Direct Detection and Identification of the Most Common Bacteria and Fungi Causing Otitis Externa by a Stepwise Multiplex PCR.

Background: Considering the importance of differential diagnosis of infectious otitis externa (OE), a stepwise PCR-based assay using universal and genus- or species-specific primers for the detection/identification of the most prevalent bacterial and fungal OE was developed and evaluated on the ear aspiration specimens of clinically suspected patients. Methods and Materials: A total of 120 ear aspiration specimens with otomycosis suspicion were subjected to manual DNA extraction using phenol-chloroform extraction after tissue digestion with a lysis buffer. The multiplex PCR was initially performed using pan-fungal and bacterial homemade primers. Pseudomonas and Staphylococcus specific primers were simultaneously used in one reaction mixture to identify the bacterial genera. Furthermore, for the identification of fungal agents, Candida species-specific multiplex primers targeting the most clinically important Candida species causing OE (i.e., C. albicans, C. parapsilosis, and C. auris), as well as Aspergillus related multiplex PCR identifying the most prevalent Aspergillus species were used in two separate reaction mixtures. All the results of multiplex PCR were interpreted based on the amplicon size. Results: The overall multiplex PCR-based detection rate of bacterial (n = 88; 73.3%) and fungal (n = 97; 81%) OE was documented to be 100% along with and complete consistency with the results of direct examination and Giemsa staining. Double amplicon bands of bacterial and fungal pathogens were evidenced in 76 specimens (63.3%). Moreover, the positivity rate of pan-fungal PCR was higher than that of the culture result. Out of 88 pan-bacterial positive PCR specimens, 66 and 47 ones were positive for Staphylococcus and Pseudomonas, respectively. In addition, 30 samples exhibited mixed infection of both, and five specimens remained negative. Out of 97 pan-fungal positive PCR specimens, 67 and 51 ones contained Candida and Aspergillus species, respectively. It should be noted that dual amplicon bands of Candida and Aspergillus-related multiplex PCR were yielded in 30 specimens. Conclusion: The stepwise multiplex PCR assay proved to be more sensitive, more rapid, as well as less cumbersome in detection and identification of fungal and bacterial OE, compared to culture.

Otomycosis Due to the Rare Fungi Talaromyces purpurogenus, Naganishia albida and Filobasidium magnum.

Otomycosis is a common finding in otorhinolaryngology clinics and is usually caused by species of Candida and Aspergillus, particularly black aspergilli. Meanwhile, other fungi can give rise to this infection, and the identification of these requires accurate methods. Here, we report three cases of otomycosis due to rare fungal pathogens. All the patients were young females, and manipulation of the ear canal was identified as a common potentially predisposing factor. In direct examination, filamentous fungal elements (in one case) and yeast cells (in two other cases) were seen. Culture was positive in all cases. Based on PCR-sequencing of internal transcribed spacers and ß-tubulin (for mold isolate), the isolated fungi were identified as Talaromyces purpurogenus, Naganishia albida and Filobasidium magnum. By susceptibility testing of the isolates to fluconazole, itraconazole, voriconazole and amphotericin B, the lowest minimum inhibitory concentration values were observed for amphotericin B followed by voriconazole. Patients were successfully treated by a combination of antifungals and corticosteroids with no relapse over the next year, except for the case due to F. magnum, in which, despite partial recovery, a course of relapse was reported in the 1-year follow-up call.

Molecular epidemiology of otomycosis in Isfahan revealed a large diversity in causative agents.

PURPOSE: To elucidate the clinical and microbial epidemiology of otomycosis in Isfahan, Iran. METHODOLOGY: From January 2016 to January 2017 all patients clinically suspected of otomycosis at Al-Zahra Hospital, Isfahan, Iran were recruited. Specimens were taken using sterile swabs by an otorhinolaryngologist and subjected to culture and microscopy using potassium hydroxide and Giemsa stain. Isolated fungi were identified based on morphological and molecular characteristics. RESULTS: Otomycosis was confirmed in 97/120 patients (80.8 %). Females (72.2 %) and patients aged 30-39 years (33 %) were more commonly affected than others. Manipulation of ear canal (62.9 %) was the most common predisposing factor. Pruritus was observed in 84.54  % of the patients followed by hearing impairment (81.4 %), and most episodes were detected over the summer (50.5 %). Culture was positive for 81 (83.5  %) of confirmed cases and molds were the most prevalent causative agents (n=51, 63 %) followed by yeasts (n=19, 23.4 %) and yeast/mold mixes (n=11, 13.6 %). For the 16 remaining patients, no growth was seen in culture despite a positive result on direct examination. In total, 92 isolates (63 molds and 29 yeasts) were recovered in culture. Application of molecular methods showed 18 fungal species and the vast majority of them belonged to Aspergillus (n=53, 57.6 %) and Candida genus. Among the species involved, Candida parapsilosis (n=22, 22.7 %) and Aspergillus tubingensis (n=15, 15.5 %) were the most encountered species. CONCLUSION: Outcomes from this study showed a different picture of prevalence, where C. parapsilosis and A. tubingensis but not Aspergillus niger were the most species encountered from patients suffering from otomycosis.

