ABSTRACT
OBJECTIVE: Mucin is an important parameter for detection and assessment in studies of airway disease including asthma and cystic fibrosis. Histochemical techniques are often used to evaluate mucin in tissues sections. Periodic acid Schiff (PAS) is a common technique to detect neutral mucins in tissue, but this technique also detects other tissue components including cellular glycogen. We tested whether depletion of glycogen, a common cellular constituent, could impact the detection of mucin in the surface epithelium of the trachea. RESULTS: Normal tissues stained by PAS had significantly more staining than serial sections of glycogen-depleted tissue with PAS staining (i.e. dPAS technique) based on both quantitative analysis and semiquantitative scores. Most of the excess stain by the PAS technique was detected in ciliated cells adjacent to goblet cells. We also compared normal tissues using the Alcian blue technique, which does not have reported glycogen staining, with the dPAS technique. These groups had similar amounts of staining consistent with a high degree of mucin specificity. Our results suggest that when using PAS techniques to stain airways, the dPAS approach is preferred as it enhances the specificity for airway mucin.
Subject(s)
Glycogen/metabolism , Liver/metabolism , Mucins/metabolism , Periodic Acid-Schiff Reaction/methods , Respiratory Mucosa/metabolism , Staining and Labeling/methods , Trachea/metabolism , Animals , Female , Male , Mice , Periodic Acid-Schiff Reaction/standards , Sensitivity and Specificity , Staining and Labeling/standards , SwineABSTRACT
PURPOSE: To establish the strength of the association between routine tear function tests and conjunctival impression cytology (CIC) and to determine whether they simulate the morphological and cytological changes that occur on the ocular surface in dry eye. What are the sensitivity, specificity and positive predictive values of these tests when CIC is considered the gold standard? METHODS: The tear film profile included tear film break up time (TBUT), Schirmer's-1, Rose Bengal scores (RBS), and impression cytology. CIC samples were obtained from the inferior bulbar conjunctiva and stained with periodic acid-Schiff and counter stained with hematoxylin and eosin. RESULTS: The mean Schirmer's value was 11.66 ± 5.90 in patients and 17.17 ± 2.97 in controls (p < 0.001). The mean TBUT in participants was 8.88 ± 3.54 and 13.53 ± 2.12 in controls (p < 0.001). Patients had a mean goblet cell density (GCD) of 490 ± 213, while the value for controls was 1,462 ± 661 (p < 0.001). Abnormal CIC was observed in 46.7% cases of dry eye and in 32.8% of controls. The correlation coefficient (L) for Schirmer's was 0.2 and 0.24 for participants and controls, respectively, while TBUT values were 0.26 and 0.38, RBS were 0.5 and 0.5, and GCD was 0.8 and 0.6 in cases and controls, respectively. CONCLUSIONS: GCD, RBS, and TBUT were better predictors of morphological and cytological changes in the conjunctiva than Schirmer's in dry eye syndrome. The sensitivity of tear function tests in diagnosing dry eye was TBUT > Schirmer's > RBS, and the specificity was Schirmer's > TBUT > RBS in decreasing order when CIC was considered the gold standard.
Subject(s)
Diagnostic Techniques, Ophthalmological/standards , Dry Eye Syndromes/diagnosis , Dry Eye Syndromes/pathology , Ophthalmology/standards , Tears , Adolescent , Adult , Case-Control Studies , Child , Conjunctiva/pathology , Eosine Yellowish-(YS) , Female , Goblet Cells/pathology , Hematoxylin , Humans , Male , Middle Aged , Periodic Acid-Schiff Reaction/standards , Predictive Value of Tests , Rose Bengal , Sensitivity and Specificity , Young AdultABSTRACT
PURPOSE: To establish the strength of the association between routine tear function tests and conjunctival impression cytology (CIC) and to determine whether they simulate the morphological and cytological changes that occur on the ocular surface in dry eye. What are the sensitivity, specificity and positive predictive values of these tests when CIC is considered the gold standard? METHODS: The tear film profile included tear film break up time (TBUT), Schirmer's-1, Rose Bengal scores (RBS), and impression cytology. CIC samples were obtained from the inferior bulbar conjunctiva and stained with periodic acid-Schiff and counter stained with hematoxylin and eosin. RESULTS: The mean Schirmer's value was 11.66 +/- 5.90 in patients and 17.17 +/- 2.97 in controls (p Schirmer's > RBS, and the specificity was Schirmer's > TBUT > RBS in decreasing order when CIC was considered the gold standard.
Subject(s)
Adolescent , Adult , Child , Female , Humans , Male , Middle Aged , Young Adult , Case-Control Studies , Conjunctiva/pathology , Diagnostic Techniques, Ophthalmological/standards , Dry Eye Syndromes/diagnosis , Eosine Yellowish-(YS) , Goblet Cells/pathology , Hematoxylin , Ophthalmology/standards , Periodic Acid-Schiff Reaction/standards , Predictive Value of Tests , Rose Bengal , Sensitivity and Specificity , TearsABSTRACT
Screening of blood films for the presence of periodic acid-Schiff (PAS)-positive lymphocyte vacuoles is sometimes used to support the diagnosis of Pompe disease, but the actual diagnostic value is still unknown. We collected peripheral blood films from 65 untreated Pompe patients and 51 controls. Lymphocyte vacuolization was quantified using three methods: percentage vacuolated lymphocytes, percentage PAS-positive lymphocytes, and a PAS score depending on staining intensity. Diagnostic accuracy of the tests was assessed using receiver operating characteristic (ROC) curves. All three methods fully discerned classic infantile patients from controls. The mean values of patients with milder forms of Pompe disease were significantly higher than those of controls, but full separation was not obtained. The area under the ROC curve was 0.98 for the percentage vacuolated lymphocytes (optimal cutoff value 3; sensitivity 91%, specificity 96%) and 0.99 for the percentage PAS-positive lymphocytes and PAS score (optimal cutoff value 9; sensitivity 100%, specificity 98%). Our data indicate that PAS-stained blood films can be used as a reliable screening tool to support a diagnosis of Pompe disease. The percentage of PAS-positive lymphocytes is convenient for use in clinical practice but should always be interpreted in combination with other clinical and laboratory parameters.
Subject(s)
Glycogen Storage Disease Type II/diagnosis , Lymphocytes/pathology , Mass Screening/methods , Periodic Acid-Schiff Reaction/methods , Vacuoles/pathology , Adult , Aged , Child , Child, Preschool , Enzyme Replacement Therapy , Eosine Yellowish-(YS) , Female , Glycogen/metabolism , Glycogen Storage Disease Type II/drug therapy , Humans , Infant , Infant, Newborn , Lymphocytes/metabolism , Male , Mass Screening/standards , Methylene Blue , Middle Aged , Periodic Acid-Schiff Reaction/standards , ROC Curve , Sensitivity and Specificity , Vacuoles/metabolism , Young AdultABSTRACT
A project group working under the European Confederation of Laboratory Medicine (ECLM) presents recommendations for standardized procedures for the Feulgen-Rossenbeck-Schiff and the periodic acid-Schiff (PAS) reactions on cytological and histological material. The advantages and disadvantages of such standardized procedures are presented here in a preamble. Both users and manufacturers are encouraged to give their opinions with a view to achieving consensus on these procedures and on how further work on these lines may proceed.
