Effect of Insulin, Epinephrine, Hydrocortisone and ACTH on the Catheptic Activity of Rat Spleen

In our experiments under in vivo conditions we followed ·the response of spleen on injected insulin, epinephrine, hydrocortisone and ACTH. Insulin caused a decrease in spleen weight, a decrease of protein nitrogen and an increase of catheptic activity 30 min. after application. Afterwards catheptic activity was significantly above control value in 60 min. (expressed in E. U./g. tissue) although· decreasing tendency was observed. 30 and 60 min. after epinephrine application spleen weight and protein nitrogen were increased whereas catheptic activity remained under the control value (expressed in E. U./mg.N). After 3 hrs the spleen weight was reduced (reduced protein nitrogen) whereas catheptic activity was significantly increased, specially if expressed in E. U./g. tissue. Hydrocortisone caused a decrease in spleen weight and protein nitrogen. Catheptic activity was significantly above control value 30 min. and 3 hrs after application, if expressed in enzyme unit per gram tissue. Two hours after ACTH administration protein nitrogen was increased, whereas catheptic activity was under the control value (expressed in E. U.jmg. N)

Effect of Insulin, Epinephrine, Hydrocortisone and ACTH on the Catheptic Activity of Rat Spleen

In our experiments under in vivo conditions we followed ·the response of spleen on injected insulin, epinephrine, hydrocortisone and ACTH. Insulin caused a decrease in spleen weight, a decrease of protein nitrogen and an increase of catheptic activity 30 min. after application. Afterwards catheptic activity was significantly above control value in 60 min. (expressed in E. U./g. tissue) although· decreasing tendency was observed. 30 and 60 min. after epinephrine application spleen weight and protein nitrogen were increased whereas catheptic activity remained under the control value (expressed in E. U./mg.N). After 3 hrs the spleen weight was reduced (reduced protein nitrogen) whereas catheptic activity was significantly increased, specially if expressed in E. U./g. tissue. Hydrocortisone caused a decrease in spleen weight and protein nitrogen. Catheptic activity was significantly above control value 30 min. and 3 hrs after application, if expressed in enzyme unit per gram tissue. Two hours after ACTH administration protein nitrogen was increased, whereas catheptic activity was under the control value (expressed in E. U.jmg. N)

Anticathepsin D Antibody-Sepharose Chromatography of Human Cathepsin D

Cathepsin D was isoilated from human tissues by anticatheipsin D antibo:dy-Serpharose 4B .chromatoigraphy. Caithepsin D, rele1ased from the immunoaffinity column formed one precipitin line with specif.ic antibody in immwnod:Lffusion and in immunoe1ectropholl1esis. The isolated proteinase is shown to be prure cathepsin D by actiivity and by inhibition with peipstatin. The quanititative determinati.on o:f cathepsin D in human tissue, taiken by bio.psy from sco,Uotic patients 1rl ivery small amount, is desert.bed. It was demonstrated that cathepsin D from human muscle and human gingival fluid was indistinguishable in immunodiffus ion from the human liver catheps iirl D

THE IMPORTANCE OF THE STANDARD SAMPLE FOR ACCURATE ESTIMATION OF THE CONCENTRATION OF NET ENERGY FOR LACTATION IN FEEDS ON THE BASIS OF GAS PRODUCED DURING THE INCUBATION OF SAMPLES WITH RUMEN LIQUOR

