A standardized method for measuring intercellular spaces in esophageal biopsies in patients with suspected gastroesophageal reflux disease (the intercellular space ratio).

OBJECTIVE: Dilated intercellular space (DIS) in esophageal biopsies is regarded as a possible early sign of mucosal injury in gastroesophageal reflux disease (GERD). This study presents a standardized approach of intercellular space measurement. MATERIAL AND METHODS: Distal and proximal esophageal biopsies were taken from 19 patients with suspected GERD, and examined with TEM. A grid containing 150 line-crossing points was applied upon each photomicrograph. The number of points falling on the intercellular space was divided by the total number of points of the grid, thereby creating a ratio called the intercellular space ratio (ISR). The ISR method was validated with regard to intra- and interobserver agreement, and was compared to a widely used method for measuring intercellular space diameter developed by Tobey et al. (Tobey NA, Carson JL, Alkiek RA, Orlando RC. Dilated intercellular spaces: a morphological feature of acid reflux-damaged human esophageal epithelium. Gastroenterology 1996;111(5):1200-1205). The ISR was also compared to other markers for GERD. Results. Pearson's correlation coefficients for intra- and interobserver agreement were 0.91 (p < 0.001) and 0.82 (p < 0.001), respectively. The Pearson's correlation coefficient between the ISR and the intercellular space diameter according to Tobey et al., measured in the same micrographs, was 0.32 (p < 0.001). The proximal ISR correlated significantly with the distal ISR (Spearman's rho = 0.57, p = 0.010), and with heartburn symptom score (Spearman's rho = 0.50, p = 0.028). CONCLUSIONS: The ISR showed a high intraobserver and interobserver agreement. It also displayed good external validity when compared to other markers for gastroesophageal reflux. A rather poor correlation was however found between the ISR and the intercellular space diameter measured as described by Tobey et al.

Validation of the Reflux, Airway and Sleep Questionnaire (RASQ).

BACKGROUND: Airway symptoms and sleeplessness in patients with gastroesophageal reflux disease (GERD) may be of importance. This study validates a new questionnaire dealing with such symptoms. MATERIAL AND METHODS: The Reflux, Airway and Sleep Questionnaire (RASQ) is self-administered, asks about 18 symptoms or diagnoses possibly related to GERD answered on a seven-point Likert scale and with a 1-year recall period. There are questions about heartburn/regurgitation, sleeplessness, snoring, pneumonia, and upper airway infections, as well as various bronchial and laryngeal symptoms. The study included 305 patients diagnosed with GERD (n = 65), laryngitis (n = 32), asthma (n = 30), chronic obstructive pulmonary disease (n = 45), acute bronchitis (n = 39), pneumonia (n = 42), or upper airway infection (n = 52) during the last year, and 708 matched healthy controls. Concurrent validity was based on comparisons between patients and controls. Convergent validity for sleeplessness and snoring were tested by comparing the RASQ with the Pittsburgh Sleep Quality Index (PSQI) and the Epworth Sleepiness Scale (ESS). Test-retest reliability was examined in patients with GERD with stable symptoms (n = 29). RESULTS: Response rates were 64% for patients and 55% for controls. Scores for RASQ as a total and all subscales were significantly higher in patients than in controls. Sleeplessness scores correlated significantly to the global PSQI score. Snoring correlated significantly with both the ESS and relevant aspects of the PSQI. Test-retest reliability and Cronbach's alpha were satisfactory, with coefficients ranging between 0.65-0.95 and 0.88-0.92, respectively. CONCLUSION: The RASQ appears to be well suited for measuring typical reflux symptoms as well as airway symptoms and sleep disturbances.

Airway symptoms and sleeping difficulties in operated and non-operated patients with gastroesophageal reflux disease.

INTRODUCTION: Anti-reflux treatment studies have not succeeded in proving a causal relationship between gastroesophageal reflux disease (GERD), airway symptoms and sleeping difficulties. In a recent follow-up study we showed that patients operated for GERD have significantly less heartburn/acid regurgitation symptoms than matched non-operated patients. These two groups probably had different degrees of reflux over a long period of time. It is thus hypothesized that operated patients would report less airway symptoms and sleeping difficulties than comparable non-operated patients. A new follow-up study of the same patients was therefore conducted. MATERIAL AND METHODS: A total of 179 patients operated for GERD and 179 matched non-operated patients with confirmed GERD were sent the Reflux, Airway & Sleep Questionnaire (RASQ), which is a new, validated questionnaire dealing with heartburn/acid regurgitation, airway symptoms, and sleeping difficulties. Answers are given on a 7-point Likert scale and the assessment period is 1 year. RESULTS: Response rates were 68% in both groups. Operated patients reported significantly less reflux symptoms than non-operated patients (p < 0.001). Patients in the surgery group also reported less symptoms in two subscales of the RASQ dealing with airway symptoms: Laryngopharyngitis (p = 0.04) and Bronchitis (p = 0.01). There was a tendency toward less sleeplessness in operated patients, but this was not statistically significant. Snoring was less bothersome in operated patients (p = 0.02). CONCLUSIONS: Patients operated for GERD have less heartburn/acid regurgitation symptoms and less airway symptoms than non-operated patients. The findings lend support to the hypothesis of a causal relationship between gastroesophageal reflux, airway symptoms, and sleeping difficulties.

Activation of REG family proteins in colitis.

AIMS: To do a genome-wide gene expression study of active and inactive ulcerative colitis and Crohn's disease (inflammatory bowel disease--IBD) and examine the most differentially expressed genes. As the study showed an extreme upregulation of all regenerating islet-derived genes (REG proteins) in active IBD, we further studied the expression of REGs on protein level in active and inactive IBD, as well as in non-IBD (pseudomembranous) colitis. METHODS: Microarray analysis was done on a total of 100 pinch biopsy samples from healthy controls and patients with Crohn's disease or ulcerative colitis. Tissue samples from IBD and pseudomembranous colitis were examined with routine histology and immunohistochemical analysis for REGIα, REGIV, DEFA6, and serotonin. RESULTS: REG mRNAs were up to 83 times overexpressed in diseased mucosa compared with mucosa from healthy individuals. REGIα and REGIV were overexpressed at immunohistochemistry and located to different mucosal cell types. REGIα was expressed in basal half of crypts, REGIV in mid and outer parts of crypts and in surface epithelium and seems to be stored in, and secreted from, goblets. Pseudomembranous colitis samples showed similar staining patterns, and some IBD samples stained REG positive without inflammation on routine histology. CONCLUSIONS: All REG family mRNAs are upregulated in IBD. REGIα and REGIV have different cellular localization, possibly reflecting different biological functions. REG protein expression also in pseudomembranous colitis shows that REG family proteins are regulated in inflammatory injury and repair, not specifically for IBD as previously thought.