Medizin - Open Access LMU - Teil 08/22

1. The mechanism of post-ischaemic ectopic impulse generation in nerve is not known, and previous measurements of excitability changes in human motor axons have appeared to conflict. We have used automatic threshold tracking and different stimulus-response combinations to follow the effects on excitability of brief (5-10 min) periods of ischaemia, too short to induce motor fasciculations. Excitability changes have been compared at different sites in axons innervating hand, arm and foot muscles. 2. Threshold was determined as the percutaneous stimulus current required to excite a single motor unit, or to evoke a constant multiunit response, after rectifying and integrating the electromyogram (EMG). Three different waveforms of stimulus current were compared: short (less than or equal to 2 ms) pulses, long (100-200 ms) pulses to measure rheobase, and 100 ms current ramps. We also measured accommodation by recording the effects of subthreshold depolarizing currents on excitability. 3. Ischaemic and post-ischaemic excitability changes were greatest in the proximal parts of the longest motor axons, and greater if the sphygmomanometer cuff was inflated over, rather than proximal to, the stimulating site. 4. Using integrated EMG responses from abductor digiti minimi, the ulnar nerve stimulated above the elbow became rapidly much less excitable after ischaemia when tested with short pulses, but more excitable when tested with current ramps. The rheobase rose briefly, but then fell, often below resting level, always staying below the pulse and ramp thresholds. 5. The latency of the response to a rheobasic stimulus altered in parallel with the threshold to short current pulses, and increased dramatically after ischaemia. This latency increase was associated with a prolonged phase of 'negative accommodation', i.e. the continued increase in excitability to a maintained subthreshold depolarizing current. 6. Changes in excitability and accommodation similar to those occurring after ischaemia were recorded following high frequency trains of stimuli. They were attributed primarily to hyperpolarization by the electrogenic sodium pump, since comparable changes could be induced by passing a steady hyperpolarizing current through the stimulating electrode. 7. Threshold and latency recordings from single motor units during and after ischaemia resembled in most respects the multiunit responses, but single unit rheobase did not show a post-ischaemic fall below the resting level. Repetitive firing contributed to the low multiunit thresholds recorded with long current pulses during the post-ischaemic period. 8. We conclude that human motor nerves become simultaneously both more and less excitable than normal after 10 min of ischaemia, depending on the choice of stimulus and response.

Current recordings from single chloride channels were obtained from excised and cell-attached patches of rat and human axons. In rat axons the channels showed an outwardly rectifying current-voltage relationship with a slope conductance of 33 pS at negative membrane potentials and 65 pS at positive potentials (symmetrical 150 mM CsCl). They were measurably for cations (PNa/PCs/PCl=0.1/0.2/1). Channel currents were independent of cytoplasmatic calcium concentration. Inactivation was not observed and gating was weakly voltage dependent. Cl− channels in human axons showed similar gating behavior but had a lower conductance.

The contrast between resistance to ischemia and ischemic lesions in peripheral nerves of diabetic patients was explored by in vitro experiments. Isolated and desheathed rat peroneal nerves were incubated in the following solutions with different glucose availability: 1) 25 mM glucose, 2) 2.5 mM glucose, and 3) 2.5 mM glucose plus 10 mM 2-deoxy-D-glucose. Additionally, the buffering power of all of these solutions was modified. Compound nerve action potential (CNAP), extracellular pH, and extracellular potassium activity (aKe) were measured simultaneously before, during, and after a period of 30 min of anoxia. An increase in glucose availability led to a slower decline in CNAP and to a smaller rise in aKe during anoxia. This resistance to anoxia was accompanied by an enhanced extracellular acidosis. Postanoxic recovery of CNAP was always complete in 25 mM HCO3(-)-buffered solutions. In 5 mM HCO3- and in HCO3(-)-free solutions, however, nerves incubated in 25 mM glucose did not recover functionally after anoxia, whereas nerves bathed in solutions 2 or 3 showed a complete restitution of CNAP. We conclude that high glucose availability and low PO2 in the combination with decreased buffering power and/or inhibition of HCO3(-)-dependent pH regulation mechanisms may damage peripheral mammalian nerves due to a pronounced intracellular acidosis.

