A hepatoprotective peptide, pyroglutamyl leucine (pyroGlu-Leu), was identified in wheat gluten hydrolysate through an in vivo activity-guided fractionation approach based on D-galactosamine-induced acute hepatitis in rats and fractionation of peptides with large-scale preparative ampholine-free isoelectric focusing. The active acidic fraction predominantly consisted of pyroglutamyl peptides and free pyroglutamic acid. Pyroglutamyl peptides were derivatized with phenyl isothiocyanate after removal of a pyroglutamyl residue by pyroglutamate aminopeptidase. The derivatives were purified by reversed-phase HPLC and subjected to sequence analysis. The active fraction contained pyroGlu-Ile, pyroGlu-Leu, pyroGlu-Gln, pyroGlu-Gln-Gln, and free pyroGlu. Ingestion of pyroGlu-Leu at 20 mg/kg body weight significantly decreased serum aspartate and alanine aminotransferases to approximately 30% and 20% of those values of the vehicle group, respectively, which were near the normal levels. Thirty minutes after ingestion of pyroGlu-Leu at 20 mg/kg, the concentration of pyroGlu-Leu in portal blood plasma increased to approximately 2 μM.
It has been demonstrated that peptides in enzymatic hydrolysates of proteins can be fractionated on the basis of the amphoteric nature of the sample peptides, by a laboratory-scale isoelectric focusing apparatus, without adding a chemically synthesized carrier ampholyte. This approach is referred to as autofocusing. In the present study, a large-scale (up to 50 L) autofocusing apparatus was developed and tested. A tank (125 cm x 25 cm x 20 cm) was divided into 12 compartments by 11 plates, each with a window covered in a thin agarose gel layer supported by a nylon screen (100 mesh). The compartments at both ends were filled with 0.1 N phosphoric acid (anode) and 0.1 N NaOH (cathode), respectively, functioning as electrode compartments. The remaining compartments were used for sample compartments. Autofocusing was carried out at constant voltage according to two different methods. In method 1, all sample compartments were filled with a 1% water solution of casein or milk whey protein hydrolysates. In method 2, two compartments located in the center of the tank were filled with 5% sample solution and the others were filled with deionized water. Compositional and sequence analyses of the autofocusing fractions revealed that peptides in the two hydrolysates can be fractionated within 24 h by the present apparatus. Better fractionation was obtained by method 2, whereas enrichment of some peptides occurred by using method 1.
Five patients with melanoacanthoma are reported. The skin lesions consisted of a single, round plaque measuring several centimeters in diameter. Histologic findings were characterized by formation within an acanthotic epidermis, of many islands of small basaloid cells and highly dendritic melanocytes. Electron microscopic examination of one lesion revealed a disturbance of melanin transfer from the highly dendritic melanocytes to the neighboring keratinocytes.
The long-term therapeutic efficacy of isosorbide, an oral hyperosmotic diuretic, was clinically evaluated in 30 patients (14 males and 16 females) who were followed up in our clinic for more than 2 years. The patients ranged in age from 14 to 73 years, with a mean age of 49.2 years. Isosorbide was given orally, continuously or intermittently, for 27-297 weeks, depending on each patient’s condition. The efficacy of the drug was determined according to the evaluation criteria of the American Academy of Ophthalmology and Otolaryngology (AAOO) and the American Academy of Otolaryngology, Head and Neck Surgery (AAO-HNS) for Ménière’s disease. Definitive episodes were successfully controlled in 15 of the 20 patients (75%) who received isosorbide continuously according to both the AAOO and the AAO-HNS criteria. Of the 10 patients who were given the drug intermittently, definitive episodes were successfully controlled in 6 patients (60%) according to the AAOO and in 7 patients (70%) according to the AAO-HNS criteria. During the treatment periods, hearing improved in 6 patients (20%), and tinnitus subjectively lessened in 15 patients (50%). Although some patients complained of abdominal discomfort, the side effects of this drug were minimal. Among the patients who responded well to the treatment, the doses of isosorbide could be tapered to a daily dose of 30 ml, less than one half of the usual daily dose of this drug. The results suggest that most patients should be asked to continue this medical treatment for at least several months before a decision can be made regarding whether the medical response is good or poor. We believe that the long-term use of isosorbide is often efficacious in stabilizing the disabling symptoms of Ménière’s disease.
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