Bottlenose dolphins (Tursiops truncatus) do also cast neutrophil extracellular traps against the apicomplexan parasite Neospora caninum.

Neutrophil extracellular traps (NETs) are web-like structures composed of nuclear DNA decorated with histones and cytoplasmic peptides which antiparasitic properties have not previously been investigated in cetaceans. Polymorphonuclear neutrophils (PMN) were isolated from healthy bottlenose dolphins (Tursiops truncatus), and stimulated with Neospora caninum tachyzoites and the NETs-agonist zymosan. In vitro interactions of PMN with the tachyzoites resulted in rapid extrusion of NETs. For the demonstration and quantification of cetacean NETs, extracellular DNA was stained by using either Sytox Orange® or Pico Green®. Scanning electron microscopy (SEM) and fluorescence analyses demonstrated PMN-derived release of NETs upon exposure to tachyzoites of N. caninum. Co-localization studies of N. caninum induced cetacean NETs proved the presence of DNA adorned with histones (H1, H2A/H2B, H3, H4), neutrophil elastase (NE), myeloperoxidase (MPO) and pentraxin (PTX) confirming the molecular properties of mammalian NETosis. Dolphin-derived N. caninum-NETosis were efficiently suppressed by DNase I and diphenyleneiodonium (DPI) treatments. Our results indicate that cetacean-derived NETs represent an ancient, conserved and relevant defense effector mechanism of the host innate immune system against N. caninum and probably other related neozoan parasites circulating in the marine environment.

Molecular cloning of two west African HIV2 isolates that replicate well in macrophages.

HIV2 strains were isolated from a Gambian with neuro-AIDS (HIV2D194) and from an asymptomatic Ghanian (HIV2D205). Like HIV1 biological subtype c, both isolates grew slowly and induced few or no syncytia, but eventually produced high levels of particle-associated reverse transcriptase (RT) in cultures of fresh peripheral blood lymphocytes. Each produced even higher levels of RT in fresh human macrophages, especially HIV2D194, where maximal RT values of 1,800,000 cpm/ml supernatant of approximately 30,000 cells were measured. The viruses were molecularly cloned after a single passage in culture. Restriction site analysis showed heterogeneity within each isolate. Nucleotide sequence analysis of HIV2D194 revealed that, genetically, it is a member of the prototypic HIV2 family, displaying 12% divergence vs. HIV2ROD and HIV2NIHZ. In contrast, HIV2D205 is the most highly divergent HIV2 strain yet described: it is equidistant in relation between the known HIV2 strains and the SIVMAC isolates (23-24% nucleotide sequence divergence).

A highly divergent HIV-2-related isolate.

It has been suggested that the human immunodeficiency virus type 2 (HIV-2) and the simian immunodeficiency virus from rhesus macaques (SIVmac) evolved from the sooty mangabey virus SIVsm (ref. 1). We now describe an HIV-2-related isolate, HIV-2-D205, from a healthy Ghanaian woman that is genetically equidistant to the prototypic HIV-2 strains and to SIVsm and SIVmac. Supported by the observation that HIV-2D205 differs in a step of envelope glycoprotein processing, our data indicate that it could represent an alternative HIV-2 subtype and that viruses of the HIV-2/SIVsm/SIVmac group could have already infected humans before HIV-2 and SIVsm/SIVmac diverged.

[Study of amino acid and carbohydrate metabolism of porcine strains of Haemophilus by thin layer chromatography]. / Untersuchungen des Aminosäure- und Kohlenhydrat-Stoffwechsels porciner Haemophilus-Stämme mittels Dünnschichtchromatographie.

The aim of the present study was to investigate porcine reference and field strains of the species Haemophilus (H.) pleuropneumoniae and H. parasuis, as well as H. Taxon "minor group" and Taxon C on their amino acid and carbohydrate metabolism by thin layer chromatography. The 17 reference strains studied showed almost identical results within the different species and taxa in both, amino acid and carbohydrate metabolism patterns. Based on a few differing enzymatic reactions a reduced species- and taxon-specific reaction pattern could be established, which included L-alanine, L-citrulline and L-threonine of the amino acids as well as D-ribose, alpha-D-xylose, mannitol, trehalose, beta-melibiose and alpha-lactose of the carbohydrates. This differentiation system allowed a reliable identification of 7 field strains whereas 4 additional ones, hitherto pre-classified as H. parasuis, could not be associated with any of the above species and taxa.

