Toluidine Blue Test Research Articles

Metabolic screening and its impact in children with nonsyndromic intellectual disability.

ObjectiveThe objective of this study was to analyze the value of routine metabolic screening tests in children with an intellectual disability (ID) and its impact on improving their outcome and quality of life through appropriate intervention and treatment.Patients and methodsThis cross-sectional study was conducted in the Pediatric Neurology Clinic, Al Khafji Joint Operations Hospital, Kingdom of Saudi Arabia. A total of 150 children with nonsyndromic ID (66% males) in the age range of 5–17 years were compared with 50 apparently healthy age- and sex-matched controls. All studied groups were subjected to detailed history taking, family pedigree, thorough clinical examination, anthropometric measurements, routine laboratory investigations and urine metabolic screening tests (ferric chloride test and toluidine blue spot test and gas chromatography–mass spectrometry). Electroencephalography, IQ, psychiatric assessment and chromosomal study were done for the patient group only.ResultsPositive consanguineous marriage, older maternal or paternal age and family history of mental disabilities in other siblings were considered as risk factors for the development of mental disabilities. History of admission to neonatal intensive care unit was significantly higher among the patient group than among the controls (P<0.05). Metabolic screening tests showed that up to 35% of patients were positive for ferric chloride test, 9% of patients were positive for gas chromatography–mass spectrometry, and only 7 out of 150 (4.7%) patients were toluidine blue test positive.ConclusionMetabolic testing should be considered in the workup of individuals with nonsyndromic ID, which will need further specific investigations to confirm the diagnosis and determine the possible treatable cases.

Importance of Early Detection of Potentially Malignant Lesions in the Prevention of Oral Cancer.

Objective:The aim of this study was to assess the importance of early diagnosis of potentially malignant lesions in the prevention of oral cancer.Material and methods:In order to achieve the set objectives, we selected our participant pool. Of the 340 patients who came to the initial examination at the Clinic for Oral Medicine and Periodontology, we selected 40 patients with potentially malignant oral lesions (PMOL) of both sexes and of different ages. After an extensive anamnesis and a clinical examination of all mucous membranes, we documented potentially malignant lesions, their localization, subjective and objective symptoms, duration of pathological lesion, and bad habits (smoking and alcohol). All lesions were colored with Toluidine blue, which indicates malignancy in 75% of cases. Patients who had a positive Toluidine blue staining test were sent to biopsy and pathohistological analysis.Results:The most common potentially malignant lesions are Lichen reticularis (25%), Lichen erosivus (20%), and Leukoplakia (15%). The most common localization of lesions is the buccal mucosa bilaterally (26%), buccal mucosa unilaterally (10%), and the lower lip (10%). The staining test with Toluidine blue was positive at 23% PMOLs. In patients with positive Toluidine staining findings, biopsy and pathohistological analysis were performed, which showed that 20% of all lesions are carcinoma (CIS, Baseocellular Carcinoma, Squamocellular Carcinoma), while 14% are mucosal epithelial dysplasia. Statistical analysis confirmed the significance of positive staining of Toluidine blue and Leukoplakia (Spearman’s coefficient = 0.427, p = 0.006, N = 40), and statistically significant positive association of Squamocellular Carcinoma and Toluidine blue staining (Spearman’s coefficient = 0.619, p = 0.000, N = 40).Conclusion:Most oral cancers are caused by potentially malignant lesions, which is why a very detailed clinical examination with the visualization of pathological lesions is very important. Toluidine blue test can be an auxiliary tool for clinical diagnosis but does not replace the pathohistological finding. Biopsy and pathohistological findings are the gold standard in the diagnosis of oral cancers, as confirmed in our research.

