Immune System Diseases

All Conditions

Neoplasms

Symptoms and General Pathology

Mouth and Tooth Diseases

Rare Diseases

Hypersensitivity

Leukoplakia

Leukoplakia, Oral

Precancerous Conditions

Pathological Conditions, Anatomical

Mouth Neoplasms

Head and Neck Neoplasms

Mouth Diseases

Stomatognathic Diseases

Oral Leukoplakia

Oral Cancer

Anti-Infective Agents

All Drugs and Chemicals

Central Nervous System Depressants

Clotrimazole

Miconazole

Fluconazole

Ethanol

Antifungal Agents

Enzyme Inhibitors

Cytochrome P-450 Enzyme Inhibitors

Hormones

Hormone Antagonists

uniform, flat, thin, smooth/ wrinkled/corrugated surface throughout the white lesion.

Tab Fluconazole 100 mg as a mouthwash ( tablet dissolved in 10 ml of drinking water and used as a mouth rinse for 2 minute and swallowed) once a day for 14 days

mixture of red and white lesions with a irregularly speckled/ nodular/ verrucous surface Tab Fluconazole 100 mg as a mouthwash ( tablet dissolved in 10 ml of drinking water and used as a mouth rinse for 2 minute and swallowed) once a day for 14 days

30 healthy controls ( age and sex matched) would also be recruited from patients who are reporting for other routine dental problems.

( Tab Fluconazole 100 mg as a mouthwash ( tablet dissolved in 10 ml of drinking water and used as a rinse for 2 minute and swallowed) once a day for 14 days WHO criteria 5As and 5Rs for tobacco cessation

Shalini Gupta, MDS,FDSRCS

This study aims to determine the correlation between candida and pro- inflammatory cytokines response in Oral Leukoplakia(OL) with antifungal therapy. Ethical clearance from the Institutes ethical committee and Informed written consent from the patient will be obtained. The study group would comprise of clinically and histopathologically confirmed cases of OL (60 patients). The control group would be 30 dental patients ( age \& sex matched) who do not any malignancy, OL or any other potentially malignant disorder of oral mucosa. Patients who have any predisposing factor for oral candidiasis will be excluded from the study. Swabs will be taken from the oral lesion and cultured for candida to determine phenotypes, virulence attributes and antifungal sensitivity. Sterile PVA opthalmic sponges will be used to collect sample from the surface of oral epithelium and then processed to determine levels of pro- inflammatory cytokines (IL-6, IL-8. IL-17, TNFα). This procedure will be repeated in study group 2 weeks after a course of antifungal therapy. The results will be tested statistically at a confidence level of 95%.

Objectives

1. To determine the association of candida phenotypes, virulence attributes ( Secreted aspartyl proteinases, biofilm formation, Phospholipase) and antifungal sensitivity with clinicopathologic characteristics in oral leukoplakia
2. To determine the secretion and levels of pro-inflammatory cytokines ( IL-6, IL-8. IL-17, TNFα ) in oral leukoplakia before and after antifungal therapy.
3. To study the correlation between candida and pro-inflammatory cytokines in oral leukoplakia with antifungal therapy

Detailed methodology including study design, outcome measures, sample size and statistical analysis.

Study design:

Prospective case- control observational study

Selection of patients:

The patients in the study group and control group will be consecutively recruited from the Oral Medicine \& Radiology clinic at Centre for Dental Education \& Research AIIMS.

Sample size:

The study will include 60 OL ( 30 Homogenous OL, 30 Non- Homogenous OL) and 30 healthy controls

Clinicopathologic characteristics:

The clinical characteristics and staging/grading would be recorded as per prepared proforma for each group. Written informed consent would be taken before any further investigation. History of tobacco, betel nut, alcohol habits with frequency, duration of habit and current status, oral burning sensation with Visual Analogue Scale (VAS) scores will be Routine blood investigations would be done before incisional biopsy and histopathological confirmation of the diagnosis. The H \& E histopathological features with Periodic Acid Schiff (PAS) staining would be recorded and grading done for study group as per standard World Health Organization criteria. The clinical staging of OL will be done as per OL staging system (OLEP) ( van der Waal 2000) with photographic records of lesions by two calibrated observers.Tobacco,betel nut and alcohol cessation counselling and oral hygiene instructions will be given.

Sample collection for study:

A). Candida phenotypes, virulence and antifungal sensitivity study. Collection , transport and processing: Two oral swabs from each patient ( pre-wet with sterile normal saline) will be taken from the oral lesion and sent to the Mycology Laboratory, Department of Microbiology for further processing as per standard laboratory procedures.

Direct microscopy: One swab sample will be examined by microscopy for detection of yeast cells by preparing gram stain.

