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Inhibition of Virulence Associated Traits by β-Sitosterol Isolated from Hibiscus rosa-sinensis Flowers Against Candida albicans: Mechanistic Insight and Molecular Docking Studies
Pallvi Mohana, Atamjit Singh, Farhana Rashid, Sharabjit Singh, Kirandeep Kaur, Rupali Rana, Preet Mohinder Singh Bedi, Neena Bedi, Rajinder Kaur, Saroj Arora
J. Microbiol. 2024;62(12):1165-1175.   Published online November 6, 2024
DOI: https://doi.org/10.1007/s12275-024-00174-5
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AbstractAbstract PDF
The emerging drug resistance and lack of safer and more potent antifungal agents make Candida infections another hot topic in the healthcare system. At the same time, the potential of plant products in developing novel antifungal drugs is also in the limelight. Considering these facts, we have investigated the different extracts of the flowers of Hibiscus rosa-sinensis of the Malvaceae family for their antifungal efficacy against five different pathogenic Candida strains. Among the various extracts, the chloroform extract showed the maximum zone of inhibition (26.6 ± 0.5 mm) against the Candida albicans strain. Furthermore, the chloroform fraction was isolated, and a sterol compound was identified as β-sitosterol. Mechanistic studies were conducted to understand the mechanism of action, and the results showed that β-sitosterol has significant antifungal activity and is capable of interrupting biofilm formation and acts by inhibiting ergosterol biosynthesis in Candida albicans cells. Microscopic and molecular docking studies confirmed these findings. Overall, the study validates the antifungal efficacy of Candida albicans due to the presence of β-sitosterol which can act as an effective constituent for antifungal drug development individually or in combination.

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  • Extraction of Hibiscus rosa sinensis Flower
    Shubham Porte, Vinayak Kaushik, Geetanjali Sahu, Sharang Bali
    Asian Journal of Pharmacy and Technology.2025; : 339.     CrossRef
  • Hibiscus rosa‐sinensis: A Multifunctional Flower Bridging Nutrition, Medicine, and Molecular Therapeutics
    Hassan Raza, Muhammad Tauseef Sultan, Khalil Ahmad, Muhammad Maaz, Shehnshah Zafar, Ahmad Mujtaba Noman, Entessar Mohammad Al Jbawi
    Food Science & Nutrition.2025;[Epub]     CrossRef
Fleagrass (Adenosma buchneroides Bonati) Acts as a Fungicide Against Candida albicans by Damaging Its Cell Wall
Youwei Wu, Hongxia Zhang, Hongjie Chen, Zhizhi Du, Qin Li, Ruirui Wang
J. Microbiol. 2024;62(8):661-670.   Published online July 3, 2024
DOI: https://doi.org/10.1007/s12275-024-00146-9
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AbstractAbstract PDF
Fleagrass, a herb known for its pleasant aroma, is widely used as a mosquito repellent, antibacterial agent, and for treating colds, reducing swelling, and alleviating pain. The antifungal effects of the essential oils of fleagrass and carvacrol against Candida albicans were investigated by evaluating the growth and the mycelial and biofilm development of C. albicans. Transmission electron microscopy was used to evaluate the integrity of the cell membrane and cell wall of C. albicans. Fleagrass exhibited high fungicidal activity against C. albicans at concentrations of 0.5% v/v (via the Ras1/cAMP/PKA pathway). Furthermore, transmission electron microscopy revealed damage to the cell wall and membrane after treatment with the essential oil, which was further confirmed by the increased levels of β-1,3-glucan and chitin in the cell wall. This study showed that fleagrass exerts good fungicidal and hyphal growth inhibition activity against C. albicans by disrupting its cell wall, and thus, fleagrass may be a potential antifungal drug.
Licochalcone A Protects Vaginal Epithelial Cells Against Candida albicans Infection Via the TLR4/NF-κB Signaling Pathway
Wei Li, Yujun Yin, Taoqiong Li, Yiqun Wang, Wenyin Shi
J. Microbiol. 2024;62(7):525-533.   Published online May 31, 2024
DOI: https://doi.org/10.1007/s12275-024-00134-z
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AbstractAbstract PDF
Vulvovaginal candidiasis (VVC) is a prevalent condition affecting a significant portion of women worldwide. Licochalcone A (LA), a natural compound with diverse biological activities, holds promise as a protective agent against Candida albicans (C. albicans) infection. This study aims to investigate the potential of LA to safeguard vaginal epithelial cells (VECs) from C. albicans infection and elucidate the underlying molecular mechanisms. To simulate VVC in vitro, VK2-E6E7 cells were infected with C. albicans. Candida albicans biofilm formation, C. albicans adhesion to VK2-E6E7 cells, and C. albicans-induced cell damage and inflammatory responses were assessed by XTT reduction assay, fluorescence assay, LDH assay, and ELISA. CCK-8 assay was performed to evaluate the cytotoxic effects of LA on VK2-E6E7 cells. Western blotting assay was performed to detect protein expression. LA dose-dependently hindered C. albicans biofilm formation and adhesion to VK2-E6E7 cells. Furthermore, LA mitigated cell damage, inhibited the Bax/Bcl-2 ratio, and attenuated the secretion of pro-inflammatory cytokines in C. albicans-induced VK2-E6E7 cells. The investigation into LA's impact on the Toll-like receptor 4 (TLR4)/nuclear factor-kappa B (NF-κB) pathway revealed that LA downregulated TLR4 expression and inhibited NF-κB activation in C. albicans-infected VK2-E6E7 cells. Furthermore, TLR4 overexpression partially abated LA-mediated protection, further highlighting the role of the TLR4/NF-κB pathway. LA holds the potential to safeguard VECs against C. albicans infection, potentially offering therapeutic avenues for VVC management.

