Neutrophils phagocytosing fungal hyphae in urinary sediment / Neutrófilos fagocitando hifas no sedimento urinário

Abstract The Phagocytosis of fungal structures by neutrophils is a well-documented function of these immune cells. However, neutrophil phagocytosis of hyphal structures in the urine sediment is not usually observed during routine sample evaluation. This is a case of hyphal phagocytosis by neutrophils in the urine of a kidney allograft recipient patient.

Resumo A fagocitose de estruturas fúngicas por neutrófilos é uma função bem documentada destas células imunes. No entanto, a fagocitose de hifas por neutrófilos no sedimento urinário não é normalmente observada durante avaliação de rotina de amostras. Este é um caso de fagocitose de hifas por neutrófilos na urina de um paciente receptor de aloenxerto renal.

Presence of hyphae in chromoblastomycosis examinations: an enigma to be solved

Abstract The detection of muriform cells in direct mycological or anatomopathological examination is considered pathognomonic for chromoblastomycosis. The morphological aspect that these fungal structures acquire were called "Borelli spiders", when associated with hyphae. Reports of this association have been described for decades, initially related to more pathogenic agents of this mycosis. More recent studies have shown aspects related to the host's immunity that participate in this process, as well as an association with a worse disease prognosis. The present study discloses the findings of complementary examinations with the presence of "Borelli's spiders" in a patient diagnosed with chromoblastomycosis.

Effect of berberine hydrochloride on cell wall integrity of Candida albicans hypha / 中国中药杂志

The aim of this paper was to investigate the effect of berberine hydrochloride on the cell wall integrity of Candida albicans hypha. The minimal inhibitory concentration(MIC) of berberine hydrochloride against clinical and standard C. albicans strains was detected by micro liquid-based dilution method; the effect of berberine hydrochloride on the colony formation of C. albicans SC5314 was investigated by spot assay; the effect of berberine hydrochloride on the metabolism of C. albicans SC5314 hypha was checked by XTT reduction assay, and the viability of C. albicans SC5314 hypha was tested by fluorescent staining assay. The effect of berberine hydrochloride on the morphology of C. albicans SC5314 hypha was examined by scanning electron microscope. The changes in the cell wall of C. albicans SC5314 hypha after berberine hydrochloride treatment were detected by transmission electron microscopy. The effect of berberine hydrochloride on β-glucan from C. albicans SC5314 was detected by flow cytometry. The effect of berberine hydrochloride on hypha-specific gene ECE1 and β-glucan synthase genes FKS1 and FKS2 in C. albicans was examined by qRT-PCR. The results showed that berberine hydrochloride showed a strong inhibitory effect on both clinical and standard strains of C. albicans, and the MIC was 64-128 μg·mL~(-1). Spot assay, XTT redunction assay and fluorescent staining assay showed that with the increase of berberine hydrochloride concentration, the viability of C. albicans SC5314 gradually decreased. The transmission electron microscopy scanning assay showed that this compound could cause cell wall damage of C. albicans. The flow cytometry analysis showed the exposure degree of C. albicans β-glucan. The qRT-PCR further showed that berberine hydrochloride could significantly down-regulate hypha-specific gene ECE1 and β-glucan synthase-related gene FKS1 and FKS2. In conclusion, this compound can down-regulate C. albicans and β-glucan synthase-related gene expressions, so as to destroy the cell wall structure of C. albicans, expose β-glucan and damage the integrity of the wall.

Cladophialophora lanosa sp. nov., a New Species Isolated from Soil

Cladophialophora is characterized by simple hyphomycetes with brown hyphae that give rise to branched chains of pale brown conidia and shows affinities with the Herpotrichiellaceae. A fungal strain belonging to the genus Cladophialophora was isolated from soil in Daegu, Korea. This strain produces numerous greenish to dark black lanose aerial mycelia with hair like structures. It is morphological similar to C. chaetospira, C. inabaensis, and C. multiseptata; however, the conidiophores and conidia sizes of the newly isolated strain (KNU16-032) are clearly different from them. The novelty of the strain was also confirmed based on phylogenetic analysis using the data sets of the internal transcribed spacer region of and the partial sequence of 28S ribosomal DNA region along with the cultural characteristics. Because morphological observations and phylogenetic analysis indicated that the strain is distinct from previously known Cladophialophora species, we propose this species as a new species Cladophialophora lanosa sp. nov., and provide the detailed descriptions in this study.

