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Purification and comparison of intracellular proteinase A in Candida spp. isolates from Malaysian and Iranian patients and infected mice

Amri Saroukolaei S, Pei Pei C, Shokri H, Asadi F

J Mycol Med, 2012 Jun;22(2):149-59.
PMID: 23518017 DOI: 10.1016/j.mycmed.2012.01.002

To compare the specific intracellular proteinase A activity in clinical isolates of Candida species isolated from Iranian and Malaysian patients, the blood and kidneys of mice infected by Candida cells isolated from these human patients.
Purification and comparison of heat shock protein 90 (Hsp90) in Candida albicans isolates from Malaysian and Iranian patients and infected mice

Khalili V, Shokri H, Khosravi AR, Akim A, Amri Saroukolaei S

J Mycol Med, 2016 Jun;26(2):94-102.
PMID: 26869383 DOI: 10.1016/j.mycmed.2015.12.007

OBJECTIVE: The purposes of this study were to purify and compare the concentration ratios of heat shock protein 90 (Hsp90) in clinical isolates of Candida albicans (C. albicans) obtained from Malaysian and Iranian patients and infected mice.
MATERIALS AND METHODS: Hsp90 was extracted using glass beads and ultracentrifugation from yeast cells and purified by ion exchange chromatography (DEAE-cellulose) and followed by affinity chromatography (hydroxyapatite). Purity of Hsp90 was controlled by SDS-PAGE and its identification was realized by immunoblotting test.
RESULTS: The graphs of ion exchange and affinity chromatography showed one peak in all C. albicans isolates obtained from both Malaysian and Iranian samples, infected mice and under high-thermal (42°C) and low-thermal (25°C) shock. In immunoblotting, the location of Hsp90 fragments was obtained around 47, 75 and 82kDa. The least average concentration ratios of Hsp90 were 0.350 and 0.240mg/g for Malaysian and Iranian isolates at 25°C, respectively, while the highest average concentration ratios of Hsp90 were 3.05 and 2.600mg/g for Malaysian and Iranian isolates at 42°C, respectively. There were differences in the ratio amount of Hsp90 between Malaysian isolates (1.01±0.07mg/g) and mice kidneys (1.23±0.28mg/g) as well as between Iranian isolates (0.70±0.19mg/g) and mice kidneys (1.00±0.28mg/g) (P<0.05).
CONCLUSION: The results showed differences in all situations tested including Iranian and Malaysian isolates, samples treated with temperatures (25°C or 42°C) and before and after infecting the mice (37°C), indicating higher virulent nature of this yeast species in high temperature in human and animal models.
Metabolic adaptation via regulated enzyme degradation in the pathogenic yeast Candida albicans

Ting SY, Ishola OA, Ahmed MA, Tabana YM, Dahham S, Agha MT, et al.

J Mycol Med, 2017 Mar;27(1):98-108.
PMID: 28041812 DOI: 10.1016/j.mycmed.2016.12.002

The virulence of Candida albicans is dependent upon fitness attributes as well as virulence factors. These attributes include robust stress responses and metabolic flexibility. The assimilation of carbon sources is important for growth and essential for the establishment of infections by C. albicans. Previous studies showed that the C. albicans ICL1 genes, which encode the glyoxylate cycle enzymes isocitratelyase are required for growth on non-fermentable carbon sources such as lactate and oleic acid and were repressed by 2% glucose. In contrast to S. cerevsiae, the enzyme CaIcl1 was not destabilised by glucose, resulting with its metabolite remaining at high levels. Further glucose addition has caused CaIcl1 to lose its signal and mechanisms that trigger destabilization in response to glucose. Another purpose of this study was to test the stability of the Icl1 enzyme in response to the dietary sugars, fructose, and galactose. In the present study, the ICL1 mRNAs expression was quantified using Quantitative Real Time PCR, whereby the stability of protein was measured and quantified using Western blot and phosphoimager, and the replacing and cloning of ICL1 ORF by gene recombination and ubiquitin binding was conducted via co-immuno-precipitation. Following an analogous experimental approach, the analysis was repeated using S. cerevisiaeas a control. Both galactose and fructose were found to trigger the degradation of the ICL1 transcript in C. albicans. The Icl1 enzyme was stable following galactose addition but was degraded in response to fructose. C. albicans Icl1 (CaIcl1) was also subjected to fructose-accelerated degradation when expressed in S. cerevisiae, indicating that, although it lacks a ubiquitination site, CaIcl1 is sensitive to fructose-accelerated protein degradation. The addition of an ubiquitination site to CaIcl1 resulted in this enzyme becoming sensitive to galactose-accelerated degradation and increases its rate of degradation in the presence of fructose. It can be concluded that ubiquitin-independent pathways of fructose-accelerated enzyme degradation exist in C. albicans.
Efficacy of modified Leeming-Notman media in a resazurin microtiter assay in the evaluation of in-vitro activity of fluconazole against Malassezia furfur ATCC 14521

