Endothelial cells inhibit receptor-mediated superoxide anion production by human polymorphonuclear leukocytes via a soluble inhibitor

Confluent monolayers of bovine pulmonary artery endothelial cells (BPAE) or human umbilical vein endothelial cells (HUVE) inhibited by 80 to 90% the production of O2- by added human neutrophils (PMNs) stimulated by plasma membrane receptor-mediated activators (formylmethionylleucylphenylalanine [fMLP], opsonized zymosan, heat-killed Staphylococci), but not by non-plasma membrane receptor-mediated activators (phorbol myristate acetate and delta-hexachlorocyclohexane). Degranulation induced by fMLP was also inhibited by BPAE. Inhibition was not affected by eicosatetraynoic acid (ETYA) or indomethacin. To assess the role of cell-cell contact, 0.45-microns-pore culture plate inserts were employed to prevent PMN-endothelial cell contact during incubation. A similar amount of inhibition of stimulated PMNs superoxide production was seen as compared to PMN-endothelial incubations where contact occurred. A soluble component released by BPAE monolayers, when added to PMNs, duplicated the inhibition seen by BPAE-PMN co-incubation. Incubation of BPAE with adenosine deaminase did not reduce inhibition of O2- production compared to controls without adenosine deaminase. There was no evidence of endothelial scavenging of O2- generated by hypoxanthine-xanthine oxidase, and inhibition of endothelial superoxide dismutase did not diminish the inhibitory effort. We conclude that cell contact is not required for BPAE inhibition of fMLP-stimulated O2- production by PMN, and that scavenging of superoxide anion is not the mechanism. The inhibitor appears to be a polypeptide with an apparent molecular weight between 1,000 and 10,000 D and does not appear to be adenosine, an arachidonate metabolite, or superoxide dismutase. The mechanism may involve down-regulation of plasma membrane receptor-mediated activation of PMNs.     

Studies on the submicroscopic structure of botryoid (rhabdomyo)-sarcomas of the nasopharynx (author's transl)

Embryonal rhabdomyosarcomas and their botryoid variant are typical soft tissue sarcomas of infancy and childhood. Generally, the tumor cells are thought to be comparable with embryonal rhabdomyoblasts. Nevertheless, many authors do not require the cross-striation of the tumor cell cytoplasm as cytological marker for the diagnosis "rhabdomyosarcoma". In our study we examined the cytology of tissue samples of 4 botryoid sarcomas of nasopharynx with the electron microscope. A varying cytological picture was found. Besides undifferentiated tumor cells constituting the main part of the growths cellular elements with resemblance to fibroblasts, smooth and striated muscle cells, histiocytic cells and lipoblasts were observed. Furthermore, a vasoformative potency of the tumor tissue was occasionally seen. The conclusion is drawn from our results that, replacing the term embryonal rhabdomyosarcoma, such tumors should better be designated as embryonal sarcomas. We think it probable that the present concept of embryonal rhabdomyosarcomas must be revised.     

Detection and identification of fungi from fungus balls of the maxillary sinus by molecular techniques

The aim of this study was to find a reliable method for the detection and identification of fungi in fungus balls of the maxillary sinus and to evaluate the spectrum of fungi in these samples. One hundred twelve samples were obtained from patients with histologically proven fungal infections; 81 samples were paraffin-embedded tissue sections of the maxillary sinus. In 31 cases, sinus contents without paraffin embedding were sent for investigation. PCR amplification with universal fungal primers for 28S ribosomal DNA and amplicon identification by hybridization with species-specific probes for Aspergillus fumigatus, Aspergillus flavus, Aspergillus niger, Aspergillus terreus, Aspergillus glaucus, Pseudallescheria boydii, Candida albicans, and Candida glabrata were performed for all samples. Furthermore, PCR products were sequenced. Fresh samples were also cultivated. Fungal DNA was detected in all of the fresh samples but only in 71 paraffin-embedded tissue samples (87.7%). Sequence analysis was the most sensitive technique, as results could be obtained for 28 (90.3%) fresh samples by this method in comparison to 24 (77.4%) samples by hybridization and 16 (51.6%) samples by culture. However, sequence analysis delivered a result for only 36 (50.7%) of the paraffin-embedded specimens. Hybridization showed reliable results for A. fumigatus, which proved to be the most common agent in fungus balls of the maxillary sinus. Other Aspergillus species and other genera were rarely found.     

