Dental records: a Belgium study.

The aim of this study was to deduce the quality of the average dental record kept by Belgian dentists and to evaluate its potential use for forensic dental casework. The evaluated material originated from 598 Dutch speaking and 124 French speaking Belgian dentists who completed a questionnaire and returned it by mail or through the internet. The age of the participating dentists ranged from 22 to 72 years of age. The results of the inquiry were statistically analysed taking parameters such as language, gender, age, university and ZIP code into account. In general there was a tendency for the young dentists from the age category 22 to 34 years of age, especially those living in larger cities, to perform better on several of the questions asked such as completion of the dental record, storage of x-rays, working with digital x-rays and a digital dental record.

Macrophage subpopulations in the mouse spleen renewed by local proliferation.

A dual origin, by both immigration of blood monocytes and local proliferation has been reported for macrophages, which are heterogeneous in several aspects. Until now few studies have focussed on renewal of macrophages. Therefore, in this study macrophage renewal by local proliferation has been studied in the spleen. Spleen tissue of mice which had received BrdU intravenously was investigated immunohistochemically to detect BrdU incorporated by macrophages. BrdU incorporation by splenic macrophages was studied under steady state conditions as well as during repopulation after experimental macrophage depletion. Under steady state conditions, BrdU incorporation was found in 3.0 +/- 0.5% of the white pulp macrophages and in approximately 1% of the metallophilic macrophages only. Comparable results were found during repopulation after experimental macrophage depletion. The data suggest that renewal of macrophages by local proliferation is restricted to certain subsets in the spleen and that the macrophage subsets of the spleen are renewed by different mechanisms, or belong to different lineages of macrophage differentiation.

Effects of the modulating agent WR2721 on myelotoxicity and antitumour activity in carboplatin-treated mice.

The selective modulation of carboplatin [diammine(1,1-cyclo-butanedicarboxylato)platinum(II)]-induced myelotoxicity was investigated in mice, using the protective agent WR2721 [S-2-(3-aminopropylamino)ethyl-phosphorothioic acid, ethiofos]. In female BALB/c mice, WR2721 (200 mg/kg intraperitoneally, i.p.) partly prevented the reduction of in vitro proliferation of whole bone marrow cells and non-adherent cells when administered at different time points relative to 90 mg/kg carboplatin (i.p.). Protection was highest when WR2721 was administered 5 min prior to carboplatin. In vitro proliferation of whole bone marrow cells and non-adherent cells in liquid culture increased from 15% of control for carboplatin alone to 45% when WR2721 was administered 5 min prior to carboplatin. However, WR2721 did not significantly prevent the loss in clonogenic capacity of early hematopoietic progenitors in the bone marrow, as determined by a bilayered soft agar colony forming units assay. In nude mice, bearing well-established subcutaneous human ovarian carcinoma xenografts OVCAR-3, WR2721 (200 mg/kg i.p.) 5 min prior to intravenous carboplatin allowed a 1.5-fold increase in the maximum tolerated dose of carboplatin as determined by overall weight loss. WR2721 alone did not affect tumour growth. However, WR2721 had a potentiating effect on the tumour growth inhibition of a standard dose of carboplatin in this model. Minimal tumour volume compared to control (T/C) decreased from 9.4% with carboplatin alone to 2.2% with WR2721 5 min prior to the same dose of carboplatin. Specific growth delay (SGD) increased from 7.4 to 10.3. With the 1.5-fold increased, equitoxic dose of carboplatin in combination with WR2721, the antitumour activity was only slightly further increased (T/C = 1.4%, SGD = 10.5).

Establishment of bone marrow-derived macrophage cell lines by soft-agar cloning: regulation of macrophage heterogeneity at bone marrow level.

To study the regulation of macrophage (m phi) heterogeneity at bone marrow level, we developed a liquid culture system in which bone marrow-derived soft-agar colonies were expanded in the presence of colony-stimulating factor-1 (CSF-1). Several cloned m phi precursor cells were established as CSF-1-dependent cell lines, and were analyzed for morphological, phenotypic and functional characteristics. The continuously proliferating cell lines expressed both immature and mature m phi markers. Only minor differences between the established cell lines were detected. Thus, our results show that during long-term culture CSF-1-responsive cloned m phi precursor cells develop along identical pathways, giving rise to progeny with comparable phenotype and function in vitro.

Macrophage phenotype regulation by colony-stimulating factors at bone marrow level.

