Mustard trypsin

Karyotyping has advanced dramatically since it was discovered that certain stains stain individual chromosomes in characteristic ways. Caspersson (Caspersson et al., 1970a,b, 1971) showed that quinacrine mustard and quinacrine dihydrochloride (Fig. 7.7) produced a characteristic banding pattern (Q-banding), and later it was found that complementary bands could be formed with Giesma (G-bands) if the chromosomes are treated first with trypsin or mild alkali (Patil et al., 1971 Seabright, 1971 Wang and Federoff, 1972). 

The mustard family (Brassicaceae) PIPs are 7 kDa proteins with 8 cysteines in highly conserved positions that form 4 disulphide linkages in a particular pattern of connectivity. The mustard PIPs are variously potent inhibitors of serine proteases such as trypsin, chymotrypsin, thrombin, plasmin and blood clotting factors Xa and Xlla [515, 520] (Table 15). 

The mustard (Brassicaceae) 7 kDa trypsin inhibitors have molecular masses of about 7 kDa and have 8 cysteines (i.e. 4 disulphide links). For other details see the legend to Table 4. 

Brassica oleraceae (cabbage) (Brassicaceae) [seed] Thrombin inhibitor (-10 kDa possible member of mustard TI family) Trypsin (0.2 pM), Thrombin, Xa, XHa, Plasmin [519]... 

Higuchi, K., Kajiki, A., Nakamura, M., Harada, S., Pula, P.J., Scott, A.L., Dannenberg, A.M., Jr. (1988). Proteases released in organ culture by acute dermal inflammatory lesions produced in vivo in rabbit skin by sulfur mustard hydrolysis of synthetic peptide substrates for trypsin-like and chymotrypsin-like enzymes. Inflammation 12 311-34. 

Bradykinin 82-84 Carrageenan Formalin Brewer s yeast Silver nitrate Turpentine Trypsin Kaolin Mustard Compound 48/80 Potassium cyanide Monoiodoacetic acid Sodium fluoride Hyaluronidase 81, 90-92 77, 79, 80, 81, 85, 86, 93-96 55,81,91,97-101 102 86 55 103 81, 104 18, 76, 77 85 85 85 21, 77, 87... 

In all these determinations, there is a very important point on which writers do not often insist enough, and which renders their results but little comparable, namely, the strength of the trypsin used. Achalme recommends a maceration of 5 to 7 per cent of pancreatin made in a physiological solution for 18 to 24 hours at 35°, in the presence of several drops of essence of mustard. The liquid is filtered and distributed in sterilized and sealed pipettes, which are kept in the dark. This trypsin is then tested with respect to its effect on the serum of an adult guinea pig. By using the method of Achalme, just described, a trypsin should be obtained, o.oi c.c. to 0.015 of which (or o.i c.c. to 0.15 c.c. of a i-io dilution) is neutralized by about o.oi c.c. (or o.i c.c. of i-ioo dilution) of guinea-pig serum, or again by 0.015 c.c. of rabbit serum. 

LC-MS is a powerful method used to detect and quantify CWAs. The use of LC-MS for CWA and hydrolysis product detection has been reviewed [7,20,26]. LC-MS methods are often used to detect CWA hydrolysis/ degradation products instead of the active agents [27-28]. LC-MS serves as a bioanalytical method for CWA detection in living systems and its contributions have also been reviewed [7, 26, 29]. A LC-MS method using an on-line trypsin digestion is used to identify GB and sulfur mustard adducts with proteins and enzymes like human butyryl cholinesterase [30]. This technique, along with similar techniques, could be applied to confirm CWA exposure when illness is suspected from an unknown toxin. 

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