Comparative study on the efficacy of tissue autofluorescence (visually enhanced lesion scope) and toluidine blue as a screening method in oral potentially malignant and malignant lesions

Objective and Study Design: Early identification of high-risk disease could greatly reduce both mortality and morbidity due to oral cancer. Hence, screening of such lesions and their early detection could improve prognosis. Metachromatic dye toluidine blue, which stains nuclear material in dysplastic epithelium, is widely used for screening of potentially malignant lesions. Visually enhanced lesion scope (VELscope) that identifies reduction in tissue fluorescence in dysplasia can also prove to be effective in screening for potentially malignant lesions. Hence, this study was done to compare the reliability of VELscope and toluidine blue as a screening tool in the diagnosis of malignant and potentially malignant lesions as compared to histopathological diagnosis which is gold standard. Materials and Methods: 90 patients with tobacco associated hyperkeratotic red and white lesion, ulcerative lesion, and frank malignancy included in study. Patients were subject to Tissue Autofluorescence (Visually Enhanced Lesion Scope) and Toluidine Blue stain tests. Nature of Autofluorescence and Toluidine Blue staining of tissue was recorded. Based on findings tissue was biopsied and examined under microscope from confirmatory diagnosis. Results: In this study, ninety patients with malignant and potentially malignant lesions were studied, VELscope showed 85.36% sensitivity and 75% specificity as compared to toluidine blue which showed 83.13% sensitivity and 87.5% specificity. The results were statistically significant (P < 0.001) with a mean deviation of ± 3. Conclusion: From the study, we concluded that VELscope was more effective as a screening tool for the detection of oral malignant lesions as compared to toluidine blue.

The effect of heracleum persicum (Golpar) oil and alcoholic extracts on sperm parameters and chromatin quality in mice

Evaluating the significance and the effects of plant-derived drugs on laboratory animal's fertility was recognized. There was antioxidant activity reported from Heracleum persicum (Golpar). Current study aims to study the antioxidant effect of Golpar extracts on sperm parameters and chromatin quality in mice. Eighteen adult male mice were divided to 3 groups (10 wk old, 35 gr weight): group1 received hydro alcoholic extract (1000 mg/kg, ip), group 2 received oil extract (200 mg/kg, ip) and group 3 serving as the sham control group that received sterile water. Finally, left cauda epididymis of each animal was dissected and sperm analysis was done accordingly. To asses sperm chromatin and DNA quality, we used aniline blue (AB), toluidine blue (TB), chromomycin A3 (CMA3) and acridine orange (AO) staining. Progressive and non-progressive sperm motility were significantly increased in group 1 in comparison with group 3 (p=0.032). There was an increasing trend in progressive sperm motility and decreasing trend in non-progressive sperm motility in group 2 in comparison with group 3, but the differences were not significant (p=0.221 and p=0.144, respectively). According to the sperm chromatin quality, the results of TB and AO tests revealed significant differences (p=0.004, p=0.000, respectively) between those groups and showed that the extracts of Golpar cause DNA damage, but no differences can be observed between them in AB and CMA3 staining (p>0.05). The results showed that Heracleum persicum extracts may improve sperm motility. Also, it has harmful effects on sperm chromatin condensation and DNA integrity in mice.

Value of videoroscopy in the detection of alterations of actinic cheilitis and the selection of biopsy areas.

BackgroundTo demonstrate the value of videoroscopy in identifying lesions and alterations not seen by oroscopy and to select the area for biopsy.Material and MethodsEighty patients were subjected to anamnesis, physical exam, videoroscopy exam, toluidine blue test and biopsy. A diagram of the lips was created to record the exact location where the lesion was found.ResultsPhysical exam identified 287 lesions, and videoroscopy identified 587 lesions; erythema and white lesions were the most common lesions associated with actinic cheilitis. Of the 59 performed biopsies, 32 (52.4%) cases were identified by videoroscopy that showed lesions that were not detected during physical examination.ConclusionsThe establishment of a diagram of the lip permitted registration of the precise location of the lesion. Videoroscopy was effective in locating lesions not seen by oroscopy. Both videoroscopy and the diagram of the lips allowed for better and earlier diagnosis and better patient follow-up for those with actinic cheilitis. Key words: Actinic cheilitis, potentially malignant disorder, videoroscopy, dermatoscopy, lip. oroscopy, diagram of lip.

Comparison of chemiluminescence and toluidine blue in the diagnosis of dysplasia in leukoplakia: a cross-sectional study.

