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07-14/P11 THE EFFECTS OF THE SERA OF INJURED PATIENTS ON THE GROWTH OF THE HUMAN PERIPHERAL BLOOD MONONUCLEAR CELLS: POSSIBLE INVOLVEMENT OF THE HEPARIN BINDING HUMORAL FACTORS IN THE PHENOMENON OF ENHANCED OSTEOGENESIS IN THE PATIENTS WITH TRAUMATIC BRAIN INJURY R. midburger 2 , S. Borović 1 , N. Žarković 1 , A. Meinitzer 2 , W. Petek 2 , M. Jurin Ruđer Bošković Institute, Zagreb; University Clinic of Traumatologa Graz, Austria Among the most provocative phenomena is the phenomenon of enhanced osteogenesis in patients with traumatic brain injury (TBI). In these patients very early consolidation of bone fractures is followed by hypertrophic callus formation or with heterotopic ossifications. Although the mechanism of this phenomenon is not clarified, it seems to be based on certain, not yet identified, humoral factors influencing tissue regeneration. Thus, the aim of the study was to get some information about the possible influence of such humoral factors on the immune system and, hence, indication on their nature and the involvement of the immune system in the phenomenon itself. We have analysed the growth of the normal human peripheral blood mononuclear cells (PBM) incubated in the presence of sera of differently injured patients and the same sera after heparin-affinity chromatography. Four experimental groups comprised subjects with the following: 1) fractures (FRA) of long bones or large joints (n = 9), 2) traumatic brain injury (TBI) only (n = 11), 3) combined traumatic brain injury and the bone fractures (TBI + FRA) (n = 10), and also 4) healthy volunteers. Effects of the sera prepared as pooled samples obtained weekly during the first four posttraumatic weeks on the growth of the PBM in the presence of 2,5 g/ml PHA-L or in the absence of the mitogen, determined by the intensity of the H-thymidine incorporation, were compared with the effects of the same concentration (1%) of the normal human pooled (n = 10) serum. In ali samples of injured patients sera, decreased PHA reactivity was observed with the tendency of recovery in the fourth week after injury except in TBI + FRA group of patients where there was no recovery during the whole period. Removal of heparin binding humoral factors from the sera caused changes in PBM reactivity on mitogen. In ali BIOCHEMIA MEDICA god. 6, br. 1, 1996 179
Sažeci 2. Hrvatskog kongresa medicinske biokemije POSTERSKI RADOVI groups there was stimulation in comparison with original sera samples of the same group, but there were differences between different group of patients. Removal of heparin binding humoral factors can cause stimulaton in PBM reactivitv on mitogen in FRA, or inconsistent behaviour in TBI, but there still is suppression in PBM reactivitv on mitogen in TBI + FRA group of patients. Spontaneous 3 H-TdR incorporation can switch from normal values (FRA, TBI) to normal or slightlv suppressed (FRA), suppression (TBI) or may remain suppressed (TBI + FRA). It was supposed that some humoral factors can be related with phenomenon of enhanced osteogenesis in patients with TBI + FRA. It seems that such humoral factors remain in the sera after removal of heparin binding compounds and that some humoral factors acting as suppressors are removed from the sera. Therefore, further investigation of heparin binding humoral factors seems very useful. 07-14/P12 PROGRAMIRANA SMRT (3-STANICA LANGERHANSOVIH OTOČIĆA U ŠEĆERNOJ BOLESTI OVISNOJ O INZULINU L. Rumora, M. Hadžija, E. Pape-Medvidović, I. Pavlić-Renar, Z. Metelko, D. Juretić, M. Slijepčeuić Institut "Ruđer Bošković", Farmaceutsko-biokemijski fakultet; Zagreb Značajka je šećerne bolesti neovisne o inzulinu (NIDDM), u velikom broju slučajeva, povišena koncentracija glukoze u krvi i povišena koncentracija inzulina. U tih bolesnika je također povišena koncentracija amilina u serumu. Amilin je polipeptid od 37 aminokiselina i sastavni je dio (3-stanica Langerhansovih otočića (Lo). Svojim djelovanjem uzrokuje inzulinsku rezistenciju i smanjenje glikogena u mišićima, a u izravnom kontaktu amilinska vlakna uzrokuju fragmentaciju DNA i programiranu smrt (3-stanica Lo. U našim pokusima ispitali smo uzrokuje li serum bolesnika oboljelih od NIDDM, koji sadrži amilin, fragmentaciju DNA i programiranu smrt - apoptozu Lo pankreasa. U pokusima su korišteni izolirani Lo štakora. Otočići, izolirani s pomoću enzima kolagenaze, su inkubirani 48 sati na 22°C s različitim koncentracijama seruma bolesnika oboljelih od NIDDM. Vijabilnost -stanica Lo određena je pod fluorescentnim mikroskopom nakon bojenja s propidijevim jodidom. Stanična DNA je izolirana kloroform/fenolskom 180 BIOCHEMIA MEDICA god. 6, br. 1, 1996
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