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The human leukocyte test system. V. DNA synthesis and mitoses in PHA-stimulated 3-day cultures.

In human leukocyte cultures st up with TC medium 199,DNA synthesis and mitotic indices were analysed by means of 3H-thymidine autoradiography and cell counting. DNA synthesis starts at around 28 hrs. The frequencies of labelled cells rise slowly and reach a maximum of around 24%. The first mitoses appear at around 38 hrs but up to 49 hrs only very few mitoses can be seen. After that time the mitotic indices rise and reach values of up to 11% cultivation in the presence of BudR for 72 hrs and staining with Hoechst 33258 stain revealed that first, second and third mitoses occur together in the cultures at this time. Irradiation of whole blood and cultivation for 72 hrs leads to mitoses containing dicentric and ring chromosomes with and without fragments, to interphases with micronuclei, to premature chromosome condensations (PCC) and to polyploid mitoses indicating that at this time first and further mitoses are present.

Autoradiography↗

PHA storage from several substrates by different morphological types in an anoxic/aerobic SBR.

An activated sludge was cultivated on a mixture of several soluble substrates (acetate, ethanol, glucose, glutamic acid, peptone, Tween 80, starch, yeast extract) in an anoxic/aerobic SBR. Highly dynamic conditions in the SBR (feast famine regime) caused fast removal of most COD in the anoxic phase (in particular acetate, ethanol, glutamic acid and glucose were totally removed) and relevant contribution of storage. In spite of that, filament abundance was always high, as is typical of bulking sludges. Filaments which developed in the reactor were characterized on a morphological basis and on the basis of their ability to grow and to store polyhydroxyalkanoates (PHAs). Three main filaments prevailed in the biocenosis, whose relative abundance was varyng with time: Nostocoida limicola II, (two different morphological types), Haliscomenobacter hydrossis and an unidentified one. It was found that maximum growth rate was higher for flocformers than for filaments on each of the tested substrates. Epifluorescence showed that storage ability was more widespread among flocformers than in the filaments. Only one type of Nostocoida limicola II was able to store PHAs. The obtained data show that aerobic growth on the little residual fraction of COD from the anoxic phase was enough to support high abundance of filamentous microorganisms.

Bioreactors↗

Cytokine release of whole blood from adult female donors challenged with mistletoe lectin-1 standardised mistletoe extract and E. coli endotoxin or phytohaemagglutinin (PHA).

Cytokine (TNF, alpha, interleukin-6) release of whole blood from healthy donors was challenged with mistletoe extract standardised for mistletoe lectin-1 (sML) and control substances (E. coli endotoxin; phytohaemagglutinin). The rationale of this investigation was non-proven warnings that pro-inflammatory cytokines induce by the application of standardised mistletoe lectins may induce tumor cell proliferation. These investigations provided evidence that non-cytotoxic concentrations of sML did not induced enhanced TNFa or interleukin-6 secretion compared to non-challenged control cells. Cytotoxic concentrations of sML, however, induced significantly higher cytokine levels than the control, obviously due to non-physiological stimuli. Immunomodulation with clinically relevant, low-dose sML incubation did not induce proinflammatory cytokine secretion in vitro.

Adult↗

[Comparative study on lymphocyte stimulation by PHA and bacterial endotoxins, particularly in malignant tumor and allergy patients].

In a group of 37 tumor patients with predominantly advanced disease, LPS stimulation (LPS of Sh. sonnei and Sh. flexneri) resulted in a significant increase of the transformation rates caused, most probably, by the high proportion of B cells. On the other hand, allergics (drug allergies) did not differ from controls. It seems possible that B cell stimulation by bacterial endotoxins can be used for the assessment of the reactivity of the organism.

Adult↗

[An evaluation of the scope of the circulation of Salmonella among the workers of commercial poultry- and meat-packing enterprises based on serological study data in the PHA test].

Serum samples from 641 workers of large poultry and meat-packing plants were studied in the passive hemagglutination test with the use of Salmonella complex and serogroup diagnostica. A specific increase in the level of anti-Salmonella antibodies in 60.7% of poultry plant workers and in 9.8% of meat-packing plant workers was established. Among the workers of the poultry plants the most pronounced immune shifts were detected in persons having contacts with sick poultry and pathological material and among the employees of the meat-packing plant, in those who ate raw sausage meat. A high level of antibodies in the professional groups under study was observed as early as in the first year of work at the plant and persisted over the whole period of this work. Under the conditions of constant contamination of the workers of poultry and meat-packing plants with small doses of salmonellae specific immunity to this infection was seemingly induced, which inhibited the development of the manifest forms of infection, but did not prevent the formation of chronic carrier state.

Adult↗