONCLUSION: PMD and clonal hematopoiesis are both common in patients with AML in first remission. These findings demonstrate the importance of baseline testing for accurate interpretation of mutation-based tumor monitoring assays for patients with AML and highlight the need for clinical trials to determine whether these complex mutation patterns correlate with clinical outcomes in AML.

Design, molecular dynamics simulation, and investigation of the mechanical behavior of DNA origami nanotubes with auxetic and honeycomb structures.

Numerous applications of DNA origami nanotubes for load-bearing purposes necessitate the improvement of properties and mechanical behavior of these types of structures, as well as the use of innovative structures such as metamaterials. To this end, the present study aims to investigate the design, molecular dynamics (MD) simulation, and mechanical behavior of DNA origami nanotube structures consisting of honeycomb and re-entrant auxetic cross-sections. The results revealed both structures kept their structural stability. In addition, DNA origami based-nanotube with auxetic cross-section exhibits negative Poisson's ratio (NPR) under tensile loading. Furthermore, MD simulation results demonstrated that the values of stiffness, specific stiffness, energy absorption, and specific energy absorption in the structure with an auxetic cross-section are higher than that of a honeycomb cross-section, similar to their behavior in macro-scale structures. The finding of this study is to propose re-entrant auxetic structure as the next generation of DNA origami nanotubes. In addition, it can be utilized to aid scientists with the design and fabrication of novel auxetic DNA origami structures.Communicated by Ramaswamy H. Sarma.

Rapid discrimination of Candida species based on optical diffraction pattern.

Candidiasis occurs mainly in immunocompromised patients. Rapid detection of Candida species can play a major role in the successful management of fungal infections and treatment monitoring. Detection and discrimination of five common strains of Candida species was performed using the optical elastic scattering diffraction pattern of their colonies. Using laser light with 632 nm wavelength and the designed optical system, optical diffraction patterns of C. albicans (ATCC12261), C. tropicalis (ATCC20336), C. glabrata (15545), C. guilliermondii (20216), and C. parapsilosis (22019) were recorded, processed and analyzed. The results of our study show that based on the different structure of Candida species and dissimilar structure of their colonies, the difference between acquired diffraction patterns are recognizable. In addition, through extraction of statistical feature of the diffraction pattern images and using classification techniques, the detection and discrimination could be performed in a semi-automatic way. The analysis of the colonies of five different Candida species by the optical diffraction patterns generated from the interaction of the laser with colonies' structures demonstrated that the technique had the potential to be applied for the detection and discrimination of various species.

An Analysis of the Capturing and Passing Ability of a DNA Origami Nanocarrier with the Aid of Molecular Dynamics Simulation.

The present article aims to investigate the ability of a DNA origami nanocarrier to successfully capture a cargo using molecular dynamics (MD) simulation. In addition, the passage of the cargo through the nanocarrier was analyzed by steered molecular dynamics (SMD) simulation. The proposed DNA origami nanocarrier is a nanotube that consists of six double helices in which a positively charged nanocargo was placed. Since the stability of the nanocarrier has been considered one of the obstacles to nanocargo transportation, different cross-sectional areas of the nanocarrier were considered as measures to analyze its structural stability. The results eventually showed that the proposed nanocarrier is able to retain the cargo while maintaining its structural stability. The analysis also revealed that the presence of the cargo increases the structural stability in parts of the nanocarrier. SMD simulation demonstrated that a feasible amount of force is required to separate the cargo and pass it through the nanocarrier, which can provide useful information in the field of smart drug delivery.

Comparison of acidifying agents and clotrimazole for treatment of otomycosis: a comprehensive one-way mini-review.