AcuD Gene Knockout Attenuates the Virulence of Talaromyces marneffei in a Zebrafish Model

Talaromyces marneffei is the only dimorphic species in its genus and causes a fatal systemic mycosis named talaromycosis. Our previous study indicated that knockdown of AcuD gene (encodes isocitrate lyase of glyoxylate bypass) of T. marneffei by RNA interference approach attenuated the virulence of T. marneffei, while the virulence of the AcuD knockout strains was not studied. In this study, T. marneffei-zebrafish infection model was successfully established through hindbrain microinjection with different amounts of T. marneffei yeast cells. After co-incubated at 28°C, the increasing T. marneffei inoculum doses result in greater larval mortality; and hyphae generation might be one virulence factor involved in T. marneffei-zebrafish infection. Moreover, the results demonstrated that the virulence of the ΔAcuD was significantly attenuated in this Zebrafish infection model.

Inhibitory effect of extract of Coptidis Rhizoma on invasion of Candida albicans hyphae in vitro / 中国中药杂志

The aim of this paper was to investigate the inhibitory effect of extract of Coptidis Rhizoma(ECR) on invasion of Candida albicans hyphae in vitro.XTT reduction method was used to evaluate the metabolic activity of C.albicans.The colony edge growth of C.albicans was observed by solid medium.The growth of C.albicans hyphae was determined on semi-solid medium.The morphology and viability changes of C.albicans hyphae were assessed by scanning electron microscope and fluorescence microscope.qRT-PCR method was used to detect the ALS3 and SSA1 expression of C.albicans invasin genes.The results showed that the metabolic viability by XTT method detected that the activity of C.albicans was gradually decreased under the intervention of 64,128 and 256 mg·L-1 of ECR respectively.128,256 mg·L-1 of ECR significantly inhibited colony folds and wrinkles on solid medium and the hyphal invasion in semi-solid medium.Scanning electron microscopy and fluorescence microscopy showed that 128,256 mg·L-1 of ECR could inhibit the formation of C.albicans hyphae.qRT-PCR results showed that the expression of invasin gene ALS3 and SSA1 was down-regulated,and especially 256 mg·L-1 of ECR could down-regulate the two genes expression by 4.8,1.68 times respectively.This study showed that ECR can affect the invasiveness of C.albicans by inhibiting the growth of hyphae and the expression of invasin.

Single Dose Topical Application of Clotrimazole for the Treatment of Otomycosis: Is This Enough?

BACKGROUND AND OBJECTIVES: To determine the efficacy of filling the external auditory meatus with 1% clotrimazole at a single visit for the treatment of otomycosis. SUBJECTS AND METHODS: This prospective study included 40 patients who were referred to our clinic with complaints of ear itching, pain, and fullness, and were diagnosed with unilateral otomycosis. After cleaning the mycotic hyphae from the external auditory meatus, the ear canal was filled with 1% clotrimazole, using an intravenous catheter and syringe. The patients received follow-up examinations on post-treatment days 7, 15, and 45. RESULTS: The follow-up otomicroscopic examinations revealed that 95% of the ear canals were entirely clean and that all symptoms had resolved. The post-treatment scores of pain, aural fullness and itching were significantly lower than the pre-treatment scores (p < 0.01). CONCLUSIONS: Filling the external auditory meatus with 1% clotrimazole at a single visit is an easy, efficient, and cost-effective treatment for otomycosis. Additionally, high patient compliance makes this treatment superior to long-term topical therapy.

A new coumarin derivative, 4-acetatecoumarin, with antifungal activity and association study against Aspergillus spp

Abstract Fungal infections have become a concern for health professionals, and the emergence of resistant strains has been reported for all known classes of antifungal drugs. Among the fungi causing disease, we highlight those that belong to the genus Aspergillus. For these reasons, the search for new antifungals is important. This study examines the effects of a coumarin derivative, 4-acetatecoumarin (Cou-UMB16) both alone and together with antifungal drugs, and its mode of action against Aspergillus spp. Cou-UMB16 was tested to evaluate its effects on mycelia growth, and germination of Aspergillus spp. fungal conidia. We investigated its possible action on cell walls, on the cell membrane, and also the capacity of this coumarin derivative to enhance the activity of antifungal drugs. Our results suggest that Cou-UMB16 inhibits Aspergillus spp. virulence factors (mycelia growth and germination of conidia) and affects the structure of the fungal cell wall. When applying Cou-UMB16 in combination with azoles, both synergistic and additive effects were observed. This study concludes that Cou-UMB16 inhibits mycelial growth and spore germination, and that the activity is due to its action on the fungal cell wall, and that Cou-UMB16 could act as an antifungal modifier.

