Papers by Shivaprakash M R
Intensive Care Medicine, Jan 21, 2015
Frontiers in Cellular and Infection Microbiology, Sep 2, 2022
Evaluation of environmental Mucorales contamination in and around the residence of COVID-19-assoc... more Evaluation of environmental Mucorales contamination in and around the residence of COVID-19-associated mucormycosis patients.
PLOS ONE, Jul 12, 2022
This study was designed to understand the molecular mechanisms of azole resistance in Candida tro... more This study was designed to understand the molecular mechanisms of azole resistance in Candida tropicalis using genetic and bioinformatics approaches. Thirty-two azole-resistant and 10 azole-susceptible (S) clinical isolates of C. tropicalis were subjected to mutation analysis of the azole target genes including ERG11. Inducible expression analysis of 17 other genes potentially associated with azole resistance was also evaluated. Homology modeling and molecular docking analysis were performed to study the effect of amino acid alterations in mediating azole resistance. Of the 32 resistant isolates, 12 (37.5%) showed A395T and C461T mutations in the ERG11 gene. The mean overexpression of CDR1,
Diagnostics
Background: The prevalence of aspergillus sensitization (AS) and allergic bronchopulmonary asperg... more Background: The prevalence of aspergillus sensitization (AS) and allergic bronchopulmonary aspergillosis (ABPA) in asthmatic children remains unclear. Objective: To systematically review the literature to estimate the prevalence of AS and ABPA in children with bronchial asthma. Methods: We searched the PubMed and Embase databases for studies reporting the prevalence of AS or ABPA in pediatric asthma. The primary outcome was to assess the prevalence of AS, while the secondary outcome was to evaluate the prevalence of ABPA. We pooled the prevalence estimates using a random effects model. We also calculated the heterogeneity and publication bias. Results: Of the 11,695 records retrieved, 16 studies with 2468 asthmatic children met the inclusion criteria. Most studies were published from tertiary centers. The pooled prevalence of AS in asthma (15 studies; 2361 subjects) was 16.1% (95% confidence intervals [CI], 9.3–24.3). The prevalence of AS was significantly higher in prospective stud...
Molecules
Delineation of clinical complications secondary to fungal infections, such as cryptococcal mening... more Delineation of clinical complications secondary to fungal infections, such as cryptococcal meningitis, and the concurrent emergence of multidrug resistance in large population subsets necessitates the need for the development of new classes of antifungals. Herein, we report a series of ring-modified histidine-containing short cationic peptides exhibiting anticryptococcal activity via membrane lysis. The N-1 position of histidine was benzylated, followed by iodination at the C-5 position via electrophilic iodination, and the dipeptides were obtained after coupling with tryptophan. In vitro analysis revealed that peptides Trp-His[1-(3,5-di-tert-butylbenzyl)-5-iodo]-OMe (10d, IC50 = 2.20 μg/mL; MIC = 4.01 μg/mL) and Trp-His[1-(2-iodophenyl)-5-iodo)]-OMe (10o, IC50 = 2.52 μg/mL; MIC = 4.59 μg/mL) exhibit promising antifungal activities against C. neoformans. When administered in combination with standard drug amphotericin B (Amp B), a significant synergism was observed, with 4- to 16-fo...
Journal of Fungi
Serum 1,3-β-d-glucan(BDG) is a broad fungal biomarker for invasive fungal disease (IFD). More dat... more Serum 1,3-β-d-glucan(BDG) is a broad fungal biomarker for invasive fungal disease (IFD). More data is still required to support the Fujifilm Wako assay as a valuable alternative to the widely used Fungitell assay. We included archived serum samples from 157 individuals (97 cases; 33-IA, 64-IC, and 60 controls) for the comparative performance evaluation of the Fungitell assay and the Fujifilm Wako assay for IFD diagnosis. The BDG value was significantly higher in patients with IFD vs. controls (70.79 pg/mL vs. 3.03 pg/mL, p: 0.0002). An area under the curve (AUC) for the IFD, IC, and IA diagnosis was 0.895, 0.910, and 0.866, respectively, for the Fujifilm Wako assay. Based on the highest Youden’s index (0.667), a cutoff of 5 pg/mL was selected as the optimum for the Fujifilm Wako assay with good sensitivity (79.4%), specificity (88.3%) and agreement (84.7%, Cohen’s k:0.691) with the Fungitell assay. The mean run-time of the Fujifilm Wako assay was 70.12 min, and real-time observation...