Culture: The second swab will be inoculated on Sabouraud dextrose agar (SDA) medium with gentamicin and will be kept at 37°C for incubation for 48 hours. Gram staining will be performed on any growth morphologically resembling to that of yeast.

Phenotypic identification: Identification of isolates was performed by using germ tube, morphology on corn meal agar, colour on triphenyl tetrazolium chloride and CHROM agar medium Determination of virulence factors of Candida spp Various virulence factors like phospholipase activity, Secreted aspartyl proteinase (SAP) and Biofilm formation (BF) will be determined as per the standard protocol.

Determination of Phospholipase (PL) activity Phospholipase activity will be measure by growing cells on egg yolk agar medium and measuring the size of the zone of precipitation. A cell suspension of 106 yeast cells/ml in saline will be prepare and 5μl will be place on the surface of the egg yolk medium. The culture will then incubated at 37°C for 7-8 days, after which the diameter of the precipitation zone around the colony will be determine. Phospholipase activity (Pz) will be measure by dividing colony diameter by the diameter of the precipitation zone (pz) around the colony formed on the plate. Each isolate will be tested in duplicate. The C. albicans SC 5314 strain will be used as positive control.

Determination of Secreted aspartyl proteinase (SAP) activity All isolates will be tested for their ability to grow and produce a clear zone of hydrolysis in bovine serum albumin (BSA) agar. . A 5μl of 1x106 cells /ml will be placed on solid medium and will be incubated at 37°C for 3-4 days. Subsequently, clearing of the opacity by hydrolysis of precipitated albumin will be recorded. The C. albicans SC 5314 strain will be used as positive control.

Determination of Biofilm formation (BF) A 100μl volume of 1 x 106 cells /ml suspension will be place on sterile, polystyrene, flat-bottom 96-well microtitre plates and will be incubated for 48 h at 37°C for adherence and biofilm. After 48 hr wells will be washed with PBS and a semi-qualitative measure of biofilm will be detected by XTT \[2,3-bis (2-methoxy-4nitro-5-sulfo-phenyl) - 2H - tetra-zolium-5-carboxanilide\] -reduction assay A colorimetric change in the XTT-reduction assay, a direct correlation of the metabolic activity of the biofilm, will then be measure in a microtiter plate reader at 490 nm after 2 hr of incubation.

Antifungal susceptibility testing of isolates Antifungal susceptibility for voriconazole fluconazole and clotrimazole will be performed by disk diffusion method as per the CLSI M44-A2 recommendation on the yeast isolates.

B). Pro- Inflammatory cytokines study Sample collection of oral secretions from lesion Four Sterile PVA ophthalmic sponges (Merocel) will be used to collect samples of oral secretions for four pro-inflammatory cytokine study from the oral lesion (other than biopsy site). This is a non- invasive method and makes it site -specific which is an advantage over other serum and saliva estimations. In addition, assessment of local cytokine levels via longitudinal sampling in patients undergoing antifungal therapy in OL may allow for an understanding of the temporal nature of cytokine elevation and clinical response to antifungals.

This procedure will be done for both the study group and control group. The subjects will be asked to abstain from eating, drinking and rinsing the mouth at least 2 hours prior to sampling. The sponges will be pre-wet with sterile normal saline, kept in contact with the lesion without rubbing or movement or 1 minute and then immediately stored in sterile containers at -80 °C till further analysis in Biochemistry department AIIMS. A photographic and clinical record of the sample site will be kept with the patient proforma for future reference. This procedure will be repeated at the same site after antifungal therapy for study group and a photographic record will be kept for comparison. Ethical clearance from Institute ethics board will be taken. Informed written consent will be obtained and information regarding the study would be given to the study subjects and controls prior sample collection.

Estimation of IL-6, IL-8, IL-17 and TNFα levels by ELISA The opthalmic sponges (Merocel ) will be thawed at room temperature for 10 min. Sponges will then be inserted into a microcentrifuge tube containing a 0.2 μm filter (SpinX centrifuge tube), equilibrated by adding 300 μl of extraction buffer and incubated for 30 min at 4 °C, followed by centrifugation at 4 °C for 30 min at 14,000 rpm. After centrifugation, the resultant supernatant will be collected for the estimation of IL-6, IL-8, IL-17 and TNFα levels by ELISA method with commercially available kits. The supernatant will be stored at -80 °C until use.

