Novel C5a receptor antagonists regulate neutrophil functions in vitro and in vivo.

Novel recombinant human C5a receptor antagonists were discovered through modification of the C terminus of C5a. The C5a1-71T1M,C27S,Q71C monomer, (C5aRAM; CGS 27913), was a pure and potent functional antagonist. The importance of a C-terminal cysteine at position 71 to antagonist properties of C5aRAM was confirmed by studying C5a1-71 derivatives with replacements of Q71, C5a derivatives of various lengths (70-74) with C-terminal cysteines, and C5a derivatives of various lengths (71-74) with Q71C replacements. The majority of C5a1-71Q71 derivatives were agonists (C5a-like) in the human neutrophil C5a-induced intracellular calcium mobilization assay. The C5a1-71Q71C derivative was an antagonist. C5a derivatives of lengths 73 and 74 with C-terminal cysteines were agonists, while lengths 70 to 72 were antagonists. C5a derivatives of lengths 72, 73, and 74 with Q71C replacements were agonists, while, again, C5a1-71Q71C was an antagonist. C5aRAM and its adducts, including its dimer, C5aRAD (CGS 32359), were pure antagonists. Additionally, CSaRAM and CSaRAD inhibited binding of 125I-labeled recombinant human C5a to neutrophil membranes (Ki = 79 and 2 pM, respectively), C5a-stimulated neutrophil intracellular calcium mobilization (8 and 13 nM), CD11b integrin up-regulation (10 and 1 nM), superoxide generation (182 and 282 nM), lysozyme release (1 and 2 microM), and chemotaxis (11 and 7 microM). In vivo, intradermal injection of C5aRAM inhibited C5a-induced dermal edema in rabbits. Furthermore, a 5-mg/kg i.v. bolus of C5aRAD significantly inhibited C5a-induced neutropenia in micropigs when challenged with C5a 30 min after C5aRAD administration. C5aRAM and C5aRAD are novel, potent C5a receptor antagonists devoid of agonist or proinflammatory activity with demonstrated efficacy in vitro and in vivo.

[Possibilities and limits of medical graphology. Determination of current status and perspectives (I)]. / Möglichkeiten und Grenzen der medizinischen Graphologie. Standortbestimmung und Perspektiven (Teil I).

As is generally known, the writing movement reacts in a sensitive manner to numerous diseases, medications, stimulants and environmental poisons, as well as to psychic and physical influences. This reaction can occur quite early in the process. Therefore, laboratory data and the physical examination can be supplemented by the handwriting. The graphological analysis is particularly helpful in the following ways: early diagnosis, documenting the course of the disease, evaluating treatment, psychological relationships as well as the problems of old age. This review, presented in three parts, intends to demonstrate to the physician simple and time-saving possibilities, as well as analytical methods, that are most likely to promise success. This needs to be done by a competent graphologist.-Prejudices against graphological methods and exaggerated expectations will be reduced if proper information is presented.

A rapid microtiter plate method for the detection of lysozyme release from human neutrophils.

An improved method was devised to measure lysozyme secreted from human neutrophils [polymorphonuclear leukocyte (PMN)] using a microtiter plate reader capable of analyzing enzyme kinetics. The assay is an adaptation of the classical photometric method which detects changes in the turbidity of a bacterial suspension, Micrococcus lysodeikticus, caused by the enzymatic activity of lysozyme. A standard curve using chicken egg white lysozyme was generated, and activity was detectable between the range of 1 and 100 ng/ml. Leukotriene B4 (LTB4)-induced lysozyme release from human PMN was comparable in both the standard assay and the microtiter plate adaptation with EC50 values of 6.5 and 7.2 nM, respectively. Other select stimuli and their receptor antagonists were also used to evaluate the method. Dose-response curves for chemotactic hexapeptide (CHP), recombinant human C5a (rhC5a), and platelet-activating factor (PAF) resulted in EC50 values of 0.14, 0.80, and 542.00 nM, respectively. Inhibition of lysozyme release was studied using receptor antagonists N-t-Boc-L-methionyl-L-leucyl-L-phenylalanine (N-t-Boc), LY223982, and protamine, which are putative inhibitors of formyl peptides (i.e., CHP), LTB4, and C5a, respectively. N-t-Boc inhibited CHP-induced (0.2 nM) enzyme release with an IC50 of 2 microM; LY223982 blocked LTB4-induced (20 nM) release resulting in an IC50 of 52 nM; and protamine inhibited rhC5a-induced (1.5 nM) release with an IC50 of 2 microM. Further studies revealed that CHP, LTB4, and rhC5a were selectively inhibited by their respective antagonists, albeit LY223982 and protamine were also weak inhibitors of CHP and LTB4, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)