Medical Mycology
Poster session 1, September 21, 2022, 12:30 PM - 1:30 PM Objective Candida tropicalis and Candi... more Poster session 1, September 21, 2022, 12:30 PM - 1:30 PM Objective Candida tropicalis and Candida glabrata account for 41.6% and 7.08% of total Candidaemia cases in India. Echinocandins are the first-line treatment option for these infections. Resistance to Echinocandins is rare with Candida sp. However, in recent years, has been noted across many centers. We determined the mechanism of echinocandin resistance in C. tropicalis and C. glabrata. Methods C. tropicalis and C. glabrata isolated from Candidaemia patients over a period of 3 years (August 2016-July 2019), identified by MALDI-TOF-MS were used in this study. Antifungal susceptibility testing was done following CLSI broth microdilution reference method (M 27A). FKS1 gene was sequenced using species-specific primers for the presence of any mutation. To determine any changes in the cell wall chitin and glucan contents, expression fold changes of chitin synthase (CHS1, CHS2, and CHS3), and glucan synthase genes upon caspofungin...
Medical Mycology
Poster session 1, September 21, 2022, 12:30 PM - 1:30 PM Objectives To study the drug susceptib... more Poster session 1, September 21, 2022, 12:30 PM - 1:30 PM Objectives To study the drug susceptibility profile of Candida albicans isolated from bloodstream infection (BSI) against azole anti-fungal agents (fluconazole, itraconazole, voriconazole, and posaconazole) in a tertiary care hospital over 5 years. Methods All blood culture bottles from patients attending our tertiary care hospital between January 2017 to December 2021 and growing Candida albicans were included in the analysis. All blood culture bottles were continuously monitored automatically inside BACTEC 9240 (BD Becton Dickinson, USA). Beep-positive bottles were further processed by preparation of Gram stain smears, sub-culture onto 5% sheep blood agar and MacConkey agar and spot inoculation onto Sabouraud's Dextrose agar (SDA). Isolates were identified as C. albicans using MALDI-TOF MS (Bruker Daltoniks, Bremen, Germany). Broth microdilution (BMD) was performed to determine minimum inhibitory concentration (MIC) of...
P394 Evaluation of diagnostic potential of recombinant secretory aspartyl proteinase 2 (Sap2) protein from Candida parapsilosis for use in systemic candidiasis
Medical Mycology
Poster session 3, September 23, 2022, 12:30 PM - 1:30 PM Objectives Systemic candidiasis is the... more Poster session 3, September 23, 2022, 12:30 PM - 1:30 PM Objectives Systemic candidiasis is the fourth most common bloodstream infection in ICU patients worldwide. Along with Candida albicans, infections caused by non-albicans Candida (NAC) species are increasingly becoming more prevalent globally along with the emergence of drug resistance. The diagnosis of systemic candidiasis is difficult due to the absence of significant clinical symptoms in patients. Since conventional diagnostics methods for candidiasis show less sensitivity and specificity, novel immunodiagnostic techniques are needed for early diagnosis. We investigated the diagnostic potential of recombinant secreted aspartyl proteinase 2 (rSap2) from C. parapsilosis for the detection of Candida infection. Methods Genomic DNA was isolated from C. parapsilosis, followed by PCR amplification of Sap2 gene using designed gene-specific primers. Sap2 protein expression and purification was performed using Ni-NTA affinity chroma...
