RNAi-mediated silencing of ARV1 in Setaria digitata impairs in-vitro microfilariae release, embryogenesis and adult parasite viability.

Setaria digitata is a nematode that resides in the peritoneal cavity of ruminants causing cerebrospinal nematodiasis disease affecting livestock and inflicting significant economic forfeitures in Asia. Further, this nematode can infect humans, causing abscesses, allergic reactions, enlarged lymph nodes, eye lesions and inflammation of the lungs. The 'ARE2 required for viability1' (ARV1) encodes for putative lipid transporter localized in the endoplasmic reticulum (ER) and Golgi complex membrane in humans and yeast. In the present study, the functional role of S. digitata ARV1 (SD-ARV1) was investigated using RNA interference (RNAi) reverse genetic tool. The targeted silencing SD-ARV1 transcripts by siRNA mediated RNAi resulted in a dramatic reduction of SD-ARV1 gene and protein expressions in S. digitata, which in turn modulated the parasitic motility, its production of eggs and microfilaria viability. Further, the same silencing caused severe phenotypic deformities such as distortion of eggs and embryonic development arrest in the intrauterine stages of adult female S. digitata. These results suggest that SD-ARV1 plays a pivotal role in worm embryogenesis, adult parasite motility and microfilariae viability. Finally, the ubiquitous presence of ARV1 in human filarial nematodes, its crucial functional roles in nematode biology and its remarkable diversity in primary protein structure compared to homologues in their hosts warrants further investigations to ascertain its candidacy in anthelmintic drug development.

Denture-induced stomatitis and associated factors in a group of patients attending a university dental hospital in Sri Lanka.

AIM: The aim of the present study was to investigate factors associated with denture-induced stomatitis (DIS) among denture wearers and their oral and hand carriage of Candida. METHODS: Sociodemographics and denture and oral hygiene practices were investigated using an interviewer-administered questionnaire in denture wearers with DIS (n = 60) and without DIS (n = 60) selected using a convenience sampling of patients attending the Department of Prosthetic Dentistry, University of Peradeniya, Sri Lanka. Denture and oral hygiene practices and denture quality were scored. DIS and denture hygiene were classified. Oral and hand carriage of Candida was assessed. RESULTS: Middle-aged females were more prone to DIS. The most common DIS pattern was Newton type ΙΙ. The mean age of existing denture of DIS patients was significantly higher than the control. Denture hygiene practice was significantly low in DIS patients. Poor denture hygiene practice and sleep wear of dentures were significantly associated with DIS. The risk of Candida hand carriage in a DIS patient carrying oral Candida was significantly higher than the risk of Candida hand carriage in a healthy denture wearer who carried oral Candida. CONCLUSION: Old dentures, poor denture hygiene, and sleep wear of dentures might contribute to DIS. Oral Candida in denture wearers could lead to Candida hand carriage.

Extracellular phospholipase production of oral Candida albicans isolates from smokers, diabetics, asthmatics, denture wearers and healthy individuals following brief exposure to polyene, echinocandin and azole antimycotics

Abstract Objective Candida albicans is the primary causative agent of oral candidosis, and one of its key virulent attributes is considered to be its ability to produce extracellular phospholipases that facilitate cellular invasion. Oral candidosis can be treated with polyenes, and azoles, and the more recently introduced echinocandins. However, once administered, the intraoral concentration of these drugs tend to be sub-therapeutic and rather transient due to factors such as the diluent effect of saliva and cleansing effect of the oral musculature. Hence, intra-orally, the pathogenic yeasts may undergo a brief exposure to antifungal drugs. We, therefore, evaluated the phospholipase production of oral C. albicans isolates following brief exposure to sub-therapeutic concentrations of the foregoing antifungals. Materials and methods Fifty C. albicans oral isolates obtained from smokers, diabetics, asthmatics using steroid inhalers, partial denture wearers and healthy individuals were exposed to sub-therapeutic concentrations of nystatin, amphotericin B, caspofungin, ketoconazole and fluconazole for one hour. Thereafter the drugs were removed and the phospholipase production was determined by a plate assay using an egg yolk-agar medium. Results The phospholipase production of these isolates was significantly suppressed with a percentage reduction of 10.65, 12.14, 11.45 and 6.40% following exposure to nystatin, amphotericin B, caspofungin and ketoconazole, respectively. This suppression was not significant following exposure to fluconazole. Conclusions Despite the sub-therapeutic, intra oral, bioavailability of polyenes, echinocandins and ketoconazole, they are likely to produce a persistent antifungal effect by suppressing phospholipase production, which is a key virulent attribute of this common pathogenic yeast.