Biological balance of sodium and potassium

Summary The rhythm of renal sodium and potassium excretion was measured in 4-h-intervals in 12 subjects. Each person exhibited clear circadian variations of each variable with a maximum between 8 a.m. and 4 p.m. In each subject and for both circadian rhythms the oscillation mean was correlated to the range of oscillation (amplitude).Increase in sodium or potassium excretion during 1 day resulted in an increase of oscillation range. The oscillation means of sodium and potassium periodicity did not correlate.The properties of biological control systems with oscillating correcting variables are comparable to those of technical control systems. The significance of circadian rhythm for the control of electrolyte balance is indicated.     

Influenza virus infection of mouse lymphoblasts alters the binding affinity of anti-H-2 antibody: Requirement for viral neuraminidase

The requirement for histocompatibility in the T lymphocyte killing of virus-infected cells has led us to investigate the effect of influenza virus infection on mouse cell surface histocompatibility (H-2) antigens. Monoclonal anti-H-2 antibody made it possible to develop equilibrium binding conditions for the assay of H-2 antigen-antibody interactions on intact cells. Scatchard analysis of anti-H-2 binding with normal and virus-infected cells yielded linear curves indicating homogeneity of the interaction at varied concentrations of antibody through saturating levels. The estimated number of 2 × 10⁵ - 5 × 10⁵ H-2 antigenic sites per mouse lymphoblast does not appear to change during the course of influenza virus infection. However, the K_a (binding affinity constant) of anti-H-2 binding is rapidly elevated by virus infection (“0” time), continues to increase for 3 h post infection, then decreases. Control cells, treated with normal egg allantoic fluid, show no change in K_a during similar incubation. This change in K_a requires the presence of active viral neuraminidase. Thermal denaturation of the neuraminidase of the virus particles abolishes their ability to induce K_a alteration, even though hemagglutinin activity is retained. Treatment of cells with neuraminidase of bacterial origin led to an elevation of K_a, but did not mimic the viral effect in time dependence and magnitude of peak responses. The time-dependent lowering of K_a from peak values appeared to relate to virus replication, since UV light-inactivated virus-induced K_a elevation, but did not produce the typical K_a decline at 4-5 h post infection. The changes in K_a of anti-H-2 binding during influenza infection reflects a virus- induced alteration of the H-2 molecule or its environment in the host cell membrane. The molecular basis of this change and its relation to H-2-restricted recognition of influenza virus-infected cells by cytotoxic T cells requires further study.     

Assessing the effect of posture change on tactile inhibition-of-return

If a peripheral target follows an ipsilateral cue with a stimulus-onset-asynchrony (SOA) of 300 ms or more, its detection is delayed compared to a contralateral-cue condition. This phenomena, known as inhibition-of-return (IOR), affects responses to visual, auditory, and tactile stimuli, and is thought to provide an index of exogenous shifts of spatial attention. The present study investigated whether tactile IOR occurs in a somatotopic vs an allocentric frame of reference. In experiment 1, tactile cue and target stimuli were presented to the index and middle fingers of either hand, with the hands positioned in an uncrossed posture (SOA 500 or 1,000 ms). Speeded target detection responses were slowest for targets presented from the cued finger, and were also slower for targets presented to the adjacent finger on the cued hand than to either finger on the uncued hand. The same pattern of results was also reported when the index and middle fingers of the two hands were interleaved on the midline (experiment 2), suggesting that the gradient of tactile IOR surrounding a cued body site is modulated by the somatotopic rather than by the allocentric distance between cue and target.     

Mapping and ranking of potential cytotoxic T epitopes in the p53 protein: effect of mutations and polymorphism on peptide binding to purified and refolded HLA molecules

