γ/δ Receptor-expressing T-cell clones from a cutaneous T-cell lymphoma suppress hematopoiesis

Summary Recently we described a cutaneous T-cell lymphoma expressing the / T-cell receptor [5]. The patient suffering from this lymphoma showed low numbers of myeloid and T cells in peripheral blood, while B and NK cells were relatively increased. In vitro culture of the patient's bone marrow (BM) cells revealed a significant suppression of myeloid/monocyte colony formation (GM-CFU) compared with normal controls. This was not due to infiltration of the BM with lymphoma cells. We speculated that a soluble factor either secreted or induced by the lymphoma cells might be responsible for the marked suppression of hematopoiesis in this patient. From a skin biopsy with infiltrating / T-lymphoma cells we established T-cell clones bearing the / T-cell receptor and resembling the phenotype of the lymphoma cells. The supernatant (SN) of these / T-cell clones reduced the number of colonies in a CFU-GM assay (using normal control BM) in comparison to SN of / T-cell clones established from the same biopsy. This suppression was seen mainly on day 7 of culture and was not neutralized by the addition of placenta-CM. The main mediator of this suppression seems to be IFN-,since it was detectable in high amounts in the SN of these / T-cell tumor clones as well as in the serum of the patient. In addition, anti-IFN- antibodies can reverse the T-cell SN-mediated suppression of CFU-GM. We conclude that high serum levels of interferon-, which is secreted in high amounts from / T-cells grown from a biopsy of a cutaneous lymphoma, can suppress hematopoiesis.Abbreviations TCR T-cell receptor - IFN- interferon- - SN supernatant - placenta CM placenta conditioned medium - BM bone marrow - CFU-GM myeloid/monocyte colony formation - NK cells natural killer cells - Ab antibody M. Wilhelm was supported by theDeutsche Forschungsgemeinschaft (DFG Wi 728-2)     

Survival of two patients with severe δ-aminolaevulinic acid dehydratase deficiency porphyria

The course of -aminolaevulinic acid dehydratase activity was studied over the 23 years in erythrocytes of two male patients. The enzyme activity was originally 1–2%, which then increased to 8% of normal levels several years after clinical manifestation of the acute hepatic porphyria syndrome. Urinary excretions of -aminolaevulinic acid and coproporphyrin III were excessively increased in the two patients with compound-heterozygous -aminolaevulinic acid dehydratase deficiency porphyria.     

The ability of heat stress and metabolic preconditioning to protect primary rat cardiac myocytes

Primary rat cardiocytes were subjected to either thermal preconditioning for 30 min at 43°C or 20 min metabolic preconditioning (10 mM deoxyglucose, 20 mM lactate, pH 6.5). Eighteen hours later cells were analysed either for hsp 70i expression or subjected to a subsequent lethal heat stress or simulated ischaemia (10 mM deoxyglucose, 20 mM lactate, 0.75 mM sodium dithionite, 12 mM potassium chloride, pH 6.5) for 2 hours and assessed for survival by trypan blue exclusion.Hsp 70i was induced over 100 fold by thermal preconditioning and 30 fold by metabolic preconditioning (p<0.001, p<0.05), hsp 90 was induced 2.71 fold and 2.24 fold (p<0.001, p<0.001) by thermal and metabolic preconditioning respectively, while hsp 60 was not induced by either treatment. Preconditioned cultures had improved survival against subsequent lethal heat stress or simulated ischaemia: Thermal preconditioning reduced death from 69.22% to 52.46% upon subsequent lethal heat stress and from 49.13% to 36.66% upon subsequent lethal simulated ischaemia. Metabolic preconditioning reduced cell death from 51.29% to 33.8% against subsequent lethal heat stress, and from 69.09% to 55.61% upon subsequent lethal simulated ischaemia. A second marker of cell death, the release of lactate dehydrogenase activity into the culture media, was reduced to 65% and 60% of control values for thermally preconditioned cells subjected to lethal heat or lethal simulated ischaemia respectively. Metabolically preconditioned cells demonstrated lactate dehydrogenase activity of 59% and 51% that of control values, when subjected to lethal heat or lethal simulated ischaemia respectively.Abbreviations hsp heat stress protein - hsp 70i inducible 70 kDa heat stress protein - LDH lactate dehydrogenase - PBS phosphate buffered saline     