Extracellular phospholipase production of oral Candida albicans isolates from smokers, diabetics, asthmatics, denture wearers and healthy individuals following brief exposure to polyene, echinocandin and azole antimycotics.

OBJECTIVE: Candida albicans is the primary causative agent of oral candidosis, and one of its key virulent attributes is considered to be its ability to produce extracellular phospholipases that facilitate cellular invasion. Oral candidosis can be treated with polyenes, and azoles, and the more recently introduced echinocandins. However, once administered, the intraoral concentration of these drugs tend to be sub-therapeutic and rather transient due to factors such as the diluent effect of saliva and cleansing effect of the oral musculature. Hence, intra-orally, the pathogenic yeasts may undergo a brief exposure to antifungal drugs. We, therefore, evaluated the phospholipase production of oral C. albicans isolates following brief exposure to sub-therapeutic concentrations of the foregoing antifungals. MATERIALS AND METHODS: Fifty C. albicans oral isolates obtained from smokers, diabetics, asthmatics using steroid inhalers, partial denture wearers and healthy individuals were exposed to sub-therapeutic concentrations of nystatin, amphotericin B, caspofungin, ketoconazole and fluconazole for one hour. Thereafter the drugs were removed and the phospholipase production was determined by a plate assay using an egg yolk-agar medium. RESULTS: The phospholipase production of these isolates was significantly suppressed with a percentage reduction of 10.65, 12.14, 11.45 and 6.40% following exposure to nystatin, amphotericin B, caspofungin and ketoconazole, respectively. This suppression was not significant following exposure to fluconazole. CONCLUSIONS: Despite the sub-therapeutic, intra oral, bioavailability of polyenes, echinocandins and ketoconazole, they are likely to produce a persistent antifungal effect by suppressing phospholipase production, which is a key virulent attribute of this common pathogenic yeast.

In vitro Impact of Limited Exposure to Subtherapeutic Concentrations of Chlorhexidine Gluconate on the Adhesion-Associated Attributes of Oral Candida Species.

OBJECTIVE: Candida albicans and its non-albicans counterparts, such as C. tropicalis, C. krusei, C. glabrata and C. dubliniensis, are the major etiological agents of oral candidosis. Their adherence to buccal epithelial cells (BEC), denture acrylic surfaces (DAS) and cell surface hydrophobicity (CSH) are attributes associated with yeast colonization and infection. Chlorhexidine gluconate (CG) is a widely used antiseptic in dentistry. When administered, the diluent effect of saliva and the cleansing effect of the oral musculature reduce its bioavailability, compromising its efficacy. Hence, intraorally, Candida undergoes a transient exposure to high CG concentrations, and thereafter it is likely to be subtherapeutic. Therefore, the impact of CG on adhesion to BEC, DAS and CSH of different oral Candida species was investigated following brief exposure to three subtherapeutic concentrations of CG. MATERIALS AND METHODS: Ten oral isolates of each of the above five Candida species obtained in Kuwait from oral rinse samples were exposed to 0.00125, 0.0025 and 0.005% CG for 30 min. Subsequently, the yeast adhesion to BEC, DAS and CSH was determined. The data were analyzed using ANOVA Dunnett's t tests. RESULTS: Exposure to the lowest dilution (0.00125%) of CG did not elicit a noteworthy collective suppression on all three adhesion traits evaluated. Exposure to 0.0025% CG curtailed the adhesion to BEC, DAS and CSH of Candida species by 50.89, 40.79 and 24.58%, respectively (p < 0.001). Exposure to the highest concentration (0.005%) of CG reduced the adhesion to BEC, DAS and CSH of Candida species by 64.68, 54.59 and 50%, respectively (p < 0.001). CONCLUSIONS: Brief exposure to subtherapeutic concentrations of CG suppressed the adhesion to BEC, DAS and CSH of oral Candida species, indicating probable pharmacodynamics that may potentiate its antiseptic properties.