The renal response to atrial natriuretic factor is blunted in cirrhosis with ascites. This might be due to alterations of renal receptors for atrial natriuretic factor. Therefore density and affinity of glomerular atrial natriuretic factor binding sites of bile duct-ligated rats with ascites (n = 10) and of sham-operated controls (n = 10) were determined. Glomerular atrial natriuretic factor binding sites were identified to be of the B-(biologically active) and C-(clearance) receptor type. Discrimination and quantitative determination of B and C receptors for atrial natriuretic factor were achieved by displacement experiments with atrial natriuretic factor(99-126) or des(18-22)atrial natriuretic factor(4-23), an analogue binding to C receptors only. Density of total glomerular atrial natriuretic factor binding sites was significantly increased in bile duct-ligated rats (3,518 ± 864 vs. 1,648 ± 358 fmol/mg protein; p < 0.05). This was due to a significant increase of C-receptor density (3,460 ± 866 vs. 1,486 ± 363 fmol/mg protein; p < 0.05), whereas density of B receptors was not significantly different in bile duct-ligated rats (58 ± 11 vs. 162 ± 63 fmol/mg protein). Affinity of atrial natriuretic factor to its glomerular binding sites did not differ significantly between both groups. These data suggest that an altered glomerular atrial natriuretic factor receptor density could be involved in the renal resistance to atrial natriuretic factor in cirrhosis with ascites.

Binding sites for atrial natriuretic factor (ANF) were determined on isolated rat glomeruli as well as on glomerular membranes. To define optimal conditions, binding of ANF was investigated varying incubation time, temperature and protein concentration. Binding conditions were found to be best at 4°C for 5 hours with 15 μg of glomerular protein. Saturation and affinity cross-linking experiments confirmed the presence of two distinct receptor subtypes - the B-receptor (130 kDa) and the C-receptor (65 kDa). Quantitative differentiation of both ANF binding sites was achieved by competitive displacement with two different unlabeled ANF ligands: a) rANF(99-126) (homologous displacement), b) des(18-22)rANF(4-23)NH2(heterologous displacement). Intact glomeruli and glomerular membranes did not differ significantly in receptor density for the B-receptor (71 ± 37 vs. 94 ± 53 fmol/mg protein) or the C-receptor (976 ± 282 vs. 966 ± 167 fmol/mg protein) or in affinity constants for the B-receptor (43 ± 36 vs. 52 ± 44 pM) or the C-receptor (876 ± 377 vs. 307 ± 36 pM). Glomerular membranes compared to glomeruli showed less nonspecific binding and less intra-assay variation of measuring points done in triplicates. This method of selective displacement should allow to study the influence of various physiological and pathophysiological conditions on the binding properties of B-and C-receptors for ANF.

Atrial natriuretic factor is a hormone intimately involved in water and salt homeostasis. The heart constitutes the major but not exclusive site of synthesis of this hormone. Among other functions, the gastrointestinal tract has endocrine functions, plays an important role in volume regulation of the body, and seems to be a target organ for atrial natriuretic factor. Therefore, the presence of atrial natriuretic factor was investigated in the human gut. Immunoreactive atrial natriuretic factor was found in intraoperatively obtained samples of normal human colon. Acidic extracts of human large intestine contained about 0.4 pmol/g wet wt of atrial natriuretic factor. Analysis of atrial natriuretic factor immunoreactivity by gel-filtration and reverse-phase high-performance liquid chromatography showed that about 65% of the immunoreactivity corresponded to the atrial natriuretic factor phohormone and about 35% corresponded to the C-terminal ANF99-126. Immunohistochemistry showed atrial natriuretic factor prohormone location in enterochromaffin cells of the colon mucosa. Altogether, these findings show the presence of atrial natriuretic factor prohormone in enterochromaffin cells of the human large intestine and may suggest this organ as a site of atrial natriuretic factor synthesis in humans.

Tue, 1 Jan 1991 12:00:00 +0100 https://epub.ub.uni-muenchen.de/5921/1/5921.pdf Schöps, P.; Lindner, Jörg; Arendt, Rainer M.; Gerbes, Alexander L.; Knorr, H. J.; Schnizer, W.