Far FE, Al-Obaidi MMJ, Desa MNM

J Mycol Med, 2018 Sep;28(3):486-491.
PMID: 29753721 DOI: 10.1016/j.mycmed.2018.04.007

BACKGROUND: Malassezia furfur is lipodependent yeast like fungus that causes superficial mycoses such as pityriasis versicolor and dandruff. Nevertheless, there are no standard reference methods to perform susceptibility test of Malassezia species yet.
AIMS: Therefore, in this study, we evaluated the optimized culture medium for growth of this lipophilic yeast using modified leeming-Notman agar and colorimetric resazurin microtiter assay to assess antimycotic activity of fluconazole against M. furfur.
RESULTS: The result showed that these assays were more adjustable for M. furfur with reliable and reproducible MIC end-point, by confirming antimycotic activity of fluconazole with MIC of 2μg/ml.
CONCLUSION: We conclude that this method is considered as the rapid and effective susceptibility testing of M. furfur with fluconazole antifungal activity.
In vitro antidermatophytic activity and cytotoxicity of extracts derived from medicinal plants and marine algae

Sit NW, Chan YS, Lai SC, Lim LN, Looi GT, Tay PL, et al.

J Mycol Med, 2018 Sep;28(3):561-567.
PMID: 30060991 DOI: 10.1016/j.mycmed.2018.07.001

OBJECTIVES: This study was conducted to evaluate the antidermatophytic activity of 48 extracts obtained from medicinal plants (Cibotium barometz, Melastoma malabathricum, Meuhlenbeckia platyclada, Rhapis excelsa, Syzygium myrtifolium, Vernonia amygdalina) and marine algae (Caulerpa sertularioides, Kappaphycus alvarezii) against Trichophyton rubrum and Trichophyton interdigitale (ATCC reference strains), and the cytotoxicity using African monkey kidney epithelial (Vero) cells. Active plant extracts were screened for the presence of phytochemicals and tested against clinical isolates of Trichophyton tonsurans.
METHODS: Six different extracts (hexane, chloroform, ethyl acetate, ethanol, methanol and water) were obtained from each plant or algae sample using sequential solvent extraction. The antidermatophytic activity for the extracts was assessed using a colourimetric broth microdilution method. The viability of Vero cells was measured by Neutral Red uptake assay.
RESULTS: All the extracts (except the water extracts of V. amygdalina, C. sertularioides and K. alvarezii) showed antidermatophytic activity against Trichophyton spp. The minimum fungicidal concentration (MFC) ranges for the plant extracts against T. rubrum and T. interdigitale are 0.0025-2.50 and 0.005-2.50mg/mL, respectively. The algae extracts exhibited lower potency against both species, showing MFC ranges of 0.08-2.50 and 0.31-2.50mg/mL, respectively. The ethanol and methanol extracts from the leaves of R. excelsa, and the methanol and water extracts from the leaves of S. myrtifolium were highly active (MFC<0.1mg/mL) and with high selectivity indices (SI>2.8) against reference strains of T. rubrum and T. interdigitale, and most of the clinical isolates of T. tonsurans. Phytochemical analysis indicates the presence of alkaloids, anthraquinones, flavonoids, saponins, tannins, phenolics and triterpenoids in the extracts.
CONCLUSIONS: The medicinal plant extracts exhibited stronger antidermatophytic activity compared to the algae extracts. The leaves of R. excelsa and S. myrtifolium are potential sources of new antidermatophytic agents against Trichophyton spp.
Tongue necrosis secondary to mucormycosis in a diabetic patient: A first case report in Malaysia

Fattah SY, Hariri F, Ngui R, Husman SI

J Mycol Med, 2018 Sep;28(3):519-522.
PMID: 30205883 DOI: 10.1016/j.mycmed.2018.06.004

Mucormycosis is a rare fungal infection and high mortality that commonly affects patients with the weakened immune system. We present an unusual case of tongue necrosis probably due to the healthcare-associated mucormycosis (HCM) in a diabetic patient. Although cannot be proved with certainty, we surmise that intubation as a risk factor in our case. The diagnosis was confirmed by histopathological examination (HPE) of the necrotic tissue specimen. The patient was responded well to lipid complex amphotericin B (250mg) regime after surgery. Subsequent follow up revealed that no signs of recurrence. Early, recognition, diagnosis, prompt treatment and awareness among clinician are representing the most effective way of managing the disease.
Histoplasmosis in cancer patients: a global scoping review (2001-2024)

Onukak AE, Nwagboso CI, Akpu BB, Etim AJ, Benjamin OE, Ereh SE, et al.

