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Inhibitory effect of heparin upon histamine release by trypsin, antigen, and proteose
8ELlXTED
AIX3TR.~CTS
189
tuberculin, the lungs show ext,ensive pneumonic lesions containing large amounts of tubercle bacilli, and the livcts and spleen are rrlativcl? normal. In the present studies, guinea pigs were injected subcutaneously with virulent human tuberculin bacilli and were skin tested at frequent intervals to determine changes in skin sensitivity. The tissues of all animals which survived six weeks or longer were studied after death. It was found that the sensitivity rose gradually during the second week after infection, increased rapidly during the third week, and then leveled off. Those animals which had spontaneously lost theil skin sensitivity to tuberculin before death showed extensive pulmonary lesions containing a large number of acid-fast bacilli but no caseation or necrosis of the alveolar walls. The spleens in the animals which had lost most of their allergy before death showed diffuse epithelioid cell Animals retaining a high degree infiltration and area.s of infarction. of skin sensitivity to tuberculin ljefore death showed few or no acidfast bacilli in t,heir pulmonary lesions. Some caseation was observetl in the lungs of these animals, but, these areas rarely contained tubercle bacilli. It was concluded that in infected guinea pigs there is a distinct relationship between t,he degree of skin sensit,ivity to tuberculin before death and the pathologic findings in the lungs. Animals which are anergic before death show severe pulmonary lesions containing large numbers of acid-fast bacilli, while those which are allergic show few or no acid-fast bacilli in their pulmonary lesions.
Inhibitory Antigen,
Effect of Heparin Upon Histamine Release by Trypsin, and Proteose. Dragstedt, C. A., and Rocha e Xilva, M. :
Proc. Sot. Exper.
Eiol. & Med. 51: 101. 194’2.
The symptoms observed it1 trypsin and I)rl)tone shock and in anaphylaxis are similar and are accompanied by a release of histamine into the plasma.. The addition of antigen, peptone, or trypsin to heparinized rabbit’s blood in vitro causes a release of histamiue from the cells to the plasma. Heparin is known to be a tryl)sin antagonist. In an effort to determine whether the mechanism of the histamine release is the same with each of the three agents, heparin in excess of the amounts required to render the blood uncoagulable was tmployecl as an inhibiting agent. Bloods obtained from normal and sensitized rabbits were mixed with sufficient heparin to prevent clotting. To normal blood samples trypsin or proteose was added. To the blood from the sensitized animals, antigen (egg white) was added. In addition. heparin was added to each mixture. Adequate controls were employed. The mixtures were centrifuged, and t,he supernatant plasmas were assayed for histamine
190
THE .JOl!RT.\T, OF AI,I,EK(;\’
‘l’he results itldicatetl tllat 111~’adclitiotr 01’ heparitl itr iIlll()lltttS in excess of that, ~*cqniretl to 1~rtlr7~ the I~loo~~ill~(~il$tIlal)le inhihitetl the release of histalninc I+IY)IIIthtl carlIs itllo the I)lilst~l;l ill all sanll~le~. regardless of the CRIIW of histalrlitlcl l*elease. I I~IIc~. ii \VRS ~*~~~clutl~l that the mechanism of histanrillca please was prol~ahly the same in trypsin and pel)tone shock antI in atlaphylasis. It is suggested that heparin, when used in sufficient quantity. map inhibit anaghylactic shock in intact allimals. cYmterlt.
The Manufacture of Antibodies in Vitro. 1). H. : ;r. &per. Med. 76: 211. 1 CM.
l’auling.
I,.. and C’an~phell.