The authors tried to differentiate malignancy-related from nonmalignant ascites with a sequence of sensitive followed by specific ascitic-fluid parameters. There were four results of this study. First, of nine parameters investigated in a first series of 48 patients, 28 with nonmalignant and 20 with malignancy-related ascites, ascitic-fluid cholesterol and fibronectin yielded the best negative predictive value of 92% each. Carcinoembryonic antigen (CEA) and cytologic examination both showed a positive predictive value of 100%. Second, combining cytologic examination (sensitivity, 70%) and CEA determination (sensitivity, 45%) increased the sensitivity to 80%. Third, cytologic findings were negative in all ascitic-fluid samples with a cholesterol concentration below the cutoff value of 45 mg/100 ml. Fourth, based on the results of the first series of 48 patients, the diagnostic sequence with cholesterol as a sensitive parameter, followed by the combination of cytologic examination and CEA determination as specific parameters, was tested in a second series of 71 patients, 37 with nonmalignant and 34 with malignancy-related ascites. Again cytologic examination was negative in all samples with cholesterol levels below 45 mg/100 ml. In the total of 119 patients, this diagnostic sequence did not identify 9% of patients with malignancy-related ascites, and 82% of samples classified as malignancy related by cholesterol levels above 45 mg/100 ml were confirmed by positive cytologic examination and/or CEA level above 2.5 ng/ml. Thus, a diagnostic sequence with ascitic-fluid cholesterol determination, followed by cytologic examination and CEA determination, in samples with cholesterol levels above 45 mg/100 ml should permit a cost-efficient routine differentiation of malignancy-related from nonmalignant ascites.

In 11 patients with decompensated cirrhosis and deteriorating renal function, the effect of the vasoconstrictor substance 8-ornithin vasopressin (ornipressin; POR 8; Sandoz, Basel, Switzerland) on renal function, hemodynamic parameters, and humoral mediators was studied. Ornipressin was infused at a dose of 6 IU/h over a period of 4 hours. During ornipressin infusion an improvement of renal function was achieved as indicated by significant increases in inulin clearance (+65%), paraaminohippuric acid clearance (+49%), urine volume (+45%), sodium excretion (+259%), and fractional elimination of sodium (+130%). The hyperdynamic circulation was reversed to a nearly normal circulatory state. The increase in systemic vascular resistance (+60%) coincided with a decrease of a previously elevated renal vascular resistance (-27%) and increase in renal blood flow (+44%). The renal fraction of the cardiac output increased from 2.3% to 4.7% (P less than 0.05). A decline of the elevated plasma levels of noradrenaline (2.08-1.13 ng/mL; P less than 0.01) and renin activity (27.6-14.2 ng.mL-1.h-1; P less than 0.01) was achieved. The plasma concentration of the atrial natriuretic factor increased in most of the patients, but slightly decreased in 3 patients. The decrease of renal vascular resistance and the increase of renal blood flow and of the renal fraction of cardiac output play a key role in the beneficial effect of ornipressin on renal failure. These changes develop by an increase in mean arterial pressure, the reduction of the sympathetic activity, and probably of an extenuation of the splanchnic vasodilation. A significant contribution of atrial natriuretic factor is less likely. The present findings implicate that treatment with ornipressin represents an alternative approach to the management of functional renal failure in advanced liver cirrhosis.

Tue, 1 Jan 1991 12:00:00 +0100 https://epub.ub.uni-muenchen.de/5994/1/Gerbes_Alexander_5994.pdf Gerbes, Alexander L. ddc:610, Medizin

Out of a random population of 4208 non-diabetic pupils without a family history of Type I diabetes 44 (1.05%) individuals had islet cell antibody (ICA) levels greater or equal to 5 Juvenile Diabetes Foundation (JDF) units. 39 of these ICA-positives could be repeatedly tested for circulating insulin autoantibodies (CIAA) using a competitive radiobinding assay. The results were compared with the insulin responses in the intravenous glucose tolerance tests (IVGTT) and with HLA types. Six pupils were positive for CIAA. All of them had complement-fixing ICA, and 5 of them were HLA-DR4 positive. Three of the 6 showed a first-phase insulin response below the first percentile of normal controls. Our data indicate that in population-based studies CIAA can be considered as a high risk marker for impaired beta-cell function in non-diabetic ICA-positive individuals.