The early diagnosis and treatment of oral cancer at a potentially-malignant stage would reduce morbidity and improve the survival rates and quality of life of patients. Leukoplakia is the most common potentially-malignant disorder. Clinical examination alone cannot differentiate between dysplastic and non-dysplastic leukoplakia. Chemiluminescence and toluidine blue are adjuncts to conventional examination for the early detection of dysplasia. The present study was conducted to compare the efficacies of chemiluminescence and toluidine blue for the diagnosis of dysplasia in leukoplakia. One hundred patients with leukoplakia were subjected to conventional oral examination, chemiluminescent examination with ViziLite and toluidine blue test, followed by histopathological confirmation of dysplasia. The sensitivity, specificity, positive predictive value, negative predictive value, and accuracy of chemiluminescence was 84.84%, 41.17%, 73.68%, 58.33%, and 70%, respectively. For toluidine blue it was 42.4%, 88.23%, 87.50%, 44.11%, and 58% respectively. Both the techniques have adjunctive utility in the diagnosis of dysplasia in leukoplakia, but toluidine blue was more effective in identifying the more severe grades of dysplasia, and thus it effectively discriminated high-risk from low-risk lesions.

Utility of toluidine blue test in accessing and detecting intra-oral malignancies.

In vivo staining reveals cytological details that might otherwise not be apparent. The aim of the study was to test the utility of toluidine blue test in detecting various types of malignant and premalignant lesions in early stage. Fifty patients with lesion in oral cavity having suspicion of malignancy clinically were selected. After subjecting the patients to clinical examination, the suspicious lesions were stained with 1% toluidine blue. The biopsy site was selected on the basis of clinical appearance and dye retention and in the sites where no retention of the stain occurred, clinical judgment directed the biopsy site. The sensitivity of toluidine blue in detecting premalignant or malignant lesions was found to be 97.8% and the over all specificity was found to be 100%. The positive predictive value, negative predictive value and diagnostic accuracy was reported to be 100, 80 and 90% respectively. Toluidine Blue staining is highly a reliable source for the detection of insitu and invasive carcinomas. Staining with this stains is an adjunct to clinical judgment, assist in the choice of biopsy site, follow up of premalignant lesions and marginal demarcation of the malignant lesions enabling an intervention method to be adopted earlier for the disease, which carries a high rate of morbidity and mortality.

Non-covalent synthesis of metal oxide nanoparticle-heparin hybrid systems: a new approach to bioactive nanoparticles.

Heparin has been conjugated to Fe3O4, Co3O4, and NiO nanoparticles (NPs) through electrostatic interactions, producing colloidal suspensions of hybrid metal oxide heparin NPs that are stable in water. Negative zeta potentials and retention of heparin’s ability to capture toluidine blue indicate that heparin’s negative charges are exposed on the surface of the coated NPs. IR results confirmed the formation of nanohybrids as did NMR experiments, which were also interpreted on the basis of toluidine blue tests. Transmission electron microscopy results revealed that the heparin coating does not modify the shape or dimension of the NPs. Dynamic light scattering and negative zeta potential measurements confirmed that heparin surface functionalisation is an effective strategy to prevent NP aggregation.

Basosquamous Cell Carcinoma of the Lower Lip Arising from Actinic Cheilitis: Case Report and up Date

Background: Basosquamous carcinoma (BSC) is a rare non-melanoma skin cancer, considered to be a subtype of basal cell carcinoma (BCC). BSC often produces distant metastases with a higher risk of recurrence than that of BCC which is not commonly found in the lip. Case Report: A 57-year-old white female patient presented an ulcer on her lower lip that had an ongoing development for over six months. Physical examination, photo documentation, videoroscopy, scraped cytology, toluidine blue test, and biopsy of the ulcer were carried out. Results: Upon physical examination we observed an actinic cheilitis associated with the ulcer. Videoroscopy revealed the presence of fissures and erosion that had not been seen by oroscopy. Toluidine blue test was only positive for the region of the ulcer. Cytological analysis revealed rare nests compatible with carcinoma. Histopathology of the biopsy revealed a carcinoma with nests lined by basal cells associated with areas of squamous differentiation. The patient was then referred to surgery for the removal of the BCC. Analysis of the specimen showed free surgical margins and the immunohistochemical panel did not confirm the initial diagnosis of BCC, indicating a subtype of BSC. After surgery, the patient has been followed by periodic consultations. She is well and without further complications. Coments: BSC is considered to be an aggressive and rare tumor affecting mainly upper face and primarily affects men over 60 years of age. Since our patient is a woman presenting the lesion in the lower lip, this highlights the unusual and interesting presentation of this case report.