Impact of brief and sequential exposure to nystatin, amphotericin B, ketoconazole, and fluconazole in modulating adhesion traits of oral Candida dubliniensis isolates.

AIM: Candida adherence is implicated in the pathogenesis of oral candidosis. Adhesion to buccal epithelial cells (BEC), germ tube (GT) formation, and relative cell surface hydrophobicity (CSH) are colonization attributes of candidal pathogenicity. Candida dubliniensis (C. dubliniensis) is allied with recurrent oral candidosis, which can be treated with nystatin, amphotericin B, ketoconazole, and fluconazole. Due to the diluent effect of saliva and the cleansing effect of the oral musculature in the oral cavity C. dubliniensis isolates undergo brief and sequential exposure to antifungal agents during therapy. Thus, in the present study, we evaluated the adhesion to BEC, GT formation, and the CSH of oral isolates of C. dubliniensis following brief and sequential exposure to nystatin, amphotericin B, ketoconazole, and fluconazole. METHODS: After determining the minimum inhibitory concentration (MIC) of the aforementioned drugs, 20 oral isolates of C. dubliniensis were briefly (1 h), and sequentially (10 days) exposed to subcidal concentrations of these drugs. Following drug removal, adhesion to BEC, GT formation, and CSH of these isolates were determined. RESULTS: The percentage reduction of adhesion to BEC, GT formation, and CSH of the isolates following exposure to antifungal agents were as follows: nystatin: 53.55%, 33.98%, and 29.83% (P < 0.001); amphotericin B: 53.84%, 36.23%, and 28.97% (P < 0.001); ketoconazole: 37.43%, 20.51%, and 16.49% (P < 0.001); and fluconazole: 8.93% (P < 0.001), 1.6%, and 0.63% (P > 0.05). CONCLUSIONS: Brief and sequential exposure of C. dubliniensis to antifungal agents would continue to wield an antifungal effect by altering its adhesion attributes, and elucidate possible pharmacodynamics by which antifungal agents might operate in modulating candidal adherence.

Impact of short-term exposure of antifungal agents on hemolysin activity of oral Candida dubliniensis isolates from Kuwait and Sri Lanka.

OBJECTIVE: Ability to produce hemolysin by Candida species is an important determinant of its pathogenicity. Candida dubliniensis is implicated in the causation of oral candidosis, which can be treated with polyene, echinocandin, and azole groups of antifungal agents as well as chlorhexidine. After oral application, however, the concentrations of these agents tend to decrease quickly to subtherapeutic levels due to the peculiarity of the oral environment. In this study, we have evaluated the effect of short-term exposure of sublethal concentrations of these drugs on hemolysin production by oral C. dubliniensis isolates obtained from two different geographical locale. MATERIALS AND METHODS: Twenty C. dubliniensis oral isolates obtained from Kuwait and Sri Lanka were exposed to sublethal concentrations of nystatin, amphotericin B, caspofungin, ketoconazole, fluconazole, and chlorhexidine for 1 h. Thereafter, the drugs were removed by dilution and the hemolysin production determined by a previously described plate assay. RESULTS: Hemolysin production of these isolates was significantly suppressed with a percentage reduction of 17.09, 16.45, 17.09, 11.39, 8.23 and 12.03 following exposure to nystatin, amphotericin B, caspofungin, ketoconazole, fluconazole, and chlorhexidine, respectively. CONCLUSION: Brief exposure to sublethal concentrations of drugs with antifungal properties appears to reduce the pathogenic potential of C. dubliniensis isolates by suppressing hemolysin production.

In vitro postantifungal effect, adhesion traits and haemolysin production of Candida dubliniensis isolates following exposure to 5-fluorocytosine.