A Case Of Cavernous Sinus Syndrome and Mutifocal Cerebral Infarction Related To Mucormycosis Of Sphenoid Sinus / 고신대학교의과대학학술지

A 54-year-old man, suffering from severe headache and ophthalmoplegia after undergoing endoscopic sinus surgery was referred to a tertiary hospital. Computed tomography (CT) revealed soft tissue density lesions in the left sphenoid sinus. The internal carotid artery was shown to be occluded in brain magnetic resonance imaging (MRI) scans without any other cerebral lesion. Endoscopic view of left nasal cavity shows whitish hyphae in the ethmoid and the sphenoid sinuses. We diagnosed him with cavernous sinus syndrome caused by mucormycosis and conducted endoscopic sinus surgery to remove remaining lesions and decompress orbit and optic nerves. After the revision surgery the patient's headache and ophthalmoplegia were improved. However, multifocal cerebral infarctions were newly discovered in a postoperative CT scan. We experienced a case of mucormycosis of sphenoid sinus resulting in occlusion of internal carotid artery and multifocal cerebral infarction, and report it with a brief review of these disease entities.

Abducens Nerve Palsy and Optic Perineuritis Caused by Fungal Sphenoidal Sinusitis

PURPOSE: To report a case of abducens nerve palsy and optic perineuritis caused by fungal sphenoidal sinusitis. CASE SUMMARY: A 48-year-old male visited emergency department for retrobulbar pain, decreased vision, and horizontal diplopia for 3 days. He reported that previous medical history was non-specific, however, blood glucose level was 328 mg/dL (70–110). He had experienced severe headache for 7 days. The best corrected visual acuity was 20/20 at right eye and 20/25 at left eye. The pupil of left eye did not have relative afferent pupillary defect. Left mild proptosis was noted. The extraocular examination showed 30 prism diopters left esotropia at primary gaze and −4 abduction limitation of left eye. The left eye showed mild optic disc swelling and inferior field defect by field test. Brain magnetic resonance imaging showed enhancement of sphenoidal sinus, ethmoidal sinus, and around optic nerve at left eye. Three days after antibiotics treatment, the vision of left eye deteriorated to 20/40 and periorbital pain developed. The drainage and biopsy of sphenoidal sinus were performed. The histopathologic examination showed hyphae consistent with aspergillosis. The ocular symptoms were improved with anti-fungal treatment. Follow-up magnetic resonance imaging performed 1 month after treatment showed improvement of lesion at left orbit. Five months after surgery, the visual acuity of left eye was improved to 20/25. The patient showed orthotropia at primary gaze without limitation. CONCLUSIONS: The abducens nerve palsy and optic perineuritis can be caused by fungal sphenoidal sinusitis. The early diagnosis and appropriate treatment can lead to favorable outcome.

Liquid-Based Cytology of the Cerebrospinal Fluid in a Case of Cryptococcal Meningitis

Cryptococcus neoformans is the most common microorganism found in cerebrospinal fluid (CSF) cytology and causes life-threatening infections in immunocompromised hosts. Although its cytomorphologic features in conventional smear cytology have been well described, those in liquid-based cytology have rarely been. A 73-year-old woman with diffuse large B-cell lymphoma presented with mental confusion and a spiking fever. To rule out infectious conditions, CSF examination was performed. A cytology slide that was prepared using the ThinPrep method showed numerous spherical yeast-form organisms with diameters of 4–11 μm and thick capsules. Occasional asymmetrical, narrow-based budding but no true hyphae or pseudohyphae were observed. Gomori methenamine silver staining was positive. Cryptococcosis was confirmed in blood and CSF through the cryptococcal antigen test and culture. Liquid-based cytology allows for a clean background and additional slides for ancillary testing, facilitating the detection of microorganisms in CSF specimens, particularly when the number of organisms is small.