Background and Purpose: This review aimed to compare the efficacy of acidifying agents and clotrimazole in the treatment of patients with otomycosis. Materials and Methods: The databases, including Research Gate, Google Scholar, ScienceDirect, Embase, Medline, Scopus, Cochrane, and library databases of clinical trials were searched in this study. The keywords were "Fungal Ear Infection" and "Otitis External" for otomycosis, "Clotrimazole", Lotrimin", "Mycelex", "Desenex", and "Clotrimaderm Mycoderm" for clotrimazole, and "Boric Acid Alcohol", "Alcohol-vinegar solution", Burow solution (Domeboro), "Isopropyl Alcohol", "VoSol" and "Acetic Acid" for acidifying agents. Regarding search strategy, a total of 53 studies were collected, 11 of which were maintained for assessment. Almost all studies were published after 1990. These articles discussed the role of clotrimazole and acidifying compounds in the treatment of otomycosis. Moreover, the route of administration, dosage, and side effects of these medications were highlighted. Results: Eight studies had similar results and claimed that clotrimazole has the best or most significant effect on the treatment of otomycosis for patients suffering from pain, itching, swelling, and hearing loss. Conclusion: Although all medications appear effective, there is a paucity of evidence to fully support the decision to choose between clotrimazole or acidifying agents for the treatment of otomycosis in terms of both efficacy and safety. However, in the biomedical field, the re-emerging investigation attention is due to the statements of a number of mechanisms defending the use of acidifying agents to treat mycosis (of antifungal-resistant species).

Meyerozyma guilliermondii species complex: review of current epidemiology, antifungal resistance, and mechanisms.

Meyerozyma guilliermondii has been accepted as a complex composed of Meyerozyma guilliermondii, Meyerozyma carpophila, and Meyerozyma caribbica. M. guilliermondii is a saprophyte detected on human mucosa and skin. It can lead to serious infections in patients with risk factors like chemotherapy, immunodeficiency, gastrointestinal or cardiovascular surgery, and oncology disorders. Most deaths related to M. guilliermondii infections occur in individuals with malignancy. In recent decades, incidence of M. guilliermondii infections is increased. Sensitivity of this microorganism to conventional antifungals (e.g., amphotericin B, fluconazole, micafungin and anidulafungin) was reduced. Prophylactic and empirical uses of these drugs are linked to elevated minimal inhibitory concentrations (MICs) of M. guilliermondii. Drug resistance has concerned many researchers across the world. They are attempting to discover appropriate solution to combat this challenge. This study reviews the most important mechanisms of resistance to antifungals developed by in M. guilliermondii species complex.

Anti-biofilm properties of eucalyptol in combination with antifungals against Candida albicans isolates in patients with hematological malignancy.

Oral candidiasis is a fungal infection caused mainly by Candida albicans and it is a major problem among hematologic malignancy patients. Biofilm formation is an attributable factor to both virulence and drug resistance of Candida species. The aim of the study was to evaluate the biofilm-producing ability of oral C. albicans isolates and to evaluate the inhibitory activity of eucalyptol on Candida biofilm, alone and in combination with antifungal agents. Samples were collected from the oral cavity of 106 patients with hematologic malignancy. The isolated yeasts were identified by PCR-sequencing. Then C. albicans isolates were analyzed for their biofilm-producing ability by crystal violet staining and MTT assay. The minimum biofilm inhibition concentrations (MBIC) of eucalyptol, amphotericin B, itraconazole, and nystatin and the in vitro interaction of eucalyptol with these drugs were tested according to CLSI-M-27-A3 protocol and checkerboard methods, respectively. From 106 patients, 50 (47.2%) were confirmed for oral candidiasis [mean ± SD age 39 ± 14 years; female 31 (62%) and male 19 (38%)]. C. albicans was isolated from 40 of 50 (80%) patients. From 40 C. albicans isolates, 24 (60%) and 16 (40%) were moderate and weak biofilm producer, respectively. The geometric mean MBIC of amphotericin B, itraconazole, nystatin and eucalyptol were 3.93 µg/mL, 12.55 µg/mL, 0.75 µg/mL and 798 µg/mL, respectively. Eucalyptol interacted synergistically with amphotericin B, itraconazole and nystatin against 12.5, 10, and 22.5% of isolates, respectively. Eucalyptol demonstrated promising activity against biofilm of C. albicans when tested alone or combined with antifungal drugs.