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  • New Curcumin Analogue (PAC) Inhibits Candida albicans Virulence, Restricts Its Adhesion Potential, and Relieves Oral Epithelial Cell Inflammation and Defense Mechanisms
    Ghazoua Mezni, Hawraa Issa, Manal Dahdah, Anaïs Poulin, Adam Daïch, Abdulaziz Alamri, Mahmoud Rouabhia, Abdelhabib Semlali
    Antibiotics.2025; 14(5): 495.     CrossRef
  • Therapeutic potential of Licochalcone A in dermatological diseases: from basic to clinical research
    Deming Liu, Xue Jiang, Fujin Yang, Jingjing Zhou, Yanxi Li, Hua Yang
    Frontiers in Pharmacology.2025;[Epub]     CrossRef
Those Nematode‑Trapping Fungi That are not Everywhere: Hints Towards Soil Microbial Biogeography
Wei Deng , Fa Zhang , Davide Fornacca , Xiao-Yan Yang , Wen Xiao
J. Microbiol. 2023;61(5):511-523.   Published online April 6, 2023
DOI: https://doi.org/10.1007/s12275-023-00043-7
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AbstractAbstract PDF
The existence of biogeography for microorganisms is a raising topic in ecology and researchers are employing better distinctions between single species, including the most rare ones, to reveal potential hidden patterns. An important volume of evidence supporting heterogeneous distributions for bacteria, archaea and protists is accumulating, and more recently a few efforts have targeted microscopic fungi. We propose an insight into this latter kingdom by looking at a group of soil nematode-trapping fungi whose species are well-known and easily recognizable. We chose a pure culture approach because of its reliable isolation procedures for this specific group. After morphologically and molecularly identifying all species collected from 2250 samples distributed in 228 locations across Yunnan province of China, we analyzed occurrence frequencies and mapped species, genera, and richness. Results showed an apparent cosmopolitan tendency for this group of fungi, including species richness among sites. However, only four species were widespread across the region, while nonrandom heterogeneous distributions were observed for the remaining 40 species, both in terms of statistical distribution of species richness reflected by a significant variance-to-mean ratio, as well as in terms of visually discernible spatial clusters of rare species and genera on the map. Moreover, several species were restricted to only one location, raising the question of whether endemicity exists for this microbial group. Finally, environmental heterogeneity showed a marginal contribution in explaining restricted distributions, suggesting that other factors such as geographical isolation and dispersal capabilities should be explored. These findings contribute to our understanding of the cryptic geographic distribution of microorganisms and encourage further research in this direction.

Citations

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  • Linking watershed formation with the phylogenetic distribution of a soil microscopic fungus in Yunnan Province, China
    Davide Fornacca, Wei Deng, Yaoquan Yang, Fa Zhang, Xiaoyan Yang, Wen Xiao
    BMC Microbiology.2024;[Epub]     CrossRef
  • Analysis of Nuclear Dynamics in Nematode-Trapping Fungi Based on Fluorescent Protein Labeling
    Liang Zhou, Zhiwei He, Keqin Zhang, Xin Wang
    Journal of Fungi.2023; 9(12): 1183.     CrossRef
Microbial co-occurrence network in the rhizosphere microbiome: its association with physicochemical properties and soybean yield at a regional scale
Sarbjeet Niraula , Meaghan Rose , Woo-Suk Chang
J. Microbiol. 2022;60(10):986-997.   Published online September 27, 2022
DOI: https://doi.org/10.1007/s12275-022-2363-x
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AbstractAbstract PDF
Microbial communities in the rhizosphere play a crucial role in determining plant growth and crop yield. A few studies have been performed to evaluate the diversity and co-occurrence patterns of rhizosphere microbiomes in soybean (Glycine max) at a regional scale. Here, we used a culture-independent
method
to compare the bacterial communities of the soybean rhizosphere between Nebraska (NE), a high-yield state, and Oklahoma (OK), a low-yield state. It is well known that the rhizosphere microbiome is a subset of microbes that ultimately get colonized by microbial communities from the surrounding bulk soil. Therefore, we hypothesized that differences in the soybean yield are attributed to the variations in the rhizosphere microbes at taxonomic, functional, and community levels. In addition, soil physicochemical properties were also evaluated from each sampling site for comparative study. Our result showed that distinct clusters were formed between NE and OK in terms of their soil physicochemical property. Among 3 primary nutrients (i.e., nitrogen, phosphorus, and potassium), potassium is more positively correlated with the high-yield state NE samples. We also attempted to identify keystone communities that significantly affected the soybean yield using co-occurrence network patterns. Network analysis revealed that communities formed distinct clusters in which members of modules having significantly positive correlations with the soybean yield were more abundant in NE than OK. In addition, we identified the most influential bacteria for the soybean yield in the identified modules. For instance, included are class Anaerolineae, family Micromonosporaceae, genus Plantomyces, and genus Nitrospira in the most complex module (ME9) and genus Rhizobium in ME23. This research would help to further identify a way to increase soybean yield in low-yield states in the U.S. as well as worldwide by reconstructing the microbial communities in the rhizosphere.