The utility of sperm DNA damage assay using toluidine blue and aniline blue staining in routine semen analysis.

ObjectiveThe aim of the present study was to examine the relationship among male age, strict morphology, and sperm chromatin structure and condensation.MethodsSperm samples from a total of 100 men underwent semen analysis, and sperm chromatin structure and condensation were assessed with toluidine blue (TB) and aniline blue (AB) tests.ResultsPrevalence of strict morphology of less than 4%, and abnormal sperm chromatin structure and condensation did not show any statistically significant differences according to male age (p=0.605, p=0.235, and p=0.080). No significant correlation was demonstrated among age of male partners, strict morphology, and abnormal sperm chromatin structure using TB and AB tests. However, abnormal sperm chromatin condensation was positively associated with sperm chromatin structure (r=0.594, p=0.000) and showed negative correlation with strict morphology (r=-0.219, p=0.029).ConclusionThe tests for sperm chromatin condensation showed a significant association with strict morphology. Further study is needed to elucidate the relationship between clinical outcome and sperm chromatin tests.

Evaluation of chemiluminescence, toluidine blue and histopathology for detection of high risk oral precancerous lesions: A cross-sectional study

BackgroundEarly detection holds the key to an effective control of cancers in general and of oral cancers in particular. However, screening procedures for oral cancer are not straightforward due to procedural requirements as well as feasibility issues, especially in resource-limited countries.MethodsWe conducted a cross-sectional study to compare the performance of chemiluminescence, toluidine blue and histopathology for detection of high-risk precancerous oral lesions. We evaluated 99 lesions from 55 patients who underwent chemiluminescence and toluidine blue tests along with biopsy and histopathological examination. We studied inter-as well as intra-rater agreement in the histopathological evaluation and then using latent class modeling, we estimated the operating characteristics of these tests in the absence of a reference standard test.ResultsThere was a weak inter-rater agreement (kappa < 0.15) as well as a weak intra-rater reproducibility (Pearson's r = 0.28, intra-class correlation rho = 0.03) in the histopathological evaluation of potentially high-risk precancerous lesions. When compared to histopathology, chemiluminescence and toluidine blue retention had a sensitivity of 1.00 and 0.59, respectively and a specificity of 0.01 and 0.79, respectively. However, latent class analysis indicated a low sensitivity (0.37) and high specificity (0.90) of histopathological evaluation. Toluidine blue had a near perfect high sensitivity and specificity for detection of high-risk lesions.ConclusionIn our study, there was variability in the histopathological evaluation of oral precancerous lesions. Our results indicate that toluidine blue retention test may be better suited than chemiluminescence to detect high-risk oral precancerous lesions in a high-prevalence and low-resource setting like India.

Utilização do teste com o azul de toluidina como método auxiliar no diagnóstico de lesões orais

Clinical evaluation is not always sufficient for a conclusive diagnosis. The objective of this study is to evaluate the validity of toluidine blue test for the diagnosis of oral lesions. The sample was comprised of 110 patients with oral lesions. Following the established protocol, the toluidine blue test was applied in all lesions. One hundred and five lesions were biopsied and histopathology considered the gold standard diagnosis. The toluidine blue test was positive in 31 out of the 110 lesions, in 20 focal and 11 diffuse positivity. It was conclude that the toluidine blue test is a useful tool as an auxiliary method for the diagnosis of oral lesions.

Assessment of oral mucosa in normal, precancer and cancer using chemiluminescent illumination, toluidine blue supravital staining and oral exfoliative cytology