Successful Treatment of Invasive Gastric Mucormycosis in a Kidney Transplant Recipient / 대한이식학회지

Mucormycosis is an extremely rare but potentially life-threatening fungal infection. Gastrointestinal (GI) mucormycosis is very rare and occurs primarily in highly malnourished patients, especially in infants and children. A 55-year-old man with end-stage renal disease due to diabetic nephropathy, who had undergone deceased donor kidney transplantation 2 years prior, complained of abdominal pain and distension with a 3-day duration. Computed tomography revealed diffuse gastric wall thickening, and a huge amount of grey colored necrotic debris surrounded by erythematous erosive mucosa was observed at the antrum to upper body by GI endoscopy. The microscopic examination obtained from a GI endoscopic specimen demonstrated peptic detritus with numerous non-septate mucor hyphae in the mucosa and submucosa. Mucormycosis was diagnosed based on the clinical findings and morphological features. A total gastrectomy was performed and an antifungal agent was administered. A microscopic examination of the surgical specimen demonstrated invasive mucormycosis with numerous fungal hyphae with invasion into the mucosa to subserosa. The patient and graft were treated successfully by total gastrectomy and antifungal therapy.

Biocontrol of Orchid-pathogenic Mold, Phytophthora palmivora, by Antifungal Proteins from Pseudomonas aeruginosa RS1

Black rot disease in orchids is caused by the water mold Phytophthora palmivora. To gain better biocontrol performance, several factors affecting growth and antifungal substance production by Pseudomonas aeruginosa RS1 were verified. These factors include type and pH of media, temperature, and time for antifungal production. The results showed that the best conditions for P. aeruginosa RS1 to produce the active compounds was cultivating the bacteria in Luria-Bertani medium at pH 7.0 for 21 h at 37 °C. The culture filtrate was subjected to stepwise ammonium sulfate precipitation. The precipitated proteins from the 40% to 80% fraction showed antifungal activity and were further purified by column chromatography. The eluted proteins from fractions 9–10 and 33–34 had the highest antifungal activity at about 75% and 82% inhibition, respectively. SDS-PAGE revealed that the 9–10 fraction contained mixed proteins with molecular weights of 54 kDa, 32 kDa, and 20 kDa, while the 33–34 fraction contained mixed proteins with molecular weights of 40 kDa, 32 kDa, and 29 kDa. Each band of the proteins was analyzed by LC/MS to identify the protein. The result from Spectrum Modeler indicated that these proteins were closed similarly to three groups of the following proteins; catalase, chitin binding protein, and protease. Morphological study under scanning electron microscopy demonstrated that the partially purified proteins from P. aeruginosa RS1 caused abnormal growth and hypha elongation in P. palmivora. The bacteria and/or these proteins may be useful for controlling black rot disease caused by P. palmivora in orchid orchards.

Allergic bronchopulmonary mycosis – pathophysiology, histology, diagnosis, and treatment

Allergic bronchopulmonary mycosis (ABPM) develops mainly in patients with asthma or cystic fibrosis via types I and III hypersensitivity reactions to filamentous fungi. Aspergillus spp., especially Aspergillus fumigatus, is the major causative fungus because of its small conidia, thermophilic hyphae, and ability to secrete serine proteases. The cardinal histological feature of ABPM is allergic (eosinophilic) mucin-harboring hyphae in the bronchi, for which the formation of extracellular DNA trap cell death (ETosis) of eosinophils induced by viable fungi is essential. Clinically, ABPM is characterized by peripheral blood eosinophilia, increased IgE levels in the serum, IgE and IgG antibodies specific for fungi, and characteristic radiographic findings; however, there are substantial differences in the clinical features of this disease between East and South Asian populations. Systemic corticosteroids and/or antifungal drugs effectively control acute diseases, but recurrences are quite common, and development of novel treatments are warranted to avoid adverse effects and emergence of drug-resistance due to prolonged treatment with corticosteroids and/or antifungal drugs.

Death due to Aspergillus Tracheobronchitis: An Autopsy Case / 대한법의학회지

Aspergillus infection is the most common cause of death due to fungi in immunocompromised hosts. Aspergillus tracheobronchitis is an uncommon but severe form of invasive pulmonary aspergillosis, which is limited entirely or mainly to the tracheobronchial tree and can often be life-threatening. We report a case of a 54-year-old man who died from Aspergillus tracheobronchitis without an underlying disease. Autopsy revealed an extensive yellowish plaque adhering to the trachea and bronchial wall. The microscopic examination of the trachea and bronchus revealed septate branching hyphae of Aspergillus.

