ABSTRACT
A novel chymotrypsin inhibitor, which detected in the seed of wild emmer wheat (Triticum dicoccoides), was purified by ion-exchange chromatography, affinity chromatography and Ultracentrifugation. On the basis of its specificity, this inhibitor was named WeCI (wild emmer chymotrypsin inhibitor). SDS-PAGE analysis displayed that the purified WeCI is a single chain polypeptide with a molecular weight of approximately 13kDa. The inhibition constants (Ki) for amylase and bovine pancreatic chymotrypsin were 1.12×10-9M and 2.41×10-9M, respectively. Automated sequencing and mass spectrometry analyses revealed that WeCI is a neutral monomeric protein consisting of 119 residues. In vitro, WeCI strongly suppressed bovine pancreatic chymotrypsin as well as chymotrypsin-like activities separated from the midgut of the beet armyworm Spodoptera exigua. No inhibitory activities were found against bovine pancreatic trypsin, bacterial subtilisin, or porcine pancreatic elastase. The primary structure of WeCI was markedly similar (46-95%) to those of several proteins belonging to the wheat crop chymotrypsin/α-amylase inhibitor superfamily and displayed the typical sequence motif of the α-amylase inhibitor-seed storage protein group. WeCI significantly inhibited the growth and development of Spodoptera exigua, dependent on inhibitor concentration. WeCI significantly increased the mortality rate of Spodoptera exigua and caused a significant decrease in its fertility.
Subject(s)
Chymotrypsin/antagonists & inhibitors , Enzyme Inhibitors/chemistry , Enzyme Inhibitors/isolation & purification , Plant Extracts/chemistry , Plant Extracts/isolation & purification , Spodoptera/drug effects , Triticum/chemistry , Animals , Chymotrypsin/metabolism , Electrophoresis, Polyacrylamide Gel , Enzyme Inhibitors/pharmacology , Israel , Pancreatic Elastase/antagonists & inhibitors , Pancreatic Elastase/chemistry , Plant Extracts/pharmacology , Seeds/chemistry , Spodoptera/chemistry , Spodoptera/enzymology , Spodoptera/growth & development , SwineABSTRACT
Objective To observe the effect of acupuncture at the muscle region of meridians plus tuina on facial nerve function in peripheral facial paralysis. Method Sixty patients with peripheral facial paralysis (recovery stage) were randomized into a treatment group (intervened by acupuncture at the muscle region of meridians plus tuina) and a control group (intervened by conventional acupuncture), 30 cases in each group. Result The facial nerve electromyography (EMG) showed that the action potential latency and amplitude were changed significantly after the treatment in both groups (P<0.05), indicating that the two methods were both effective in recovering facial nerve function, while acupuncture at the muscle region of meridians plus tuina produced a more significant treatment effect compared to conventional acupuncture. In the treatment group, same treatment with different intervention times produced significantly different effects on the Toronto facial grading system (TFGS) (P<0.05), indicating that the earlier the intervention, the more significant the therapeutic efficacy. Conclusion Acupuncture at the muscle region of meridians plus tuina can produce a more significant effect in recovering facial nerve function in peripheral facial paralysis compared to conventional acupuncture.