Context:Carcinoma in an early stage of development is hard to detect clinically because the lesion may not be palpable and color of the lesional tissue is not necessarily different from the color of the surrounding mucosa. In order to improve the efficacy of the diagnosis, techniques are being developed to complement clinical examination and to facilitate the identification of initial carcinomas.Aims:To find out the efficacy of chemiluminescent illumination (ViziLite™) for the diagnosis in precancer and cancer patients and compare this result to toluidine blue staining and oral exfoliative cytology.Materials and Methods:This study was done in 3 groups. Each group consists of 10 cases. Group I consists of normal appearing mucosa. Group II and III consist of clinically diagnosed pre-cancer and clinically suggestive of cancer respectively. Chemiluminescent illumination, toluidine blue supravital staining, oral exfoliative cytology and biopsy were performed in all cases.Statistical analysis used:SPSS version 10.05 was used to calculate positive and negative predictive values.Results:In Group I, all 10 patients showed negative result to ViziLite™. 8 patients showed positivity and 2 patients showed negativity to ViziLite™ test in Group II. 9 patients were positive and one patient was negative for ViziLite™.Conclusions:Chemiluminescent illumination test was sensitive for precancerous and cancerous lesions, which presented as keratotic lesions and red-white lesions. It showed negative result to erosive lesions. Toluidine blue staining test was reliable in precancerous and cancerous lesions, which present as erosive and red-white lesions. It showed negative result to keratotic lesions. Oral exfoliative cytology has diagnostic value in cancer patients than in precancer patients. These Results indicate that chemiluminescent illumination test is relatively reliable and accurate than toluidine blue staining test and useful chair side diagnostic test.

Evaluation of the Frequency of Micronuclei in Exfoliated Cells from Oral Lesions Previously Identified by Toluidine Blue

Objective: Patients using a removable prosthesis are susceptible to a variety of oral lesions that may progress to cancer. Toluidine blue (TB) staining is used to identify premalignant lesions, but the results are still controversial. Since micronuclei (MN) are a biomarker of genetic instability, the objective of this study was to determine the frequency of MN in white lesions of the oral mucosa and to compare the results with those of the TB test. Study Design: The study included 20 removable prosthesis users with white lesions that were previously classified as toluidine positive or negative. The frequency of MN was evaluated in exfoliated cells from lesions and normal mucosa. Nuclear anomalies were also registered. Results: A significant increase (p < 0.05) in the frequency of MN was observed in exfoliated cells from lesions compared to normal mucosal cells, and no relationship was seen with TB staining. Lifestyle factors or gender did not influence the results. Conclusions: The frequency of MN is a sensitive biomarker and can be used to predict genomic instability in white oral lesions. The MN assay may serve as a good parameter in the battery of tests used to identify high-risk individuals, contributing to the identification of the biological conditions of oral lesions.

The use of toluidine blue in the detection of pre-malignant and malignant oral lesions

The purpose of this study was to evaluate the efficacy of the toluidine blue (TB) test as a diagnostic tool in the detection of malignant and dysplastic lesions of the oral cavity. This study was carried out because of a lack of consensus among different authors on the utility of TB, as well as to determine useful adjuncts to detect oral pre-cancer and cancer. The study included 160 patients with oral mucosal disorders that included suspicious or malignant lesions detected at clinical visual examination, confirmed by histopathological evaluation. All lesions were submitted to TB staining. The sensitivity and specificity for the detection of malignant or dysplastic lesions by this test were 65.5% and 73.3%, respectively. Overall, the detection rate with TB (sensitivity) was slightly lower compared with those reported by other authors but the specificity was comparable to several reports. Positive predictive value (35.2%) was also lower than previous studies, whereas negative predictive value (90.6%) was similar. The simplicity of the test procedure and the validity of derived values suggest TB staining can be a valuable adjunct to the diagnostic process, as long as it is carefully correlated with the clinical characteristics of the mucosal disorder and histopathological diagnosis.

Evaluation of male fertility potential by Toluidine Blue test for sperm chromatin structure assessment

We have previously suggested that the Toluidine Blue (TB) test can be used for sperm chromatin structure assessment. In this study, we wished to evaluate the clinical applicability of the TB test in assessing male fertility potential using well-defined groups of fertile and infertile men. Sixty-three fertile and 79 infertile men were tested. Infertility thresholds for the proportion of sperm with abnormal [TB dark cells (TBDCs)] and normal [TB light cells (TBLCs)] chromatin structure were set by the ROC curve analysis. Thresholds of 45% TBDC and 20% TBLC were highly predictive for infertility (specificity of the test: 92 and 90%, respectively), but they were poor predictors of the fertility (sensitivity of the test: 42 and 32%, respectively). Odds ratio for infertility was 7.5 [95% confidence interval (CI): 2.7-20.8] when the 45% TBDC threshold was used and 4.4 (95% CI: 1.7-11.6) when the 20% TBLC threshold was used. The TB test can be suggested for clinical use as a complementary test for standard semen analysis to diagnose male infertility.