Citations

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  • Interaction of soil microbial communities with soil nutrients, enzymes activity, and allelopathic compounds in flue-cured tobacco continuous monocropping field
    Yihong Pan, Tiyuan Xia, Qiulian Peng, Kun Huang, Lijuan Zhou, Guohuan Xiong
    Physiological and Molecular Plant Pathology.2026; 141: 103032.     CrossRef
  • Soybean productivity can be enhanced by understanding rhizosphere microbiota: evidence from metagenomics analysis from diverse agroecosystems
    Honglei Ren, Huilong Hong, Bire Zha, Sobhi F. Lamlom, Hongmei Qiu, Yongqiang Cao, Rujian Sun, Haorang Wang, Junkui Ma, Hengbin Zhang, Liping Sun, Qing Yang, Changjun Zhou, Xiulin Liu, Xueyang Wang, Chunlei Zhang, Fengyi Zhang, Kezhen Zhao, Rongqiang Yuan,
    Microbiome.2025;[Epub]     CrossRef
  • Impact of Bacterial Pustule Disease Occurrence on Bacterial and Fungal Communities Within Vegetable Soybean Plants
    Choosak Khaengraeng, Wuttichai Mhuantong, Usawadee Chaiprom, Sawita Suwannarat, Nattakorn Kuncharoen, Nutjarin Haewou, Warapon Bunkoed, Tiyakhon Chatnaparat
    Plant Pathology.2025; 74(5): 1228.     CrossRef
  • A molecular conveyor belt-associated protein controls the rotational direction of the bacterial type 9 secretion system
    Abhishek Trivedi, Jacob A. Miratsky, Emma C. Henderson, Abhishek Singharoy, Abhishek Shrivastava, Clay Fuqua
    mBio.2025;[Epub]     CrossRef
  • Discontinuation of glacial meltwater input reshapes the diversity and stability of eukaryotic planktonic microbial communities in glacial lakes
    Xiaodong Li, Qianggong Zhang, Peng Zhang, Qing Yang, Zijian Lu, Yanan Zhou, Meiqi Huang, Guangli Mu, Zhao Xue, Linyuan Lu, Yindong Tong
    Environmental Research.2025; 286: 122785.     CrossRef
  • Rice-microbiome-metabolite crosstalk: Deciphering the defence signatures of traditional rice landraces against blast (Magnaporthe oryzae)
    R. Karan, M.K. Prasannakumar, N. Vamsidharreddy, J. Harish, H.B. Mahesh, C. Manjunatha, K.N. Pallavi, Pramesh Devanna, Selva babu Selvamani, Aditya Narayan Sarangi
    Plant Stress.2025; 18: 101084.     CrossRef
  • The rhizosphere microbiome of 51 potato cultivars with diverse plant growth characteristics
    Benoit Renaud Martins, Viviane Radl, Krzysztof Treder, Dorota Michałowska, Karin Pritsch, Michael Schloter
    FEMS Microbiology Ecology.2024;[Epub]     CrossRef
  • Response of Soil Microorganisms and Phenolic to Pseudostelariae heterophylla Cultivation in Different Soil Types
    Yingying Liu, Dan Wu, Yongjun Kan, Li Zhao, Chang Jiang, Wensheng Pang, Juan Hu, Meilan Zhou
    Eurasian Soil Science.2024; 57(3): 446.     CrossRef
  • Analysis of the rhizosphere bacterial diversity of Angelica dahurica var. formosana from different experimental sites and varieties (strains)
    Meiyan Jiang, Fei Yao, Yunshu Yang, Yang Zhou, Kai Hou, Yinyin Chen, Dongju Feng, Wei Wu
    PeerJ.2023; 11: e15997.     CrossRef
  • Long-term fertilization coupled with rhizobium inoculation promotes soybean yield and alters soil bacterial community composition
    Wanling Wei, Dawei Guan, Mingchao Ma, Xin Jiang, Fenliang Fan, Fangang Meng, Li Li, Baisuo Zhao, Yubin Zhao, Fengming Cao, Huijun Chen, Jun Li
    Frontiers in Microbiology.2023;[Epub]     CrossRef
Complete gammaproteobacterial endosymbiont genome assembly from a seep tubeworm Lamellibrachia satsuma
Ajit Kumar Patra , Yong min Kwon , Youngik Yang
J. Microbiol. 2022;60(9):916-927.   Published online August 1, 2022
DOI: https://doi.org/10.1007/s12275-022-2057-4
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AbstractAbstract PDF
Siboglinid tubeworms thrive in hydrothermal vent and seep habitats via a symbiotic relationship with chemosynthetic bacteria. Difficulties in culturing tubeworms and their symbionts in a laboratory setting have hindered the study of host-microbe interactions. Therefore, released symbiont genomes are fragmented, thereby limiting the data available on the genome that affect subsequent analyses. Here, we present a complete genome of gammaproteobacterial endosymbiont from the tubeworm Lamellibrachia satsuma collected from a seep in Kagoshima Bay, assembled using a hybrid approach that combines sequences generated from the Illumina and Oxford Nanopore platforms. The genome consists of a single circular chromosome with an assembly size of 4,323,754 bp and a GC content of 53.9% with 3,624 protein-coding genes. The genome is of high quality and contains no assembly gaps, while the completeness and contamination are 99.33% and 2.73%, respectively. Comparative genome analysis revealed a total of 1,724 gene clusters shared in the vent and seep tubeworm symbionts, while 294 genes were found exclusively in L. satsuma symbionts such as transposons, genes for defense mechanisms, and inorganic ion transportations. The addition of this complete endosymbiont genome assembly would be valuable for comparative studies particularly with tubeworm symbiont genomes as well as with other chemosynthetic microbial communities.

Citations

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  • Comparative metagenomic analysis reveals the adaptive evolutionary traits of siboglinid tubeworm symbionts
    Jinyi Liu, Yingli Zhou, Jingchun Feng, Chaofeng Cai, Si Zhang
    Frontiers in Microbiology.2025;[Epub]     CrossRef
  • Genomic and transcriptomic analyses illuminate the molecular basis of the unique lifestyle of a tubeworm, Lamellibrachia satsuma
    Taiga Uchida, Yuki Yoshioka, Yu Yoshida, Manabu Fujie, Ayuta Yamaki, Akira Sasaki, Koji Inoue, Chuya Shinzato
    DNA Research.2023;[Epub]     CrossRef
Effect of biostimulation and bioaugmentation on hydrocarbon degradation and detoxification of diesel-contaminated soil: a microcosm study
Patricia Giovanella , Lídia de Azevedo Duarte , Daniela Mayumi Kita , Valéria Maia de Oliveira , Lara Durães Sette
J. Microbiol. 2021;59(7):634-643.   Published online May 15, 2021
DOI: https://doi.org/10.1007/s12275-021-0395-2
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AbstractAbstract PDF
Soil contamination with diesel oil is quite common during processes of transport and storage. Bioremediation is considered a safe, economical, and environmentally friendly approach for contaminated soil treatment. In this context, studies using hydrocarbon bioremediation have focused on total petroleum hydrocarbon (TPH) analysis to assess process effectiveness, while ecotoxicity has been neglected. Thus, this study aimed to select a microbial consortium capable of detoxifying diesel oil and apply this consortium to the bioremediation of soil contaminated with this environmental pollutant through different bioremediation approaches. Gas chromatography (GC-FID) was used to analyze diesel oil degradation, while ecotoxicological bioassays with the bioindicators Artemia sp., Aliivibrio fischeri (Microtox), and Cucumis sativus were used to assess detoxification. After 90 days of bioremediation, we found that the biostimulation and biostimulation/ bioaugmentation approaches showed higher rates of diesel oil degradation in relation to natural attenuation (41.9 and 26.7%, respectively). Phytotoxicity increased in the biostimulation and biostimulation/bioaugmentation treatments during the degradation process, whereas in the Microtox test, the toxicity was the same in these treatments as that in the natural attenuation treatment. In both the phytotoxicity and Microtox tests, bioaugmentation treatment showed lower toxicity. However, compared with natural attenuation, this approach did not show satisfactory hydrocarbon degradation. Based on the microcosm experiments results, we conclude that a broader analysis of the success of bioremediation requires the performance of toxicity bioassays.