Analysis of lymphocyte subsets in patients with aplastic anemia before and during immunosuppressive therapy

Summary To define the contribution of T-lymphocyte subsets in the development of aplastic anemia (AA), T-cell subpopulations including T cells, T cells, and TCS1-positive T cells, were analyzed by cytophotometry in the peripheral blood (PB) and bone marrow (BM) of patients with AA before and after 6 weeks of therapy with anti-lymphocyte globulin (ALG), methylprednisolone, and cyclosporin A (CSA). In nine patients with AA a significant decrease of PB- and BM-derived T cells was observed after 6 weeks of therapy as compared with normal controls. At diagnosis, the CD4/CD8 ratio in PB and BM of the patients did not differ from the ratio in the control population; however, a reversed ratio (< 1) was present in PB as well as in BM after weeks of therapy. Interestingly, lymphocytes expressing the T-cell receptor (TCR) were significantly decreased both before (PB 1.2±0.1%; BM 0.8±0.1%) and after 6 weeks of therapy (PB 0.7±0.1%; BM 0.7±0.1%) as compared with healthy controls (PB 2.4±0.2%; BM 2.3±0.2%). However, the proportion of the -T-cell subpopulation expressing the TCS1 phenotype was markedly increased before (PB 42±3.5%; BM 31±3%) and especially after 42 days of therapy (PB 77±12%; BM 45±2%) as compared with that in normal subjects (PB 19±2%; BM 9.7±0.8%). At present, follow-up is under evaluation to correlate these findings with hematological response. The pathophysiological significance of the observed alterations within the T-cell subsets and especially the T-cell populations will require further functional analyses, in particular since TCS1-positive T cells exhibit autoimmunological capacity.Presented at the annual meeting of the German Society for Hematology and Oncology, 4–7 October 1992, Berlin     

Detection of clonality by polymerase chain reaction in childhood B-lineage acute lymphoblastic leukemia

Summary DNA-based PCR with various sets of primers for TCR /, and Ig heavy chain (IgH) genes were used to study clonality in childhood B-lineage acute lymphoblastic leukemia. Amplification of the IgH CDR-III was observed in 75 of 120 analyzed cases (62.5%). From all analyzed groups, the IgH gene rearrangement was most often observed in pre-B ALL (85.7%) and was rather rare in null-ALL (34.5%). TCR delta gene rearrangement was the most common, and was observed in 77 patients (64.2%). The typical pattern of rearrangements was defined as anincomplete V2 to D3, V2 to D2, or D3 to D3 to D2 recombination product. Rearrangements of TCR gamma gene we observed in 61 cases (50.8%). TCR gamma gene rearrangements were detected predominantly in null-ALL and early B-ALL (55.2% and 60%, respectively) and were rather rare in other groups. Of all eight V segments of VI group, the most frequent gene usage concerns regions V2, V4, and V7. We have confirmed that IgH gene amplification, together with TCR gamma and delta gene amplification, provides a rapid, sensitive approach to assessing clonality in ALL almost in 100% of cases.This work was financed by KBN grants 4.0551.91.01 and 6.6346.92.03     

Changes in expression levels of genes involved in fatty acid metabolism: upregulation of all three members of the PPAR family (α, γ, δ) and the newly described adiponectin receptor 2, but not adiponectin receptor 1 during neonatal cardiac development of the rat