Specific antibody was rnanufacturetl in vitro from norlnal serunl globulin. Methyl 1)luc was enlployetl as the antigen and was ltlisetl with the gamma globulin fraction of fresh beef serum. Sodium hydroxide was then added as a denaturing agent. ;ifter standing for from fifteen to thirty nlinutes, thr> mixture was dialyzed at b” (‘. against a stream of phosphate buffer of pH 7.5. ‘l’hus the alkali-drnatured protein molecules slowI> assumed config77rations, stable it7 neutral solution while in contacdt, with antigen. The pH of the proteilrantigen solution ehanyetl front 11 .O to 7.5, ant1 small amounts of protein-dye precipitate were obtwinrtl which were f01717d to I)e the resnli Tt was impossihlc, howc\-er, to separal e of antigen-antil)ody reaction. the dye from the tlye-protein 1)laeeil)itate to obtain a specific pt*ecil)itill fo1* the dye. Antil)odirs hon~o~opous to the ;ixopheliylat,sonic groups and to pneunlo~oceus polysacch;ll.itle type ITI were also nlanufnctut~ecl in vitro from Tarions serum l)rotcins hy tlenatu~atioii-~*eiiati~t,atioll processes. Wheel ~~r~eu~~~ococcus l)olysacchaI.ide Iyl)c 1 I I was cnll)loye~l as the antigen, the resulting antisera I)rccipit:itcti type II I polysac.charitle and agglutillittetl tyl)e II I ~)neun7ococci. ‘I’ypes 1 ntld \‘1 I I I)olysacchariclr were not ~)recil)ilntetl, and pneuniococci types r ;iml I I were not agglnt inated by tile ll~illlll~~l~~tl77~~tl atltisrrult~. It LVilS (‘OIIc*ludetl that normal scrutI1 glolmliti Van he convet~trfl i77to anlil)otly it1 vitro when it is denatured ant1 Iaenaturcd ill the presence of antigel]. l)ei7aturntion can he ac~com~~lishetl l)y heat or c~helriic*alaction.
Pediatrics A Series of Substitutes for Milk in the Treatment of Allergies. I,. Z., and Silverstone, P. C.: .J. Pediat. 21: 635, 1942.
\Volpe.
?;ine milk substitute formulas are offered and an analysis of their composition given. ‘l’hese fornlulas are constructed from oat, barley. soy, lima bean, pea, taro, rice, rye. or corn flours with the addition of cottonseed, olive, sesame, corn, peanut, or soy oil, and gelatin, dextrose.
AIX3TR.~CTS
189
tuberculin, the lungs show ext,ensive pneumonic lesions containing large amounts of tubercle bacilli, and the livcts and spleen are rrlativcl? normal. In the present studies, guinea pigs were injected subcutaneously with virulent human tuberculin bacilli and were skin tested at frequent intervals to determine changes in skin sensitivity. The tissues of all animals which survived six weeks or longer were studied after death. It was found that the sensitivity rose gradually during the second week after infection, increased rapidly during the third week, and then leveled off. Those animals which had spontaneously lost theil skin sensitivity to tuberculin before death showed extensive pulmonary lesions containing a large number of acid-fast bacilli but no caseation or necrosis of the alveolar walls. The spleens in the animals which had lost most of their allergy before death showed diffuse epithelioid cell Animals retaining a high degree infiltration and area.s of infarction. of skin sensitivity to tuberculin ljefore death showed few or no acidfast bacilli in t,heir pulmonary lesions. Some caseation was observetl in the lungs of these animals, but, these areas rarely contained tubercle bacilli. It was concluded that in infected guinea pigs there is a distinct relationship between t,he degree of skin sensit,ivity to tuberculin before death and the pathologic findings in the lungs. Animals which are anergic before death show severe pulmonary lesions containing large numbers of acid-fast bacilli, while those which are allergic show few or no acid-fast bacilli in their pulmonary lesions.
Inhibitory Antigen,
Effect of Heparin Upon Histamine Release by Trypsin, and Proteose. Dragstedt, C. A., and Rocha e Xilva, M. :
Proc. Sot. Exper.
Eiol. & Med. 51: 101. 194’2.