P205 Uncommon Aspergillus species encountered in human infections
Medical Mycology
Poster session 2, September 22, 2022, 12:30 PM - 1:30 PM Objective Aspergillus species are ubiq... more Poster session 2, September 22, 2022, 12:30 PM - 1:30 PM Objective Aspergillus species are ubiquitously found in environment and have propensity to infect both immunocompromised as well as immunocompetent individuals. Though the most common species implicated in human illness include Aspergillus fumigatus and A. flavus, many uncommon Aspergillus species are increasingly being reported. Moreover, these Aspergillus species have the potential to afflict varied organ systems and can thereby, be implicated in a wide array of infections. Here, we describe three patients infected by rare Aspergillus species viz. A. sclerotium, A. cristatus, and A. japonicum causing brain abscess, corneal ulcer and chronic dacrocystitis. Methods Uncommon Aspergillus species were isolated from clinical samples (nasal scraping, corneal scraping, and brain biopsy). Samples were inoculated on Sabouraud dextrose agar (incubated at 37°C and 25°C) and brain heart infusion agar (incubated at 25°C), followed by sl...
Medical Mycology
Poster session 3, September 23, 2022, 12:30 PM - 1:30 PM Objective Rhytidhysteron spp. are emer... more Poster session 3, September 23, 2022, 12:30 PM - 1:30 PM Objective Rhytidhysteron spp. are emerging dematiaceous fungi implicated in subcutaneous infections in immunocompromised as well as immunocompetent individuals. Diagnosis of Rhytidhysteron spp is delayed owing to non-sporulation and misidentification. The present study was conducted to characterize Rhytidhysteron spp. isolated from India by molecular techniques. Method We retrieved a total of eight isolates of Rhytidhysteron spp from our National Culture Collection for Pathogenic Fungi (NCCPF) that were received from various centers across India. Colony morphology on Sabouraud's dextrose agar (SDA) and potato dextrose agar (PDA) was recorded. Molecular identification was performed by sequencing internal transcribed spacer (ITS) and 28 s region of rDNA. Phylogenetic tree was constructed using the Neighbor-Joining method along with sequences of standard strains retrieved from the NCBI database. Results Rhytidhysteron spp w...
Medical Mycology
Poster session 2, September 22, 2022, 12:30 PM - 1:30 PM Objectives Aspergillus flavus accounts... more Poster session 2, September 22, 2022, 12:30 PM - 1:30 PM Objectives Aspergillus flavus accounts for ∼ 10% of bronchopulmonary aspergillosis and is the second leading etiological agent of invasive aspergillosis worldwide. It is the commonest cause of fungal rhinosinusitis and ocular mycoses in tropical countries including India. We report amplified fragment length polymorphism genotyping of a set of clinical and environmental isolates to unravel the genetic diversity of A. flavus in India and to further determine correlation between isolate genotype and the source/clinical phenotype of the Aspergillus infection. Methods Two sets of morphologically identified isolates of A. flavus from clinical (n = 71) and environmental sources (n = 22) were included in the study using a stratified random sampling method. Clinical strains were isolated from lower respiratory tract specimens (n = 22), sinus and sino-nasal biopsies (n = 25), corneal samples (n = 12), and others (n = 12). Environmenta...
Medical Mycology
Poster session 3, September 23, 2022, 12:30 PM - 1:30 PM Objectives The evaluation of serum β-D... more Poster session 3, September 23, 2022, 12:30 PM - 1:30 PM Objectives The evaluation of serum β-D-glucan, galactomannan, and Aspergillus fumigatus specific IgG in the diagnosis and clinical management of patients with chronic pulmonary aspergillosis. Methods Consecutive patients with a past history of pulmonary tuberculosis (PTB) with clinical suspicion of chronic pulmonary aspergillosis (CPA) attending the outpatient services of PGIMER, Chandigarh were included from August 2019 to August 2021, and CPA was defined as described by Denning et al. including both radiological and mycological evidence. Age and sex-matched post-PTB individuals selected from a population attending the chest clinic were enrolled as controls. Serum samples were obtained from all participants and testing for serum β-D-glucan (Fungitell, CapeCod), galactomannan (Platelia, BioRad) and Aspergillus fumigatus specific IgG (ImmunoCAP, Phadia) was performed as per manufacturer's instructions. Results A total of ...