Viral antibodies were tested in a cohort of 44 isletcell antibody-positive individuals age 7–19 years, and 44 of their islet cell antibody-negative age and sex-matched classmates selected from a population study of 4208 pupils who had been screened for islet cell antibodies. Anti-coxsackie B1-5 IgM responses were detected in 14 of 44 (32%) of the islet cell antibody-positive subjects and in 7 of 44 (16%) control subjects. This difference did not reach the level of statistical significance. None of the islet cell antibody-positive subjects had specific IgM antibodies to mumps, rubella, or cytomegalovirus. There was also no increase in the prevalence or the mean titres of anti-mumps-IgG or IgA and anti-cytomegalovirus-IgG in islet cell antibody-positive subjects compared to control subjects. These results do not suggest any association between islet cell antibodies, and possibly insulitis, with recent mumps, rubella or cytomegalo virus infection. Further studies are required to clarify the relationship between islet cell antibodies and coxsackie B virus infections.

Tue, 1 Jan 1991 12:00:00 +0100 https://epub.ub.uni-muenchen.de/6055/1/6055.pdf Scherbaum, Werner A.; Trucco, Massimo; Usadel, Klaus-Henning; Rudert, William A.; Schmidt, K.; Thomas, H.; Manfras, Burkhard J.; Glück, Michael; Seißler, Jochen; Böhm, Bernhard Otto

Islet cell antibodies (ICAs) were determined in a large cohort of white nondiabetic schoolchildren (n = 4287) from a homogenous population in southern Germany. The prevalence of ICA levels greater than or equal to 5 Juvenile Diabetes Foundation (JDF) U was 1.05% (95% confidence interval 0.8-1.4%). Analysis of HLA-DR beta and -DQ beta alleles revealed that the specificities found to be increased in insulin-dependent (type I) diabetic subjects with the same ethnic background were also associated with ICA positivity in the nondiabetic schoolchildren. HLA-DR3 (P less than 0.01) and -DR4 (P less than 0.01) phenotypes and absence of Asp residue (P less than 0.01) at codon 57 of the HLA-DQ beta-chain were significantly increased in ICA+ compared with control subjects. High levels of ICAs, which were categorized as either greater than or equal to 17 or greater than or equal to 30 JDF U, were found to be associated with amino acids other than Asp at position 57 of the HLA-DQ beta-chain. No association of ICA level was found for HLA-DR phenotypes.

Tue, 1 Jan 1991 12:00:00 +0100 https://epub.ub.uni-muenchen.de/6064/1/6064.pdf Gerbes, Alexander L. ddc:610, Medizin

Tue, 1 Jan 1991 12:00:00 +0100 https://epub.ub.uni-muenchen.de/6111/1/6111.pdf Ten Bruggencate, Gerrit; Sutor, Bernd

Tue, 1 Jan 1991 12:00:00 +0100 https://epub.ub.uni-muenchen.de/6128/1/6128.pdf Kellerer, Albrecht M.

Tue, 1 Jan 1991 12:00:00 +0100 https://epub.ub.uni-muenchen.de/6129/1/6129.pdf Kellerer, Albrecht M. ddc:610, Medi

Multifractionation isoeffect data are commonly analysed under the assumption that cell survival determines the observed tissue or tumour response, and that it follows a linear-quadratic dose dependence. The analysis is employed to derive the α/β ratios of the linear-quadratic dose dependence, and different methods have been developed for this purpose. A common method uses the so-called Fe plot. A more complex but also more rigorous method has been introduced by Lam et al. (1979). Their method, which is based on numerical optimization procedures, is generalized and somewhat simplified in the present study. Tumour-regrowth data are used to explain the nonparametric procedure which provides α/β ratios without the need to postulate analytical expressions for the relationship between cell survival and regrowth delay.

Tue, 1 Jan 1991 12:00:00 +0100 https://epub.ub.uni-muenchen.de/6144/1/6144.pdf Vollmar, Angelika M.; Gerbes, Alexander L. ddc:610, Medizin

Tue, 1 Jan 1991 12:00:00 +0100 https://epub.ub.uni-muenchen.de/6173/1/6173.pdf Kellerer, Albrecht M. ddc:610, Medizin 0

Tue, 1 Jan 1991 12:00:00 +0100 https://epub.ub.uni-muenchen.de/6192/1/6192.pdf Zaider, M.; Spencer, L. V.; Rösch, W. C.; Kellerer, Albrecht M.; Berger, M. J.; Alsmiller, R. G.; Rossi, H. H. ddc:610, Medizin 0