During neonatal cardiac development, the heart changes its substrate preference from glucose to fatty acids. The aim of this study was to investigate the changes in mRNA expression levels of genes involved in the control of cardiac fatty acid metabolism in the transition from neonatal to adult life. Methods mRNA expression levels for peroxisome proliferator activated receptor (PPAR) , and , PPAR co–factor 1 and (PGC–1 and ), 9–cis retinoc–acid–activated receptor , and (RXR , , ), 5–AMP activated protein kinase (AMPK) 1 and 2, adiponectin receptor 1 and 2 (AR 1 and AR 2) were measured in heart tissue of neonatal 0–day, 7–day and 21– day old rats. Results mRNA expression of all three members of the PPAR family were upregulated significantly from day 0 to day 21 ( +117%, +133%, +203%). In addition, m–RNA expression of all RXR isoforms increased from day 0 to day 7 ( +125%, +69%; +41%). AR 2 exhibited a small but significant increase in mRNA expression (+ 46%). Conclusions We were able to demonstrate for the first time that in addition to PPAR, also PPAR and , as well as all RXR isoforms and AR 2 are upregulated in the heart during neonatal development.Drs. Steinmetz and Quentin contributed equally to this publication     

Characterization of an acute T lymphoblastic leukemia expressing the γ/δ T-cell receptor

Summary Leukemic cells of a 20 year old patient, suffering from acute lymphoblastic leukemia, were characterized by surface marker and functional analysis. A significant cell population within this type of leukemia expresses concomitantly the CD4 and CD8 antigen on the same cell and might represent a new differentiation stage of T-cells with the / receptor. The leukemic cells show a distinct pattern of growth response to mitogens and lymphokines, which might correlate to their differentiation stage. Moreover, a natural killer-like activity can be induced in these cells by IL-2.Abbreviations FITC fluorescein isothiocyanate - PE phycoerythrin - IL-2 interleukin 2; - / TCR gamma/delta T cell receptor - NK natural killer - PBL peripheral blood lymphocytes - T-ALL acute T lymphoblastic leukemia - ConA concanavalin A - PMA phorbol myristate acetate - BM bone marrow - IL-2R IL-2 receptor - TdT terminal deoxynucleotidyl transferase Supported by the Deutsche Forschungsgemeinschaft (DFG Wi-728/3-1)     

A monoclonal antibody-linked immunoassay for hemoglobin H disease

Summary A murine monoclonal antibody (mAb) was generated that recognizes hemoglobin (Hb) H, the tetrameric form (₄) of human -globin chains. The antibody ₄-1 (1, ) does not react with Hbs A, F, Bart's, or isolated chains, indicating that the antibody recognizes an epitope comprised of multiple chains. A simple, rapid, and sensitive enzyme immunoassay was established to detect and quantitate Hb H in hemolysates from subjects with Hb H disease. The globin level in these patients was also measured using the monoclonal antibody -1, which is specific for chains of Hb A₂. With these assays, 20 hemolysates from subjects with Hb H disease' ten from normal adults and ten from newborn babies were analyzed. The percent of Hb H ranged from 1.5% to 25% in Hb H patients. There was a significant average reduction (32%) in chains in these samples as compared with the normal average adult value. The decreased expresion of chains thus results in a reduction of the levels of normal Hbs A and A₂ and accumulation of ₄, causing Hb H disease.Work supported by NIH grant HLB-41544     