Effects of back exercises versus transcutaneous electric nerve stimulation on relief of pain and disability in operating room nurses with chronic non-specific LBP: a randomized clinical trial.

BACKGROUND: Low back pain (LBP) is one of the most common musculoskeletal disorders related to working. Due to the nature of nursing work, this problem is often seen in nurses, including those who work in the operating rooms. Depending on the cause, there are various surgical and non-surgical methods to treat LBP. The present study was aimed to compare the effect of two therapeutic methods of back exercises and transcutaneous electrical nerve stimulation (TENS) on the disability and pain of operating room nurses with LBP. METHODS: In this clinical trial forty-four eligible operating room nurses (30 women, 14 men, mean age: 37.86 ± 6.74) with chronic nonspecific LBP were randomly assigned to back exercises (including the strengthening and stretching exercise (n = 22)) or TENS (n = 22) groups by permuted block randomization method. These interventions were performed in both groups three sessions of 15 min per week for 6 weeks. The McGill pain questionnaire for back pain and the Oswestry disability questionnaire for disability assessment were completed immediately before and after the interventions. RESULTS: After 6 weeks, the mean of pain and disability decreased significantly in both groups compared to the baseline. Based on the results, significant decreases in the pain score (mean difference (95% CI): - 8.95 (- 12.77 to - 5.14); P-value < 0.001) and disability score (mean difference (95% CI): - 8.73(- 12.42 to - 5.03); P-value < 0.001) were revealed in the back exercises group after the intervention compared to the baseline. In addition, after the intervention in TENS group, the mean pain intensity and disability showed significant decrease, respectively (mean difference (95% CI): - 16.18 (- 19.81 to - 12.55); P-value < 0.001; mean difference (95% CI): - 15.82 (- 19.24 to - 12.40); P-value < 0.001). After adjusting for the baseline values, the TENS group had a significantly higher pain score reduction than the back exercises group (mean difference (95% CI): - 4.23 (- 8.03 to - 0.44); P-value =0.030; Cohen's d = 0.81). In addition, TENS led to a significant more decrease in the disability scores compared to the back exercises (mean difference (95% CI): - 3.99 (- 7.35 to - 0.64); P-value =0.021; Cohen's d = 0.73). Furthermore, a statistically significant time by group interaction effect on pain and disability score was found (interaction p < 0.001). CONCLUSION: Pain and disability were improved in both groups following 18 intervention sessions. However, pain and disability were improved to a greater extent in the TENS group than in the back exercises group. TRIAL REGISTRATION: The trial was retrospectively registered in the Iranian Registry of Clinical Trials ( www.irct.ir ) on 03/02/2019 as IRCT20180408039227N1 .

The Functionality of Apigenin as a Novel Cardioprotective Nutraceutical with Emphasize on Regulating Cardiac Micro RNAs:.

Cardiovascular diseases (CVDs) are considered the most common disorder and the leading cause of mortality globally. The etiology of CVDs depends on a variety of genetic and acquired parameters. Nowadays, a dramatic surge appeared in published reports to find the association between microRNAs (miRNAs) and CVDs in order to understand the cause of the disease, rapid diagnosis with the introduction of valid biomarkers, and target as a therapeutic approach. Apigenin is a novel nutraceutical flavonoid that cardioprotective properties are suggested. The current review aimed to evaluate the beneficial features of this phytochemical against CVDs with an emphasis on its ability to regulate the miRNAs. The findings demonstrated that Apigenin could regulate cardiac miRNAs, including miR-103, miR-122-5p, miR-15b, miR-155, and miR-33. Consequently, preventing CVDs is possible through different effects such as the promotion of cholesterol efflux, prevention of hyperlipidemia, alteration in ATP Binding Cassette Subfamily A Member 1 (ABCA1) levels, reducing of cardiocytes apoptosis, and retarding myocytes fibrosis. Also, it can regulate signaling pathways, protect against endothelial dysfunction, maintain oxidative balance, and decrease inflammatory factors and reactive oxygen species. Hence, apigenin regulatory characteristics affecting miRNAs expression could introduce this flavonoid as a novel cardioprotective phytochemical against different CVDs.