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  • Heavy fuel oil-contaminated soil remediation by individual and bioaugmentation-assisted phytoremediation with Medicago sativa and with cold plasma-treated M. sativa
    Jūratė Žaltauskaitė, Rimas Meištininkas, Austra Dikšaitytė, Laima Degutytė-Fomins, Vida Mildažienė, Zita Naučienė, Rasa Žūkienė, Kazunori Koga
    Environmental Science and Pollution Research.2024; 31(20): 30026.     CrossRef
  • Soil Corrosivity Under Natural Attenuation
    Larissa O. da Silva, Sara H. de Oliveira, Rafael G. C. da Silva, Magda R. S. Vieira, Ivanilda R. de Melo, Severino L. Urtiga Filho
    Materials Research.2024;[Epub]     CrossRef
  • Updating risk remediation-endpoints for petroleum-contaminated soils? A case study in the Ecuadorian Amazon region
    Daniel Hidalgo-Lasso, Karina García-Villacís, Jeaneth Urvina Ulloa, Darwin Marín Tapia, Patricio Gómez Ortega, Frederic Coulon
    Heliyon.2024; 10(9): e30395.     CrossRef
  • Recent advances in the development and applications of luminescent bacteria–based biosensors
    Yingying Li, Yuankun Zhao, Yiyang Du, Xuechun Ren, He Ding, Zhimin Wang
    Luminescence.2024;[Epub]     CrossRef
  • Oil biodegradation studies with an immobilized bacterial consortium in plant biomass for the construction of bench-scale bioreactor
    Rachel M. Ferreira, Bernardo D. Ribeiro, Danielle.M.A. Stapelfeldt, Rodrigo P. do Nascimento, Maria de.F.R. Moreira
    Cleaner Chemical Engineering.2023; 6: 100107.     CrossRef
  • Application of Luminescent Bacteria Bioassay in the Detection of Pollutants in Soil
    Kai Zhang, Meng Liu, Xinlong Song, Dongyu Wang
    Sustainability.2023; 15(9): 7351.     CrossRef
  • Salicylate or Phthalate: The Main Intermediates in the Bacterial Degradation of Naphthalene
    Vasili M. Travkin, Inna P. Solyanikova
    Processes.2021; 9(11): 1862.     CrossRef
Review
Microbial source tracking using metagenomics and other new technologies
Shahbaz Raza , Jungman Kim , Michael J. Sadowsky , Tatsuya Unno
J. Microbiol. 2021;59(3):259-269.   Published online February 10, 2021
DOI: https://doi.org/10.1007/s12275-021-0668-9
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AbstractAbstract PDF
The environment is under siege from a variety of pollution sources. Fecal pollution is especially harmful as it disperses pathogenic bacteria into waterways. Unraveling origins of mixed sources of fecal bacteria is difficult and microbial source tracking (MST) in complex environments is still a daunting task. Despite the challenges, the need for answers far outweighs the difficulties experienced. Advancements in qPCR and next generation sequencing (NGS) technologies have shifted the traditional culture-based MST approaches towards culture independent technologies, where communitybased MST is becoming a method of choice. Metagenomic tools may be useful to overcome some of the limitations of community-based MST methods as they can give deep insight into identifying host specific fecal markers and their association with different environments. Adoption of machine learning (ML) algorithms, along with the metagenomic based MST approaches, will also provide a statistically robust and automated platform. To compliment that, ML-based approaches provide accurate optimization of resources. With the successful application of ML based models in disease prediction, outbreak investigation and medicine prescription, it would be possible that these methods would serve as a better surrogate of traditional MST approaches in future.

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  • Beyond borders: A systematic review and meta-analysis of human-specific faecal markers across geographical settings
    Leah R. Barrett, Paris Beasy, Yussi M. Palacios Delgado, John D. Boyce, Karin Leder, David T. McCarthy, Rebekah Henry
    Critical Reviews in Environmental Science and Technology.2025; 55(7): 447.     CrossRef
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    Abhay B. Fulke, Srilakshmi Mamidala, Pradnya Nikalje
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    Junxia Yuan, Jinlu Yang, Yue Sun, Yaqi Meng, Ziwei He, Wende Zhang, Lei Dang, Yan Song, Kanyan Xu, Na Lv, Ziding Zhang, Pei Guo, Hong Yin, Wenyu Shi
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    Sandra L. McLellan, Anthony Chariton, Annachiara Codello, Jill S. McClary-Gutierrez, Melissa K. Schussman, Ezequiel M. Marzinelli, Judith M. O’Neil, Eric J. Schott, Jennifer L. Bowen, Joe H. Vineis, Lois Maignien, Clarisse Lemonnier, Morgan Perennou, Kare
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    Science of The Total Environment.2024; 912: 168565.     CrossRef
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    Camila Duitama González, Riccardo Vicedomini, Téo Lemane, Nicolas Rascovan, Hugues Richard, Rayan Chikhi
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    Zeyou Chen, Yujing Duan, Lichun Yin, Ying Chen, Yingang Xue, Xiaolong Wang, Daqing Mao, Yi Luo
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    Katalin Demeter, Rita Linke, Elisenda Ballesté, Georg Reischer, René E Mayer, Julia Vierheilig, Claudia Kolm, Margaret E Stevenson, Julia Derx, Alexander K T Kirschner, Regina Sommer, Orin C Shanks, Anicet R Blanch, Joan B Rose, Warish Ahmed, Andreas H Fa
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    Job, O.S., Bala, J.D., Abdulraham, A.A., Friday, N.N., Ibekie, S.A., Tsebam, C.J, Abudullahi, D.
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    GeoHealth.2023;[Epub]     CrossRef
  • Humans and Hoofed Livestock Are the Main Sources of Fecal Contamination of Rivers Used for Crop Irrigation: A Microbial Source Tracking Approach
    Constanza Díaz-Gavidia, Carla Barría, Daniel L. Weller, Marilia Salgado-Caxito, Erika M. Estrada, Aníbal Araya, Leonardo Vera, Woutrina Smith, Minji Kim, Andrea I. Moreno-Switt, Jorge Olivares-Pacheco, Aiko D. Adell
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  • Interfacing Machine Learning and Microbial Omics: A Promising Means to Address Environmental Challenges
    James M. W. R. McElhinney, Mary Krystelle Catacutan, Aurelie Mawart, Ayesha Hasan, Jorge Dias
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    Renu Kumari, Benu Dhawan
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    Jang-Cheon Cho
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Journal Articles
Genetic linkage map construction and quantitative trait loci mapping of agronomic traits in Gloeostereum incarnatum
Wan-Zhu Jiang , Fang-Jie Yao , Li-Xin Lu , Ming Fang , Peng Wang , You-Min Zhang , Jing-Jing Meng , Jia Lu , Xiao-Xu Ma , Qi He , Kai-Sheng Shao
J. Microbiol. 2021;59(1):41-50.   Published online November 17, 2020
DOI: https://doi.org/10.1007/s12275-021-0242-5
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AbstractAbstract PDF
Gloeostereum incarnatum is an edible medicinal mushroom widely grown in China. Using the whole genome of G. incarnatum, simple sequence repeat (SSR) markers were developed and synthetic primers were designed to construct its first genetic linkage map. The 1,048.6 cm map is composed of 10 linkage groups and contains 183 SSR markers. In total, 112 genome assembly sequences were anchored, representing 16.43 Mb and covering 46.41% of the genome. Selfing populations were used for quantitative trait loci (QTL) targeting, and the composite interval mapping method was used to co-localize the mycelium growth rate (potato dextrose agar and sawdust), growth period, yield and fruiting body length, and width and thickness. The 14 QTLs of agronomic traits had LOD values of 3.20–6.51 and contribution rates of 2.22– 13.18%. No linkage relationship was found between the mycelium growth rate and the growth period, but a linkage relationship was observed among the length, width and thickness of the fruiting bodies. Using NCBI’s BLAST alignment, the genomic sequences corresponding to the QTL regions were compared, and a TPR-like protein candidate gene was selected. Using whole-genome data, 138 candidate genes were found in four sequence fragments of two SSR markers located in the same scaffold. The genetic map and QTLs established in this study will aid in developing selective markers for agronomic traits and identifying corresponding genes, thereby providing a scientific basis for the further gene mapping of quantitative traits and the marker-assisted selection of functional genes in G. incarnatum breeding programs.