Vorbestehendes Risikoprofil und Dekubitalulzera im intensivmedizinischen Bereich

Zusammenfassung In diesem Beitrag werden die Risikofaktoren zu Beginn des Krankhausaufenthaltes bei Patienten, die auf Intensivstationen ein Dekubitalulkus entwickelten, mit denen der Patienten verglichen, die auf Normalstationen ein Dekubitalulkus entwickelten.Im Rahmen einer prospektiven Erhebung wurde durch das Pflegepersonals von April 2003 bis April 2004 bei jedem Patienten am Tag der Aufnahme ein 29 Punkte umfassendes Risikoprofil dokumentiert. Ingesamt umfasst die Auswertung 49 904 Behandlungsfälle, von denen 5073 (10,2%) mindestens einen Tag auf einer Intensivstation verbrachten. Insgesamt entwickelten 94 Patienten während eines Intensivaufenthaltes ein neues Dekubitalulkus und 186 Patienten ohne einen Aufenthalt auf einer Intensivstation.Patienten, die auf einer Intensivstation ihr Druckulkus entwickelten, unterschieden sich im Alter, der Geschlechterverteilung und der Krankenhausverweildauer nicht von den Patienten, die ihr Druckulkus ohne Intensivaufenthalt entwickelten. Der Anteil der operierten Patienten war mit 72% nur gering höher als mit 60% bei den nicht intensivmedizinisch betreuten Patienten (p=0,046). Die mittlere Anzahl von Risikofaktoren zu Beginn des Krankenhausaufenthaltes war mit 10,0±5,7 im Vergleich zu 7,5±4,9 bei den Patienten, die ihr Druckulkus auf Normalstation entwickelten, erhöht (p=0,001).Bei den Intensivpatienten waren bereits bei Aufnahme die Risikofaktoren stark sedierende Medikamente, gefäßverengende Medikamente, Op-Dauer >60 Minuten, Fieber, Sepsis, Stuhl- und Urininkontinenz, Druckgefährdung durch Ableitungssysteme oder Fixierung, Störung des Druck-, Schmerz- oder Temperaturempfindens häufiger als bei Patienten, die unabhängig von einem Intensivaufenthalt ein Druckulkus entwickelten.Die vorgestellten Daten zeigen, dass Patienten, die während eines Aufenthalts auf einer Intensivstation ein Dekubitalulkus entwickelten, schon bei Aufnahme ins Krankenhaus ein erhöhtes Risiko aufweisen.für das Interdisziplinäre Dekubitus-Projekt     

Celiac Disease Without Villous Atrophy

Current diagnostic criteria of celiac disease require small bowel villous atrophy, although the damage develops gradually. We therefore searched for evidence of disease in 10 adults suspected to have celiac disease, but evincing only minor mucosal inflammation and increase in ⁺ cells without villous atrophy. Twenty untreated celiac and 27 nonceliac patients served as biopsy controls. CD3⁺, ⁺, and ⁺ cells were increased in patients with only minor mucosal lesions, but less than in celiac patients. The inflammation resolved on gluten-free diet, and abdominal symptoms were alleviated. Eight of 10 had positive endomysial, seven gliadin, and nine tissue transglutaminase antibodies; all normalized on diet. Eight patients had osteopenia; HLA DQ2 was found in all. Minor mucosal lesions with an increase in ⁺ intraepithelial lymphocytes were suggestive of celiac disease. Our patients showed a clinical, histological, and serological recovery on diet; risk of osteopenia speaks in favor of dietary treatment.     

Further identification of protein kinase C isozymes in mouse epidermis

In the current study, the protein kinase C (PKC) isozymes present in mouse epidermis have been identified using immunological and chromatographic methods. Six PKC isozymes, PKC, PKC, PKC, PKC, PKC, and PKC, were identified in unfractionated epidermal preparations by protein immunoblotting. The subcellular distribution and presence of these isozymes was further verified by hydroxyapatite (HA) chromatography with the exception of PKE, which could not be detected following HA chromatography. The five PKC isozymes recovered following HA chromatography were detected in both epidermal cytosol and particulate fractions, although PKC was found in a much higher proportion relative to the other PKC isozymes in the particulate fraction using histone H1 as the substrate. The biochemical properties of the epidermal PKC isozymes partially purified by HA chromatography agreed with those reported for other tissues and further supported their immunological identification in epidermal preparations. The activities of HA chromatography peaks corresponding to PKC, PKC, and PKC were found to be dependent on both Ca²⁺ and phosphatidylserine (PtdSer), whereas, the activities of HA peaks corresponding to PKC and PKC were Ca²⁺-independent but PtdSer-dependent. The HA peak corresponding to PKC also displayed a characteristic biphasic modulation by arachidonic acid (activation at low, inactivation at high concentrations) and inactivation by preincubation with PtdSer. PKC activity was also characteristic, in that it was dependent on PtdSer and was not increased by the phorbol ester, 12-O-tetradecanoylphorbol 13-acetate. Some differences in substrate specificity were also observed between the epidermal PKC isozymes. The presence of multiple isozymes of PKC in mouse epidermis suggests that the different isozymes may play distinct roles in signal transduction and tumor promotion in this tissue.Abbreviations PKC protein kinase C - HA hydroxyapatite - PtdSer phosphatidylserine - TPA 12-O-tetradecanoylphorbol 13-acetate This work was supported by USPHS grants CA 38871 (J.D.), CA 57596 (J.D.) and core grant CA 16672     

