The strict separation of clinical and academic budgets: an analysis at a German medical university department of anaesthesia.

In the past, cross-funding between medical and academic budgets has been commonly practiced at university medical departments in Germany. Recent developments have however, generated pressures to enforce the separation of funding for both areas, as intended by law, and to distribute them on the basis of performance. A pilot project at Münster University Hospital, initiated by the German Federal Ministry for Research and Education in 1998, is described here. The main aim of the project was to find ways of distinguishing between research and teaching areas, on the one hand, and health care on the other, in terms of documenting performance and itemizing costs.

Transendothelial migration of 27E10+ human monocytes.

The myeloid-related proteins MRP8 (S100A8) and MRP14 (S100A9), two members of the S100 family of calcium-binding proteins, are co-expressed and form a cell-surface and cytoskeleton-associated heterodimer upon calcium mobilization which is recognized by the mAb 27E10. The heterodimer is abundantly expressed in the cytoplasm of granulocytes and a subpopulation of blood monocytes. Previously, we and others demonstrated endothelium-associated MRP8/14 in inflamed tissues in the vicinity of transmigrating leukocytes, suggesting a function of the proteins in this process. Here, we demonstrate that 27E10(+) cells represent a fast-migrating monocyte subpopulation which preferentially utilizes an ICAM-1-dependent mechanism. The following observations imply a function of MRP8/14 in the transmigration process: (i) higher secretion of MRP8/14 from 27E10(+) monocytes compared to 27E10(-) monocytes after interaction with activated endothelium, (ii) higher expression of CD11b on 27E10(+) compared to 27E10(-) monocytes, (iii) up-regulation of CD11b on 27E10(-) monocytes in the presence of MRP14 or MRP8/14 heterodimers but not MRP8 and (iv) active participation of MRP14 but not of MRP8 in transmigration as shown by blocking with respective antibodies. We show that the interaction of 27E10(+) monocytes with activated endothelium leads to MRP8/14 release which may account for the high MRP8/14 concentrations in body fluids of patients with acute or chronic inflammatory diseases. Released MRP8/14 may serve a function by enhancing CD11b expression and/or affinity in human monocytes and by participating in the transendothelial migration mechanism. Thus, MRP8/14 substantially contributes to the recruitment of monocytes to an inflammatory site.

In experimental leishmaniasis deficiency of CD18 results in parasite dissemination associated with altered macrophage functions and incomplete Th1 cell response.

We investigated experimental leishmaniasis in CD18-deficient mice. Whereas wild-type (WT) CD18-/- mice (129SV/C57BL/6) were resistant to infection, CD18-/- mice revealed increasing visceral dissemination of parasites. Unlike in other susceptible strains, infected footpads of CD18-/- mice did not ulcerate, due to an abolished recruitment of granulocytes. In vitro, CD18-/- macrophages were able to phagocytose opsonized Leishmania major despite absence of CR3, albeit phagocytosis rate was 50% lower than in WT macrophages. We found that uptake was partially mediated by scavenger receptors. As infected CD18-/- macrophages showed impaired ability to produce NO and to eliminate parasites, CD18 is one mediator of NO production. CD18 is also involved in reduction of IL-12 release by L. major-infected macrophages, as uptake of opsonized parasites (via CR3) decreased IL-12 release only in WT, but not in CD18-/- macrophages. When T cells from infected CD18-/- mice were restimulated with antigen-presenting cells (APC), they released no IL-2 or IL-4, but a little IFN-gamma, associated with lack of proliferation. This deficiency was linked to absence of CD18 on T cells, but not on APC. Substitution with IL-2 specifically restored a Th1-like response with proliferation and release of IFN-gamma. Thus, while impaired phagocytosis, NO production, and recruitment of granulocytes in CD18-/- mice may not reverse resistance, and while unrestricted IL-12 release supports development of Th1 cells, the failure of T cells to release IL-2 and to proliferate causes susceptibility.

Murine leukocytes with ring-shaped nuclei include granulocytes, monocytes, and their precursors.