The symptoms observed it1 trypsin and I)rl)tone shock and in anaphylaxis are similar and are accompanied by a release of histamine into the plasma.. The addition of antigen, peptone, or trypsin to heparinized rabbit’s blood in vitro causes a release of histamiue from the cells to the plasma. Heparin is known to be a tryl)sin antagonist. In an effort to determine whether the mechanism of the histamine release is the same with each of the three agents, heparin in excess of the amounts required to render the blood uncoagulable was tmployecl as an inhibiting agent. Bloods obtained from normal and sensitized rabbits were mixed with sufficient heparin to prevent clotting. To normal blood samples trypsin or proteose was added. To the blood from the sensitized animals, antigen (egg white) was added. In addition. heparin was added to each mixture. Adequate controls were employed. The mixtures were centrifuged, and t,he supernatant plasmas were assayed for histamine
190
THE .JOl!RT.\T, OF AI,I,EK(;\’
‘l’he results itldicatetl tllat 111~’adclitiotr 01’ heparitl itr iIlll()lltttS in excess of that, ~*cqniretl to 1~rtlr7~ the I~loo~~ill~(~il$tIlal)le inhihitetl the release of histalninc I+IY)IIIthtl carlIs itllo the I)lilst~l;l ill all sanll~le~. regardless of the CRIIW of histalrlitlcl l*elease. I I~IIc~. ii \VRS ~*~~~clutl~l that the mechanism of histanrillca please was prol~ahly the same in trypsin and pel)tone shock antI in atlaphylasis. It is suggested that heparin, when used in sufficient quantity. map inhibit anaghylactic shock in intact allimals. cYmterlt.
The Manufacture of Antibodies in Vitro. 1). H. : ;r. &per. Med. 76: 211. 1 CM.
l’auling.
I,.. and C’an~phell.
Specific antibody was rnanufacturetl in vitro from norlnal serunl globulin. Methyl 1)luc was enlployetl as the antigen and was ltlisetl with the gamma globulin fraction of fresh beef serum. Sodium hydroxide was then added as a denaturing agent. ;ifter standing for from fifteen to thirty nlinutes, thr> mixture was dialyzed at b” (‘. against a stream of phosphate buffer of pH 7.5. ‘l’hus the alkali-drnatured protein molecules slowI> assumed config77rations, stable it7 neutral solution while in contacdt, with antigen. The pH of the proteilrantigen solution ehanyetl front 11 .O to 7.5, ant1 small amounts of protein-dye precipitate were obtwinrtl which were f01717d to I)e the resnli Tt was impossihlc, howc\-er, to separal e of antigen-antil)ody reaction. the dye from the tlye-protein 1)laeeil)itate to obtain a specific pt*ecil)itill fo1* the dye. Antil)odirs hon~o~opous to the ;ixopheliylat,sonic groups and to pneunlo~oceus polysacch;ll.itle type ITI were also nlanufnctut~ecl in vitro from Tarions serum l)rotcins hy tlenatu~atioii-~*eiiati~t,atioll processes. Wheel ~~r~eu~~~ococcus l)olysacchaI.ide Iyl)c 1 I I was cnll)loye~l as the antigen, the resulting antisera I)rccipit:itcti type II I polysac.charitle and agglutillittetl tyl)e II I ~)neun7ococci. ‘I’ypes 1 ntld \‘1 I I I)olysacchariclr were not ~)recil)ilntetl, and pneuniococci types r ;iml I I were not agglnt inated by tile ll~illlll~~l~~tl77~~tl atltisrrult~. It LVilS (‘OIIc*ludetl that normal scrutI1 glolmliti Van he convet~trfl i77to anlil)otly it1 vitro when it is denatured ant1 Iaenaturcd ill the presence of antigel]. l)ei7aturntion can he ac~com~~lishetl l)y heat or c~helriic*alaction.
Pediatrics A Series of Substitutes for Milk in the Treatment of Allergies. I,. Z., and Silverstone, P. C.: .J. Pediat. 21: 635, 1942.
\Volpe.
?;ine milk substitute formulas are offered and an analysis of their composition given. ‘l’hese fornlulas are constructed from oat, barley. soy, lima bean, pea, taro, rice, rye. or corn flours with the addition of cottonseed, olive, sesame, corn, peanut, or soy oil, and gelatin, dextrose.