Effect of cigarette smoke on "Candida albicans" growth and its interaction with human gingival fibroblasts

L’objectif principal de cette étude est d’étudier l’effet de la fumée de cigarette (FC) sur C. albicans et sur son interaction avec les fibroblastes gingivaux humains. Nous avons démontré que Candida se multiplie présence FC, en adoptant la forme hyphe. La FC rende C. albicans sensible au H2O2, mais résistant aux la chaleur et au NaCl. La FC augmente la production de chitine par C. albicans. Préincubé avec le FC, C. albicans adhère beaucoup plus aux cellules gingivales en monocouche, prolifère plus et adopte la forme hyphe plus facilement. Les fibroblastes quant à eux ils montrent une réduction de leur croissance, mais produisent plus de l’IL-1b. En conclusion: le CFC module d’un côté C. albicans et de l’autre côté les cel-lules de l’hôte. Ceci suggère un négatif important de la fumée de cigarette favorisant les candidoses orales chez les fumeurs.The main objective of this study was to investigate the effect of cigarette smoke (CS) on C. albicans and its interaction with human gingival fibroblasts. We have shown that Candida multiplies presence CS, by adopting the hyphae form. CS makes C. albicans sensitive H2O2, but resistant to heat and NaCl. CS in-creases chitin production by C. albicans. Preincubated with CS, C. albicans adheres more to gingival cells in monolayer, proliferating more and adopts the hyphae form more easily. Fibroblasts show a reduction of their growth, but produce more IL-1b. CSC modulates C. albicans and on the other side also CS modulates the host cells. This suggests a significant negative cigarette smoke promotes oral candidiasis in smokers

The effects of electronic cigarettes on human gingival cells and Candida albicans