Contact lens-related bilateral and simultaneous Acremonium keratitis / Ceratite bilateral e simultânea por Acremonium relacionada ao uso de lentes de contato

Abstract This is a case of bilateral and simultaneous Acremonium keratitis related to intermittent and alternating eye soft contact lens use, which has a delayed diagnose, presented amphotericin B resistance with persistent hypopyon and had a positive response to topical natamycin. Besides the unusual presentation, there was no history of trauma or contact with vegetable matter, usually associated to the majority of cases of keratomicosys by filamentous fungi.

Resumo Trata-se de um caso de ceratite bilateral e simultânea por Acremonium relacionada ao uso intermitente e sem respeitar a lateralidade do uso de lentes de contato gelatinosas. Houve diagnóstico tardio, apresentando resistência a anfotericina B e hipópio persistente, com resposta positiva a natamicina tópica. Além da forma incomum de apresentação, não houve histórico de trauma ou contato com material vegetal, associado à maioria dos casos de ceratite por fungos filamentosos.

A Case of Aspergillosis and Actinomycosis of Each Side of Maxillary Sinuses / 대한이비인후과학회지

Aspergillosis or actinomycosis is a very rare disease of paranasal sinuses. It is an infectious bacterial disease caused by actinomyces species, which is a gram-positive, anaerobic bacillus. Although actinomyces species are part of the normal flora commensal in the oral cavity or GI tract, they, in rare cases, they can invade the mucosa and form a fistula or an abscess once the mucous membrane is damaged by inflammation or trauma. Aspergillosis is a fungal infection, and the frequency of its outbreaks has been increasing because diagnostic techniques have been improved, and the use of antibiotic, steroid and anticancer medicines have increased as well as the growth of incidence of metabolic diseases. The diagnosis of Aspergillosis is to be confirmed by pathologic findings after surgery. Sulfur granules are histologically observed for actinomycosis, and 45° branched hyphae are to be found for Aspergillosis. This describes the first case reported in Korea where actinomycosis and Aspergillosis was diagnosed separately at each side of maxillary sinuses.

A Case of Tinea Incognito Mimicking Folliculitis in Atopic Dermatitis Patient / 대한의진균학회지

A 27-year-old male with underlying history of atopic dermatitis presented with skin lesion on the trunk and upper extremity, occurring a month ago. Scattered erythematous multiple tiny maculopapules and patches were noticed. The lesion was first diagnosed as aggravated atopic dermatitis with folliculitis based on the clinical manifestation, but had failed to improve with antibiotics and medications for atopic dermatitis. KOH examinations were performed on his upper arm's lesion, and with detection of hyphae and isolation of Trichophyton rubrum by fungus culture, he was successfully treated with oral itraconazole 200 mg/day and topical flutrimazole. Tinea incognito refers to dermatophyte infections with atypical clinical presentation caused by previous treatment with steroid. Nowadays, the cases of tinea incognito have been increasing due to the increased usage of steroid and other immunosuppressants. Therefore, it is essential to include fungal infection in the differential diagnosis when cutaneous lesions are noticed in patients taking steroid and other immunosuppressants. Herein, we present a tinea incognito that clinically mimicked folliculitis in atopic dermatitis patient.

A Case of Laryngeal Candidiasis Presented as Laryngeal Leukoplakia

Candida exists in the oral cavity as normal flora, which is cultured in 7% of the population. And the development of candidiasis is usually related to the systemic or local immunosuppressed status such as diabetes, long-term antibiotics, steroid, radiation therapy or chemotherapy. However, isolated laryngeal candidiasis in immunocompetent patients is a rare entity with fewer than 40 cases reported in the world. Symptoms of laryngeal candidiasis are variable such as hoarseness, dysphagia, or odynophagia according to its extent, but it has clinical importance because of its resemblance with laryngeal premalignant or malignant lesions. Diagnosis is made by biopsy under direct laryngoscopy with special staining to identify the hyphae. In this article, we report a case of laryngeal candidiasis presented as leukoplakia localized on vocal fold with literature reviews.