Macrophages (m phi s) can be divided into several subpopulations, which differ in phenotype, function, and localization patterns. However, little is known about the mechanisms that regulate this heterogeneity. We investigated whether m phi heterogeneity is regulated by colony-stimulating factors (CSFs) at the bone marrow level. By clonal expansion of bone marrow-derived precursor cells in the presence of CSF-1, granulocyte-macrophage CSF or multi-CSF (interleukin-3), phenotypic heterogeneity was observed between m phi colonies. Heterogeneity was found especially when different CSF culture conditions were used but also between m phi colonies derived under the same CSF culture condition. Our results illustrate that CSFs from the bone marrow hemopoietic microenvironment are important for the induction of phenotypic heterogeneity within the progeny of cloned m phi precursor cells during maturation and differentiation in vitro.

Milky spots in the mouse omentum may play an important role in the origin of peritoneal macrophages.

In the milky spots, precursors of cells belonging to the mononuclear phagocyte system (MPS), such as monoblasts, can be found on the basis of ultrastructural endogenous peroxidase cytochemistry. Therefore, in the present study, we investigated the milky spots using a panel of monoclonal antibodies, especially antibodies (ER-MP) that recognize macrophage precursor antigens. Early macrophage precursor antigens ER-MP12 and ER-MP58 were detected only on cells localized inside the milky spots. On the other hand, an antigen which disappears late in the course of macrophage differentiation, ER-MP20, was detected in high amounts on cells both inside and around the milky spots. This clearly indicates that macrophage precursors are centrally localized inside the milky spots, while more differentiated cells are found in peripheral areas. Moreover, long-term culture of milky spot tissue resulted in the forming of a monolayer of stromal cells which supported macrophage proliferation in vitro. In conclusion, both in situ and in vitro studies demonstrated that mouse milky spots have a microenvironment in which precursor cells of the MPS can home and proliferate, illustrating that milky spots play a role as a source of local macrophage generation, e.g. that of the free peritoneal macrophages.

Chronic inflammation induces the expression of dendritic cell markers not related to functional antigen presentation on peritoneal exudate macrophages.

Phenotypic heterogeneity between macrophages in situ has been reported, using different macrophage-specific monoclonal antibodies. Microenvironmental factors, which may play an important role in inducing this heterogeneity were studied in a local inflammatory response in vivo. Our results show that peritoneal exudate macrophages, formed during Thioglycollate induced chronic inflammation, acquire expression of the dendritic cell markers NLDC-145 and 6D2. In contrast to these phenotypic characteristics, NLDC-145+/6D2+ exudate cells do not function like dendritic cells in an allogeneic-MLR, but are suppressive when compared with steady state peritoneal cells. The expression of these membrane markers on exudate cells is discussed.

Different epitopes on the dendritic cell-associated NLDC-145 molecule during ontogeny.

A monoclonal antibody, 6D2, is described that recognizes a different epitope on the NLDC-145 dendritic cell associated molecule in the mouse. During ontogeny the epitope appears on interdigitating cells in lymphoid organs only around birth, whereas the NLDC-145 antigen can be detected as early as day 16 of gestation. No differences can be observed in the expression of the two antigenic determinants on the epithelial cells of the thymus during ontogeny. Evidence is presented that the two antibodies recognize different epitopes on the same molecule.

Functional and molecular characterization of B cell line derived interleukin-1 alpha.

