An insight into the role of protein kinases as virulent factors, regulating pathogenic attributes in Candida albicans.

Candida albicans is a polymorphic, opportunistic pathogen, member of normal human microbiome causing candidiasis. It causes wide range of infections from superficial skin infections to life-threatening systemic infections. The pathogenicity in C. albicans attributes through several morphological characteristics and virulence factors. These morphological features are regulated by various molecules among which kinases are the most important. Several kinases and kinase signaling cascades play a well established role in Candidiasis. In this review we present an update on our current understanding of the pathogenicity attributes which is regulated by kinases as virulence factors.

Partial purification and characterization of a thermophilic and alkali-stable laccase of Phoma herbarum isolate KU4 with dye-decolorization efficiency.

Production of an extracellular thermophilic and alkali stable laccase from Phoma herbarum isolate KU4 was reported for the first time, both in submerged fermentation (SmF, highest 1590 U/mL) and solid state fermentation (SSF, highest 2014.21 U/mL) using agro-industrial residues. The laccase was partially purified to 7.93 fold with the apparent molecular weight of 298 kDa. The enzyme had pH optimum at 5.0 and temperature optimum at 50 °C, with maximum stability at pH 8.0. It showed activity towards various phenolic and non-phenolic compounds. The kinetic parameters, Km, Vmax and Kcat of the laccase for DMP were 0.216 mM, 270.27 U/mg and 506.69 s-1, respectively. Laccase activity was inhibited by various metal ions and conventional inhibitors, however, it was slightly increased by Zn2+. The laccase showed good decolorization efficiency towards four industrial dyes, namely, methyl violet (75.66%), methyl green (65%), indigo carmine (58%) and neutral red (42%) within 24 h. FTIR analysis of the decolorized products confirmed the degradation of the dyes. The decolorization efficiency of the enzyme suggests that the partially purified laccase could be used to decolorize synthetic dyes present in industrial effluents and for waste water treatments. The thermophilic and alkali stable laccase may also have wider potential industrial applications.

Evaluation of cancer awareness, cancer education, and prevention intervention techniques among university-level students in the United States and India.

BACKGROUND: Cancer is one of the leading causes of death globally. A considerable number of different cancer types may be preventable, using primary intervention techniques, such as health education, cancer awareness, behaviors and lifestyle modifications. The present study conducted a comparative assessment of cancer awareness among undergraduate students of the United States and India. MATERIALS AND METHODS: Students from an Institution in India (KC) (55 females, 33 males), and an Institution in the United States of America (SHU) (226 female, 58 male) during 2019-2020 participated in this study. Participants (n = 372) across all majors and all years (first through fourth year) completed an online questionnaire and answered the questions on their demographic characteristics (e.g., gender, age, and location), academic status (e.g., year of study, major), multiple-choice questions about cancer knowledge, and opinion questions (e.g., "where would you find info," "should therapies be free"). Student responses were collected using Qualtrics Survey Software. Excel was used to analyze responses. We conducted statistical Χ2 tests for independence to determine whether there is a statistically significant difference between the expected frequencies and the observed frequencies in one or more categories of a contingency table, with a significance of ɑ = 0.01. While small sizes due to the small institutions and the response pool, we note that we achieved the necessary "n" for all tests reported. RESULTS: Our research shows a few important statistically significant differences, including knowledge of cancer and breast lumps is dependent on location, ranking of global cancer deaths is dependent on location, and that cancer knowledge is dependent on the information source. All for Χ2 tests with P < 0.001. CONCLUSIONS: Further encouragement of education for young people in various aspects of cancer and cancer prevention, as well as information facility and sources of reliable data, could be helpful for improving the overall health and primary prevention. A thorough assessment is needed to understand the responsible factors for the observed cancer knowledge variations among students of two different places.

In silico analyses of Wnt1 nsSNPs reveal structurally destabilizing variants, altered interactions with Frizzled receptors and its deregulation in tumorigenesis.

Wnt1 is the first mammalian Wnt gene, which is discovered as proto-oncogene and in human the gene is located on the chromosome 12q13. Mutations in Wnt1 are reported to be associated with various cancers and other human diseases. The structural and functional consequences of most of the non-synonymous SNPs (nsSNPs), present in the human Wnt1 gene, are not known. In the present work, extensive bioinformatics analyses are used to screen 292 nsSNPs of Wnt1 for predicting pathogenic and harmless polymorphisms. We have identified 10 highly deleterious nsSNPs among which 7 are located within the highly conserved areas. These 10 nsSNPs are also predicted to affect the post-translational modifications of Wnt1. Further, structure based stability analyses of these 10 highly deleterious nsSNPs revealed 8 variants as highly destabilizing. These 8 highly destabilizing variants were shown to have high BC score and high RMSIP score from normal mode analyses. Based on the deformation energies, obtained from the normal mode analyses, variants like G169A, G169S, G331R and G331S were found to be unstable. Molecular Dynamics (MD) simulations revealed structural stability and fluctuation of WT Wnt1 and its prioritized variants. RMSD remained fluctuating mostly between 4 and 5 Å and occasionally between 3.5 and 5.5 Å ranges. RMSF in the CTD region (residues 330-360) of the binding pocket were lower compared to that of WT. Studying the impacts of nsSNPs on the binding interface of Wnt1 and seven Frizzled receptors have predicted substitutions which can stabilize or destabilize the binding interface. We have found that Wnt1 and FZD8-CRD is the best docked complex in our study. MD simulation based analyses of wild type Wnt1-FZD8-CRD complex and the 8 prioritized variants revealed that RMSF was higher in the unstructured regions and RMSD remained fluctuating in the region of 5 Å ± 1 Å. We have also observed differential Wnt1 gene expression pattern in normal, tumor and metastatic conditions across different tissues. Wnt1 gene expression was significantly higher in metastatic tissues of lungs, colon and skin; and was significantly lower in metastatic tissues of breast, esophagus and kidney. We have also found that Wnt1 deregulation is associated with survival outcome in patients with gastric and breast cancer. Furthermore, these computationally screened highly deleterious nsSNPs of Wnt1 can be analyzed in population based genetic studies and may help understand the Wnt1 associated diseases.