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    Kameron T Wittmeyer, Sara J Oppenheim, Keith R Hopper, J Hesselberth
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[Protocol]Rapid method for chromatin immunoprecipitation (ChIP) assay in a dimorphic fungus, Candida albicans
Jueun Kim , Jung-Shin Lee
J. Microbiol. 2020;58(1):11-16.   Published online June 11, 2019
DOI: https://doi.org/10.1007/s12275-020-9143-2
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AbstractAbstract PDF
A chromatin immunoprecipitation (ChIP) assay is a method to identify how much a protein of interest binds to the DNA region. This method is indispensable to study the mechanisms of how the transcription factors or chromatin modifications regulate the gene expression. Candida albicans is a dimorphic pathogenic fungus, which can change its morphology very rapidly from yeast to hypha in response to the environmental signal. The morphological change of C. albicans is one of the critical factors for its virulence. Therefore, it is necessary to understand how to regulate the expression of genes for C. albicans to change its morphology. One of the essential methods for us to understand this regulation is a ChIP assay. There have been many efforts to optimize the protocol to lower the background signal and to analyze the results accurately because a ChIP assay can provide very different results even with slight differences in the experimental procedure. We have optimized the rapid and efficient ChIP protocol so that it could be applied equally for both yeast and hyphal forms of C. albicans. Our method in this protocol is also comparatively rapid to the method widely used. In this protocol, we described our rapid method for the ChIP assay in C. albicans in detail.

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β-1,3-Glucan/CR3/SYK pathway-dependent LC3B-II accumulation enhanced the fungicidal activity in human neutrophils
Ding Li , Changsen Bai , Qing Zhang , Zheng Li , Di Shao , Xichuan Li
J. Microbiol. 2019;57(4):263-270.   Published online February 5, 2019
DOI: https://doi.org/10.1007/s12275-019-8298-1
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AbstractAbstract PDF
Since molecular genotyping has been established for the Candida species, studies have found that a single Candida strain (endemic strain) can persist over a long period of time and results in the spread of nosocomial invasive candidiasis without general characteristics of horizontal transmissions. Our previous study also found the existence of endemic strains in a cancer center in Tianjin, China. In the current study, we performed further investigation on endemic and non-endemic Candida albicans strains, with the aim of explaining the higher morbidity of endemic strains. In an in vivo experiment, mice infected with endemic strains showed significantly shorter survival time and higher kidney fungal burdens compared to mice infected with non-endemic strains. In an in vitro experiment, the killing percentage of neutrophils to endemic strains was significantly lower than that to non-endemic strains, which is positively linked to the ratio of LC3B-II/I in neutrophils. An immunofluorescence assay showed more β-1,3-glucan exposure on the cell walls of nonendemic strains compared to endemic strains. After blocking the β-glucan receptor (CR3) or inhibiting downstream kinase (SYK) in neutrophils, the killing percent to C. albicans (regardless of endemic and non-endemic strains) and the ratio of LC3B-II/I of neutrophils were significantly decreased. These data suggested that the killing capability of neutrophils to C. albicans was monitored by β-1,3-glucan via CR3/SYK pathway-dependent LC3B-II accumulation and provided an explanation for the variable killing capability of neutrophils to different strains of C. albicans, which would be beneficial in improving infection control and therapeutic strategies for invasive candidiasis.

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Reviews
REVIEW] Plasma membrane organization promotes virulence of the human fungal pathogen Candida albicans
Lois M. Douglas , James B. Konopka
J. Microbiol. 2016;54(3):178-191.   Published online February 27, 2016
DOI: https://doi.org/10.1007/s12275-016-5621-y
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AbstractAbstract PDF
Candida albicans is a human fungal pathogen capable of causing lethal systemic infections. The plasma membrane plays key roles in virulence because it not only functions as a protective barrier, it also mediates dynamic functions including secretion of virulence factors, cell wall synthesis, invasive hyphal morphogenesis, endocytosis, and nutrient uptake. Consistent with this functional complexity, the plasma membrane is composed of a wide array of lipids and proteins. These components are organized into distinct domains that will be the topic of this review. Some of the plasma membrane domains that will be described are known to act as scaffolds or barriers to diffusion, such as MCC/eisosomes, septins, and sites of contact with the endoplasmic reticulum. Other zones mediate dynamic processes, including secretion, endocytosis, and a special region at hyphal tips that facilitates rapid growth. The highly organized architecture of the plasma membrane facilitates the coordination of diverse functions and promotes the pathogenesis of C. albicans.

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REVIEW] Hgc1-Cdc28–how much does a single protein kinase do in the regulation of hyphal development in Candida albicans?
Yue Wang
J. Microbiol. 2016;54(3):170-177.   Published online February 27, 2016
DOI: https://doi.org/10.1007/s12275-016-5550-9
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AbstractAbstract PDF
The fungal human pathogen Candida albicans can cause invasive infection with high mortality rates. A key virulence factor is its ability to switch between three morphologies: yeast, pseudohyphae and hyphae. In contrast to the ovalshaped unicellular yeast cells, hyphae are highly elongated, tube-like, and multicellular. A long-standing question is what coordinates all the cellular machines to construct cells with distinct shapes. Hyphal-specific genes (HSGs) are thought to hold the answer. Among the numerous HSGs found, only UME6 and HGC1 are required for hyphal development. UME6 encodes a transcription factor that regulates many HSGs including HGC1. HGC1 encodes a G1 cyclin which partners with the Cdc28 cyclin-dependent kinase. Hgc1- Cdc28 simultaneously phosphorylates and regulates multiple substrates, thus controlling multiple cellular apparatuses for morphogenesis. This review is focused on major progresses made in the past decade on Hgc1’s roles and regulation in C. albicans hyphal development and other traits important for infection.