The cytokine secreted by a human hybrid B cell line (STS 25) obtained by fusion of the B lymphoblastoid cell line WI-L2-729-HF2 with neoplastic B cells from a patient with B cell non-Hodgkin's lymphoma (B-NHL) was characterized as IL-1 alpha. STS 25 cells express the idiotypic (Id+) immunoglobulin (Ig) specific for the neoplastic B cells of the B-NHL patient. STS 25 cells are weakly positive for surface mu delta kappa and in addition express the surface markers CD19, CD20, CD23, HLA class I and II, and the 4F2 activation antigen. STS 25 cells are also Epstein-Barr nuclear antigen positive but do not secrete viral particles. Serum-free culture supernatant from STS 25 cells (STS 25 SUP) does not show activity in assays for interleukin-2 (IL-2), -4 (IL-4), -6 (IL-6), interferon or tumor necrosis factor, but is active in the thymocyte costimulation assay and the D10.G4.1 T helper clone proliferation assay for interleukin-1 (IL-1). The IL-1 character of the STS 25 SUP activity was confirmed in inhibition studies with three different poly- or monoclonal anti-IL-1 antibodies (31, 88, and 94% inhibition in thymocyte costimulation assay, respectively). Furthermore, complete blocking of D10.G4.1 cell proliferation mediated by STS 25 SUP was observed by including anti-IL-1 alpha specific antibody in the assay, whereas anti-IL-1 beta antibody had no effect. These results indicate that this STS 25 SUP activity can be attributed to the presence of IL-1 alpha in the supernatant. Northern blot analysis of total STS 25 cellular RNA using IL-1 alpha or IL-1 beta specific probes revealed the constitutive expression of IL-1 alpha messenger RNA by STS 25 cells. In contrast, no IL-1 beta message was detectable, not even after treatment of the cells with phorbol ester or cycloheximide, which resulted in approximately 5-fold enhancement of IL-1 alpha mRNA expression. Binding studies with radiolabeled recombinant (r) IL-1 alpha indicated the presence of high numbers of IL-1 receptors on STS 25 cells (1,170 per cell, Kd = 392 pM). Although both IL-1 alpha and IL-1 beta bound to these IL-1 receptors, no indication was found for IL-1 mediated regulation of STS 25 cell growth.(ABSTRACT TRUNCATED AT 400 WORDS)

HLA expression and function in single and double HLA-B27-transgenic mice.

The expression and function of HLA antigens in mice single transgenic for HLA-B27.2 (sTGM-B27.2) or double transgenic (dTGM) for HLA-B27.2 and human beta 2-microglobulin (h beta 2m) were compared. B27.2 could be well detected on the cell membrane of lymphocytes of sTGM. However, the expression in sTGM was much lower than in dTGM mice. Nevertheless, also in sTGM mice, the B27-transgene product possessed all functional properties of a class I HLA molecule. This was shown by the recognition and induction of antibodies and cytotoxic T cells, by the induction of "allo"-immunity, including skin graft rejection, and by the ability to present viral antigens. In dTGM, the expression of B27 on peripheral blood lymphocytes, spleen and lymphnode cells was comparable to H-2. However, on thymocytes, a relatively lower expression of HLA than H-2 was observed. This low expression of B27 on thymocytes is in concert with the observation that B27 is expressed only in the medulla of the thymus and not detectable in the cortex.

Recognition of xeno-(HLA, SLA) major histocompatibility complex antigens by mouse cytotoxic T cells is not H-2 restricted: a study with transgenic mice.

Cytotoxic T lymphocytes (CTLs) recognize antigens in the context of major histocompatibility complex (MHC) class I gene products. The T-cell receptor (TCR) that mediates this MHC-restricted antigen recognition recognizes short peptide fragments rather than the intact antigen. Presentation of peptides to the TCR may thus be a major function of the MHC. An intriguing question emerging from this model is whether peptide presentation also applies to foreign MHC antigens and which of the available MHC molecules can present preferentially the peptides of the foreign MHC molecule. Allo- and xenoreactive CTLs might either recognize native MHC class I molecules or peptides presented by self MHC or by the foreign class I MHC itself. The finding that synthetic peptides corresponding to MHC class I regions are recognized by allo- and xenoreactive CTLs suggests that recognition of foreign MHC by CTLs might involve degraded fragments presented by syngeneic class I molecules. We used MHC transgenic mice as a tool to study these questions. The CTL responses against human (HLA) antigen B27 were analyzed by using HLA-B27 transgenic mice with various H-2 haplotypes. We report here that mouse xeno-MHC-specific (anti-B27) CTLs are perfectly able to kill human and mouse cells expressing the appropriate xenoantigen and that in primary and secondary responses to xeno-MHC, the mouse T-cell repertoire does not use self-H-2 as a restriction element. Absence of H-2 restriction was confirmed by the lack (less than 1/10(6] of H-2-restricted HLA-specific CTL precursors. Therefore, H-2-restricted recognition of xeno-MHC antigens cannot be generalized as part of a classical MHC class I-specific response. These results indicate that xenoreactive CTLs usually recognize intact MHC molecules or MHC peptides preferentially presented by their native MHC molecule. We suggest the latter possibility.

Anorexia nervosa with transient hypopituitarism.

A case is presented with sudden onset of cachexia and anorexia with hypopituitarism, starting early and progressing gradually. After about 15 months the patient recovered; first he lost the anorexia, then the endocrine functions and growth became normal. Conflicting reports about hypopituitarism in anorexia nervosa and similar syndromes may be due to lack of longitudinal observations and the transient nature of the endocrine disorders.