An auto-anti-B in an A1B person. Serological studies

Summary The serum of a patient (Mr. Lat) with the regular blood group A₁B contains an anti-B reacting with all cells having a B antigen except b_x and cis AB. The anti-B reacts at 4° C and occasionally at room temperature as shown by agglutination, absorption-elution and by thermo-dynamic assays. The antibody is regarded as an irregular autoantibody belonging to the group of the so called suppressed or latent antibodies.

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Zusammenfassung Das Serum eines Patienten (Mr. Lat) mit der normalen Blutgruppe A₁B enthält Anti-B, das mit allen Zellen reagiert, die ein B-Antigen aufweisen, ausgenommen B_x und cis AB. Das Anti-B reagiert bei 4° C und gelegentlich bei Zimmertemperatur, was durch Agglutination, Absorption-Elution und durch thermodynamische Unter-suchungen gezeigt wurde. Der Antikörper wird als irregulärer Autoantikörper angesehen, der zur Gruppe der supprimierten oder latenten Antikörper gehört.

     

Mutations in the R-type pyruvate kinase gene and altered enzyme kinetic properties in patients with hemolytic anemia due to pyruvate kinase deficiency

Summary The biochemical properties of erythrocyte pyruvate kinase (PK) together with mutations found in the coding sequence of the R-PK gene in five patients with severe hemolytic anemia due to PK deficiency are described. The enzyme variants were designated PK Mosul (homozygote), PK Bukarest_1,2, PK Hamburg₁, PK Köln₁, and PK Essen (compound heterozygote). PK Mosul showed normal positive cooperative substrate binding, PK Bukarest_1,2 exhibited noncooperative behavior, and PK Hamburg₁ and PK Köln₁ displayed mixed cooperativity, whereas PK Essen was negative cooperative. PK Mosul was found to be homozygous for the mutation 1151 ACG to ATG, resulting in an amino acid substitution 384 Thr to Met. In one allele of PK Bukarest_1,2 a single nucleotide substitution GAG-TAG was found at nucleotide 721, causing a change of 241 Glu to a chain termination codon (PK Bukarest₁). Additionally, in the second allele of this patient a point mutation at position 1594 (CGG-TGG) occurs, changing 532 Arg to Trp (PK Bukarest₂). Direct sequencing showed the heterozygosity of the patient's mother (PK Bukarest₁/normal) at position 721 and of the patient's father (PK Bukarest₂ /normal) at position 1594. A point mutation at position 1529 (CGA-CAA), causing an amino acid substitution 510 Arg-Gln, was identified in PK Hamburg₁ and PK Köln₁. The second mutation in these variants was not detected. In PK Essen no mutation in the coding sequence was found at all. Screening for the mutation at position 1529 in further compound heterozygote patients and in normal subjects of Western European origin showed that this exchange is a common mutation responsible for PK deficiency in this population.Supported by theDeutsche Forschungsgemeinschaft, Grants no. La 527/1 and Ne 416/1.     