Tue, 1 Jan 1991 12:00:00 +0100 https://epub.ub.uni-muenchen.de/6193/1/6193.pdf Zaider, Marco; Spencer, Lewis V.; Rösch, William C.; Kellerer, Albrecht M.; Berger, Martin J.; Alsmiller, R. G.; Rossi, Harald H. ddc:610, Medizin

Tue, 1 Jan 1991 12:00:00 +0100 https://epub.ub.uni-muenchen.de/6195/1/6195.pdf Kellerer, Albrecht M. ddc:610, Medi

ICRP hat 1977 zum ersten Mal Risikokoeffizienten für strahlcninduzierte Krebsmortalität angegeben. Diese Zahlen stützen sich vor allem auf die Beobachtungen an den Atombomben-Überlebenden. Durch die Revision der Atombombcn-Dosimetric und die Weiterführung der Beobachtungen bis zum Jahr 1985 ergaben sich erhöhte Risikoschätzungen. Ihre Zahlenwerte und die Ursachen für die Änderung werden diskutiert. Die Risikoschätzungen für die in jungem Alter Bestrahlten sind noch unsicher. Die neuen Empfehlungen der ICRP basieren auf Berechnungen, die sich aus dem Modell des relativen Risikos ergeben, das im Widerspruch steht zu dem beobachteten Trend abnehmender Proportionalitätsfaktoren des Exzeßrisikos mit zunehmender Zeit nach Bestrahlung. Ein modifiziertes Modell des relativen Risikos, bei dem die Proportionalitätsfaktoren nur vom erreichten Alter abhängen, führt für Bestrahlungen im Alter von weniger als 30 Jahren zu deutlich geringeren Projektionen in die Zukunft. Gcmittclt über alle Altersstufen erhält man Risikowertc, die etwa halb so hoch sind wie die Schätzungen der ICRP. Mit dem modifizierten Modell erhält man ohne den hypothetischen Reduktionsfaktor zur Extrapolation auf kleine Dosen etwa die gleichen Risikokoeffizienten, wie sie von ICRP angegeben werden.

Tue, 1 Jan 1991 12:00:00 +0100 https://epub.ub.uni-muenchen.de/6245/1/6245.pdf Berghaus, Alexander ddc:610,

Tue, 1 Jan 1991 12:00:00 +0100 https://epub.ub.uni-muenchen.de/6246/1/6246.pdf Jovanovic, S.; Berghaus, Alexander ddc

Tue, 1 Jan 1991 12:00:00 +0100 https://epub.ub.uni-muenchen.de/6247/1/6247.pdf Berghaus, Alexander

Tue, 1 Jan 1991 12:00:00 +0100 https://epub.ub.uni-muenchen.de/6248/1/6248.pdf Kippenhahn, K.; Berghaus, Alexander ddc:610, Medizin 0

Tue, 1 Jan 1991 12:00:00 +0100 https://epub.ub.uni-muenchen.de/6263/1/6263.pdf Brunnee, T.; Berghaus, Alexander; Jovanovic, S.; Dokic, D. ddc:610, Medizin

Tue, 1 Jan 1991 12:00:00 +0100 https://epub.ub.uni-muenchen.de/6265/1/6265.pdf Berghaus, Alexander; Jovanovic, S. ddc:610, M

Tue, 1 Jan 1991 12:00:00 +0100 https://epub.ub.uni-muenchen.de/6315/1/6315.pdf Barclay, David; Chmelevsky, Danielle; Stock, Gabriele; Kellerer, Albrecht M.

Tue, 1 Jan 1991 12:00:00 +0100 https://epub.ub.uni-muenchen.de/6474/1/6474.pdf Eisenmenger, Wolfgang; Tutsch-Bauer, E. ddc:610, Medizin

Tue, 1 Jan 1991 12:00:00 +0100 https://epub.ub.uni-muenchen.de/6475/1/6475.pdf Eisenmenger, Wolfgang ddc:610, Medizin

Tue, 1 Jan 1991 12:00:00 +0100 https://epub.ub.uni-muenchen.de/6477/1/6477.pdf Wilske, J.; Eisenmenger, Wolfgang ddc:610, Medizin

