Establishment of methods to analyze the structural and functional integrity of the quail (Coturnix coturnix japonica) sperm plasma membrane.

1. The objectives of this study were to establish the use of the fluorophores Hoechst 33342 and propidium iodide for the evaluation of sperm plasma membrane integrity and to identify an adequate hypoosmotic solution for the evaluation of sperm membrane functionality in quails.2. Sperm samples were collected from the vas deferens of nine quails. After initial evaluation, the samples were subjected to a flash-frozen assay. Three treatments with the following proportions of fresh sperm and sperm subjected to flash freezing were prepared as follows: 100:0 (T100), 50:50 (T50), and 0:100 (T0). The hypoosmotic swelling test used distilled water (0 mOsm/l) and fructose solutions (50, 100, and 200 mOsm/l).3. Immediately after recovery, the samples showed 75.6 ± 5.0% motility with vigour of 3.7 ± 0.3 and 96.1 ± 0.5% of the sperm appeared normal. The membrane integrity test showed 62.2 ± 5.2% intact sperm at T100, 29.0 ± 4.1% at T50 and 0.1 ± 0.1% at T0. Moreover, a greater number of reactive sperm (74.7 ± 6.7%) were observed when incubated in distilled water (0 mOsm/l) in comparison to other solutions (P < 0.05).4. The association of fluorescent probes composed of Hoechst 33342 and propidium iodide provided an efficient assessment of the integrity of the plasmatic membrane of quail spermatozoa. However, the study identified that the hypoosmotic swelling test has little predictive value regarding sperm membrane functionality in this species.

Pyricularia graminis-tritici, a new Pyricularia species causing wheat blast.

Pyricularia oryzae is a species complex that causes blast disease on more than 50 species of poaceous plants. Pyricularia oryzae has a worldwide distribution as a rice pathogen and in the last 30 years emerged as an important wheat pathogen in southern Brazil. We conducted phylogenetic analyses using 10 housekeeping loci for 128 isolates of P. oryzae sampled from sympatric populations of wheat, rice, and grasses growing in or near wheat fields. Phylogenetic analyses grouped the isolates into three major clades. Clade 1 comprised isolates associated only with rice and corresponds to the previously described rice blast pathogen P. oryzae pathotype Oryza (PoO). Clade 2 comprised isolates associated almost exclusively with wheat and corresponds to the previously described wheat blast pathogen P. oryzae pathotype Triticum (PoT). Clade 3 contained isolates obtained from wheat as well as other Poaceae hosts. We found that Clade 3 is distinct from P. oryzae and represents a new species, Pyricularia graminis-tritici (Pgt). No morphological differences were observed among these species, but a distinctive pathogenicity spectrum was observed. Pgt and PoT were pathogenic and highly aggressive on Triticum aestivum (wheat), Hordeum vulgare (barley), Urochloa brizantha (signal grass), and Avena sativa (oats). PoO was highly virulent on the original rice host (Oryza sativa), and also on wheat, barley, and oats, but not on signal grass. We conclude that blast disease on wheat and its associated Poaceae hosts in Brazil is caused by multiple Pyricularia species. Pyricularia graminis-tritici was recently found causing wheat blast in Bangladesh. This indicates that P. graminis-tritici represents a serious threat to wheat cultivation globally.

Reduction of interleukin 1 and interleukin 2 production by patients with advanced cancer.

1. The concentrations of interleukin 1 (IL-1) and interleukin 2 (IL-2) produced in the supernatants of peripheral blood mononuclear cell cultures from patients with advanced cancer were measured to identify some of the causes of the immunological impairment characteristic of malignant disease. 2. Mononuclear cells obtained from 19 cancer patients were stimulated to produce IL-1 and IL-2 and compared with those of healthy controls. A severe reduction of both IL-1 and IL-2 activity was observed. 3. There was no correlation between the lower number of OKT4+ cells observed in these patients and the levels of IL-2 production. The removal of monocytes did not bring IL-2 levels to normal. Impaired IL-2 production could not be restored to normal by addition of IL-1. 5. These results suggest that exogenous IL-1 and IL-2 may be useful in cancer immunotherapy.

Reduction of interleukin 1 and interleukin 2 production by patients with advanced cancer

The concentrations of interleukin 1 (IL-1) and interleukin 2 (IL-2) produced in the supernatants of peripheral blood mononuclear cell cultures from patients with advanced cancer were measured to identify some of the causes of the immunological impairment characteristic of malignant disease. Mononuclear cells obtained from 19 cancer patients were stimulated to produce IL-1 and IL-2 and compared with those of healthy controls. A severe reduction of both IL-1 and IL-2 activity was observed. There was no correlation between the lower number of OKT4+ cells observed in these patients and the levels of IL-2 production. The removal of monocytes did not bring IL-2 levels to normal. Impaired IL-2 production could not be restored to normal by addition of IL-1. These results suggest that exogenous IL-01 and IL-2 may be useful in cancer immunotherapy