Interpretation of blood cultures yielding staphylococcus aureus

Summary Forty-eight patients with blood cultures positive for Staphylococcus aureus were classified according to clinical criteria in three groups: definite, possible, and doubtful septicemia. Using traditional blood culture sets with two bottles (thioglycollate and tryptic soy broths), we found that patients with definite septicemia always showed more than one positive bottle per day if more than one set was drawn, that the mean detection time was 1.7 days, and that 95% of the first positive bottles and 92% of all positive bottles grew within two days of incubation. Patients with doubtful septicemia were more often (88%) positive in one bottle only, the mean detection time for all bottles was 3.7 days, and only 35% of the first positive bottles and 33% of all positive bottles yielded growth within two days. Possible cases took a position between these two extremes but tended more towards the doubtful cases. The implications of these findings for the interpretation of blood cultures with S. aureus are discussed.

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Interpretation von Blutkulturen mit Staphylococcus aureus

Zusammenfassung 48 Patienten, aus deren Blutkulturen Staphylococcus aureus gezüchtet worden war, wurden nach klinischen Kriterien in drei Gruppen eingeteilt: unzweifelhafte, mögliche und zweifelhafte Septikämie. Bei Verwendung eines traditionellen Blutkultursystems mit zwei Flaschen pro Kultur (Thioglykolat- und Tryptic Soy-Bouillons) ergab sich, daß Patienten mit unzweifelhafter Septikämie stets mehr als eine positive Flasche pro Tag aufwiesen — sofern mehr als eine Kultur pro Tag entnommen worden war —, daß die mittlere Bebrütungsdauer bis zur Positivität 1,7 Tage betrug, und daß 95% der ersten positiven Flaschen und 92% aller positiven Flaschen innerhalb von zwei Tagen Wachstum von S. aureus zeigten. Patienten mit zweifelhafter Septikämie zeigten häufiger (in 88%) Wachstum in nur einer Flasche, die mittlere Bebrütungsdauer betrug 3,7 Tage, und nur 35% der ersten positiven Flaschen und 33% aller positiven Flaschen ergaben Wachstum innerhalb von zwei Tagen. Mögliche Septikämien nahmen eine Zwischenstellung ein, tendierten jedoch mehr nach der zweifelhaften Kategoric. Folgerungen für die Interpretation von Blutkulturen mit S. aureus werden diskutiert.

     

Einsatz von β-Blockern bei kardiovaskulären Erkrankungen und Asthma bronchiale/COPD

Zusammenfassung Zahlreiche groe randomisierte Studien zeigen die signifikante Reduktion der Gesamtmortalität bei Myokardinfarkt und chronischer Herzinsuffizienz unter -Blockertherapie. Patienten mit COPD bzw. Asthma bronchiale und akutem Myokardinfarkt oder chronischer Herzinsuffizienz wird diese lebensverlängernde Therapie allerdings häufig vorenthalten. Dabei profitieren Studien zufolge gerade COPD-Patienten aufgrund ihres hohen kardiovaskulären Risikoprofils von einer solchen Therapie. Aktuelle Metaanalysen belegen zudem die gute klinische Verträglichkeit einer kardioselektiven -Blockertherapie ohne wesentliche Beeinträchtigung der Lungenfunktion. Neuere Arbeiten legen nahe, dass die Mortalitätsreduktion durch eine -Blockertherapie die potenziellen Risiken bei COPD und möglicherweise auch bei leichten Formen des Asthma überwiegt. Kontraindikationen für eine -Blockertherapie sind neben akuten Exazerbationen schwere Formen von COPD, mittelgradig bis schweres Asthma sowie eine Dauertherapie mit -2-Sympatomimetika. Die Therapie mit einem -Blocker sollte nach Abwägung des Nutzen-Risiko-Verhältnisses individuell auf den einzelnen Patienten mit initial niedriger Dosierung eingestellt werden. Dabei sollten vorrangig kardioselektive Substanzen mit nachgewiesenem Mortalitätsbenefit bei Myokardinfarkt und chronischer Herzinsuffizienz gewählt werden.     

