Impact of Pronase, Sodium Thiosulfate, and Methylene Blue Combinations on Development and Survival of Sodium Hypochlorite Surface-Disinfected Zebrafish (Danio rerio) Embryos.

Embryo surface disinfection is utilized in aquaculture to decrease the risk of pathogen introduction into established colonies. Zebrafish embryos are commonly disinfected with unbuffered sodium hypochlorite at 25-50 ppm for 10 min with or without concurrent treatment with chemicals, including pronase (Pron), sodium thiosulfate, and/or methylene blue; however, the impact of these chemicals on embryo survival and development has not been evaluated. In this study, AB and casper embryos were exposed to disinfection protocols that used Pron, sodium thiosulfate, and/or methylene blue (given alone, in various combinations, or all three combined) with 50 and 100 ppm sodium hypochlorite performed 6 and 24 h postfertilization (HPF). All groups were evaluated for survival, hatching, and malformations at 5 days postfertilization. Maximal survival (69%-97%) and hatching rates (66%-94%) were generally observed with sodium hypochlorite disinfection followed by exposure to both Pron and sodium thiosulfate and maintenance in standard embryo medium without methylene blue. Methylene blue had variable effects on survival and hatching. Higher survival and hatching rates were seen in AB embryos disinfected at 6 HPF and casper embryos disinfected at 24 HPF. Susceptibility to sodium hypochlorite toxicity differed by strain, emphasizing the need to test disinfection protocols on small embryo cohorts.

Sensitization to Empynase(pronase B) in exposed hospital personnel and identification of the Empynase allergen.

BACKGROUND: Empynase is a proteolytic enzyme that is widely used as an anti-inflammatory drug in Korea. We evaluated the prevalence of sensitization to Empynase in association with respiratory allergy symptoms in exposed hospital personnel, and identified the IgE-binding components in the Empynase extract, using sera with high levels of specific IgE antibodies. METHODS: A total of 154 hospital personnel (135 nurses and 19 pharmacists) who worked in a university hospital and 123 unexposed healthy control subjects were enrolled. A questionnaire was administered that addressed demographics, job category, history of atopic diseases, diverse symptoms including nasal and lower respiratory symptoms, and the association of symptoms with work. Skin prick tests (SPTs) to common aeroallergens and Empynase extract were performed. Empynase-specific IgE antibody was detected by ELISA, and ELISA inhibition tests were conducted. IgE-binding components were identified by SDS-PAGE and IgE immunoblotting. RESULTS: Forty-two subjects (27.3%) complained of work-related respiratory symptoms (WRRS). Five nurses (3.7%) and one pharmacist (5.3%) had work-related asthma symptoms, and 34 nurses (25.2%) and six pharmacists (31.6%) had work-related rhinitis symptoms. The prevalence of sensitization to Empynase on SPTs was 20.1%, and tended to be higher in pharmacists (31.6%) than in nurses (18.5%). It was estimated that 3.9-8.4% of hospital personnel had WRRS attributable to Empynase. The duration of exposure was longer in positive SPT responders than in negative responders (51.9+/-27.5 vs. 39.2+/-27.3 months, respectively; P<0.05), and the prevalence of Empynase-positive SPTs was significantly higher in subjects with asthma than in those without asthma (57.1% vs. 18.4%, respectively; P<0.05). The levels of Empynase-specific IgE antibodies were significantly higher in pharmacists (76.1+/-83.4 OD units) and nurses (56.3+/-103.0 OD units) than in normal controls (39.8+/-12.7 OD units; P<0.05). Seven subjects (two pharmacists and five nurses) had high serum levels of Empynase-specific IgE antibodies; six of these subjects had WRRS. ELISA inhibition tests were performed with the sera of these six subjects, revealing significant inhibition only with the addition of Empynase. Four strongly staining protein bands (sizes: 36, 33, 16, and 10 kDa) from Empynase extract were observed to bind to the IgE antibodies of sensitized subjects. Conclusion Exposure to Empynase powder may cause rhinitis and asthma in hospital personnel, and the pathogenic mechanism appears to be IgE mediated.

A 1-h topical therapy for the treatment of Helicobacter pylori infection.

OBJECTIVE: A novel topical therapeutic methodology for the treatment of Helicobacter pylori infection was developed and studied in 25 patients with H. pylori to evaluate safety and efficacy. METHODS: The patients had been given lansoprazole (30 mg, hs) orally and pronase (18,000 tyrosine units, b.i.d.) for the 2 days before topical therapy. One hundred milliliters of solution with 80 ml of 7% sodium bicarbonate and 20 ml of contrast medium meglumine sodium amidotrizoate containing bismuth subnitrate (1 g), amoxicillin (2 g), metronidazole (1 g), and pronase (36,000 tyrosine units) were instilled into the stomach through a nasally introduced 16-Fr intestinal tube with a balloon at its radiopaque tip, which was inflated with approximately 25 ml of air and lodged postbulbarly at the superior duodenal angle under fluoroscopy, thus preventing leakage of the solution distally into the jejunum. The solution was kept in the stomach for 1 h, and the patient's position was changed every 15 min from the sitting to the supine, prone, and right lateral position to expose the entire gastric mucosa. The solution was suctioned at the end of the procedure. RESULTS: H. pylori infection was successfully cured in 24 (96%) patients, confirmed 4 wk after the therapeutic procedure by negative smear, culture, and histology of the antral and corpus biopsy specimens. No side effects were observed except for loose stools in one case. CONCLUSION: This 1-h topical therapy is a safe, effective, and well tolerated procedure for the treatment of H. pylori infection. With further improvements and modifications of the method itself, as well as of the drug regimens, this method may become a highly efficient modality for anti-H. pylori therapy.