Genotyping of Candida albicans and Comparison of its Antifungal Resistance Pattern in the South Indian Region

Kumar, K.P. Prasanna; Tejashree, A.

doi:10.22207/jpam.16.3.69

Cited by 3 publications

(2 citation statements)

References 35 publications

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“…DNA extraction was performed using QIAamp DNA Mini Kit #51306 (Qiagen, Germantown, MD, United States) according to manufacturer instructions and previously published protocol ( Gerasimidis et al, 2016 ; Panek et al, 2018 ; Mousa et al, 2021 ; Kumar and Tejashree, 2022 ). Approximately 20 mg of tissue was cut into small pieces and subjected to overnight lysis.…”

Section: Methodsmentioning

confidence: 99%

A scalable corneal xenograft platform: simultaneous opportunities for tissue engineering and circular economic sustainability by repurposing slaughterhouse waste

Wang

Shakeel

Salih

et al. 2023

Front. Bioeng. Biotechnol.

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Introduction: Corneal disease is a leading cause of blindness globally that stems from various etiologies. High-throughput platforms that can generate substantial quantities of corneal grafts will be invaluable in addressing the existing global demand for keratoplasty. Slaughterhouses generate substantial quantities of underutilized biological waste that can be repurposed to reduce current environmentally unfriendly practices. Such efforts to support sustainability can simultaneously drive the development of bioartificial keratoprostheses.Methods: Scores of discarded eyes from the prominent Arabian sheep breeds in our surrounding region of the United Arab Emirates (UAE) were repurposed to generate native and acellular corneal keratoprostheses. Acellular corneal scaffolds were created using a whole-eye immersion/agitation-based decellularization technique with a widely available, eco-friendly, and inexpensive 4% zwitterionic biosurfactant solution (Ecover, Malle, Belgium). Conventional approaches like DNA quantification, ECM fibril organization, scaffold dimensions, ocular transparency and transmittance, surface tension measurements, and Fourier-transform infrared (FTIR) spectroscopy were used to examine corneal scaffold composition.Results: Using this high-throughput system, we effectively removed over 95% of the native DNA from native corneas while retaining the innate microarchitecture that supported substantial light transmission (over 70%) after reversing opacity, a well-established hallmark of decellularization and long-term native corneal storage, with glycerol. FTIR data revealed the absence of spectral peaks in the frequency range 2849 cm−1 to 3075 cm−1, indicating the effective removal of the residual biosurfactant post-decellularization. Surface tension studies confirmed the FTIR data by capturing the surfactant’s progressive and effectual removal through tension measurements ranging from approximately 35 mN/m for the 4% decellularizing agent to 70 mN/m for elutes highlighting the effective removal of the detergent.Discussion: To our knowledge, this is the first dataset to be generated outlining a platform that can produce dozens of ovine acellular corneal scaffolds that effectively preserve ocular transparency, transmittance, and ECM components using an eco-friendly surfactant. Analogously, decellularization technologies can support corneal regeneration with attributes comparable to native xenografts. Thus, this study presents a simplified, inexpensive, and scalable high-throughput corneal xenograft platform to support tissue engineering, regenerative medicine, and circular economic sustainability.

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Section: Methodsmentioning

confidence: 99%

A scalable corneal xenograft platform: simultaneous opportunities for tissue engineering and circular economic sustainability by repurposing slaughterhouse waste

Wang

Shakeel

Salih

et al. 2023

Front. Bioeng. Biotechnol.

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show abstract

“…DNA extraction was performed using QIAamp DNA Mini Kit #51306 (Qiagen, Germantown, MD, USA) according to manufacturer instructions and previously published protocol (21)(22)(23)(24). Approximately 20 mg of tissue was cut into small pieces and subjected to overnight lysis.…”

Section: Dna Quantificationmentioning

confidence: 99%

A scalable corneal xenograft platform: simultaneous opportunities for tissue engineering and circular economic sustainability by repurposing slaughterhouse waste

Wang¹,

Shakeel²,

Salih³

et al. 2023

Preprint

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High-throughput platforms that can generate copious quantities of corneal grafts will be invaluable in addressing the existing global demand for keratoplasty. Slaughterhouses generate substantial underutilized biological waste that can be repurposed to reduce current environmentally unfriendly practices. Such efforts to support ecological sustainability can simultaneously drive the development of transplantable ovine corneas and patient-specific bioartificial keratoprostheses. This study presents a dataset highlighting techniques that pioneered the development of a simplified, inexpensive, and high-throughput corneal xenograft platform. To the best of our knowledge, this is the first dataset to be generated in the United Arab Emirates devised on repurposing slaughterhouse waste in a manner that supports circular economic practices. These efforts outline the first step in developing a platform that can produce dozens of native ovine xenografts and acellular corneal scaffolds that effectively preserve ocular transparency, transmittance, and ECM components, and can be used to potentially create individualized and transplantable grafts. Such native grafts can then be genetically modified to reduce rejection and retrovirus transmission in recipients. Analogously, decellularization technologies can support corneal regeneration with key attributes comparable to native xenografts.

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Phenotypic and genotypic characterization of Candida species from the oral cavity of healthy individuals in Lublin province, Poland

Andrzejuk,

Tokarska-Rodak,

Dyrda

et al. 2024

Journal of Oral Microbiology

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Background Candida spp., particularly C. albicans , are commonly isolated fungi in the oral cavity. However, their prevalence in healthy participants and their genotype–phenotype relation remains elusive. Aim This study aimed to update the information on Candida species colonizing the oral cavity of healthy population, identify the most common species, and characterize the intraspecific diversity to determine the genotype-phenotype relationship. Methods Oral swabs of healthy participants who declared an absence of oral infection were analyzed. Microbiological methods: chromogenic media, sugar assimilation tests, drug susceptibility, filamentation tests, temperature tolerance analysis, and assessment of biofilm formation ability. Genotyping methods: PCR amplification of the internal transcribed spacer (ITS) region with MspI restriction enzyme digestion and 25S rDNA region. Results Of the 500 individuals tested, 130 harbored C. albicans in 77%, C. dubliniensis in 12%, Pichia kudriavzevii (previously C. krusei ) in 8%, and Nakaseomyces glabrata (previously C. glabrata ) in 3%. The microbiological tests yielded conflicting results. Analysis of the 25S rDNA transposable intron region contributed to the identification of individual Candida spp. and intraspecific identification of C. albicans genotypes. Genotype A accounted for 70% ( n = 100) of C. albicans isolates, whereas genotypes B, C, and D ( C. dubliniensis ) accounted for 17%, 9%, and 4% of the isolates, respectively. Conclusion The results indicate a complex genotype–phenotype relationship in Candida spp. and recommends combining microbiological and molecular methods for the efficient typing of Candida spp.

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Genotyping of Candida albicans and Comparison of its Antifungal Resistance Pattern in the South Indian Region

Cited by 3 publications

References 35 publications

A scalable corneal xenograft platform: simultaneous opportunities for tissue engineering and circular economic sustainability by repurposing slaughterhouse waste

A scalable corneal xenograft platform: simultaneous opportunities for tissue engineering and circular economic sustainability by repurposing slaughterhouse waste

A scalable corneal xenograft platform: simultaneous opportunities for tissue engineering and circular economic sustainability by repurposing slaughterhouse waste

Phenotypic and genotypic characterization of Candida species from the oral cavity of healthy individuals in Lublin province, Poland

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