TCR1+ large granular lymphocyte proliferation in rheumatoid arthritis

The T-lymphoproliferative syndrome is characterized by a proliferation of large granular lymphocytes (LGL). It is often associated with neutropenia, and in 30% of cases with rheumatoid arthritis (RA). Phenotypic analysis has demonstrated that in most cases of RA with T-proliferative disease, the LGL represent T cells with a clonal rearrangement of the / T cell receptor (TCR2). Here, three patients with / TCR1⁺ LGL proliferation suffering from long-standing arthritis and neutropenia are described. The first patient with RA showed an expansion of a heterogeneous CD2⁺ CD16⁺ CD56^- LGL population, of which 30% coexpressed TCR1 with V1 rearrangement. The second patient with ankylosing spondylitis and RA was suffering from proliferation of TCR1⁺ (V9^-, V1^-), CD2⁺ CD16^- CD56^- LGL with low coexpression of CD8. The third patient with RA was suffering from a proliferation of TCR1⁺ (V1⁺, V9^-) CD4^- CE8^- CD16^- CD56^- lymphocytes. On the basis of these unusual findings, the pathogenetic role of TCR1⁺ T cells in RA is discussed.     

Effect of gold therapy on CD5+ B-cells and TCR γδ + T-cells in patients with rheumatoid arthritis

Summary Circulating CD5⁺ B-cell levels in 15 patients with rheumatoid arthritis (RA) not receiving remittive therapy was significantly increased when compared to 17 normal controls (mean±SE: RA, 19.7±2.85%; controls, 11.6±1.67%; P<0.02). In contrast, 24 patients with RA receiving gold sodium thiomalate therapy (GST) had similar CD5⁺ B-cell levels (11.88±1.65) when compared to controls and significantly reduced levels when compared to the RA group not receiving remittive agents (P<0.01). Furthermore, TCR ⁺ T-cell levels were also assessed in these patients groups. These values were not significantly different between any of the groups (controls, 4.46±1.36%; GST, 6.88±1.73%; RA, 2.73±0.55%), although 42% of the GST treated group had ⁺ T-cell levels higher than the entire untreated RA group. No correlation was observed between the levels of TCR ⁺ T-cells and CD5⁺ B-cells in any of these groups. These results suggested that therapy does influence the level of CD5⁺ B-cells and ⁺ T-cells in these patients.     

Modulation of pacemaker activity in sheep cardiac Purkinje fibers by stimulation of β-adrenoceptor subtypes

The electrophysiological effects mediated by ₁- and ₂- in spontaneously active sheep cardiac Purkinje fibers were investigated using the non-selective agonist (–)-isoproterenol (IPN) and the selective agonists (–)-noradrenaline (₁) and procaterol (₂) in the absence and presence of the selective antagonists bisoprolol (₁) and ICI 118,551 (₂).IPN (0.01 mol/l) increased the spontaneous rate by 54% and the slope of diastolic depolarization by 68% of the respective control values. Further, IPN increased the action potential duration at –20 mV (APD –20 mV) from 96 to 154 ms, reduced the APD –70 mV by 17% and the duration of the diastole by 39% and slightly hyperpolarized the maximum diastolic potential. These effects were partially inhibited by ICI 118,551 (0.03 mol/l), diminished by bisoprolol (0.1 mol/l) and almost completely blocked by the combination of both antagonists. Concentration response curves of IPN were influenced by the selective antagonists as follows: ICI 118,551 (0.03 mol/l) shifted the curves to the right by 0.2–0.4 log units and increased the slope factor. Bisoprolol (0.1 mol/l) induced a greater shift to the right by 1.1–1.5 log units. Combination of bisoprolol with ICI 118,551 shifted the curves to the right by 1.5–1.7 log units.Noradrenaline (0.3 mol/l) elicited similar actions as IPN. Bisoprolol (0.1 mol/l) shifted the concentration response curves of noradrenaline to the right by 1.1–1.9 log units. Actions of procaterol (0.1 mol/l) were weak, attained only 15–35% of the maximal effects of IPN and could be blocked by ICI 118,551 (0.03 mol/l).These results show that the increase of pacemaker activity induced by catecholamines in sheep cardiac Purkinje fibers is predominantly mediated by stimulation of ₁. However, contribution of ₂ mediated effects could be demonstrated.Supported by Ministerium für Wissenschaft und Forschung, Nordrhein-Westfalen, Projekt-Nr, 40008786.     