The phenomenon of postantifungal effect (PAFE), which is the suppression of candidal growth following brief exposure to antifungal agents, is linked with candidal pathogenicity. Adhesion to buccal epithelial cells (BEC), germ tube (GT) formation and relative cell surface hydrophobicity (CSH) are all adhesion traits of candidal pathogenicity. Ability to produce haemolysin by Candida species is also a determinant of its pathogenicity. There is no information on either the PAFE or its impact on adhesion traits and haemolysin production of oral Candida dubliniensis isolates following exposure to 5-fluorocytosine (5-FC). Hence, the focus of this investigation was to research the in vitro PAFE, adhesion to BEC, GT formation, relative CSH and haemolysin production on 20 C. dubliniensis isolates following exposure to 5-FC. Following obtaining the minimum inhibitory concentration (MIC) of 5-FC, isolates of C. dubliniensis were exposed to sub-lethal concentrations (×3 MIC) of 5-FC for 1 h. After this brief exposure, the antimycotic was removed and PAFE, adhesion to BEC, GT formation, relative CSH and haemolysin production was determined by formerly described in vitro methods. MIC (µg/ml) of C. dubliniensis isolates to 5-FC ranged from 0.002 to 0.125. The mean PAFE (hours) elicited by 5-FC on C. dubliniensis isolates was approximately 1 h. Exposure to 5-FC suppressed the ability of C. dubliniensis isolates to adhere BEC, GT formation, relative CSH and haemolysin activity by a mean percentage reduction in 50.98%, 29.51%, 36.79% and 12.75% (P < 0.001 for all) respectively. Therefore, brief exposure of C. dubliniensis isolates to 5-FC appears to exert an antifungal effect by subduing its growth, adhesion traits as well as haemolysin production.

In vitro adhesion of oral Candida dubliniensis isolates to acrylic denture surfaces following brief exposure to sub-cidal concentrations of polyenes, azoles and chlorhexidine.

OBJECTIVES: We aimed to investigate the effect of brief exposure to sub-cidal concentrations of nystatin, amphotericin B, ketoconazole, fluconazole and chlorhexidine gluconate on the adhesion of oral Candida dubliniensis isolates to the surface of acrylic dentures. METHODS: After determining the minimum inhibitory concentration of each drug, 20 oral isolates of C. dubliniensis were exposed to sub-cidal concentrations of the drugs for 1 h. The drugs were then removed by dilution, and the adhesion of the isolates to denture acrylic strips was assessed by an in vitro adhesion assay. RESULTS: Compared to the controls, exposure to nystatin, amphotericin B, ketoconazole, fluconazole and chlorhexidine gluconate suppressed the ability of C. dubliniensis isolates to adhere to acrylic denture surfaces with a reduction of 74.68, 74.27, 57.31, 44.57 and 56.53% (p < 0.001 for all drugs), respectively. CONCLUSIONS: Brief exposure to sub-cidal concentrations of anti-mycotics suppressed the adhesion of C. dubliniensis oral isolates to acrylic denture surfaces.

Assessing Oral Cancer Knowledge Among Undergraduate Dental Students in Kuwait University.

Lack of general dental practitioner oral cancer knowledge has been shown to be a major factor to delays in referral and treatment. Dentists' competence and confidence in detecting oral cancer may be strongly influenced by their dental school training. Therefore, the aim of this study was to assess dental student's awareness and knowledge of prevention and early detection of oral cancer. Through the use of questionnaires, the students' knowledge of oral cancer risk factors, diagnostic concepts, and clinical care of patients was assessed. An excellent response rate (97.3 %) was observed in this study. Overall, the students' knowledge of diagnostic items was significantly higher (p ≤ 0.001) than their knowledge of risk factors. A high percentage of students identified tobacco (98.6 %) and alcohol (76.7 %) as the principal risk factors and would offer advice regarding modification of these habits (75.3 %). Also, their knowledge of non-risk factors was significantly lower than their knowledge of proven risk factors (p ≤ 0.001). Only one fifth of the students regarded visual inspection to be an effective screening method. Only 32.9 % stated that all suspicious lesions should be biopsied, and as low as 2.7 % had assisted in taking a biopsy. This study highlights the need for a more structured teaching program with greater emphasis on the early signs and risk factors of oral cancer, performing routine oral examination, referral for biopsy, and appropriate early management of suspicious oral lesions.

HIV/AIDS: dental assistants' self-reported knowledge and attitudes in Kuwait.