J Mycol Med, 2024 Dec;34(4):101511.
PMID: 39500230 DOI: 10.1016/j.mycmed.2024.101511

Although classified as an AIDS-defining illness, several reports show histoplasmosis also affects patients living with cancers including haematological malignancies and solid tumours. However, reviews describing cases of histoplasmosis in malignancies are lacking in the literature. We identified a total of thirty-four cases with twenty (58.8 %) cases reported from the USA, four from Brazil (11.8 %), three from India (8.8 %), and one each from Singapore (2.9 %), France (2.9 %), Netherlands (2.9 %), Colombia (2.9 %), Canada (2.9 %), Morocco (2.9 %), and Malaysia (2.9 %). 82.4 % (n = 28) of the cases were adults. Presenting symptoms were majorly fever (61.7 %), lymphadenopathy (50.0 %) and weight loss (29.4 %). Essential haematologic findings were pancytopaenia (n = 7, 20.6 %), neutropenia (n = 2, 5.9 %) and anaemia (n = 5, 14.7 %). The associated cancers were predominantly haematological and comprised 73.5 % (n = 25) of all cases. The diagnosis of histoplasmosis was via histopathology (n = 23, 67.6%), culture (n = 13, 38.2%), Histoplasma antigen assay (n = 13, 38.2%), anti-Histoplasma antibody assay (n = 5, 14.7%), PCR and sequencing (n = 2, 5.9%), peripheral blood film/direct microscopy (n = 4, 11.8%) and cytology (n = 1, 2.9%). Of the thirty-four cases, twenty-four (70.6%) had favourable outcomes, eight (23.5%) died, one (2.9%) was lost to follow-up and in one (2.9%) case, the outcome was not stated. Histoplasmosis is not an uncommon opportunistic disease complicating malignancies but is paradoxically underdiagnosed in Africa given the huge burden of cancers in that region. Besides following chemotherapy and the use of steroids, tumour necrosis factor-α antagonists therapy, hematopoietic stem cell transplantation and environmental exposure were factors associated with Histoplasma infection in patients with malignancies. A resolution to promptly screen suspected or confirmed cases of malignancies for histoplasmosis will improve diagnosis and clinical outcomes.
Ibrexafungerp for the treatment of vulvovaginal candidiasis: A systematic review and meta-analysis of randomized placebo-controlled trials

Kow CS, Ramachandram DS, Hasan SS, Thiruchelvam K

J Mycol Med, 2025 Mar;35(1):101534.
PMID: 39892065 DOI: 10.1016/j.mycmed.2025.101534

INTRODUCTION: Vulvovaginal candidiasis (VVC) is a prevalent fungal infection affecting millions of women globally, primarily caused by Candida species, most notably Candida albicans. Ibrexafungerp emerges as a promising candidate in the treatment arsenal against VVC, presenting a novel approach to combating this prevalent fungal infection.
METHODS: A systematic literature search was conducted across major databases, including PubMed, EMBASE, and the Cochrane Library, to identify relevant randomized controlled trials (RCTs) evaluating the efficacy and safety of ibrexafungerp in the treatment of VVC. Following rigorous methodology, data extraction, risk of bias assessment using Cochrane's RoB 2 tool, and meta-analysis were conducted.
RESULTS: Four RCTs were included in the analyses. The ibrexafungerp regimen utilized across the studies were 300 mg administered twice daily for one day. Meta-analysis revealed that ibrexafungerp was associated with significantly higher clinical cure rates compared to placebo in patients with VVC (pooled odds ratio (OR) 2.32; 95 % confidence interval (CI) 1.80 to 2.98). Complete symptom resolution was achieved in a greater proportion of participants receiving ibrexafungerp (pooled OR 2.76; 95 % CI 1.62 to 4.71). Analysis of treatment-emergent adverse events revealed a significant higher incidence of at least one treatment-emergent adverse event with ibrexafungerp compared to placebo (pooled OR 2.83; 95 % CI 2.06 to 3.88).
CONCLUSION: This study provides robust support for the efficacy of ibrexafungerp in the treatment of VVC. While the safety profile of ibrexafungerp appears favorable with mostly mild adverse events reported, decision-making in the clinical context should be guided by individual patient factors.