Growth Hormone (GH) Secretion in Primary Adrenal Insufficiency: Effects of Cortisol Withdrawal and Patterned Replacement on GH Pulsatility and Circadian Rhythmicity

We studied the effects of cortisol withdrawal and patterned replacement upon spontaneous GH secretion and circadian rhythmicity in 7 patients with Addison's disease. Hydrocortisone was administered in physiological daily total dosages, and all resulting plasma cortisol values were 2–15 g/dl. It was given in 3 pulsatile modes: simulating physiological rhythm, reverse diurnal rhythmicity and continuous pulsatility. All modes of cortisol administration increased mean 24h, GH pulse amplitude and interpulse GH levels. During saline infusions circadian GH rhythmicity was preserved, with GH being at its highest between 2400–0400 h. Administration of hydrocortisone in any mode did not modify circadian GH rhythmicity. We conclude: Cortisol replacement in physiological daily doses increases GH output in patients with Addison's disease by augmenting GH pulse amplitude and interpulse levels. This is likely due to the attenuation of hypothalamic somatostatin (SRIF) secretion by physiologic levels of cortisol. By inference, it implies that cortisol deficiency leads to diminution of GH output with low GH pulse amplitude, likely as a result of an augmented hypothalamic somatostatin secretion. However, circadian rhythmicity of GH secretion is glucocorticoid-independent.     

Über Beziehungen zwischen Steroiden und krebserzeugenden Verbindungen (V.)

Zusammenfassung Suspensionen von 2-acetamino-^1.3.5(10)-oestratrien und 3-Acetamino-^1.3.5(10)-oestratrien in Tricaprylin und solche in Glycerin wurden Mäusen subcutan injiziert. Tumoren an der Applikationsstelle wurden nicht beobachtet, doch wurde eine gegenüber den Kontrollen vermehrte Bildung von Tumoren entfernt vom Applikationsort beobachtet.—Unwirksam erwies sich 3-Amino-^1.3.5(10)-oestratrienol-(17) im Pinselungstest an der Maus; im Fütterungstest nach Huggins an der weiblichen Sprague-Dawley-Ratte entstand bei 1 von 10 Tieren ein Mammacarcinom. Die bisher erzielten Ergebnisse sollten dazu anregen, Vertreter dieser Verbindungsklasse auf breiterer Basis zu prüfen.—Eine hypothetische Reaktionsfolge für die Umwandlung von natürlichen Oestrogenen über ihre p-Chinole in die Amino-Analoge (Transaminierung mit Pyridoxamin-5-phosphat) wird diskutiert.

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Relations between steroids and carcinogenic compounds (V.)2-Acetamino- and 3-Acetamino-^1.3.5(10)-estratriene and 3-Amino-^1.3.5(10)-estratrienol-(17)

Summary Suspensions of 2-acetamino-^1.3.5(10);-estratriene and 3-acetamino-^1.3.5(10)-estratriene in tricapryline and in glycerol were injected subcutaneously into mice. Tumors at the site of application were not observed; however, remote from the site of application an increased formation of tumors was observed. 3-Amino-^1.3.5(10)-estratrienol-(17) proved to be ineffective in the skinpainting-test on the mouse; in the feeding-test according to Huggins with female Sprague-Dawley-rats one mammary carcinoma appeared in 1 out of 10 animals. The results obtained so far should stimulate the investigation of representations of this class of compounds on a broader basis.—A hypothetical reaction sequence for the conversion of natural estrogens through their p-quinols into the amino-analogs (transamination with pyridoxamine-5-phosphate) is discussed.

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IV. Mitteilung s. Neumann und Thomas (1967).

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Die histologischen Untersuchungen wurden mit Unterstützung der Deutschen Forschungsgemeinschaft durchgeführt.