Medical Mycology
Poster session 2, September 22, 2022, 12:30 PM - 1:30 PM Objectives Lasiodiplodia theobromae is... more Poster session 2, September 22, 2022, 12:30 PM - 1:30 PM Objectives Lasiodiplodia theobromae is a dematiaceous fungus, rarely reported to cause infections in humans. This case-series was conducted to identify the potential risk-factors and spectrum of diseases caused by L. theobromae. Methods We performed a retrospective analysis of all cases from which the isolates of the L. theobromae were accessioned at the National culture collection of pathogenic fungi (NCCPF), over the last 10 years (January 2012- March 2022). The isolates were retrieved and identified by conventional (colony morphology, color, and microscopic appearances) and molecular (sequencing of ITS1 region of ribosomal DNA) methods. In vitro antifungal susceptibility testing (AFST) was performed by microbroth dilution recommended by the Clinical and Laboratory Standards Institute (CLSI). (M38-A2). The clinical features, demographic details and outcome were recorded. Results In 10 years, a total of 20 patients were ide...
Background: Increased occurrence of mucormycosis in India during the second wave of the COVID-19 ... more Background: Increased occurrence of mucormycosis in India during the second wave of the COVID-19 pandemic in early 2021 in India subsequently prompted us to undertake a multi-site case-control investigation. The objectives were to examine the monthly trend of Covid-19 Associated Mucormycosis (CAM) cases among in-patients and to identify factors associated with it.Methods: Eleven study sites were involved across India and archived records since 1st January till 30th September, 2021 were used for trend analysis. The cases and controls were enrolled during 15th June 2021 to 30th September 2021. Data were collected using a semi-structured questionnaire. Among 1211 enrolled participants, 336 were CAM cases and 875 were COVID-19 positive non-mucormycosis controls. Results: Admitted CAM-case number reached highest point in May 2021 after a month of peak admission for COVID-19. Odds of developing CAM increased with the history of working in a dusty environment (adjusted odds ratio; aOR 3.24...
Mycopathologia
Background In experimental models, the expression of glucose-regulated protein 78 (GRP78) in endo... more Background In experimental models, the expression of glucose-regulated protein 78 (GRP78) in endothelial cells played a role in the pathogenesis of mucormycosis. However, the role of GRP78 in COVID-19-associated mucormycosis (CAM) has not been studied. We hypothesized that serum GRP78 levels are elevated in subjects with CAM. Objective To compare the serum GRP78 levels in subjects with CAM and COVID-19 controls without mucormycosis. Design And Setting We performed a hospital-based, case-control study between 1 April 2021 and 31 May 2021. Participants We enrolled 24 subjects each of CAM and COVID-19 subjects without mucormycosis. We also measured serum GRP78 levels in ten healthy controls. Exposure The primary exposure studied was serum GRP78 concentration, estimated using a commercially available ELISA kit in stored serum samples. Results We found the mean ± standard deviation (SD) serum GRP78 levels significantly higher (p = 0.0001) among the CAM (374.3 ± 127.3 pg/ mL) than the COVID-19 (246.4 ± 67.0 pg/mL) controls. The proportion of subjects with an abnormal GRP78 level ([ mean [184.8 pg/mL] plus two SD [23.2 pg/mL] of GRP78 from healthy participants) was 87.5% and 45.8% in the CAM group and COVID-19 controls, respectively. Serum GRP78 level was independently associated with CAM (odds ratio 1.011;
Diabetology, 2022
Recent Mucorales-mediated outbreaks of infections and an association of fungal infection with COV... more Recent Mucorales-mediated outbreaks of infections and an association of fungal infection with COVID-19 cases, as observed for COVID-19-associated mucormycosis (CAM), have posed new challenges for the management of patients in critical care units. Diabetes and hyperglycemia are integrally linked to the severity of COVID-19, and uncontrolled diabetes mellitus and COVID-19 have recently been (independently or in combination) associated with the emergence of aggressive mucormycosis due to attendant defects in innate immune recognition pathways. Therefore, the identification of novel global cellular stressors upregulated during diabetes to understand the contribution of diabetes-associated metabolic vulnerabilities can help build a Metabolic-Stress-Associated Interactome (MSAI). This interactome can help reshape the metabolic inflammation (meta-inflammation) underlying the clinical manifestations of COVID-19 to facilitate the rational design of effective therapies for COVID-19 and CAM. A...
