sepharose and Psoriasis

Topics: Animals; Climatic Processes; Denial, Psychological; Female; Fraud; Humans; Keratins; Male; Motivation; Peptide Hydrolases; Psoriasis; Sepharose; Skin; Space Flight; United States National Aeronautics and Space Administration

Topics: Animals; Climatic Processes; Denial, Psychological; Female; Fraud; Humans; Keratins; Male; Motivation; Peptide Hydrolases; Psoriasis; Sepharose; Skin; Space Flight; United States National Aeronautics and Space Administration

Keratin-agarose plate and keratin-polyacrylamide enzymography methods were developed to demonstrate proteolytic digestion of epidermal keratin. By applying these methods, keratin hydrolase was purified from Tris-buffered saline extract of psoriatic scales by 50% ammonium sulfate precipitation, passage through a lysine-Sepharose column, DEAE-Sepharose, Sephacryl S-200, high-performance cation-exchange chromatography on Mono S, and aprotinin-Sepharose affinity chromatography. The final preparation demonstrated a single protein band at molecular weight 30,000 judged by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Furthermore, in keratin-polyacrylamide slab gels, the purified enzyme preparation showed a translucent band at molecular weight 30,000, indicating keratin digestion. Keratin hydrolase digested reassembled epidermal keratin as well, whereas it had no effect on guinea pig hair keratin. The enzyme demonstrated a high level of hydrolytic activity on Ile-Pro-Arg-p-nitroanilide and other peptidyl arginine substrates, while it showed a low level of activity on Val-Leu-Lys-p-nitroanilide, and no activity on Arg-Pro-Tyr-p-nitroanilide, Glu-Pro-Val-p-nitroanilide, or Ala-Ala-Ala-p-nitroanilide. The keratin hydrolase was a serine proteinase, inactivated by diisopropylfluorophosphate, phenylmethylsulfonyl fluoride, tosyl-lysyl-chloromethyl ketone, antipain, leupeptin, soybean trypsin inhibitor, aprotinin, and p-aminobenzamidine. The keratinolytic activity was not detected in normal epidermal extract.

Topics: Adult; Animals; Electrophoresis, Polyacrylamide Gel; Female; Guinea Pigs; Humans; Keratins; Male; Middle Aged; Peptide Hydrolases; Protease Inhibitors; Psoriasis; Sepharose; Skin

The random and active migration of neutrophil leukocytes in agarose was studied in sixty-one patients with psoriasis and in healthy controls. E. coli filtrate was used as chemoattractant. The random as well as the active migration was increased in psoriasis. The increase was most pronounced in those cases with widespread psoriasis. In twenty patients who were followed for 1-3 years, the neutrophil chemotactic activity remained essentially unchanged.

Topics: Adult; Aged; Chemotaxis, Leukocyte; Female; Humans; Male; Middle Aged; Neutrophils; Psoriasis; Sepharose