Although several studies have been conducted to assess dentists' knowledge of and attitudes towards human immunodeficiency virus (HIV)/acquired immune-deficiency syndrome (AIDS), few have targeted dental assistants. The main aims of this study were to assess the knowledge of and attitudes towards HIV/AIDS among dental assistants in Kuwait and to compare the knowledge and attitudes of dental assistants at Kuwait University Dental Center (KUDC) with those of dental assistants in Ministry of Health (MoH) hospitals. The secondary objective was to determine if any intervention was needed to provide more information to dental assistants on HIV/AIDS. A cross-sectional study was conducted by distributing questionnaires. The study sample included 85 dental assistants from each of KUDC and the MoH. The questionnaire included questions to assess the assistants' knowledge and attitude towards HIV/AIDS. Statistical data analysis was conducted using SPSS 20.0. Qualitative data were analysed using the Pearson chi-square text for any association or the Z-test for proportion to test the significance of differences. A total of 167 questionnaires were completed, returned and analysed. KUDC dental assistants were found to have significantly more knowledge about HIV/AIDS than their MoH counterparts, whereas the assistants at the MoH clinics displayed a more positive attitude towards patients with HIV/AIDS (P < 0.05). Although dental assistants at KUDC were more knowledgeable than those at the MoH clinics, there are still some misconceptions that need to be addressed, in addition to the negative attitudes displayed by some of the respondents. It would therefore be beneficial to increase awareness about HIV/AIDS patients through lectures, seminars and workshops targeting dental assistants.

Post-antifungal effect and adhesion to buccal epithelial cells of oral Candida dubliniensis isolates subsequent to limited exposure to amphotericin B, ketoconazole and fluconazole.

AIM: The post-antifungal effect (PAFE) of Candida and its adherence to oral mucosal surfaces are important determinants of candidal pathogenicity. Candida dubliniensis is allied with recurrent oral candidosis. Oral candidosis can be treated with amphotericin B, ketoconazole and fluconazole. There is no information on the PAFE and its impact on adhesion to oral buccal epithelial cells (BEC) of oral C. dubliniensis isolates. Therefore, the main objective was to reconnoiter the PAFE and adhesion to BEC of 20 C. dubliniensis isolates following brief exposure to aforementioned antimycotics. METHODS: After determining the minimum inhibitory concentration (MIC), C. dubliniensis isolates were exposed to sub-lethal concentrations of these drugs for 1 h. Following subsequent drug removal, the PAFE and adhesion to BEC, was determined by a turbidometric method, and an adhesion assay, respectively. RESULTS: Minimum inhibitory concentration (µg/mL) to amphotericin B, ketoconazole and fluconazole, ranged from 0.002 to 0.125, 0.002 to 0.012 and 0.016 to 0.38, respectively. Amphotericin B and ketoconazole induced mean PAFE (hours) were 2.21 and 0.6, respectively. Fluconazole failed to produce a detectable PAFE. Compared to controls, amphotericin B, ketoconazole and fluconazole suppressed the ability to adhere to BEC with a mean percentage reduction of 74.31%, 49.80% (P < 0.0001) and 29.36% (P < 0.05), respectively. CONCLUSIONS: Brief exposure to sub-lethal concentrations of aforementioned drugs would exert an antifungal effect by modifying the growth and adhesion of C. dubliniensis isolates.

Impact of brief and sequential exposure to nystatin on the germ tube formation and cell surface hydrophobicity of oral Candida albicans isolates from human immunodeficiency virus-infected patients.

OBJECTIVE: To evaluate the impact of brief and sequential exposure to nystatin on the germ tube formation and cell surface hydrophobicity of oral isolates of Candida albicans obtained from patients infected with human immunodeficiency virus (HIV). MATERIALS AND METHODS: After determining the minimum inhibitory concentration of nystatin, 10 oral isolates of C. albicans from 10 different HIV-infected patients were briefly (1 h) and sequentially (10 days) exposed to subtherapeutic concentrations of nystatin. Following a subsequent drug removal, the germ tube formation and cell surface hydrophobicity of these isolates were determined via a germ tube induction assay and an aqueous hydrocarbon assay, respectively. The data obtained from these assays for the control (unexposed to nystatin) and nystatin-exposed isolates were analyzed using Student's t tests. RESULTS: The mean percentage reduction in the germ tube formation and cell surface hydrophobicity of the nystatin-exposed isolates compared to the controls was 30.12 ± 1.99 (p < 0.001) and 29.65 ± 2.33 (p < 0.001), respectively. CONCLUSION: These data elucidate the possible pharmacodynamic mechanisms by which nystatin might operate in vivo in the modulation of candidal virulence.