Namen tega dela je bil preveriti, ali je uporaba standardnega vzorca pri izvajanju Hohenheimskega plinskega preizkusa sploh potrebna. V obdobju treh let smo inkubirali z vampnim sokom in vitro 24 serij vzorcev voluminozne krme. Poleg vzorcev krme smo v poskus vključili tudi standardni vzorec sena, ki so nam ga poslali z univerze v Hohenheimu (HFT-99). Polovico serij smo inkubirali z vampnim sokom krav, polovico z vampnih sokom ovc. Merili smo količino plina, ki se razvije med 24 urno inkubacijo in jo primerjali z deklarirano vrednostjo vzorca HFT-99. Poleg HFT-99 smo v poskus vključili tudi 25 preizkusnih vzorcev krme z znanimi in vivo določenimi prebavljivostnimi koeficienti. Na podlagi tvorbe plina pri HFT-99 smo ugotovili, da vrsta živali (govedo ali ovce) ni vplivala na aktivnost vampnega soka (41,4 vs. 42,2 ml plina na 200 mg sušine, p>0,1). Tudi razlike med leti niso bile značilne (41,9, 41,2 in 42,3 ml plina na 200 mg sušine, p>0,1). Variabilnost med serijami je bila približno 10%, od 38,9 do 43,7 ml plina na 200 mg sušine). Tvorba plina pri HFT-99 je bila v tem poskusu približno 6 % manjša od vrednosti, ki so jo dobili na Univerzi Hohenheim (41,8 vs. 44,43 ml na 200 mg sušine). To nakazuje, da obstajajo sistematična odstopanja med laboratoriji. V primeru petindvajsetih preizkusnih vzorcev je korekcija na podlagi standardnega vzorca zmanjšala povprečno razliko in vitro ocenjenih vsebnosti neto energije za laktacijo (NEL) od in vivo določenih vrednosti. Sklepamo, da je glede na variabilnost med serijami in glede na sistematične razlike v aktivnosti vampnega soka med laboratorijema, uporaba standardnega vzorca pri izvajanju Hohenheimskega plinskega preizkusa nujna.The aim of this work was to examine the necessity of using the standard sample at the Hohenheim gas test. During a three year period, 24 runs of forage samples were incubated with rumen liquor in vitro. Beside the forage samples also the standard hay sample provided by the Hohenheim University (HFT-99) was included in the experiment. Half of the runs were incubated with rumen liquor of cattle and half with the rumen liquor of sheep. Gas produced during the 24 h incubation of standard sample was measured and compared to a declared value of sample HFT-99. Beside HFT-99, 25 test samples with known digestibility coefficients determined in vivo were included in the experiment. Based on the gas production of HFT-99, it was found that donor animal (cattle or sheep) did not significantly affect the activity of rumen liquor (41.4 vs. 42.2 ml of gas per 200 mg dry matter, P>0.1). Neither differences between years (41.9, 41.2 and 42.3 ml of gas per 200 mg dry matter, P>0.1) were significant. However, a variability of about 10% (from 38.9 to 43.7 ml of gas per 200 mg dry matter) was observed between runs. In the present experiment, the gas production in HFT-99 was about 6% lower than the value obtained by the Hohenheim University (41.8 vs. 44.43 ml per 200 mg dry matter). This indicates a systematic error between the laboratories. In the case of twenty-five test samples, correction on the basis of the standard sample reduced the average difference of the in vitro estimates of net energy for lactation (NEL) from the in vivo determined values. It was concluded that, due to variation between runs and systematical differences in rumen liquor activity between two laboratories, the results of Hohenheim gas test have to be corrected on the basis of standard sample

THE EFFECT OF NITROGEN FERTILIZATION OF MAIZE ON PROTEIN CONCENTRATION AND IN VITRO FEMENTABILITY OF GRAIN

Z in vitro metodo, ki temelji na merjenju plina, ki se razvije pri inkubaciji vzorcev z vampnim sokom, smo preučevali vpliv gnojenja koruze z dušikom na fermentabilnost koruznega zrnja v vampu. Nastajanje plina smo merili kontinuirano do 72 ur, kumulativno nastajanje plina pa opisali z Gompertzovo enačbo Y=A*exp(- exp(-d*(t-tm))). Primerjali smo sedem postopkov, od katerih je bil eden negnojen, ostali pa gnojeni s 100 do 250 kg N ha–1. Pridelek zrnja in vsebnost surovih beljakovin (SB) v njem, sta se z gnojenjem povečevala linearno. Na kg dodanega N se je pridelek zrnja povečal za 25 kg sušine (SS) ha–1, vsebnost SB pa za 0,13 g kg–1 SS. Vsebnost SB v zrnju, ki se je gibala od 83 do 115 g kg–1 SS, je bila v tesni povezavi z dinamiko nastajanja plina. Največja stopnja nastajanja plina (MPR) je bila v negativni povezavi z vsebnostjo SB v zrnju (R2 = 0,53; p < 0,06), čas MPR (tm) pa v pozitivni povezavi s skupno količino dodanega N z gnojenjem (R2 = 0,74; p < 0,05) in z vsebnostjo SB v zrnju (R2 = 0,88; p < 0,01). Izgleda, da intenzivno gnojenje koruze z N omejuje vampno prebavljivost koruznega škroba. Zaradi premestitve prebave škroba iz vampa v nižji del prebavnega trakta lahko pričakujemo pri koruznem zrnju intenzivno gnojene koruze boljše izkoriščanje energije kot pri zrnju koruze, ki je pomanjkljivo oskrbljena z N.The effect of nitrogen fertilization of maize on fermentability of maize grain in the rumen was studied by means of in vitro method based on the measurement of gas produced during the incubation of samples with rumen liquor. Gas production was recorded continuously up to 72 h incubation time and cumulative gas production was described by the Gompertz equation Y=A*exp(-exp(-d*(t-tm))). Seven treatments, one of them unfertilized and others fertilized with 100 to 250 kg N ha–1, were compared. Grain yield and concentration of crude protein (CP) in grain increased linearly with nitrogen fertilization. Grain yield increased for 25 kg dry matter (DM) ha–1 and CP concentration for 0.13 g kg–1 DM per each additional kg of N. Concentration of CP in grain, which varied from 83 to 115 g kg–1 DM, was closely related to the dynamics of gas production. The maximal gas production rate (MPR) was negatively related to CP concentration in the grain (R2 = 0.53; p < 0.10) and the time of MPR (tm) was positively related to the amount of added N (R2 = 0.74; p < 0.05) and concentration of CP in the grain (R2 = 0.88; p < 0.01). It is likely that intensive N fertilization of maize limits ruminal digestion of maize starch. Due to the shift of starch digestion from the rumen to lower gastrointestinal tract better utilization of energy can be expected in maize grain of extensively fertilized maize than in the grain of maize, in which supply of N is sub-optimal