While it is generally accepted that cholesterol supersaturation of bile is of key importance in the rapid formation of cholesterol crystals, the role of total biliary protein and pH in the pathogenesis of cholesterol gallstones is less well understood. The relation of cholesterol saturation, total protein, and pH was studied in 73 gallbladder bile samples with and 35 gallbladder bile samples without cholesterol crystals. In samples containing crystals, a trend to higher values of cholesterol and to a higher cholesterol saturation index was observed. However, significantly (P = 0.02) higher concentrations of total protein were found in samples with crystals [0.80 +/- 0.40 g/dL (8.0 +/- 4.0 g/L)] than in samples without crystals [0.63 +/- 0.26 g/dL (6.3 +/- 2.6 g/L)]. Moreover, of 22 bile samples with total protein concentrations greater than 10.0 g/L, cholesterol crystals were detected in all but 2. Total lipids, bile acids, phospholipids, and pH values were not significantly different in the two groups of bile samples. It was concluded that high biliary protein concentrations are frequently associated with cholesterol crystals and may, therefore, be a possible risk factor in the pathogenesis of cholesterol gallstones.

Tue, 1 Jan 1991 12:00:00 +0100 https://epub.ub.uni-muenchen.de/6514/1/6514.pdf Eisenmenger, Wolfgang ddc:610, Medizin

Tue, 1 Jan 1991 12:00:00 +0100 https://epub.ub.uni-muenchen.de/6517/1/6517.pdf Eisenmenger, Wolfgang; Penning, R.; Betz, P.

Tue, 1 Jan 1991 12:00:00 +0100 https://epub.ub.uni-muenchen.de/6544/1/6544.pdf Jüngst, Dieter

Tue, 1 Jan 1991 12:00:00 +0100 https://epub.ub.uni-muenchen.de/6546/1/6546.pdf Paumgartner, Gustav; Jüngst, Dieter; Ritter, Christoph von ddc:610, Medizin

The regulatory immediate-early (IE) protein pp89 of murine cytomegalovirus induces CD8+ T lymphocytes that protect against lethal murine cytomegalovirus infection. The IE1 epitope is the only epitope of pp89 that is recognized by BALB/c cytolytic T lymphocytes (CTL). Using synthetic peptides, the optimal and minimal antigenic sequences of the IE1 epitope have been defined. To evaluate the predictive value of data obtained with synthetic peptides, recombinant vaccines encoding this single T-cell epitope were constructed using as a vector the hepatitis B virus core antigen encoded in recombinant vaccinia virus. In infected cells expressing the chimeric proteins, only IE1 epitope sequences that were recognized as synthetic peptides at concentrations lower than 10(-6) M were presented to CTL. Vaccination of mice with the recombinant vaccinia virus that encoded a chimeric protein carrying the optimal 9-amino-acid IE1 epitope sequence elicited CD8+ T lymphocytes with antiviral activity and, furthermore, protected against lethal disease. The results thus show for the first time that recombinant vaccines containing a single foreign nonameric CTL epitope can induce T-lymphocyte-mediated protective immunity.

The immediate-early gene ie2 of murine cytomegalovirus was characterized. The 1.75-kb ie2 transcript is spliced from three exons, of 78, 124, and 1,283 nucleotides, which are separated by introns of 1,245 and 364 nucleotides. An ATG codon located in the third exon leads into an open reading frame of 391 codons. Immediate-early expression of the predicted polypeptide was confirmed by immunoprecipitation of a 43-kDa protein by using an antiserum raised against a bacterial fusion protein. The predicted IE2 amino acid sequence has regions with similarity to amino acid sequences of members of the human cytomegalovirus US22 family.

The effect of murine cytomegalovirus on myelopoiesis was studied in long-term bone marrow culture to find an in vitro correlate for the lethal virus interference with bone marrow reconstitution (W. Mutter, M. J. Reddehase, F. W. Busch, H.-J. Bühring, and U. H. Koszinowski, J. Exp. Med. 167:1645-1658, 1988). The in vitro generation of granulocyte-monocyte progenitors (CFU-GM) discontinued after infection of the stromal cell layer, whereas the proliferation and differentiation of CFU-GM to granulocyte-monocyte colonies remained unaffected. A protocol was established to probe the functional integrity of earlier hematopoietic cells. Pre-CFU-GM (the progenitors of the CFU-GM) could be recovered from an infected bone marrow donor culture by transfer onto an inductive recipient stromal cell layer. Thus, at least in vitro, infection of bone marrow stroma appears to be the only cause of the defect in myelopoiesis.