Effect of Plasma Pretreatment on Heparin Immobilization on Polymer Sheet

The effect of plasma pretreatment on immobilization of heparin on polymer surface was investigated using argon and/or ammonia surface-wave plasmas. Heparin was immobilized by binding with the amino groups introduced by the ammonia plasma on the polymer surface and was confirmed by toluidine blue test and X-ray photoelectron spectroscopy (XPS). The ratio of sulfur contained in heparin significantly increased with the contents of amino groups after the heparin immobilization process. It suggests that most heparins are covalently bound to the amino groups introduced on the polyurethane surfaces.

Collins test in patients with vulvar pruritus

E-mail address: sicantal3@yahoo.com. Pruritus is the most common symptom of vulvar disorders. The prevalence of vulvar pruritus in the female population is unknown, as is the prevalence of most of the conditions responsible for the itching [1]. It is important to recognize that vulvar pruritus is a symptom and not a disease, and optimal management should be to search for the underlying cause of the problem so that appropriate treatment can be given. In particular, it is important to differentiate between benign, premalignant, and malignant conditions [1]. Biopsy is the primary diagnostic tool for vulvar lesions [2]. The important question here is to accurately determine the site of the biopsy. To increase the sensitivity and specificity of vulvar biopsy, Collins toluidine blue test can be used to define more accurately the areas of vulvar abnormalities [1]. The present study was conducted at Al-Yarmouk Teaching Hospital and Al-Habibiya Maternity Hospital in Baghdad, Iraq, between September 2004 and June 2005. A total of 100 women aged between 40 and 60 years suffering from vulvar pruritus of variable duration were included in the study. All women gavewritten consent for participation. Each patient had a full history taken and underwent a complete physical examination. Exclusion criteria were infections, contact dermatitis, psoriasis, seborrheic dermatitis, use of barrier contraception, ulcer or mass of the vulva, or any visible lesions of the vagina and cervix. To identify areas of vulvar abnormalities, toluidine blue was applied to the vulva. Dark blue colored areas that might represent abnormal skinwere biopsied. If therewas no abnormal staining, a biopsywas taken from the most pruritic areas of the vulva to detect false-negative cases. All biopsies were examined by one histopathologist.

Increased sperm ubiquitination correlates with abnormal chromatin integrity

Ubiquitin, a 8.5 kDa peptide that marks other proteins for proteasomal degradation, tags defective spermatozoa during epididymal passage and is proposed as a biomarker for sperm quality. The present study was designed to evaluate the relationships between sperm ubiquitination, sperm chromatin integrity and semen parameters. Semen samples from 63 couples were collected and analysed according to World Health Organization criteria. Each sample was evaluated for sperm ubiquitination by the direct immunofluorescence method, using anti-ubiquitin antibodies. Chromatin integrity of the same samples was analysed using acridine orange (AO) and toluidine blue (TB) tests. A positive correlation was found between ubiquitinated spermatozoa and the percentage of spermatozoa with abnormal chromatin (AO: r = 0.58, P < 0.001 and TB: r = 0.48, P < 0.001). Negative correlations were obtained between sperm ubiquitination and: sperm count ( r = −0.2, P = 0.048), sperm morphology ( r = –0.36, P = 0.003), rapidly progressive motility ( r = –0.25, P = 0.044) and slow progressive motility ( r = −0.28, P = 0.022). Sperm ubiquitination was positively correlated with the percentage of immotile spermatozoa. These results show that among semen parameters, chromatin abnormality is more closely associated with sperm ubiquitination and further validate sperm ubiquitination as a suitable marker for sperm quality.

Evaluation of fertility potential by toluidine blue test and the sperm chromatin structure assay

Sperm chromatin integrity is important for mammalian fertilization. The sperm chromatin structure assay (SCSA) using flow cytometry is frequently used for the assessment of fragmented sperm DNA. On the other hand, Toluidine blue (TB) assay is a simple alternate test for sperm chromatin assessment. The objective of our study was to evaluate the Toluidine blue test and SCSA for the assessment of sperm chromatin damage in proven and unproven donors.