Plusieurs alternatives ont été mises au point pour réduire les effets de la cigarette sur la santé buccale et générale. La plus récente de ces initiatives est la cigarette électronique. Plusieurs études montrent que la cigarette électronique contient moins de produits toxiques comparativement à la cigarette standard. Ces études concluent que la cigarette électronique est moins nocive pour la santé. Cependant, d’autres études émettent des doutes sur l’innocuité de la cigarette électronique étant donné la présence de multiples produits chimiques. Ces derniers peuvent interagir négativement avec plusieurs parties du corps, dont la cavité buccale. Les objectifs de cette étude sont (i) d’évaluer les effets d’expositions répétées (1, 2 ou 3 fois) au condensé de cigarette électronique sur la morphologie, la croissance, la migration et l’apoptose des fibroblastes gingivaux humains (ii) d’évaluer les effets de la vapeur de la cigarette électronique sur la croissance, la production de chitine et l’expression de certains gènes codant pour des protéines de la famille des "secreted aspartyl proteinases (SAP par C. albicans avec des temps d’exposition de 15 min, deux fois par jour, pendant 2 et 3 jours. (iii) d’évaluer l’interaction des cellules épithéliales gingivales avec C. albicans préalablement exposé à la cigarette électronique. Nous avons utilisé différentes techniques de biologie cellulaire, de biologie moléculaire et de microbiologie. Nos travaux montrent que les fibroblastes exposés au condensé de cigarette électronique ont une morphologie anormale (cellules plus grosses, vacuolées,.) et un taux de prolifération plus faible comparativement aux cellules non exposées. Ces observations sont consolidées par un taux plus élevé de cellules apoptotiques comparativement aux cellules non exposées. L’analyse de la migration cellulaire montre que le condensé de cigarette électronique réduit de façon significative la capacité de migration des fibroblastes. Il est à noter que les effets sont plus importants avec la cigarette, suivi de la cigarette électronique contenant la nicotine, puis celle sans nicotine. Les effets de la cigarette électronique sont moins importants que ceux du condensé de cigarette, mais plus sérieux comparativement aux cellules non exposées. Nos études montrent que l’exposition de C. albicans à la cigarette électronique entraîne une augmentation de sa croissance. Cette observation est supportée par un taux plus élevé de chitine produite par C. albicans exposé à la cigarette électronique. L’analyse de la transformation montre des formes hyphes plus longues après l’exposition à cigarette électronique. Nous avons aussi observé que la cigarette électronique augmente l’expression des gènes SAP2, SAP3 et SAP9 par C. albicans comparativement au contrôle (non exposé). L’exposition de C. albicans à la cigarette électronique favorise l'adhésion de la levure aux cellules épithéliales, augmente le taux de tyransformation de la levure. L’exposition de C. albicans à la cigarette électronique, puis son contact avec les cellules épithéliales cause une libération importante de la lactate deshydrogénase (LDH), et la différenciation des cellules épithéliales, mais réduit le taux de croissance de ces cellules gingivales. Les résultats globaux indiquent que les cigarettes électroniques peuvent interagir avec le microbiome buccal de l’utilisateur. Parce que les cigarettes électroniques réduisent la croissance des cellules gingivales et augmentent l'apoptose cellulaire, cela peut diminuer l'immunité innée dans la cavité buccale, ce qui pourrait augmenter le risque d'infections buccales, telles que la candidoseElectronic cigarettes (e-cigarettes) were designed to replace regular cigarette smoking and to contribute to smoking cessation. E-cigarettes require the use of vaping liquid that contains propylene glycol (PG) and vegetable glycerin (VG) as well as nicotine in various concentrations and flavours. Several studies comparing ecigarettes to conventional cigarettes show that e-cigarettes contain lower levels of toxic compounds and for this reason are deemed safer. However, a growing body of evidence shows that e-cigarettes contain many chemicals including formaldehyde, acetaldehyde, acrolein, and toluene, which may have adverse effects on different body parts, including the oral cavity. The first objective of this study was to investigate the impact of repeated exposures (1, 2, or 3 times) to e-cigarette condensates with or without nicotine on normal human gingival fibroblast morphology, proliferation, migration, and apoptosis. The second objective was to evaluate the effect of e-cigarettes vapors on the growth changes of C. albicans from blastospore to hyphal form and the expression of secreted aspartic proteinases (SAPs) SAP2, SAP3, and SAP9 genes by C. albicans, with exposure times of 15 min twice a day for 2 and 3 days. The third objective was to shed light on the interaction between e-cigarette-exposed C. albicans and gingival epithelial cells. Various cell biology, molecular biology, and microbiology protocols were deployed. Results show that exposure of gingival fibroblasts to nicotine-rich e-cigarette condensate altered both cell morphology and proliferation rate. Exposure to the ecigarette condensate also increased the levels of apoptotic fibroblasts. Fibroblast migration was delayed after culture scratches were exposed to e-cigarette condensate. Although e-cigarettes are considered to be less harmful than are conventional cigarettes, e-cigarettes significantly harmed the fibroblasts compared to non-exposed cells. E-cigarette exposure also increased C. albicans growth and hyphal length. The exposed C. albicans produced high levels of chitin and expressed high mRNA levels of SAP2, SAP3, and SAP9 genes. When in contact with gingival epithelial cells, e-cigarette-exposed C. albicans adhered better compared to the controls. Indirect communication between e-cigarette-exposed C. albicans and gingival epithelial cells led to epithelial cell differentiation, reduced cell growth, and increased lactate dehydrogenase (LDH) activity. Overall results indicate that e-cigarettes may interact with the user’s oral microbiome. Because e-cigarettes reduce gingival cell growth and increase cell apoptosis, this may decrease the innate immunity in the oral cavity, which could increase the risk of oral infections, such as candidiasis

E-Cigarettes Increase <i>Candida albicans</i> Growth and Modulate its Interaction with Gingival Epithelial Cells

Electronic cigarette (e-cigarette) vapor comes in contact with the different constituents of the oral cavity, including such microorganisms as Candida albicans. We examined the impact of e-cigarettes on C. albicans growth and expression of different virulent genes, such as secreted aspartic proteases (SAPs), and the effect of e-cigarette vapor-exposed C. albicans on gingival epithelial cell morphology, growth, and lactate dehydrogenase (LDH) activity. An increase in C. albicans growth was observed with nicotine-rich e-cigarettes compared with non-exposed cultures. Following exposure to e-cigarette vapor, C. albicans produced high levels of chitin. E-cigarettes also increased C. albicans hyphal length and the expression of SAP2, SAP3, and SAP9 genes. When in contact with gingival epithelial cells, e-cigarette-exposed C. albicans adhered better to epithelial cells than the control. Indirect contact between e-cigarette-exposed C. albicans and gingival epithelial cells led to epithelial cell differentiation, reduced cell growth, and increased LDH activity. Overall, results indicate that e-cigarettes may interact with C. albicans to promote their pathogenesis, which may increase the risk of oral candidiasis in e-cigarette users