The gene regulatory immediate early protein, pp89, of murine cytomegalovirus interacts with both DNA-associated and isolated histones in vitro. We characterized the histone-binding region of pp89 and its cellular localization during cell division to examine the possible interaction between pp89 and chromatin. pp89 expressed constitutively in cell line BALB/c 3T3 IE1 does not interact with condensed chromatin. As observed in infected cells, pp89 is localized within the nucleus of cells during interphase but spreads throughout the cell plasma following degradation of the nuclear membrane during early mitosis. In late telophase, pp89 is reorganized within the nucleus. Analysis of pp89 deletion mutants and of fragments generated by cleavage at pH 2·5 revealed that the regions responsible for association with histone are located between amino acids 71 and 415, and are not identical with the domain that shows homology to histone H2B or the highly acidic carboxy-terminal region. A potential gene-activating role of the high affinity of pp89 for isolated histones and the low affinity for DNA-associated histones is discussed.

Processing of endogenously synthesized proteins generates short peptides that are presented by MHC class I molecules to CD8 T lymphocytes. Here it is documented that not only the sequence of the presented peptide but also the residues by which it is flanked in the protein determine the efficiency of processing and presentation. This became evident when a viral sequence of proven antigenicity was inserted at different positions into an unrelated carrier protein. Not different peptides, but different amounts of the antigenic insert itself were retrieved by isolation of naturally processed peptides from cells expressing the different chimeric proteins. Low yield of antigenic peptide from an unfavorable integration site could be overcome by flanking the insert with oligo-alanine to space it from disruptive neighboring sequences. Notably, the degree of protection against lethal virus disease related directly to the amount of naturally processed antigenic peptide.

Tue, 1 Jan 1991 12:00:00 +0100 https://epub.ub.uni-muenchen.de/6728/1/Koszinowski_Ulrich_6728.pdf Forman, J.; Stroynowski, I.; Hood, Leroy; Koszinowski, Ulrich H.; Jones-Youngblood, S.; Hammer, R. E.; Aldrich, C. J.

Residues critical for establishing a trimolecular interaction with a major histocompatibility complex (MHC)-encoded receptor and a T cell antigen receptor (TcR) were determined for an antigenic nonapeptide. The N-terminal residue proved to be involved in binding of the peptide to both receptors and the C-terminal residue was essential for MHC binding. While substitution of either of these critical terminal residues by alanine resulted in an almost complete loss of peptide antigenicity, simultaneous substitution of both created a new functional ligand for the same MHC molecule and the same TcR. Notably, in the biterminally substituted peptide, the core residues took on new roles in the trimolecular interaction in that a residue critical in the authentic nonapeptide for TcR binding became critical for MHC binding and former spacer residues became essential to various degrees for the interaction with either receptor or both. Thus, apparently, the loss of the terminal residues' contribution was at least partially compensated by a redistribution of the roles among the remaining residues. These results reflect a cooperative contribution of all residues of an antigenic peptide to its binding to both receptors and thus challenge a static definition of agretope and epitope as MHC and TcR binding sites.

Tue, 1 Jan 1991 12:00:00 +0100 https://epub.ub.uni-muenchen.de/6780/1/6780.pdf Paumgartner, Gustav; Sauerbruch, Tilman; Berger, H.; Kirsch, C.-M.; Jüngst, Dieter; Bayerdörffer, E.

Tue, 1 Jan 1991 12:00:00 +0100 https://epub.ub.uni-muenchen.de/6791/1/6791.pdf Paumgartner, Gustav; Lang, Thomas; Jüngst, Dieter

Tue, 1 Jan 1991 12:00:00 +0100 https://epub.ub.uni-muenchen.de/6883/1/6883.pdf Fruhmann, G.; Remien, J.; Schopohl, J.; Liebl, B.; Langenmayer, Irmgard; Emslander, H.-P.; Hauck, R.; Haen, E. ddc:610, Medizin 0

Tue, 1 Jan 1991 12:00:00 +0100 https://epub.ub.uni-muenchen.de/7023/1/Reincke_Martin_7023.pdf Winkelmann, W.; Allolio, B.; Reincke, Martin ddc:610, Medizin