Enhancement of the Anti-biofilm Activity of Gold Nanoparticles- Itraconazole Conjugates in Resistant Candida glabrata.

INTRODUCTION: Onychomycosis, also called tinea unguium, is a common fungal infection affecting the nails. After dermatophytes, Candida species are recognized as second-line pathogens responsible for this infection. The treatment of onychomycosis requires a long time and is associated with high rates of recurrence. Antifungal medicines conjugated with gold (Au-NP) nanoparticle are the possible platforms for the reduction of drug resistance. METHODS: In the present study, we reported the in-vitro antifungal activity of itraconazole (ITZ) - Au conjugates, time-kill studies, and biofilm-producing ability of six ITZ-resistant C. glabrata. RESULTS: 3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium bromide (MTT) quantitative results revealed that four out of six resistant isolates studied able to form biofilms in vitro. ITZ-Au conjugates were more effective than ITZ or Au nanoparticle alone, and the time-kill tests pointed to the suitable effect of ITZ-Au conjugate. CONCLUSION: The present study concluded that ITZ-Au conjugates have an inhibitory effect on the biofilm of resistant C. glabrata isolates. Further studies are needed to compare the ex-vivo onychomycosis model.

Activities of Nanoparticles Against Fluconazole-Resistant Candida parapsilosis in Clinical Isolates.

Candida parapsilosis is a non-albicans Candida spp. associated with bloodstream infections in critically ill patients. Failure to treat it effectively due to delay in diagnosis often leads to serious illnessess. The present research aimed to investigate the antifungal activities of nanoparticles (NPs) against fluconazole-resistant C. parapsilosis strains. Ten strains were used from archived clinical isolates. Antifungal activities of NPs were examined based on the Clinical and Laboratory Standards Institute (M27-A3/S4) guideline. The morphological changes of strains exposed to each NP were observed by scanning electron microscope (SEM). The effect of NP on the membrane permeability of C. parapsilosis and the viability of the cells was assessed using the confocal laser scanning microscopy and 3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay, respectively. The cytotoxicity was evaluated against three mammalian cell lines. Minimum Inhibitory Concentration of NPs of 10 strains was in the concentration range of 0.5-4 µg/mL; these results were confirmed with the viability test. The antifungal activity of synthesized silver NPs (AgNPs) against resistant C. parapsilosis was greater in comparison with the gold NPs (AuNPs). The SEM images indicated a difference in the fungal morphology of the fungi. The propidium iodide uptake by C. parapsilosis cells showed concentration-dependent mortality in NPs treatment with a confocal laser scanning microscope. There was a notable difference (p < 0.01) in the cell viability in the concentration range of 0.5-4 µg/mL between NPs based on the MTT assay. In addition, these NPs exhibited very low toxicity for three mammalian cell lines, specially at 0.5 µg/mL. AgNPs and AuNPs had fungicidal activities against fluconazole-resistant C. parapsilosis strains. It is crucial to have knowledge based on fundamental research to find new ways to overcome resistant microorganisms.

A Study on the Bending Stiffness of a New DNA Origami Nano-Joint.