Leukocytes with ring-shaped nuclei (ring cells) are present in bone marrow (BM; approximately 50% of BM cells), in peripheral blood (PB), and in inflammatory infiltrates of mice, but also in humans during myeloproliferative disorders. They are usually referred to as polymorphonuclear cells (PMN), but we demonstrate that they additionally encompass different types of mononuclear (MNC)-like ring cells. PMN ring cells had constricted ring-shaped nuclei with a wide cytoplasmic center and were sorted among the GR-1high fraction. The MNC-like ring cells belonged to the GR-1low fraction. Their nuclei were not segmented and the cytoplasmic center of their nuclei was small. They were heterogeneous with one subgroup containing monocytes/macrophages according to ultrastructure, immunophenotype (BM8, F4/80, CD13, ER-HR3), activity of unspecific esterase, and phagocytosis of Leishmania major. A second subgroup contained myeloic precursor cells as they proliferated (Ki67), expressed ER-MP12, and showed on ultrastructure distribution patterns of peroxidase activity compatible with myelocytes, promyelocytes, or promonocytes. A third subgroup of cells had large, sometimes lobulated nuclei, was lineage marker(negative/low) (GR-1, Mac-1, B220 etc.), CD38-, but c-kit+ and sca-1+, and thus belonged to a close progeny of murine hematopoietic stem cells. In PB, ring cells encompassed mainly PMN, but also monocytes and cells with characteristics of both the granulocytic and monocytic lineage. Thus, ring cells comprise mature and precursor forms of myeloic cells. Their analysis revealed that in mice a clear distinction between the granulocytic and monocytic lineage beyond the GM-CFU stage is not always feasible.

Several new bacteriophage T4 genes, mapped by sequencing deletion endpoints between genes 56 (dCTPase) and dda (a DNA-dependent ATPase-helicase) modulate transcription.

We have analyzed DNA of wild-type T4 and of 13 independent large viable deletions isolated by Homyk and Weil (Virology 61 (1974) 505-523) and by Little (Virology 53 (1973) 47-59), by sequencing, cloning, and expression studies. The deletions can be explained by illegitimate recombination between short (4- to 15-bp) ectopic repeats. In four deletions, adjacent regions are partially homologous, and in at least one of them, the base adjacent to the overlap was deleted during recombination. The sequence 5'-GGGC, which has not been associated with T4 deletions in other map regions, occurs within three repeats, and near the repeats in four more of the 13 deletions. Five previously named genes, 69, soc, mrh, modA, and dda were mapped relative to the deletion endpoints. Nine additional ORFs were found interspersed between them. One of these shares some similarities with mrh (modulates rpoH; Frazier and Mosig, Gene 88 (1990) 7-14), and another one resembles modA (coding for an ADP-ribosyl-transferase that modifies RNA polymerase alpha subunits, Skórko et al., Eur. J. Biochem. 79 (1977) 55-66) respectively. We found that the host's heat shock sigma factor, sigma32, is phosphorylated, and that Mrh protein modulates this phosphorylation. The ORF dda.9 downstream of mrh has a patchy similarity with conserved C-terminal segments (motifs) of sigma32; therefore, we call it srh. Another ORF, dda.2 located between modA and dda, shares sequence similarity with sigma70, and we call it srd. We consider the possibility that Srh and Srd act as decoys for sigma32, or sigma70, respectively. Expression of several of the ORFs from cloned DNA appears to be toxic to the host bacteria. Mutant clones only could be constructed from gene 69 and from modA. Moreover, dda.2 (srd)-containing bacteria grow extremely slowly, and they form filaments in liquid cultures. Clones carrying mrh and srh show less severe filamentation. Our results highlight the importance of 'non-essential' genes for phage development and evolution.

Combined myxoid liposarcoma and angiolipoma of the spermatic cord.

To date, only 14 cases of myxoid liposarcoma of the spermatic cord have been reported in the literature. The growth is a rare, usually highly differentiated malignancy that tends to recur locally, rarely metastasizes, and has a good prognosis following complete removal. We describe a new case of a highly differentiated myxoid liposarcoma of the spermatic cord, combined with an angiolipoma. Left inguinal orchiectomy with high ligation of the cord was performed. Staging revealed no evidence of regional or distal metastases. By the latest visit, at 30 months, the tumor had not recurred. The diagnosis, management, and prognosis of this oncologic entity are discussed in the light of the literature.

EEG development in early treated PKU patients from birth to 6 years of age.

In 126 early treated PKU patients (type I and type II) a close EEG follow up was performed from birth up to 6 years of age. A total of 1465 EEGs were performed before and after onset of dietary treatment and on 11 more subsequent occasions. The composition of the background activity was normal up to 6 years when only a small number of the children (19) showed no dominant alpha activity. The frequency of epileptiform activity of generalised as well as focal type was low in the first 2 years of life, but afterwards slightly enhanced in comparison to normal control groups. Other findings like generalised theta paroxysms or focal slow waves were rarely observed. Under a standardised protein load at 6 months (52 patients) and at 5 years of age (42 patients) a moderate generalised slowing of the background activity but no other abnormalities were noted.