[The controversy regarding the cause of death of Wolfgang Amadeus Mozart (28 January 1756-5 May 1791)]. / Zum Meinungsstreit über die Uraschen des Todes von Wolfgang Amadeus Mozart (28. 1. 1756-5. 12. 1791).

All since two hundred years closely contested hypotheses about the causes of Wolfgang Amadeus Mozart's death are to be verified and to be made credible by the same "original sources" in spite of considerable contrasts. In the present study it is tried to indicate the reasons for the multifariousness of the opinions and to represent Mozart's death as the consequence of a multifactorial process.

[Possibilities and limitations of rapid radiologic detection of selected chemicals in acute poisoning of unknown origin]. / Möglichkeiten und Grenzen eines radiologischen Schnellnachweises ausgewählter Chemikalien bei akuten Vergiftungen unbekannter Provenienz.

Knowledge of the radiopacity of bromine-containing hypnotics and of halogenated hydrocarbons has led to numerous recommendations and speculations. Supported by own investigations the sensible application of a rapid radiological determination in the practice of clinical toxicology is discussed. This should always be carried out, if solvent residues, gastric contents or gastric lavage fluid are available, a ingestion is suspected, and the result of chemical analysis is not promptly obtainable. However, a hint at the group of halogenated hydrocarbons is only possible, if additional information about miscibility with water is accessible. Revealing the presence of radiopaque material in tablet or liquid form by radiologic studies of the abdomen may be very helpful for the rapid evaluation of symptomatology and first therapeutic consequences, but it does not allow any chemical identification of the poison.

[Etiologic and pathogenetic aspects of the demyelinating process in multiple sclerosis]. / Atiologische und pathogenetische Aspekte zum Demyelinisationsprozess bei der Multiplen Sklerose.

In our opinion there is no specific germ for multiple sclerosis. The pathogenic process is developed after a clinically "silent" period and results in a demyelinization process. Partially genetical aspects have to take into account for certain disease processes. Up to now both, the theory of infection as well as the neuroallergetic theory are of importance.

[Progress of research in the diagnosis of multiple sclerosis by study of the cerebrospinal fluid]. / Forschungsfortschritte zur Diagnose der Multiplen Sklerose durch Liquoruntersuchung.

There are neither specific cell investigations nor immunological tests for the diagnosis of MS. Various immunological alterations occur in the course of the disease, but none is unique to MS. Accurate diagnosis remains confined to the clinical field. The diagnosis can be supported by CSF analysis. Laboratory examinations can help to determine the particular stage of the disease and provide clues to the etiology and pathogenesis. The appearance of abnormal CSF levels increases the certainty of the diagnosis.

[Clinical and pathophysiologic aspects of viral infections of the CNS]. / Klinische und pathophsiologische Aspekte bei viralen Infektionen des ZNS.

The available defensive mechanisms are generally sufficient to protect the central nervous system (CNS) against the penetration of infectious particles. The blood-brain-barrier plays a decisive role in this process. The present paper represents the most important circumstances realizing that: 1. The macrophage-lymphocyte-system, 2. the "oligoclonal-CSF-IgG-reaction", and 3. the influence of psychological and physiological burdens on clinical-chemical or immunological parameters. It seems to be important to overcome the instability of the immunosystem and to reestablish the base-line.

Research approaches on the diagnosis of multiple sclerosis by CSF examination.

The diagnostic value of CSF oligoclonal IgG bands, intrathecal IgG synthesis, cytology and markers of myelin breakdown in the cerebro-spinal fluid were evaluated in multiple sclerosis. None of the various immunological alterations were unique to MS. The diagnosis depends therefore mainly on course and clinical symptoms, but can be supported by CSF analysis.