[Amino acid metabolism by thin layer chromatography for the differentiation of Bordetella and Alcaligenes species]. / Untersuchungen des Aminosäurestoffwechsels mit der Dünnschichtchromatographie zur Differenzierung von Bordetellen- und Alcaligenes-Spezies.

The amino acid metabolism of Bordetella and Alcaligenes strains was investigated for differentiation purposes. All species investigated reacted differently with a number of amino acids. The genus Bordetella could be differentiated from that of Alcaligenes by its inability to metabolize L-lysine, L-arginine and D-histidine. The recently described species Bordetella avium, as well as Achromobacter xylosoxidans (Yabuuchi), considered as Alcaligenes denitrificans subsp. xylosoxidans, presented themselves as independent species too. Bordetella odorans, now assigned to the species Alcaligenes faecalis was indistinguishable from the other strains of Alcaligenes faecalis investigated.

[Results of treatment of rhabdomyosarcomas (RMS) in children. A report of the Cooperative Soft Tissue Sarcoma Study (CWS-81) of the Society of Pediatric Oncology]. / Ergebnisse der Behandlung von Rhabdomyosarkomen (RMS) bei Kindern. Ein Bericht der Cooperativen Weichteilsarkomstudie (CWS-81) der Gesellschaft für Pädiatrische Onkologie.

This analysis refers to 129 children with RMS who were treated between 1981 and 1985 according to the protocol guidelines of the CWS-81 study. The duration of chemotherapy depended on the initial post-operative stage. Patients with stage I and IIA were not to receive any radiotherapy, and patients with primary stage III were treated according to the results of a 16-week chemotherapy treatment: either without radiation (stage Ipc), or with radiotherapy using 40 Gy (stage (IIpc) or 50 Gy (stage IIIpc). The median time of observation was 27 months (Juni 1985). The essential results of the study are as follow: RMS of the extremities in stages I and IIA need radiotherapy, contrary to all other localisations with the same stages. Patients with RMS stage III who are tumor-free after initial chemotherapy (histologically checked) do not need radiotherapy. Patients with microscopic residue (IIpc) or macroscopic residue (IIIpc) after pretreatment showed no difference in their local relapse rate, whereas metastases were found only in the group having macroscopic tumor residue up to week 16. An additional examination of this study observed tumor response during initial chemotherapy and its relationship to prognosis. This analysis showed that the degree of tumor regression per unit of time permits the most favorable prognostic statement. Patients with clinical complete remission after 7/9 weeks showed a 100% relapse-free chance of survival, independent of localisation, tumor size or histological subtype. Those with a tumor reduction of greater than 2/3 but no complete remission showed 67% chance of survival, and those with greater than 1/3-2/3 tumor reduction had 25%. Tumor response kinetics under initial chemotherapy allows better individual therapy in the future. The overall result of the study concerning localisation and stage corresponds to that of the IRS I and II studies. Patients with undifferentiated sarcomas, extraossary Ewing's sarcoma and synovial sarcomas can be treated according to the same principles as RMS, since no significant differences in prognosis could be found.

[Hyalurodinase activity of beta-hemolytic streptococci of the Lancefield group C]. / Untersuchungen zur Hyaluronidase-Aktivität beta-hämolysierender Streptokokken der Lancefield-Gruppe C.

A total of 110 strains of beta-hemolytic streptococci, belonging to serogroup C (Lancefield), isolated from horses (71 S. zooepidemicus, 27 S. equisimilis and 12 S. equi) as well as 5 reference strains were tested for their ability to produce hyaluronidase. The determinations were carried out in a culture test on agarose gel and in a liquid test system (turbidity test according to DiFerrante). The results of both methods used showed that the three Streptococcus species could be differentiated by the relative quantitative determination of hyaluronidase activity. S. equisimilis strains produce 5 to 10 times more hyaluronidase than those of S. zooepidemicus. The strains of S. equi did not show enzyme production. In addition parallel tests by determination of final N-acetyl-D-glucosamine groups and by turbidity test were done to confirm the hyaluronidase activity of S. equisimilis strains. The comparison of results (Fig. 3) obtained from these tests, showed a good correspondence as demonstrated by a correlation index of 0,93.