A Proteinaceous Alpha-Amylase Inhibitor from Moringa Oleifera Leaf Extract: Purification, Characterization, and Insecticide Effects against C. maculates Insect Larvae

The main objective of the current study was the extraction, purification, and enzymatic characterization of a potent proteinaceous amylase inhibitor from Moringa oleifera. The antimicrobial potential and insecticide effects against C. maculates insect larvae were also studied. The &alpha;-amylase inhibitor was extracted in methanol (with an inhibitory activity of 65.6% &plusmn; 4.93). Afterwards, the inhibitor &alpha;AI.Mol was purified after a heat treatment at 70 &deg;C for 15 min followed by one chromatographic step of Sephadex G-50. An apparent molecular weight of 25 kDa was analyzed, and the N-terminal sequence showed the highest identity level (89%) with the monomeric &alpha;-amylase inhibitor from Triticum dicoccoides. &alpha;AI.Mol was found to tolerate pH values ranging from 5.0 to 11.0 and showed maximal activity at pH 9.0. Thermal stability was remarkably important, since the inhibitory activity was maintained at 55% after 1 h of incubation at 70 &deg;C and at 53% after an incubation of 45 min at 80 &deg;C. The potency of the current purified inhibitor against amylases from different origins indicates that &alpha;AI.Mol seems to possess the highest affinity toward human salivary &alpha;-amylase (90% inhibitory activity), followed by the &alpha;-amylase of insects Callosobruchus maculatus and Tribolium confusum (71% and 61%, respectively). The kinetic parameters were also calculated, and the Kmax and Vmax of the digestive amylase were estimated at 185 (mmol/min/mg) and 0.13 mM, respectively. The inhibitor possesses a strong bactericidal effect against Gram+ and Gram- strains, and the MIC values were &gt;1 against B. cereus but &gt;6 against E. coli. Interestingly, the rates of survival and pupation of C. maculates insect larvae were remarkably affected by the purified &alpha;AI.Mol from Moringa oleifera

Cigarette Smoke-Exposed Candida albicans Increased Chitin Production and Modulated Human Fibroblast Cell Responses

The predisposition of cigarette smokers for development of respiratory and oral bacterial infections is well documented. Cigarette smoke can also contribute to yeast infection. The aim of this study was to investigate the effect of cigarette smoke condensate (CSC) on C. albicans transition, chitin content, and response to environmental stress and to examine the interaction between CSC-pretreated C. albicans and normal human gingival fibroblasts. Following exposure to CSC, C. albicans transition from blastospore to hyphal form increased. CSC-pretreated yeast cells became significantly (P<0.01) sensitive to oxidation but significantly (P<0.01) resistant to both osmotic and heat stress. CSC-pretreated C. albicans expressed high levels of chitin, with 2- to 8-fold recorded under hyphal conditions. CSC-pretreated C. albicans adhered better to the gingival fibroblasts, proliferated almost three times more and adapted into hyphae, while the gingival fibroblasts recorded a significantly (P<0.01) slow growth rate but a significantly higher level of IL-1β when in contact with CSC-pretreated C. albicans. CSC was thus able to modulate both C. albicans transition through the cell wall chitin content and the interaction between C. albicans and normal human gingival fibroblasts. These findings may be relevant to fungal infections in the oral cavity in smokers

Biochemical Study of Bacillus stearothermophilus Immobilized Lipase for Oily Wastewater Treatment

Traditional wastewater treatments involve expensive mechanical and physiochemical methods, so researchers have been developing cost-effective, sustainable technologies that use enzymes to produce higher quality effluents and recover more energy and nutrients from wastewater. A thermostable, alkaline, and solvent-tolerant lipase was partially purified from thermophilic Bacillus stearothermophilus. The lipase displayed maximum activity at 50 &deg;C and pH 11.0 and catalyzed both short- and long-chain triacylglycerols at similar rates. B. stearothermophilus lipase also exhibited high stability when incubated at 40 &deg;C for 1 h with anionic and non-ionic surfactants. Studies show that thermostable enzymes can be improved through immobilization and modification of other reaction conditions. Therefore, B. stearothermophilus lipase was immobilized through adsorption on CaCO3, Celite 545, and silica gel with the CaCO3 support producing the best adsorption rate (89.33%). The optimal initial lipase activity was approximately 4500 U.g&minus;1 after 60 min. Interestingly, 93% of the initial lipase activity was retained after six cycles, and almost 50% of the initial activity remained after 12 cycles. Furthermore, immobilization improved storage stability with 98.85% of the initial lipase activity retained after 60 days of storage at 4 &deg;C. The biochemical characteristics of immobilized lipase shifted toward a slightly alkaline region, reaching maximum activity at pH 12. The optimal temperature of immobilized lipase was 60 &deg;C. Immobilization also improved enzymatic stability by widening the pH range from 5&ndash;9 (for free lipase) to 4&ndash;11, and thermostability by reaching 65 &deg;C. The application of immobilized lipase in wastewater treatment was observed through oil layer biodegradation. Notably, treating wastewater for 10 days with immobilized lipase almost removed the chemical oxygen demand (COD) from 1950.1 down to 4.04 mg.L&minus;1. Similarly, lipid content was almost removed from 15,500 &plusmn; 546 mg.L&minus;1 down to 12 mg.L&minus;1. All results highlight the potential value of CaCO3-immobilized lipase as an effective biocatalyst for hydrolyzing wastewater

