The Implementation of HIV Self-Testing in Resource-Limited Settings Where the HIV Disease Burden is High.

In resource-limited settings, there is growing evidence that HIV testing is lacking among high-risk key populations such as men having sex with men, injection drug users, and transgenders largely due to stigma, discrimination, and lack of confidentiality. Findings from recent studies among high-risk key populations and the general population from various regions including resource-limited settings support the need for wider accessibility of HIV self-testing (HIV-ST) to reach those who may not otherwise have access to testing. Therefore, HIV-ST has untapped potential as a strategy to improve access to HIV testing and to increase testing frequency among key high-risk populations and their partners. Though HIV-ST has emerged as a safe, acceptable, and effective way to reach people, there are several roadblocks to implementing the HIV-ST policy, and fast-track policy implementation needs to be necessitated with newer or modified strategic plans.

Extensive ERG11 mutations associated with fluconazole-resistant Candida albicans isolated from HIV-infected patients.

BACKGROUND AND PURPOSE: Azoles are preferred antifungal agents given their inexpensiveness, limited toxicity, and potentiality of oral administration. However, the extensive use of prophylactic azole therapy for chronic infections, especially in immunocompromised patients, has led to an increase in azole resistance, thereby rising health care costs. Fluconazole resistance is associated with poor clinical outcomes and the emergence of new infections. The present study aimed to investigate the mutations of ERG11 gene in fluconazole-resistant Candida albicans isolates. MATERIALS AND METHODS: This study was conducted on 80 clinical samples collected from HIV-infected patients with suspected candidiasis in Tagore Medical College Hospital and Government Hospital of Thoracic Medicine, Chennai, India, for a period of 18 months (May 2016-December 2017). The antifungal susceptibility pattern was determined by agar diffusion and broth dilution techniques as per the Clinical and Laboratory Standards Institute guidelines. The ERG11 gene of the known fluconazole-resistant strains of C. albicans was amplified by polymerase chain reaction (PCR). In addition, the samples were subjected to sequencing and mutation analysis. RESULTS: A total of 60 Candida species were isolated from HIV patients and were speciated using standard, conventional, and molecular methods. Candida albicans comprised 28.3% (n=17) of the Candida isolates, 59% (n=10) of which were resistant to fluconazole. Sequencing of the amplified product of ERG11 C. albicans gene isolates showed that they were highly mutated and included many nonsense mutations which were not reported earlier. CONCLUSION: The molecular characterization of ERG11 gene showed many missense and nonsense mutations. Such mutations, which were unique to the geographical area under investigation, could be concluded to account for the development of resistance to fluconazole.

Molecular identification and antifungal susceptibility pattern of Candida species isolated from HIV infected Patients with candisiasis.

BACKGROUND AND PURPOSE: Opportunistic fungal infections have been on a growing trend since the last two decades. Among the opportunistic fungal agents, Candida species, Cryptococcus neoformans, and Aspergillus fumigatus account for most of the life-threatening infections in immunocompromised individuals. Regarding this, the present study aimed to investigate the molecular identification and antifungal susceptibility pattern of Candida species isolated from HIV-infected patients. MATERIALS AND METHODS: This study was conducted on 80 clinical samples collected from HIV-infected patients with suspected candidiasis referring to Tagore Medical College and Hospital, Rathinamangalam and Government Hospital of Thoracic Medicine, in Chennai, India, for 18 months (i.e., May 2016-December 2017). Phenotypic and molecular identification was accomplished using internal transcribed spacer region 1 (ITS1) and ITS4 primers. The antifungal susceptibility pattern of the isolates against four antifungal agents was also determined by both disk diffusion and broth dilution methods. RESULTS: In the present study, the prevalence of candidiasis was obtained as 75% (n=60). Candida tropicalis was the predominant identified species. All the emerging species (i.e., Kodamaea ohmeri, Hanseniaspora opuntiae, and C. orthopsilosis) were identified through molecular identification since the phenotypic identification was inconclusive. In terms of the susceptibility pattern, 63.3% and 18.3% of the isolates were resistant to fluconazole and voriconazole, respectively. Candida albicans was also found to be resistant to amphotericin B. CONCLUSION: Molecular assay led to the identification of K. ohmeri, H. opuntiae, and C. orthopsilosis, which were multidrug-resistant. This study highlighted the need for the prompt and timely identification of clinical yeast isolates given the emergence of many rare species and their capability of causing life-threatening infections and outbreaks. In the laboratories where molecular diagnostic methods are not available, alternative services of reference laboratories can be utilized as cost-effective measures. With regard to the growing prevalence of antifungal drug resistance, antifungal susceptibility testing should be made mandatory for effective patient management.

Erratum for Nagesh et al., phytochemical analysis and docking study of compounds present in a polyherbal preparation used in the treatment of dermatophytosis.

[This corrects the article DOI: 10.29252/cmm.3.4.6.].

Antifungal Activity, Gas Chromatographic-Mass Spectrometric Analysis And in silico Study of Punica Granatum Peel Extracts Against Fluconazole Resistant Strains of Candida Species.