A monoclonal antibody against the antigens (IL-6, IL-8, IL-17 and TNFα) has been pre-coated onto the wells of the microtiter strips provided. Antigens present in the sample or standard will be incubated with the plates to allow binding of antigens to the antibody. This is followed by the addition of a primary monoclonal anti- IL-6, IL-8, IL-17 and TNFα antibody respectively conjugated to biotin in respective microtiter plates. An avidin-HRP conjugated antibody specific for primary antibody will then be added to the wells. After incubation and following a wash, to remove any unbound antibody enzyme reagent, a TMB one-step substrate reagent reactive with HRP will be added to the wells. The color development will be terminated by adding acid and absorbance was measured at 450 nm. A reference curve will be obtained by plotting the different concentrations of standard samples versus absorbance and levels of the antigens in samples tested will be calculated by its standard plot.

Antifungal therapy The study group will be treated with antifungal therapy ( Tab Fluconazole 100 mg as a mouthwash ( tablet dissolved in 10 ml of drinking water and used as a rinse for 2 minute and swallowed) once a day for 14 days). The procedure will be demonstrated to each study subject before starting the treatment. The oral burning sensation and clinical characteristics of the lesions with intraoral photographs will be recorded after antifungal therapy.

Statistical analysis:

The distribution of candida phenotypes, virulence attributes and antifungal sensitivity in the study and control groups would be correlated with levels of IL-6, IL-8, IL-17 and TNFα and clinicopathological characteristics. Correlations between virulence factors and clinical characteristics will be determined. Cohens kappa statistic will be used to determine intraobserver reliability. A comparison of the levels of pro inflammatory cytokines(IL-6, IL-8, IL-17 and TNFα) before and after antifungal therapy will be done in the study group. The results would be tested for statistical significance.The Chi- square and the Mann- Whitney U tests will be used to estimate the statistical significance of difference observed between the groups.

7.Ethical Clearance Obtained

Candida Associated Cytokines in Oral Leukoplakia

Association of Candida and Antifungal Therapy With Pro-inflammatory Cytokines in Oral Leukoplakia- A Pilot Study

maximum Length( L) and width (W) L1: L/W ≤ 2 cm L2: L /W 2-4 cm L3: L/W ≥ 4cm

il6,il8,il17,TNF-alpha by ELISA before and after antifungal treatment

Scores as \[ 0- no change, 1- minimal change ( Reduction in erythema \< 1cm of oral lesion, and 2- significant change( Reduction in erythema ≥ 1cm of oral lesion)

Scores as \[ 0- no change, 1- minimal change ( Reduction in thickness \< 1cm of oral lesion, and 2- significant change( Reduction in thickness ≥ 1cm of oral lesion)

Swabs for smear or culture positive for Candida

Phospholipase activity (Pz) was measured by dividing colony diameter by the diameter of the precipitation zone (Pz) around the colony formed on the plate. A Precipitation zone (in mm) around colony 1.0 : 0 negative (-), 0.99-0.9 : 1 weak (+), 0.89-0.8: 2 poor (++), 0.79-0.7: 3 moderate (+++), \< 0.69: 4 Strong positive(++++),

Secreted aspartyl proteinase activity was measured by dividing colony diameter by the diameter of the precipitation zone (Pz) around the colony formed on the plate. A Precipitation zone (in mm) around colony 1.0 : 0 negative (-), 0.99-0.9 : 1 weak (+), 0.89-0.8: 2 poor (++), 0.79-0.7: 3 moderate (+++), \< 0.69: 4 Strong positive(++++),

Absorbance values of test wells at 490nm of XTT reduction assay- absorbance values of control

Using CLSI document M44-A for disk diffusion method for Fluconazole (25 μg), Voriconazole (1 μg) and Clotrimazole (50 μg) S: Sensitive R: Resistant I: Intermediate

Mild Dysplasia Moderate Dysplasia Severe Dysplasia PAS stain positive/ negative

Histological features of Oral Squamous Cell Carcinoma

All India Institute of Medical Sciences, New Delhi

Associate Professor

Adenocarcinoma

Inflammation

Carcinoma

Neoplasms, Glandular and Epithelial

Neoplasms by Histologic Type

The patient will be evaluated for baseline clinical and laboratory values, and counselled on very low carbohydrate diet.

The patient will be evaluated for baseline clinical and laboratory values, and counselled on normal healthy .

1 : 4 ratio of carbohydrate to fat of the total daily calories intake

Very Low Carbohydrate Diet

Vicky S Budipramana, PhD

Fransiscus Arifin, MD

This study examine the effects of very low carbohydrate diet (in which the calories requirements are mostly from fat) to the level of systemic inflammation (measured by Glasgow Prognostic Score), serum lactate and TNF Alpha levels

This study examine the effects of very low carbohydrate diet (in which the calories requirements are mostly from fat) to the level of systemic inflammation (measured by Glasgow Prognostic Score), serum lactate and TNF Alpha levels. The subjects are colorectal adenocarcinoma patients who were decided by experts to be managed by best supportive care. These patients are not deemed to be suitable for surgical intervention , chemotherapy or radiotherapy anymore. The main treatment is supportive. We would like to examined the effect of very low carbohydrate diet on these patients, in which theoretically tumtor cells of the colorectal adenocarcinoma needed glucose to replicate and increases the systemic inflammation response.