The postantifungal effect and phospholipase production of oral Candida albicans from smokers, diabetics, asthmatics, denture wearers and healthy individuals following brief exposure to subtherapeutic concentrations of chlorhexidine gluconate.

Candida albicans is the major aetiological agent of oral candidosis and one of its important virulent factors is the production of extracellular phospholipases, which can be modulated by subtherapeutic concentrations of antifungal agents thus decreasing their pathogenicity. Hence, considering that chlorhexidine gluconate (CG) is a common antimicrobial mouthwash used in dentistry and that its concentration in the mouth reaches subtherapeutic levels during dosage intervals due to the diluent effect of saliva and cleansing effect of the oral musculature, the postantifungal effect (PAFE) and the phospholipase production of oral C. albicans following brief exposure to subtherapeutic concentrations of CG was studied. Fifty C. albicans planktonic oral isolates obtained from smokers, diabetics, asthmatics using steroid inhalers, partial denture wearers and healthy individuals were exposed to three subtherapeutic concentrations of CG (0.005%, 0.0025% and 0.00125%) for 1 h. Isolates unexposed to CG was the control group. Thereafter the antiseptic was removed and the PAFE and phospholipase production was determined by a turbidometric method and a plate assay using an egg yolk agar medium respectively. Mean PAFE (hours) of 50 oral isolates of C. albicans following 1-h exposure to 0.005%, 0.0025% and 0.00125% CG was 6.97, 1.85 and 0.62 respectively. The phospholipase production of these isolates was significantly suppressed with a percentage reduction of 21.68, 18.20 and 14.04% following exposure to 0.005%, 0.0025% and 0.00125% CG respectively. Brief exposure of C. albicans isolates to subtherapeutic concentrations of CG would wield an antifungal effect by suppressing growth and phospholipase production, thereby quelling its pathogenicity.

The postantifungal effect of nystatin and its impact on adhesion attributes of oral Candida dubliniensis isolates.

The postantifungal effect (PAFE) has an impact on candidal pathogenicity. However, there is no information on either the PAFE or its impact on adhesion traits of oral Candida dubliniensis isolates. Oral candidosis can be treated topically with nystatin. Adhesion to buccal epithelial cells (BEC), germ tube (GT) formation and relative cell surface hydrophobicity (CSH) are all colonisation attributes of candidal pathogenicity. Hence, the main objective of this study was to investigate the in vitro PAFE on 20 C. dubliniensis isolates following exposure to nystatin. In addition, the impact of nystatin-induced PAFE on adhesion to BEC, GT formation and relative CSH of C. dubliniensis isolates were also evaluated. After determining the minimum inhibitory concentration (MIC) of nystatin, C. dubliniensis isolates were exposed to sublethal concentrations of nystatin for 1 h. Following this exposure, the drug was removed and PAFE, adhesion to BEC, GT formation and relative CSH were determined by a previously described turbidometric method, adhesion assay, germ tube induction assay and biphasic aqueous-hydrocarbon assay respectively. MIC (µg/ml) of C. dubliniensis isolates to nystatin ranged from 0.09 to 0.78. The nystatin-induced mean PAFE (hours) on C. dubliniensis isolates was 2.17. Compared with the controls, exposure to nystatin suppressed the ability of C. dubliniensis isolates to adhere BEC, GT formation and relative CSH by a mean percentage reduction of 74.45% (P < 0.0001), 95.92% (P < 0.0001) and 34.81 (P < 0.05) respectively. Hence, brief exposure of C. dubliniensis isolates to nystatin would continue to wield an antifungal effect by suppressing growth as well as its adhesion attributes.

Changes in germ tube formation and cell-surface hydrophobicity of oral Candida dubliniensis isolates following brief exposure to sub-cidal concentrations of polyene and azole antifungal agents.