Functional Characterization and Anti-Tumor Effect of a Novel Group II Secreted Phospholipase A₂ from Snake Venom of Saudi <i>Cerastes cerates gasperetti</i>

Secreted phospholipases A2 are snake-venom proteins with many biological activities, notably anti-tumor activity. Phospholipases from the same snake type but different geographical locations have shown similar biochemical and biological activities with minor differences in protein sequences. Thus, the discovery of a new phospholipase A2 with unique characteristics identified in a previously studied venom could suggest the origins of these differences. Here, a new Group II secreted phospholipase A2 (Cc-PLA2-II) from the snake venom of Saudi Cerastes cerastes gasperetti was isolated and characterized. The purified enzyme had a molecular weight of 13.945 kDa and showed high specific activity on emulsified phosphatidylcholine of 1560 U/mg at pH 9.5 and 50 °C with strict calcium dependence. Interestingly, stability in extreme pH and high temperatures was observed after enzyme incubation at several pH levels and temperatures. Moreover, a significant dose-dependent cytotoxic anti-tumor effect against six human cancer cell lines was observed with concentrations of Cc-PLA2 ranging from 2.5 to 8 µM. No cytotoxic effect on normal human umbilical-vein endothelial cells was noted. These results suggest that Cc-PLA2-II potentially has angiogenic activity of besides cytotoxicity as part of its anti-tumor mechanism. This study justifies the inclusion of this enzyme in many applications for anticancer drug development

Analysis of 1-Aroyl-3-[3-chloro-2-methylphenyl] Thiourea Hybrids as Potent Urease Inhibitors: Synthesis, Biochemical Evaluation and Computational Approach

Urease is an amidohydrolase enzyme that is responsible for fatal morbidities in the human body, such as catheter encrustation, encephalopathy, peptic ulcers, hepatic coma, kidney stone formation, and many others. In recent years, scientists have devoted considerable efforts to the quest for efficient urease inhibitors. In the pharmaceutical chemistry, the thiourea skeleton plays a vital role. Thus, the present work focused on the development and discovery of novel urease inhibitors and reported the synthesis of a set of 1-aroyl-3-[3-chloro-2-methylphenyl] thiourea hybrids with aliphatic and aromatic side chains 4a&ndash;j. The compounds were characterized by different analytical techniques including FT-IR, 1H-NMR, and 13C-NMR, and were evaluated for in-vitro enzyme inhibitory activity against jack bean urease (JBU), where they were found to be potent anti-urease inhibitors and the inhibitory activity IC50 was found in the range of 0.0019 &plusmn; 0.0011 to 0.0532 &plusmn; 0.9951 &mu;M as compared to the standard thiourea (IC50 = 4.7455 &plusmn; 0.0545 &mu;M). Other studies included density functional theory (DFT), antioxidant radical scavenging assay, physicochemical properties (ADMET properties), molecular docking and molecular dynamics simulations. All compounds were found to be more active than the standard, with compound 4i exhibiting the greatest JBU enzyme inhibition (IC50 value of 0.0019 &plusmn; 0.0011 &micro;M). The kinetics of enzyme inhibition revealed that compound 4i exhibited non-competitive inhibition with a Ki value of 0.0003 &micro;M. The correlation between DFT experiments with a modest HOMO-LUMO energy gap and biological data was optimal. These recently identified urease enzyme inhibitors may serve as a starting point for future research and development