Patients were randomized in to two groups, with very low carbohydrate diet and with normal diet

Care provider are not told whether the patient belongs to intervention or control group

Very Low Carbohydrate Diet Effects to GPS, Serum Lactate and TNF Alpha on Colorectal Cancer

Effect of Very Low Carbohydrate Diet to Glasgow Prognostic Score, Serum Lactate and TNF Alpha on Colorectal Cancer Patients With Best Supportive Care

A score to evaluate systemic inflammation response

Level of Tumor Necrosis Factor Alpha in the blood serum

Dr. Soetomo General Hospital

Respiratory Tract (Lung and Bronchial) Diseases

Asthma

Necrosis

Bronchial Diseases

Respiratory Tract Diseases

Lung Diseases

Lung Diseases, Obstructive

Respiratory Hypersensitivity

Hypersensitivity, Immediate

Stephen T Holgate, MD

We have developed in vitro systems including primary epithelial cell cultures and explant cultures of bronchial tissues to study the interaction between the bronchial mucosa and allergens. This approach involves culturing bronchial biopsies under optimal conditions and stimulate them with allergens thus enabling us to perform a dynamic study without the need of performing several bronchoscopies and will allow the testing of unapproved substances, which could, otherwise, not be delivered, in vivo.

We wish to apply these ex vivo bronchial culture systems to assess cytokine release in moderately severe asthmatics and evaluate the effects of blocking TNF-alpha signalling using anti TNF-alpha monoclonal antibodies.

Volunteers with moderately severe asthma will be recruited and carefully characterized in terms of lung function, asthma symptom scores, medication usage and allergen sensitivity (by blood and skin tests). If suitable, they will then be asked to withdraw their corticosteroids for a week with careful monitoring of the symptoms and peak flows before undergoing fibreoptic bronchoscopy as was carried out in the previous explant study. Bronchial brushings and biopsies will be carried out as per standard technique used in University medicine. Primary epithelial cell cultures will be established from bronchial brushings and will be used for optimising the dose of anti TNF-alpha required for inhibition of TNF-alpha induced responses. The biopsies will be placed in either culture medium alone, allergen extract or allergen extract plus anti TNF-alpha antibody and cultured for 24 hours. The biopsies will then be either snap frozen in liquid nitrogen for analysis of cytokines or processed for immunohistochemical analysis. The supernatants from each study will be recovered and analysed for cytokine (IL-5, IL-8, IL-13, MIP 1 alpha and RANTES) release by ELISA.

* bronchial brushings and biopsies
* Blood

Study title	NCT ID	Conditions	Interventions	Eligibility	Organization
Candida Associated Cytokines in Oral Leukoplakia	NCT04712929	Oral Leukoplaki...	Fluconazole	18 Years - 100 Years	All India Institute of Medical Sciences, New Delhi
Very Low Carbohydrate Diet Effects to GPS, Serum Lactate and TNF Alpha on Colorectal Cancer	NCT03221920	Diet Modificati... Colorectal Aden... Systemic Inflam... TNF Alpha Lactate	Very Low Carboh...	18 Years - 65 Years	Dr. Soetomo General Hospital
Candida Associated Cytokines in Oral Leukoplakia	NCT04712929	Oral Leukoplaki...	Fluconazole	18 Years - 100 Years	All India Institute of Medical Sciences, New Delhi
Candida Associated Cytokines in Oral Leukoplakia	NCT04712929	Oral Leukoplaki...	Fluconazole	18 Years - 100 Years	All India Institute of Medical Sciences, New Delhi
An Investigation of Tumor Necrosis Factor (TNF)-Alpha in Asthma Using Biopsy Explants and Primary Bronchial Epithelial Cell Cultures	NCT01161303	Asthma		18 Years - 65 Years	University Hospital Southampton NHS Foundation Trust

Spots Global Cancer Trial Database for tnf alpha

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Spots Global Cancer Trial Database for tnf alpha

Every month we try and update this database with for tnf alpha cancer trials from around the world to help patients and their families find trials that might be right for them. We offer this 100% free of charge and do not endorse any of the trials listed here, we hope it helps you or a loved one.

Every month we try and update this database with for tnf alpha cancer trials from around the world to help patients and their families find trials that might be right for them.
We offer this 100% free of charge and do not endorse any of the trials listed here, we hope it helps you or a loved one.