Adherence of Candida has been implicated as the initial process in the pathogenesis of oral candidosis. Candidal germ tubes and its relative cell-surface hydrophobicity (CSH) are contributory attributes. Candida dubliniensis is currently documented as an opportunistic pathogen allied with recurrent oral candidosis. Oral candidosis can be treated with polyene and azole antifungals such as amphotericin B, ketoconazole and fluconazole. However, the intraoral concentration of these drugs fluctuates and becomes sub-therapeutic because of the diluent effect of saliva and cleansing effect of the oral musculature. Hence, intraorally, the pathogenic yeast may undergo a brief exposure to antifungal drugs. The objective of this study was to investigate the effect of brief exposure to sub-lethal concentrations of these antifungals on the germ tube formation and CSH of C. dubliniensis. After determining the minimum inhibitory concentration of the drugs, 20 oral isolates of C. dubliniensis were exposed to sub-lethal concentrations of these antifungals for 1 h. Following this brief exposure, the drugs were removed, and following subsequent incubation in a germ tube inducing medium and exposure to bi-phasic hydrocarbon assay, the germ tube formation and CSH of these isolates was quantified respectively. Compared with controls, exposure to amphotericin B almost completely suppressed the ability to form germ tubes with a mean percentage reduction of 95.91% (P < 0.0001), whereas ketoconazole and fluconazole also significantly inhibited germ tube formation but to a lesser degree with a mean percentage reduction of 18.73% and 12.01% respectively (P < 0.05). Compared with controls, exposure to amphotericin B and ketoconazole elicited a significant suppression on CSH with a mean percentage reduction of 33.09% and 21.42%, respectively (P < 0.001), whereas exposure to fluconazole did not elicit a significant suppression on CSH (9.21%; P > 0.05). In clinical terms it appears that, even a short exposure to sub-lethal concentrations of these drugs, a situation all too familiar in the oral environment, would continue to exert an antifungal effect by suppressing the pathogenic potency of C. dubliniensis.

Changes in the cell surface hydrophobicity of oral Candida albicans from smokers, diabetics, asthmatics, and healthy individuals following limited exposure to chlorhexidine gluconate.

OBJECTIVE: The objective of this study was to determine the cell surface hydrophobicity of 40 oral Candida albicans isolates obtained from smokers, diabetics, asthmatics using steroid inhalers, and healthy individuals, following brief exposure to subtherapeutic concentrations of chlorhexidine gluconate. MATERIALS AND METHODS: Forty C. albicans oral isolates (10 isolates each from smokers, diabetics, asthmatics using steroid inhalers, and healthy individuals) were exposed to 3 subtherapeutic concentrations of chlorhexidine gluconate (0.00125, 0.0025, and 0.005%) for 30 min. Thereafter, the antiseptic was removed and the cell surface hydrophobicity was measured by a biphasic aqueous-hydrocarbon assay. RESULTS: Compared to the unexposed controls, the cell surface hydrophobicity of C. albicans isolates was suppressed by 5.40% (p > 0.05), 21.17% (p < 0.05), and 44.67% (p < 0.05) following exposure to 0.00125, 0.0025, and 0.005% chlorhexidine gluconate, respectively. CONCLUSIONS: A brief period of transient exposure to subtherapeutic concentrations of chlorhexidine gluconate may modulate the cell surface hydrophobicity of C. albicans isolates and thereby may reduce candidal pathogenicity.

Cell surface hydrophobicity of oral Candida dubliniensis isolates following limited exposure to sub-therapeutic concentrations of chlorhexidine gluconate.