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MINIREVIEW] Histone deacetylase-mediated morphological transition in Candida albicans
Jueun Kim , Ji-Eun Lee , Jung-Shin Lee
J. Microbiol. 2015;53(12):805-811.   Published online December 2, 2015
DOI: https://doi.org/10.1007/s12275-015-5488-3
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AbstractAbstract
Candida albicans is the most common opportunistic fungal pathogen, which switches its morphology from single-cell yeast to filament through the various signaling pathways responding to diverse environmental cues. Various transcriptional factors such as Nrg1, Efg1, Brg1, Ssn6, and Tup1 are the key components of these signaling pathways. Since C. albicans can regulate its transcriptional gene expressions using common eukaryotic regulatory systems, its morphological transition by these signaling pathways could be linked to the epigenetic regulation by chromatin structure modifiers. Histone proteins, which are critical components of eukaryotic chromatin structure, can regulate the eukaryotic chromatin structure through their own modifications such as acetylation, methylation, phosphorylation and ubiquitylation. Recent studies revealed that various histone modifications, especially histone acetylation and deacetylation, participate in morphological transition of C. albicans collaborating with well-known transcription factors in the signaling pathways. Here, we review recent studies about chromatin-mediated morphological transition of C. albicans focusing on the interaction between transcription factors in the signaling pathways and histone deacetylases.

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Research Support, Non-U.S. Gov'ts
Deletion analysis of LSm, FDF, and YjeF domains of Candida albicans Edc3 in hyphal growth and oxidative-stress response
Eung-Chul Kim , Jinmi Kim
J. Microbiol. 2015;53(2):111-115.   Published online January 28, 2015
DOI: https://doi.org/10.1007/s12275-015-4727-y
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AbstractAbstract
Candida albicans is an opportunistic fungal pathogen whose responses to environmental changes are associated with the virulence attributes. Edc3 is known to be an enhancer of the mRNA decapping reactions and a scaffold protein of cytoplasmic processing bodies (P-bodies). Recent studies of C. albicans Edc3 suggested its critical roles in filamentous growth and stress-induced apoptotic cell death. The edc3/edc3 deletion mutant strain showed increased cell survival and less ROS accumulation upon treatment with hydrogen peroxide. To investigate the diverse involvement of Edc3 in the cellular processes, deletion mutations of LSm, FDF, or YjeF domain of Edc3 were constructed. The edc3-LSmΔ or edc3-YjeFΔ mutation showed the filamentation defect, resistance to oxidative stress, and decreased ROS accumulation. In contrast, the edc3-FDFΔ mutation exhibited a wild-type level of filamentous growth and a mild defect in ROS accumulation. These results suggest that Lsm and YjeF domains of Edc3 are critical in hyphal growth and oxidative stress response.

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    Chenxing Wei, Caiyi Wen, Yuanyuan Zhang, Hongyan Du, Rongrong Zhong, Zhengzhe Guan, Mengjiao Wang, Yanhong Qin, Fei Wang, Luyang Song, Ying Zhao
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Cyclic Dipeptides from Lactic Acid Bacteria Inhibit the Proliferation of Pathogenic Fungi
Min-Kyu Kwak , Rui Liu , Min-Kyu Kim , Dohyun Moon , Andrew HyoungJin Kim , Sung-Hyun Song , Sa-Ouk Kang
J. Microbiol. 2014;52(1):64-70.   Published online January 4, 2014
DOI: https://doi.org/10.1007/s12275-014-3520-7
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AbstractAbstract PDF
Lactobacillus plantarum LBP-K10 was identified to be the most potent antifungal strain from Korean traditional fermented vegetables. The culture filtrate of this strain showed remarkable antifungal activity against Ganoderma boninense. Five fractions from the culture filtrate were observed to have an inhibitory effect against G. boninense. Also, the electron ionization and chemical ionization indicated that these compounds might be cyclic dipeptides. Of the five active fractions, two fractions showed the most significant anti-Ganoderma activity, and one of these fractions inhibited the growth of Candida albicans. These compounds were identified to be cis-cyclo(L-Val-L-Pro) and cis-cyclo(L-Phe-L-Pro), as confirmed by X-ray crystallography.

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Enhancement of Anti-candidal Activity of Endophytic Fungus Phomopsis sp. ED2, Isolated from Orthosiphon stamineus Benth, by Incorporation of Host Plant Extract in Culture Medium
Tong Woei Yenn , Chong Chai Lee , Darah Ibrahim , Latiffah Zakaria
J. Microbiol. 2012;50(4):581-585.   Published online July 21, 2012
DOI: https://doi.org/10.1007/s12275-012-2083-8
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AbstractAbstract PDF
This study examined the effect of host extract in the culture medium on anti-candidal activity of Phomopsis sp. ED2, previously isolated from the medicinal herb Orthosiphon stamineus Benth. Interestingly, upon addition of aqueous host extract to the culture medium, the ethyl acetate extract prepared from fermentative broth exhibited moderate anticandidal activity in a disc diffusion assay. The minimal inhibitory concentration of this extract was 62.5 μg/ml and it only exhibited fungistatic activity against C. albicans. In the time-kill study, a 50% growth reduction of C. albicans was observed at 31.4 h for extract from the culture incorporating host extract. In the bioautography assay, only one single spot (Rf 0.59) developed from the extract exhibited anti-candidal activity. A spot with the a similar Rf was not detected for the crude extract from YES broth without host extract. This indicated that the terpenoid anti-candidal compound was only produced when the host extract was introduced into the medium. The study concluded that the incorporation of aqueous extract of the host plant into the culture medium significantly enhanced the anti-candidal activity of Phomopsis sp. ED2.

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Review
REVIEW] Candida albicans, a Major Human Fungal Pathogen
Joon Kim , Peter Sudbery
J. Microbiol. 2011;49(2):171-177.   Published online May 3, 2011
DOI: https://doi.org/10.1007/s12275-011-1064-7
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AbstractAbstract PDF
Candida albicans is the most common human fungal pathogen (Beck-Sague and Jarvis, 1993). It is normally a harmless commensal organism. However, it is a opportunistic pathogen for some immunologically weak and immunocompromised people. It is responsible for painful mucosal infections such as the vaginitis in women and oral-pharangeal thrush in AIDS patients. In certain groups of vulnerable patients it causes severe, life-threatening bloodstream infections and it causes severe, life-threatening bloodstream infections and subsequent infections in the internal organs. There are various fascinating features of the C. albicans life cycle and biology that have made the pathogen the subject of extensive research, including its ability to grow in unicellular yeast, psudohyphal, and hyphal forms (Fig. 1A); its ability to switch between different but stable phenotypic states, and the way that it retains the ability to mate but apparently loses the ability to go through meiosis to complete the sexual cycle. This research has been greatly facilitated by the derivation of the complete C. albicans genome sequence (Braun et al., 2005), the development of a variety of molecular tools for gene manipulation, and a store of underpinning knowledge of cell biology borrowed from the distantly related model yeast Saccharomyces cerevisiae (Berman and Sudbery, 2002; Noble and Johnson, 2007). This review will provide a brief overview of the importance of C. albicans as a public health issue, the experimental tools developed to study its fascinating biology, and some examples of how these have been applied.