The thermotolerant multidrug-resistant ascomycete Candida auris rapidly emerged since 2009 and si... more The thermotolerant multidrug-resistant ascomycete Candida auris rapidly emerged since 2009 and simultaneously evolved in different geographical zones worldwide, causing superficial as well as systemic infections. The molecular events that orchestrated this sudden emergence of the killer fungus remain mostly elusive. Here, we identify centromeres in C. auris and related species, using a combined approach of chromatin immunoprecipitation and comparative genomic analyses. We find that C. auris and multiple other species in the Clavispora/Candida clade shared a conserved small regional centromere landscape lacking pericentromeres. Further, a centromere inactivation event led to karyotypic alterations in this species complex. Inter-species genome analysis identified several structural chromosomal changes around centromeres. In addition, centromeres are found to be rapidly evolving loci among the different geographical clades of the same species of C. auris. Finally, we reveal an evolutio...
The Genus Aspergillus - Pathogenicity, Mycotoxin Production and Industrial Applications, 2021
Aspergillus species are ubiquitous saprophytes and opportunistic pathogens causing wide spectrum ... more Aspergillus species are ubiquitous saprophytes and opportunistic pathogens causing wide spectrum of diseases in humans depending on the host immune status. Following pathogen entry, various soluble bronchopulmonary factors enhance conidial clearance. However, due to virulence factors and poor host immune response Aspergillus conidia bind and damage the airway epithelium. The host immune cells like neutrophils and macrophages recognise Aspergillus spp. through various pathogen recognition receptors and form reactive oxygen species which mediate conidial killing. Neutrophils also attack extracellular hyphae by oxidative attack, non-oxidative granule proteins and neutrophil extracellular traps. In case of adaptive immunity, Th1 cells are crucial sources of IFN-γ mediated protective immunity. The Th17 also display a highly pro-inflammatory which is counterbalanced by a Treg cell. B cells and antibodies also enhance fungal clearance although excessive IgE production may result in atopy. ...
Journal of Fungi, 2021
Coronavirus disease (COVID-19)-associated mucormycosis (CAM) is an emerging threat globally, espe... more Coronavirus disease (COVID-19)-associated mucormycosis (CAM) is an emerging threat globally, especially in India. More than 40,000 CAM cases have been reported in India. The emergence of CAM cases in India has been attributed to environmental, host, and iatrogenic factors. Mucorales spore burden has been reported globally; however, their presence is higher in tropical countries such as India, contributing to the emergence of CAM. Before the COVID-19 pandemic, patients with diabetes mellitus, haematological malignancies, solid organ transplants, corticosteroid therapy and neutropenia were more prone to mucormycosis, whereas in COVID-19 patients, virus-induced endothelial dysfunction, hyperglycaemia, and immune dysfunction following corticosteroid use increase the risk of acquiring mucormycosis. The interaction of Mucorales spores with the epithelial cells, followed by endothelial invasion, is a crucial step in the pathogenesis of mucormycosis. Endothelial damage and increased endothe...
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Papers by Shivaprakash M R