Kinome analyses of Candida albicans, C. parapsilosis and C. tropicalis enable novel kinases as therapeutic drug targets in candidiasis.

Candida spp. have attracted considerable attention as they cause serious human diseases in immunocompromised individuals. The genomes of the pathogenic Candida spp. have been sequenced, but systemic characterizations of their kinomes are yet to be reported. As in various eukaryotes, the protein kinases play crucial regulatory roles in pathogenicity of Candida. Increased frequency of antifungal resistance in Candida spp. requires significant attention to explore novel therapeutic molecules for their control. The present in-silico study involves novel bioinformatics strategies to identify the kinase proteins and their potential drug targets with the purpose to combat fungal infections. The study reports 103, 107 and 106 kinase proteins from 3 Candida spp., C. albicans, C. parapsilosis and C. tropicalis, respectively. Moreover, 79 common kinase proteins were identified, of which 54 proteins play essential roles in Candida spp. and 42 proteins were human non-homologues. Among the essential and human non-homologous protein kinases, 9 were found to be common essential human non-homologues, of which 6 are uniquely present in Candida. These 6 protein kinases namely, Hsl1, Npr1, Ptk2, Kin2, Ksp1 and orf19.3854 (CAALFM_CR06040WA) are involved in various molecular and cellular processes regulating virulence or pathogenicity. Further, these 6 kinases are prioritized as potential drug targets and explored for discovering new lead compounds against candidiasis. The drug repurposing approach for these 6 kinases show 13 approved drugs and investigational compounds that might play substantial inhibitory roles during combating candidiasis.

Network analysis of hyphae forming proteins in Candida albicans identifies important proteins responsible for pathovirulence in the organism.

Candida albicans causes two types of major infections in humans: superficial infections, such as skin and mucosal infection, and life-threatening systemic infections, like airway and catheter-related blood stream infections. It is a polymorphic fungus with two distinct forms (yeast and hyphal) and the morphological plasticity is strongly associated with many disease causing proteins. In this study, 137 hyphae associated proteins from Candida albicans (C. albicans) were collected from different sources to create a Protein-Protein Interaction (PPI) network. Out of these, we identified 18 hub proteins (Hog1, Hsp90, Cyr1, Cdc28, Pkc1, Cla4, Cdc42, Tpk1, Act1, Pbs2, Bem1, Tpk2, Ras1, Cdc24, Rim101, Cdc11, Cdc10 and Cln3) that were the most important ones in hyphae development. Ontology and functional enrichment analysis of these proteins could categorize these hyphae associated proteins into groups like signal transduction, kinase activity, biofilm formation, filamentous growth, MAPK signaling etc. Functional annotation analysis of these proteins showed that the protein kinase activity to be essential for hyphae formation in Candida. Additionally, most of the proteins from the network were predicted to be localized on cell surface or periphery, suggesting them as the main protagonists in inducing infections within the host. The complex hyphae formation phenomenon of C. albicans is an attractive target for exploitation to develop new antifungals and anti-virulence strategies to combat C. albicans infections. We further tried to characterize few of the most crucial proteins, especially the kinases by their sequence and structural prospects. Therefore, through this article an attempt to understand the hyphae forming protein network analysis has been made to unravel and elucidate the complex pathogenesis processes with the principal aim of systems biological research involving novel Bioinformatics strategies to combat fungal infections.

Tamarind kernel powder: a novel agro-residue for the production of cellobiose dehydrogenase under submerged fermentation by Termitomyces clypeatus.

The study investigates the potential of substitution of the conventional carbohydrate nutrient (cellulose) in media with cheap agro-residues for cellobiose dehydrogenase production by Termitomyces clypeatus (CDHtc) under submerged conditions. Different agro-residues tested for enzyme production were characterized using FTIR and XRD analysis. As CDHtc production was highest with tamarind kernel powder (TKP), it was selected for process optimizations through shake-flask fermentations. The optimized parameters were then applied to batch cultures in a 5 L bioreactor that gave enzyme yield (57.4 U mL⁻¹) similar to that obtained under shake-flask fermentations (57.05 U mL⁻¹). The study also made an attempt to predict CDHtc production with respect to time of fermentation and mycelial growth. The specific growth rate and carrying capacity of the mycelia were also determined, and the values lie in the ranges of 0.024-0.027 h⁻¹ and 7.2-7.1 mg mL⁻¹, respectively.

Interference of laccase in determination of cellobiose dehydrogenase activity of Pleurotus ostreatus (Florida) using dichlorophenol indophenol as the electron acceptor.

Pleurotus ostreatus (Florida), ITCC 3308 produces approximately 9.0 U/ml extracellular cellobiose dehydrogenase (CDH) in cellulose medium after 7 days of growth. However, no activity could be detected if the assay was done with cellobiose as the substrate and 2,6-dichlorophenol indophenol (DPIP) as the electron acceptor in absence of any laccase inhibitor. Kinetic study showed that V(max)/K(m) value was very high for rDPIP (reduced 2,6-dichlrophenol indophenol) oxidation by laccase. Oxygen consumption rate of rDPIP oxidation by the enzyme was found to be highest among all the tested substrates. The present study indicated that rDPIP was a good substrate for laccase. Therefore, caution is needed to measure CDH activity by monitoring DPIP reduction in a system where laccase is likely to be present.