The present article aims to investigate the mechanical properties of a new DNA origami nano-joint using the steered molecular dynamics (SMD) simulation. Since the analysis of mechanical properties is of great importance in bending conditions for a nano-joint, the forces are selected to achieve angular changes in the joint by the resultant torque. In this study, the nano-joint is considered as a beam in order to use mechanical equations to extract the mechanical properties of the designed nano-joint. In addition, the bending stiffness of the beam is investigated in different modes of deflection using the Euler-Bernoulli beam theory. The results revealed that the value of bending stiffness increases with increasing deflection, and the changes in the bending stiffness relative to the deflection is linear. The proposed DNA origami nano-joint can be used as a joint in nanorobots and can be effectively applied in nanorobotic systems to move different components.

Caspofungin resistance in clinical Aspergillus flavus isolates.

INTRODUCTION AND AIMS: The present study was conducted to determine the candidate genes involved in caspofungin (CAS) resistance in clinical isolates of Aspergillus flavus (A. flavus). MATERIALS AND METHODS: The antifungal susceptibility assay of the CAS was performed on 14 clinical isolates of A. flavus using the CLSI-M-38-A2 broth micro-dilution protocol. Since CAS had various potencies, the minimum effective concentration (MEC) of anidulafungin (AND) was also evaluated in the present study. The FKS1 gene sequencing was conducted to assess whether mutations occurred in the whole FKS1 gene as well as hot spot regions of the FKS1 gene of the two resistant isolates. A complementary DNA-amplified fragment length polymorphism (CDNA-AFLP) method was performed to investigate differential gene expression between the two resistant and two sensitive clinical isolates in the presence of CAS. Furthermore, quantitative real-time PCR (QRT-PCR) was utilized to determine the relative expression levels of the identified genes. RESULTS: No mutations were observed in the whole FKS1 gene hot spot regions of the FKS1 genes in the resistant isolates. A subset of two genes with known biological functions and four genes with unknown biological functions were identified in the CAS-resistant isolates using the CDNA-AFLP. The QRT-PCR revealed the down-regulation of the P-type ATPase and ubiquinone biosynthesis methyltransferase COQ5 in the CAS-resistant isolates, compared to the susceptible isolates. CONCLUSION: The findings showed that P-type ATPase and ubiquinone biosynthesis methyltransferase COQ5 might be involved in the CAS-resistance A. flavus clinical isolates. Moreover, a subset of genes was differentially expressed to enhance fungi survival in CAS exposure. Further studies are recommended to highlight the gene overexpression and knock-out experiments in A. flavus or surrogate organisms to confirm that these mentioned genes confer the CAS resistant A. flavus.

A Review on Molecular Mechanisms of Antifungal Resistance in Candida glabrata: Update and Recent Advances.

Candida glabrata is the second frequent etiologic agent of mucosal and invasive candidiasis. Based on the recent developments in molecular methods, C. glabrata has been introduced as a complex composed of C. glabrata, Candida nivariensis, and Candida bracarensis. The four main classes of antifungal drugs effective against C. glabrata are pyrimidine analogs (flucytosine), azoles, echinocandins, and polyenes. Although the use of antifungal drugs is related to the predictable development of drug resistance, it is not clear why C. glabrata is able to rapidly resist against multiple antifungals in clinics. The enhanced incidence and antifungal resistance of C. glabrata and the high mortality and morbidity need more investigation regarding the resistance mechanisms and virulence associated with C. glabrata; additional progress concerning the drug resistance of C. glabrata has to be further prevented. The present review highlights the mechanism of resistance to antifungal drugs in C. glabrata.

Comparison of Antifungal Properties of Gold, Silver, and Selenium Nanoparticles Against Amphotericin B-Resistant Candida glabrata Clinical Isolates.