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Research Support, Non-U.S. Gov'ts
The Three-Dimensional Morphology of Candida albicans as Seen by High-Resolution Scanning Electron Microscopy
Michela Isola , Raffaella Isola , Maria Serenella Lantini , Alessandro Riva
J. Microbiol. 2009;47(3):260-264.   Published online June 26, 2009
DOI: https://doi.org/10.1007/s12275-008-0212-1
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AbstractAbstract PDF
The fine structure of Candida albicans has been repeatedly described by transmission electron microscopy, whereas studies by high-resolution scanning electron microscopy (HRSEM) are rare and devoted solely to the study of its external morphology. This report describes the results of an HRSEM study on C. albicans carried out by an osmium maceration protocol modified to better retain the structural characteristics of this yeast. Thus, we visualized various intracellular structures including invaginations of cell membrane (plasmalemmasomes), nuclear envelope, mitochondria, the vacuolar system, and two additional structures that might represent a form of endoplasmic reticulum and the Golgi apparatus. The present investigation, which for the first time shows the organelles of C. albicans at the 3D level, may lead to a better understanding of its cell physiology.

Citations

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Characterization of Osh3, an Oxysterol-binding Protein, in Filamentous Growth of Saccharomyces cerevisiae and Candida albicans
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J. Microbiol. 2006;44(5):523-529.
DOI: https://doi.org/2445 [pii]
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OSH3 is one of the seven yeast homologues of the oxysterol binding proteins (OSBPs) which have the major binding affinity to the oxysterols and function as regulator of cholesterol biosynthesis in mammals. Mutational analysis of OSH3 showed that OSH3 plays a regulatory role in the yeast-to-hyphal transition through its oxysterol-binding domain in Saccharomyces cerevisiae. The OSH3 gene was also identified in the pathogenic yeast Candida albicans. Deletion of OSH3 caused a defect in the filamentous growth, which is the major cause of the C. albicans pathogencity. The filamentation defect of the mutation in the MAPK-associated transcription factor, namely cph1Δ was suppressed by overexpression of OSH3. These findings suggest the regulatory roles of OSH3 in the yeast filamentous growth and the functional conservations of OSH3 in S. cerevisiae and C. albicans.
Role of CaBud6p in the Polarized Growth of Candida albicans
Yunkyoung Song , Jeong-Yoon Kim
J. Microbiol. 2006;44(3):311-319.
DOI: https://doi.org/2381 [pii]
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Bud6p is a component of a polarisome that controls cell polarity in Saccharomyces cerevisiae. In this study, we investigated the role of the Candida albicans Bud6 protein (CaBud6p) in cell polarity and hyphal development. CaBud6p, which consists of 703 amino acids, had 37% amino-acid sequence identity with the Bud6 protein of S. cerevisiae. The homozygous knock-out of CaBUD6 resulted in several abnormal phenotypes, such as a round and enlarged cells, widened bud necks, and a random budding pattern. In hypha-inducing media, the mutant cells had markedly swollen tips and a reduced ability to switch from yeast to hypha. In addition, a yeast two-hybrid analysis showed a physical interaction between CaBud6p and CaAct1p, which suggests that CaBud6p may be involved in actin cable organization, like Bud6p in S. cerevisiae. Taken together, these results indicate that CaBud6 plays an important role in the polarized growth of C. albicans.
Journal Article
Molecular Epidemiological Analysis of Bloodstream Isolates of Candida albicans from a University Hospital over a Five-Year Period
Jong Hee Shin , Yu Gyung Og , Duck Cho , Seung Jung Kee , Myung Geun Shin , Soon Pal Suh , Dong Wook Ryang
J. Microbiol. 2005;43(6):546-554.
DOI: https://doi.org/2291 [pii]
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We assessed the genetic relations and epidemiological links among bloodstream isolates of Candida albicans, which were obtained from a university hospital over a period of five years. The 54 bloodstream isolates from the 38 patients yielded 14 different karyotypes, 29 different patterns after digestion with SfiI (REAG-S), and 31 different patterns after digestion with BssHII (REAG-B) when analyzed using three different pulsed-field gel electrophoresis (PFGE) typing methods. In 11 patients with serial bloodstream isolates, all strains from each patient had the same PFGE pattern. The dendrograms for all of the strains revealed that the distribution of similarity values ranged from 0.70 to 1.0 in the REAG-S patterns, and from 0.35 to 1.0 in the REAG-B patterns. Overall, the combination of the three different PFGE methods identified 31 distinct types, reflecting the results obtained using the REAG-B alone different. different Five PFGE types were shared among 22 isolates from 12 patients. These types of strains were more frequently associated with central venous catheter-related fungemia than the other 26 type strains (92% versus 31%; P < 0.005). Of five PFGE types, four isolates were determined to be epidemiologically related: each of these types was primarily from two or three patients who had been hospitalized concurrently within the same intensive care unit. Our results suggest that the REAG-B constitutes perhaps the most useful PFGE method for investigating C. albicans candidemia and also shows that a relatively high proportion of C. albicans candidemia may be associated with exogenous acquisition of clonal strains.
Research Support, Non-U.S. Gov't
Complete Sequence of a Gene Encoding KAR3-Related Kinesin-like Protein in Candida albicans
Min-Kyoung Kim , Young Mi Lee , Wankee Kim , Wonja Choi
J. Microbiol. 2005;43(5):406-410.
DOI: https://doi.org/2283 [pii]
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In contrast to Saccharomyces cerevisiae, little is known about the kinesin-like protein (KLP) in Candida albicans. The motor domain of kinesin, or KLP, contains a subregion, which is well conserved from yeast to humans. A similarity search, with the murine ubiquitous kinesin heavy chain region as a query, revealed 6 contigs that contain putative KLPs in the genome of C. albicans. Of these, the length of an open reading (ORF) of 375 amino acids, temporarily designated CaKAR3, was noticeably short compared with the closely related S. cerevisiae KAR3 (ScKAR3) of 729 amino acids. This finding prompted us to isolate a [lambda] genomic clone containing the complete CaKAR3 ORF, and here the complete sequence of CaKAR3 is reported. CaKAR3 is a C-terminus motor protein, of 687 amino acids, encoded by a non-disrupting gene. When compared with ScKAR3, the amino terminal region of 112 amino acids was unique, with the middle part of the 306 amino acids exhibiting 25% identity and 44% similarity, while the remaining C-terminal motor domain exhibited 64% identity and 78% similarity, and have been submitted to GeneBank under the accession number AY182242.