THE IMPORTANCE OF THE STANDARD SAMPLE FOR ACCURATE ESTIMATION OF THE CONCENTRATION OF NET ENERGY FOR LACTATION IN FEEDS ON THE BASIS OF GAS PRODUCED DURING THE INCUBATION OF SAMPLES WITH RUMEN LIQUOR

Namen tega dela je bil preveriti, ali je uporaba standardnega vzorca pri izvajanju Hohenheimskega plinskega preizkusa sploh potrebna. V obdobju treh let smo inkubirali z vampnim sokom in vitro 24 serij vzorcev voluminozne krme. Poleg vzorcev krme smo v poskus vključili tudi standardni vzorec sena, ki so nam ga poslali z univerze v Hohenheimu (HFT-99). Polovico serij smo inkubirali z vampnim sokom krav, polovico z vampnih sokom ovc. Merili smo količino plina, ki se razvije med 24 urno inkubacijo in jo primerjali z deklarirano vrednostjo vzorca HFT-99. Poleg HFT-99 smo v poskus vključili tudi 25 preizkusnih vzorcev krme z znanimi in vivo določenimi prebavljivostnimi koeficienti. Na podlagi tvorbe plina pri HFT-99 smo ugotovili, da vrsta živali (govedo ali ovce) ni vplivala na aktivnost vampnega soka (41,4 vs. 42,2 ml plina na 200 mg sušine, p>0,1). Tudi razlike med leti niso bile značilne (41,9, 41,2 in 42,3 ml plina na 200 mg sušine, p>0,1). Variabilnost med serijami je bila približno 10%, od 38,9 do 43,7 ml plina na 200 mg sušine). Tvorba plina pri HFT-99 je bila v tem poskusu približno 6 % manjša od vrednosti, ki so jo dobili na Univerzi Hohenheim (41,8 vs. 44,43 ml na 200 mg sušine). To nakazuje, da obstajajo sistematična odstopanja med laboratoriji. V primeru petindvajsetih preizkusnih vzorcev je korekcija na podlagi standardnega vzorca zmanjšala povprečno razliko in vitro ocenjenih vsebnosti neto energije za laktacijo (NEL) od in vivo določenih vrednosti. Sklepamo, da je glede na variabilnost med serijami in glede na sistematične razlike v aktivnosti vampnega soka med laboratorijema, uporaba standardnega vzorca pri izvajanju Hohenheimskega plinskega preizkusa nujna.The aim of this work was to examine the necessity of using the standard sample at the Hohenheim gas test. During a three year period, 24 runs of forage samples were incubated with rumen liquor in vitro. Beside the forage samples also the standard hay sample provided by the Hohenheim University (HFT-99) was included in the experiment. Half of the runs were incubated with rumen liquor of cattle and half with the rumen liquor of sheep. Gas produced during the 24 h incubation of standard sample was measured and compared to a declared value of sample HFT-99. Beside HFT-99, 25 test samples with known digestibility coefficients determined in vivo were included in the experiment. Based on the gas production of HFT-99, it was found that donor animal (cattle or sheep) did not significantly affect the activity of rumen liquor (41.4 vs. 42.2 ml of gas per 200 mg dry matter, P>0.1). Neither differences between years (41.9, 41.2 and 42.3 ml of gas per 200 mg dry matter, P>0.1) were significant. However, a variability of about 10% (from 38.9 to 43.7 ml of gas per 200 mg dry matter) was observed between runs. In the present experiment, the gas production in HFT-99 was about 6% lower than the value obtained by the Hohenheim University (41.8 vs. 44.43 ml per 200 mg dry matter). This indicates a systematic error between the laboratories. In the case of twenty-five test samples, correction on the basis of the standard sample reduced the average difference of the in vitro estimates of net energy for lactation (NEL) from the in vivo determined values. It was concluded that, due to variation between runs and systematical differences in rumen liquor activity between two laboratories, the results of Hohenheim gas test have to be corrected on the basis of standard sample