BACKGROUND: The present study aimed to investigate the antifungal activity of Nanoparticles (NPs) against amphotericin B-resistant Candida glabrata (C. glabrata) strains. METHODS: Twelve resistant (C. glabrata) strains were isolated from archived clinical isolates. Antifungal activity was conducted according to Clinical and Laboratory Standards Institute's (CLSI) guidelines, document M27-A3/S4. The Scanning Electron Microscope (SEM) was used to observe the morphological changes of strains exposed to each nanoparticle. RESULTS: Minimum Inhibitory Concentration (MIC) of nanoparticles of all strains was in the concentration range of 0.125 to 0.5 µg/Ml. The synthesized Ag-NPs showed superior antifungal activity against (C. glabrata) compared to Se-NPs and Au-NPs. The scanning electron microscope images revealed the difference in the fungal morphology between the untreated and treated fungi with nanoparticles. CONCLUSION: The Ag-NPs, followed by Se-NPs synthesized, revealed significant anti-fungal activity against resistance regardless of their antifungal-resistant mechanisms.

Multidrug-resistant Trichophyton mentagrophytes genotype VIII in an Iranian family with generalized dermatophytosis: report of four cases and review of literature.

BACKGROUND: The global spread of terbinafine-resistant Trichophyton mentagrophytes with point mutations in the squalene epoxidase (SQLE) gene is a big concern. AIM: The present study presents a series of unusual familial cases of generalized dermatophytosis caused by multidrug-resistant T. mentagrophytes genotype VIII. METHODS: Initially, the skin samples of each patient were taken and then subjected to direct microscopy and culture in Mycosel Agar. The molecular identification of Trichophyton species (spp.) was performed for all family members. In addition, the immunologic tests were requested, and an antifungal susceptibility test was carried out using the broth microdilution protocol based on the Clinical and Laboratory Standards Institute M38, third edition. The SQLE gene for a terbinafine-resistant T. mentagrophytes genotype VIII was sequenced and confirmed its nucleotide sequence to KU242352 as a susceptible strain. RESULTS: Based on the results of mycological examination and ITS rDNA sequencing, the etiologic agent was identified as T. mentagrophytes as a zoophilic dermatophyte. This species showed multiple drug resistance in vitro against terbinafine (minimum inhibitory concentration (MICs ≥8 µg/ml), itraconazole (MIC ≥4), and fluconazole (MIC ≥16). The SQLE gene of the isolate was subjected to sequencing for mutation, which showed a point mutation as TTC/TTA in the gene leading to Phe397Leu amino acid substitution in the enzyme. Only one of the family members responded to itraconazole and was cured after the long-term use of itraconazole. Other family members were treated with oral voriconazole with no recurrence. CONCLUSION: The transmission of this resistant T. mentagrophytes to other countries due to globalization is a serious issue to be considered.

Contribution of CTCF binding to transcriptional activity at the HOXA locus in NPM1-mutant AML cells.

Transcriptional regulation of the HOXA genes is thought to involve CTCF-mediated chromatin loops and the opposing actions of the COMPASS and Polycomb epigenetic complexes. We investigated the role of these mechanisms at the HOXA cluster in AML cells with the common NPM1c mutation, which express both HOXA and HOXB genes. CTCF binding at the HOXA locus is conserved across primary AML samples, regardless of HOXA gene expression, and defines a continuous chromatin domain marked by COMPASS-associated histone H3 trimethylation in NPM1-mutant primary AML samples. Profiling of the three-dimensional chromatin architecture in primary AML samples with the NPM1c mutation identified chromatin loops between the HOXA cluster and loci in the SNX10 and SKAP2 genes, and an intergenic region located 1.4 Mbp upstream of the HOXA locus. Deletion of CTCF binding sites in the NPM1-mutant OCI-AML3 AML cell line reduced multiple long-range interactions, but resulted in CTCF-independent loops with sequences in SKAP2 that were marked by enhancer-associated histone modifications in primary AML samples. HOXA gene expression was maintained in CTCF binding site mutants, indicating that transcriptional activity at the HOXA locus in NPM1-mutant AML cells may be sustained through persistent interactions with SKAP2 enhancers, or by intrinsic factors within the HOXA gene cluster.