AIMS: Candida species is the common cause of opportunistic fungal infections all over the world with increased mortality and morbidity especially in immunosuppressed patients. Fluconazole is the first line therapy for candidiasis. The antifungal resistance pattern in high-risk patients is a major concern. The present study was aimed to assess the anticandidal activity of Punica granatum peel against fluconazole resistant Candida species isolated from HIV patients. MATERIALS & METHODS: Ethanol, chloroform, petroleum ether and aqueous extracts of the peel of P. granatum were evaluated against standard strains of Candida spp. and fluconazole resistant clinical isolates by agar diffusion and broth dilution techniques. The GC-MS analysis of the extracts was performed to identify the phytochemicals present in it. The predominant phytochemical was subjected to molecular docking study to determine its binding efficacy with lanosterol 14-alpha demethylase. RESULTS: P. granatum peel extracts showed excellent anticandidal activity with ethanol extract exhibiting the most inhibitory activity. C. albicans and C. krusei were the most inhibited and C. parapsilosis was the least inhibited species. The GC-MS analysis of the ethanol extract identified five predominant phytochemicals. On docking studies, the five phytochemicals showed a good binding to the lanosterol 14-alpha demethylase. CONCLUSION: The present study is the first report on the antifungal activity of various extracts of P. granatum against fluconazole resistant Candida isolates. Ethanol extract of P. granatum peel showed excellent anticandidal activity against fluconazole resistant Candida spp. Hence, it can be explored further to identify a potential drug candidate.

Antifungal activity of curcumin-silver nanoparticles against fluconazole-resistant clinical isolates of Candida species.

INTRODUCTION: Candida species is the common form of opportunistic fungal infections, especially in immunosuppressed individuals. Fluconazole is the first-line therapy for candidiasis as it is affordable and readily available. However, the antifungal resistance pattern in high-risk patients is a major concern. AIM: The objective of the present study was to assess the anticandidal activity of curcumin-silver nanoparticles (C-Ag-NPs) against fluconazole-resistant Candida species isolated from HIV patients. MATERIALS AND METHODS: Ten milliliters of 0.1 M silver nitrate (AgNO3) and 3 ml curcumin solution was heated in a water bath for 1 h at 60°C. The formation of the Ag-NPs was determined by color change from light yellow to brownish. The solution was centrifuged at 9000 rpm for 15 min and was washed with ethanol and later lyophilized for 24 h to obtain the purified curcumin-Ag-NPs (C-Ag-NPs). A stock of 1 mg/ml of C-Ag-NPs was prepared in deionized water. The agar diffusion test and broth dilution tests were conducted to determine the anticandidal activity of C-Ag-NPs. RESULTS: C-Ag-NPs showed a better antifungal activity compared to curcumin and AgNO3 solution. Candida glabrata and Candida albicans were the most inhibited and Candida tropicalis was the least inhibited species. The mean zone diameter was 22.2 ± 0.8 mm, 20.1 ± 0.8 mm, and 16.4 ± 0.7 mm against C. glabrata, C. albicans and C. tropicalis respectively. Other Candida species under the study were also inhibited. Inhibitory activity was dose dependent and it increased with concentration. The minimum inhibitory concentration values for different Candida species ranged from 31.2 µg/ml to 250 µg/ml. CONCLUSION: This is the first report on the antifungal activity of C-Ag-NPs against fluconazole-resistant Candida isolates. C-Ag-NPs can be explored further to identify a potential drug candidate that can be used for the treatment of candidiasis due to fluconazole-resistant strains of Candida species.

Phytochemical analysis and docking study of compounds present in a polyherbal preparation used in the treatment of dermatophytosis.

BACKGROUND AND PURPOSE: Soleshine is a polyherbal preparation established in the market for the treatment of cracks and tinea pedis, which is applied externally. This preparation is composed of the extracts of indigenous plants, namely Azadirachta indica, Lawsonia alba, and Shorea robusta, mixed with castor oil and sesame oil. In the present study, an attempt was made to identify the constituents of soleshine and identify some potential drug-like molecules that can inhibit important drug targets of the dermatophytes using molecular docking method. MATERIALS AND METHODS: The active ingredients of polyherbal preparation were identified with the aid of gas chromatography-mass spectrometry (GC-MS). Two major compounds were selected based on the retention time and percentage of the area covered in the graph for docking study. The three-dimensional structures of 1,3-ß-glucan synthase, chitinase, fungalysin, and lumazine synthase were derived by homology modelling using MODELLER software, version 9.0. The docking of the ligand and receptor was performed using iGEMDOCK and AutodockVina software. The physicochemical properties, lipophilicity, hydrophilicity, and drug likeness properties were obtained from the Swiss ADME online server tool. RESULTS: The GC-MS analysis demonstrated the presence of different phytochemical compounds in the extract of polyherbal preparation. A total of 20 compounds were identified, among which 3,7-dimethyl-2,6-octadienaland 2-pentene-2-methyl were the major compounds. Regarding 3,7-dimethyl-2,6-octadienal, the covered area and height were 40.15% and 46.17%, respectively. These values were 31.90% and 23.33% for 2-pentene-2-methyl, respectively. These two major compounds had an excellent binding affinity and obeyed the rules for the drug likeness and lead likeness. CONCLUSION: As the findings indicated, the two major ingredients present in soleshine showed a good antifungal activity as they inhibited the enzymes responsible for the survival of fungal organism; furthermore, they were appropriate for the lead molecules.