In many cancer cells, the p53 gene displays point mutations that result in stabilization and accumulation of the p53 protein. Therefore, p53 peptides could be presented to the immune system by tumor cells; thus, p53 might be a suitable target antigen for developing an immunotherapy against tumors using cytotoxic T lymphocytes (CTL). To map candidate CTL epitopes, we synthesized 150 peptides of 8–11 residues that contained putative anchor motifs required for binding to common HLA class I molecules. They were tested for their capacity to promote the assembly of purified and refolded HLA-A1, A2, B7 and B8 molecules. The following wild-type p53 peptides were found to be reactive with the HLA molecules tested: 196–205 and 226–234 bound moderately to HLA-A1; 25–35, 65–73, 129–137, 187–197, 263–272 and 264–272 bound strongly, and 187–195 and 256–264 moderately to HLA-A2; 26–35, 63–73, 189–197, 249–257 and 321–330 bound strongly to HLA-B7; and 135–143, 210–218 and 375–383 bound weakly to HLA-B8. We also analyzed the effects of p53 mutations occurring naturally in tumors on peptide/HLA assembly. We found substitutions that enhanced, diminished or had no effect on the peptide binding to HLA molecules. Polymorphism at position 72 mainly affected peptide/HLA-B7 binding, the proline allele P72 giving a less-reactive peptide (63–73) than the arginine allele R72. We have ranked potential p53 epitopes according to their reactivity for purified HLA molecules, allowing the selection of appropriate peptides and HLA molecules to attempt CTL induction in vitro.     

An inhibitor of cytotoxic functions produced by CD8+CD57+ T lymphocytes from patients suffering from AIDS and immunosuppressed bone marrow recipients

An inhibitor of the cytotoxic functions (ICF) mediated by human immunodeficiency virus (HIV)- or HLA-specific cytotoxic T lymphocytes, natural killer and lymphokine-activated killer (LAK) cells is secreted by CD8⁺CD57^? T lymphocytes, a subset expanded during infection with HIV and after bone marrow transplantation. We previously showed an apparent molecular mass of 20–30 kDa for this soluble glycosylated concanavalin A-binding inhibitor which is distinct from known cytokines. Here, we report a characterization of the mechanism of action of this CD8⁺CD57⁺ ICF. We show that the ICF-induced inhibition of LAK cell cytolytic activity is transient, with a spontaneous recovery of cytolytic potential after 18 h. When testing interactions of ICF with a large set of cytokines we found that the ICF-mediated inhibition of cytotoxic functions is antagonized by two cytokines: recombinant interleukin (rIL)-4 and recombinant interferon (rIFN)-γ. Finally, we show that ICF acts at the level of cytolytic effector cells, where it induces a significant increase of cyclic AMP (cAMP) level. In contrast, no modification of either cell surface antigen expression or of target/effector cell conjugate formation could be evidenced. Addition of rIL-4 and rIFN-γ reverses such an increase of cAMP levels and in parallel restores the cytolytic activity. Altogether, these data demonstrate that the glycoprotein ICF produced by CD8⁺CD57⁺ cells (1) inhibits cell-mediated cytotoxicity by sensitizing cytolytic effector cells to the cAMP pathway, and (2) is part of a cytokine network controlling cell-mediated cytotoxic functions.     

Trophic action of pharmacological substances with a guanidine group on mouse neuroblastoma cells and chick ganglionic neurons in culture

A series of substances (designated CTQ compounds) with a guanidine group have been synthesized and tested for their ability to promote neuronal survival and neurite outgrowth. Mouse neuroblastoma clonal cell lines grown in serum-containing medium for 10 days as well as primary cultures of embryonic chicken ganglion neurons grown in serum-free defined medium for 1 or 2 days have been used for the experiments. Among the various CTQ compounds (CTQ1–CTQ20) tested, only CTQ8 exerted positive neurotrophic effects on these peripheral neuronal cells. At a concentration of 10⁻⁴ M, CTQ8 enhanced neuritogenesis of neuroblastoma cells. However, the most striking influence of CTQ8 was its promoting effect (6- to 10-fold) on the survival of chicken ciliary and dorsal root ganglionic neurons at concentrations ranging from 10⁻³ M to 5×10⁻⁴ M.     

Microvascular surgery as an adjunctive tool in renal transplantation

There has been a serious shortage of suitable kidneys for transplantation since this procedure became the treatment of choice for many patients with end-stage renal failure. Some harvested kidneys are discarded due to complicated or injured renal vasculature and some potential living related donors are judged unsuitable because their kidneys have multiple vessels. The authors review the basic microsurgical techniques they have used in such situations to salvage kidneys for transplantation. They emphasize the ex-vivo, "bench", microsurgical method for protecting the kidney from prolonged warm ischemia time (as with multiple complicated in-situ anastomoses). Several illustrative case reports from their recent experience are presented. The authors conclude that microvascular surgery is an important adjunct to the armamentarium of the transplant surgeon.