Purification and characterization of extracellular aspartic proteinase of candida albicans
Na, Byoung-Kuk , Lee, Seong Il , Kim, Sin Ok , Park, Young Kil , Bai, Gill Han , Kim, Sang Jae , Song, Chul Yong
J. Microbiol. 1997;35(2):109-116.
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An extracellular proteinase of Candida albicans was purified by a combination of 0-75% ammonium sulfate precipitation, DEAE Sepharose Fast Flow ion exchange chromatography, and Sephacryl S-200 HR molecular sieve chromatography. Its mlecular weight was approximately 41 kDa on SDS-PAGE and isoelectric point was 4.4. The enzyme was inhibited by pepstain A. Optimum enzyme activity ranged from pH 2.0 to 3.5 with its maximum at pH 2.5 and a temperature of 45℃. The addition of divalent cations, Ca^2+, Zn^2+ and Mg^2+, resulted in no significant inhibition of enzymatic activity. However, some inhibitory effects were observed by Fe^2+, Ag^2+ and Cu^2+. With BSA as substrate, an apparent K_m was determined to be 7 × 10^7 M and K_I, using pepstatin A as an inhibitor, was 8.05 × 10^8 M. N-terminal amino acid sequence was QAVPVTLXNEQ. Degradation of BSA and fibronectin was shown but not collagen, hemoglobin, immunoglobulin G, or lysozyme. The enzyme preferred peptides with Glu and Leu at the P₁position, but the enzyme activity was highly reduced when the P₂position was Phe or Pro. This enzyme showed antigenicity against sera of patients with candidiasis.
Protective Effects of Antoxidant Enzymes of Candida albicans against Oxidative Killing by Macrophages
Kim, Hye Jin , Na, Byoung Kuk , Kim, Moon Bo , Choi, Duk Young , Song, Chul Yong
J. Microbiol. 1999;37(2):117-122.
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Protective roles of antioxidant enzymes, copper-zinc superoxide dismutase (CuZnSOD), manganese superoxide dismutase (MnSOD), and catalase of Candida albicans against exogenous reactive oxygens and oxidative killing by macrophages were investigated. The initial growth of C. albicans was inhibited by reactive, oxygen-producing chemicals such as hydrogen peroxide, pyrogallol, and paraquat, but it was restored as the production of antioxidant enzymes were increased. The growth inhibition of C. albicans by reactive, oxygen-producing chemicals was reduced by treating the purified candidal SOD and catalase. Also, in the presence of SOD and catalase, the oxidative killing of C. albicans by macrophages was significantly inhibited. These results suggest that antioxidant enzymes, CuZnSOD, MnSOD, and catalase of C. albicans may play important roles in the protection of C. albicans not only from exogenous oxidative stress but also from oxidative killing by macrophages.
Rapid PCR Method for Detecting Candida albicans Using Primers Derived from the Integrin-like Protein Gene [alpha]INT1 of Candida albicans
Young-Hee Lim , Do-Hyun Lee
J. Microbiol. 2000;38(2):105-108.
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Oligonucleotide primers amplifying a 344 bp fragment on the integrin-like protein alpha-INT1p gene ([alpha]INT1) of Candida albicans were synthesized for screening of C. albicans from clinicalsamples by the polymerase chain reaction (PCR). The PCR specifically amplified DNA from C. albicans and none from any other Candida, fungal, or human DNA in standard strains used here. The PCR assay showed that the primers (LH1 and LH2) were specific for 26 isolates of C. albicans from clinical samples, whereas the positive fragment, 344 bp, was not amplified from 15 clinical isolates including 14 other medically important Candida species and an isolate of Saccharomyces cerevisiae. PCR was conducted on the urine samples of 20 patients and 4 samples were C. albicans positive. The detection limit of the PCR assay for C. albicans was shown to be approximately 10 cells/ml saline. The PCR system using 344 bp [alpha]INT1 as a target is more specific and rapid than the conventional culture method, and the sensitive detection method is applicable to clinical diagnosis of C. albicans infections.
Intracellular Posttranslational Modification of Aspartyl Proteinase of Candida albicans and the Role of the Glycan Region of the Enzyme
Byoung-Kuk Na , Chul-Yong Song
J. Microbiol. 2000;38(4):218-223.
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Using two drugs, tunicamycin and brefeldin A, which affect protein processing, we investigated the intracellular processing mechanism of secreted aspartyl proteinase 1 (SAP1) of Candida albicans. Three intracellular forms of SAP1 were detected by immunoblotting using monoclonal antibody (MAb) CAP1. Their molecular weights were approximately 40, 41 and 45 kDa, respectively. The 41 kDa protein is a glycoprotein and may be the same as the extracellular form judging by its molecular mass. The 40 kDa protein was the unglycosylated form and its molecular mass coincided with deglycosylated SAP1 and the 45 kDa protein was also the unglycosylated form. Neither the 40 and 45 kDa proteins were detected in the culture supernatant of C. albicans. These suggested that the 40 and 45 kDa proteins might be intracellular precursor forms of SAP1. These results show that SAP1 is translated as a 45 kDa precusor form in the endoplasmic reticulum and the 45 kDa precursor form undergoes proteolytic cleavage after translocation into the Golgi apparatus, generating the 40 kDa precursor form. This 40 kDa precursor is converted into a 41 kDa mature form through glycosylation in the Golgi apparatus. The mature form of the 41 kDa protein is sorted into secretory vesicles and finally released into the extracellular space through membrane fusion. When the glycan region of SAP1 was digested with N-glycosidase F, both stability and activity of the enzyme decreased. These results indicate that the glycan attached to the enzyme may, at least in part, be related to enzyme stability and activity.
Multiplex Polymerase Chain Reaction Assay for Simultaneous Detection of Candida albicans and Candida dubliniensis
Young-Hee Lim , Do-Hyun Lee
J. Microbiol. 2002;40(2):146-150.
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A multiplex polymerase chain reaction (PCR) assay was developed for the identification of two Candida species-albicans and dubliniensis. Three sets of primers were selected from different genomic sequences to specifically amplify a 516 bp fragment within the top2 gene, specific for several species of the genus Candida (CCL primers); a 239 bp fragment within the [alpha]INT1 gene, specific for Candida albicans (CAL primers); and a 175 bp fragment within the ALSD1 gene, specific for Candida dubliniensis (CDL primers). Using the primers in conjunction (multiplex PCR), we were able to detect both C. albicans and C. dubliniensis and to differentiate between them. The detection limit of the PCR assay was approximately 10 cells per milliliter of saline. Thus, this multiplex PCR assay can be applied for differentiation of C. albicans and C. dubliniensis from clinical specimens.

Journal of Microbiology : Journal of Microbiology
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