Candidal adhesion has been implicated as the initial step in the pathogenesis of oral candidiasis and cell surface hydrophobicity (CSH) has been implicated in adhesion to mucosal surfaces. Candida dubliniensis is an opportunistic pathogen associated with recurrent oral candidiasis. Chlorhexidine gluconate is by far the commonest antiseptic mouth wash prescribed in dentistry. At dosage intervals the intraoral concentration of this antiseptic fluctuates considerably and reaches sub-therapeutic levels due to the dynamics of the oral cavity. Hence, the organisms undergo only a limited exposure to the antiseptic during treatment. The impact of this antiseptic following such exposure on CSH of C. dubliniensis isolates has not been investigated. Hence, the main objective of this study was to investigate the effect of brief exposure to sub-therapeutic concentrations of chlorhexidine gluconate on the CSH of C. dubliniensis isolates. Twelve oral isolates of C. dubliniensis were briefly exposed to three sub-therapeutic concentrations of 0.005%, 0.0025% and 0.00125% chlorhexidine gluconate for 30 min. Following subsequent removal of the drug, the CSH of the isolates was determined by a biphasic aqueous-hydrocarbon assay. Compared with the controls, exposure to 0.005% and 0.0025% chlorhexidine gluconate suppressed the relative CSH of the total sample tested by 44.49% (P < 0.001) and 21.82% (P < 0.018), respectively, with all isolates being significantly affected. Although exposure to 0.00125% of chlorhexidine gluconate did not elicit a significant suppression on the total sample tested (7.01%; P > 0.05), four isolates of the group were significantly affected. These findings imply that exposure to sub-therapeutic concentrations of chlorhexidine gluconate may suppress CSH of C. dublinienis isolates, thereby reducing its pathogenicity and highlights further the pharmacodynamics of chlorhexidine gluconate.

Effects of subtherapeutic concentrations of chlorhexidine gluconate on germ tube formation of oral Candida.

OBJECTIVE: The main objective of this study was to investigate the effect of brief exposure to subtherapeutic concentrations of chlorhexidine gluconate on germ tube formation of Candida albicans isolates obtained from smokers, diabetics, asthmatics using steroid inhalers and healthy individuals. MATERIALS AND METHODS: Forty isolates of C. albicans were used in this study. All these isolates were quantified for germ tube formation without exposure to the drug and were used as the control group for data analysis. Isolates were also exposed to three subtherapeutic concentrations of chlorhexidine gluconate (0.00125, 0.0025 and 0.005%) for 30 min (limited exposure); the antiseptic was then removed and germ tube formation of these isolates was quantified microscopically following incubation in a germ tube-inducing medium. RESULTS: Compared with the unexposed controls, brief exposure to all concentrations of chlorhexidine gluconate suppressed the ability of the C. albicans isolates to form germ tubes in increasing order by 13.72% (p < 0.001 to p = 0.02), 46.16% (p < 0.001) and 72.46% (p <0.001). CONCLUSIONS: These findings show that brief exposure to subtherapeutic concentrations of chlorhexidine gluconate may modulate germ tube formation of C. albicans isolates, thereby suppressing their pathogenicity, and further elucidate the pharmacodynamic mechanisms by which chlorhexidine gluconate may operate in vivo.

Prevalence of Candida dubliniensis among oral Candida isolates in patients attending the Kuwait University Dental Clinic.

OBJECTIVES: The aim of this study was to determine the oral candidal carriage of patients seeking dental treatment at the Kuwait University Dental Clinic and to ascertain the Candida species composition among them. METHODS: 370 oral rinse samples were collected from patients. The germ tube test, CHROMagar Candida medium and VITEK 2 yeast identification system were used for species identification. C. dubliniensis isolates were confirmed by the production of rough colonies with hyphal fringes and chlamydospores on simplified sunflower seed agar. RESULTS: Of the 370 samples investigated, 160 (43.24%) showed Candida in culture. The isolation of Candida was significantly higher in individuals who were smokers or were under medication for either diabetics or asthma [99 (62%)] compared to healthy individuals [61 (38%)]. Of the 210 samples which did not yield Candida, 131 (62.38%) were healthy and 79 (37.62%) were associated with smoking or with usage of drugs for aforementioned conditions. Species isolated were C. albicans [102 (63.7%)], C. dubliniensis [23(14.3%)], C. krusei [13 (8.1%)], C. tropicalis [12 (7.5%)] and C. glabrata [10 (6.2%)]. CONCLUSIONS: Candida species were more prevalent in patients having predisposing factors implicated in oral candidosis, such as in smokers, diabetic patients and asthmatic patients using inhalation steroids. C. albicans was the most prevalent species isolated, followed by C. dubliniensis.