Comparing the effects of Botulinum Toxin-A and cetirizine on the treatment of allergic rhinitis.

BACKGROUND: There are few reports on the effects of intranasal Botulinum Toxin-A (BTX-A) as a treatment of allergic rhinitis (AR). In this study, we compared the efficacy of intranasal BTX-A to cetirizine in the treatment of AR. METHODS: Fifty AR patients at the age of 26.2 ± 9.1 years (64% females), were recruited to the trial according to the Allergic Rhinitis and its Impact on Asthma (ARIA) criteria. Participants randomly received either intranasal injection of BTX-A (75IU Dysport®) or cetirizine (10mg/day). Symptoms (based on the ARIA) and side effects were assessed every two weeks for two months. Quality of life was evaluated before and after the study using the Rhinasthma questionnaire. RESULTS: Total symptom severity score of patients significantly decreased (P < 0.001) and quality of life significantly improved (P < 0.001) at the same level in both groups. Side effects included nasal dryness (4%) and epistaxis (4%) in the BTX-A group. In the cetirizine group 44% sleepiness and 4% blurred vision was reported. CONCLUSIONS: Nasal injection of BTX-A shows the same therapeutic effects as cetirizine in the management of AR. Since BTX is expensive, we do not suggest it in the first line of treatment for AR. However, BTX-A is a potential treatment for patients who are resistant or not compliant to the routine medications of AR. Further studies are required to investigate implications and limitations of BTX-A in the treatment of AR.

Solitary laryngeal Kaposi sarcoma in a kidney transplant patient.

After the first description of Kaposi sarcoma in 1872, many cases of this tumor were reported worldwide. This tumor is multifocal and laryngeal involvement is considered to be as unusual site. Kaposi sarcoma is almost always are associated with classical skin lesion, and only about 5% of non-acquired immune deficiency syndrome Kaposi sarcomas are reported to be located in the larynx, and they are almost always associated with classical skin lesion. [corrected]. We report a kidney transplant recipient diagnosed with solitary laryngeal Kaposi sarcoma 21 months after transplantation, who was treated with combined surgery, chemotherapy, and immunosuppressive modification.

Low-level laser for treatment of tinnitus: a self-controlled clinical trial.

BACKGROUND: Despite the high prevalence and morbidity, tinnitus still remains an obscure symptom. We assessed the efficacy of low-level laser for treatment of tinnitus. METHODS: It was a self controlled clinical trial study on 61 outpatients with subjective tinnitus. The patients were irradiated with a 650-nm, 5-mW soft laser for twenty days and twenty minutes per day. The sensation of tinnitus was measured on a Visual Analog Scale (VAS) before and two weeks after treatment and they were compared by means of Wilcoxon signed ranktest. RESULTS: Thirty-eight (62.3%) patients were men and twenty-three (37.7%) were women. Fourteen patients (31.8%) worked in noisy environment. The VAS mean difference before and after the treatment was statistically significant (p < 0.0001). The best treatment effect was in the youngest group and there were significant differences between this group and the middle age and older groups (p = 0.018 and 0.001, respectively). The mean VAS score reduction was not statistically significant between male and female patients (p = 0.23). Also, the treatment outcome according to the noise level in patient's workplaces was not significantly different in women (p = 0.693), but it was significant in men (p = 0.029). CONCLUSIONS: Transmeatal low-level laser irradiation is effective for the treatment of tinnitus and some variables like age and job can affect the treatment outcome.

Effects of adenoidectomy on markers of endothelial function and inflammation in normal-weight and overweight prepubescent children with sleep apnea.

BACKGROUND: This trial study aimed to assess the effects of adenoidectomy on the markers of endothelial function and inflammation in normal-weight and overweight prepubescent children with obstructive sleep apnea (OSA). METHODS: This trial study was conducted in Isfahan, Iran in 2009. The study population was comprised of 90 prepubescent children (45 normal-weight and 45 overweight children), aged between 4-10 years old, who volunteered for adenoidectomy and had OSA documented by validated questionnaire. The assessment included filling questionnaire, physical examination, and laboratory tests; it was conducted before the surgery and was repeated two weeks and six months after the surgery. RESULTS: Out of the 90 children evaluated, 83 completed the 2-week evaluation and 72 patients continued with the study for the 6-month follow up. Markers of endothelial function, i.e., serum adhesion molecules including endothelial leukocyte adhesion molecule (E-selectin), intercellular cell adhesion molecule-1 (ICAM-1), and vascular cell adhesion molecule-1 (sVCAM-1), and the markers of inflammation, i.e., interleukin-6, and high-sensitive C-reactive protein (hsCRP) decreased significantly in both normal-weight and overweight children after both two weeks and six months. After six months, the total and LDL-cholesterol showed a significant decrease in the overweight children. CONCLUSIONS: The findings of the study demonstrated that irrespective of the weight status, children with OSA had increased levels of the endothelial function and inflammation markers, which improved after OSA treatment by adenoidectomy. This might be a form of confirmatory evidence on the onset of atherogenesis from the early stages of the life, and the role of inflammation in the process. The reversibility of endothelial dysfunction after improvement of OSA underscores the importance of primordial and primary prevention of chronic diseases from the early stages of the life.