The structure and evolution of the spider monkey delta-globin gene

We have isolated the delta-globin gene of the New-World spider monkey, Ateles geoffroyi, and compared its nucleotide sequence with those of other primate delta- and beta-globin genes. Among primate delta-globin genes, the rate of nonsynonymous substitutions is much less than the rate of synonymous substitutions. This suggests that primate delta- globin genes may remain under evolutionary conservation, perhaps because hemoglobin A2 has an as yet unknown physiological importance.      

Distribution and abundance of littoral benthic fauna in Canadian prairie saline lakes

The littoral benthos of 18 lakes in Alberta and Saskatchewan ranging in salinity from 3 to 126 (g1^–1 TDS) were investigated twice, in the spring and in the summer of 1986. Multiple Ekman dredge samples were taken at water depths of about 0.5, 1.0 and 2 metres in each transect. Two to three transects were used in each lake according to its estimated limnological diversity for a total of 114 stations. A total of 76 species was present varying from 29–31 species in the three lakes of lowest salinity (means of 3.1–5.55) to only 2 species in lakes exceeding 100. Species richness decreased rapidly in salinities greater than 15.Biomass maximum mean of 10.91 g m^–2 dry weight (maximum 63.0 g m^–2) occurred in culturally eutrophic Humboldt Lake (3.1) but one third as great in other low salinity lakes. However, biomass again increased to about 4.5 gm^–2 in two lakes of 15 As the salinity increased still further biomass declined steadily until a minimum of 0.0212 g m^–2 was recorded in most saline Aroma Lake (mean 119). Summer biomass (11 lakes) was greater than spring biomass (4 lakes) because some groups such as amphipods, corixids and ostracods became more abundant in summer. Wet weight biomass averaged 15.8 of dry weight biomass.Seasonality (spring or summer), sediment texture and organic matter content, water depth, pH, salinity (TDS) and the presence of aquatic plants ( plant cover) were considered in the matrix involving species dry weight biomass at each of 117 stations. TWINSPAN classification of the samples yielded a dendrogram with 18 indicator species. Successive dichotomies divided these indicator species into four main lake groups based on salinity, i.e., Group I: 3–10 (Gammarus, Glyptotendipes I, Chironomus cf. plumosus), Group II: 10–38%. (Hyalella, Enallagma,Bezzia), Group III: 38–63 (Hygrotus salinarius, Cricotopus ornatus), Group IV: >63 (Dolichopodidae, Ephydra hians). Each of these main groups was subdivided into smaller groups of lakes based on factors such as pH, seasonality (spring or summer species dominance), organic matter and plant cover. Depth of samples played no apparent role.     

Long-term effects of dexamethasone and nerve growth factor on adrenal medullary cells cultured from young adult rats

Summary Normal postnatal rat chromaffin cells and rat pheochromocytoma cells are known to show extensive Nerve Growth Factor (NGF)-induced process outgrowth in culture, and this outgrowth from the postnatal chromaffin cells is abolished by the corticosteroid dexamethasone. To determine whether adult rat chromaffin cells respond to NGF and dexamethasone, dissociated adrenal medullary cells from 3-month-old rats were cultured for 30 days in the presence or absence of these agents. Such cultures contained typical chromaffin cells, chromaffin cells with processes, and neurons. Fewer than 2 % of normal adult chromaffin cells formed processes under any of the conditions studied, and statistically significant changes in this proportion were not detectable in the presence of NGF or dexamethasone. Adrenal medullary neurons, however, were observed only in the presence of NGF, in cultures with or without dexamethasone, and thus appear to be previously unreported NGF targets which require NGF for survival or process outgrowth. Dexamethasone markedly increased total catecholamine content, total content of epinephrine, and tyrosine hydroxylase activity in cultures with or without NGF. In contrast, postnatal rat chromaffin and rat pheochromocytoma cells which have been studied in culture do not produce epinephrine under any of these conditions. It is concluded that rat adrenal chromaffin cells undergo age-related changes in both structural and functional plasticity. The in vitro characteristics of rat pheochromocytoma cells more closely resemble those of postnatal than of adult rat chromaffin cells, but may not entirely reflect the properties of the majority of chromaffin cells in either age group.     

An Evaluation of “Informed Consent” with Volunteer Prisoner Subjects

“Informed consent” sets a goal for investigators experimenting with human subjects, but little is known about how to achieve or evaluate it in an experiment. In a 3-year, double-blind study with incarcerated men, we attempted to provide a “free and informed consent” and evaluated our efforts with an unannounced questionnaire administered to subjects after they completed the experiment. At that time, approximately two-thirds had sufficient information for an informed consent, but only one-third was well informed about all key aspects of the experiment and one-third was insufficiently informed to give an informed consent. We found that institution- or study-based coercion was minimal in our experiment. From our evaluation of the questionnaire and experience at the study institution, we conclude that an experiment with human subjects should be designed to include an ongoing evaluation of informed consent, and active attempts should be made to avoid or minimize coercive inducements. Experiments with significant risk, which require a long duration and/or large sample size relative to the institution's population, should probably not be performed on prisoner subjects. The experimenter should be independent of the penal institution's power structure. Presenting and explaining a consent form to volunteers on one occasion is probably an in adequate procedure for obtaining and maintaining an informed consent.     

Lenalidomide overcomes suppression of human natural killer cell anti-tumor functions by neuroblastoma microenvironment-associated IL-6 and TGFβ1

Background

Treatment for children with high-risk neuroblastoma with anti-disialoganglioside mAb ch14.18, IL-2, and GM-CSF plus 13-cis-retinoic acid after myeloablative chemotherapy improves survival, but 40 % of patients still relapse during or after this therapy. The microenvironment of high-risk neuroblastoma tumors includes macrophages, IL-6, and TGFβ1. We hypothesized that this microenvironment suppresses anti-tumor functions of natural killer (NK) cells and that lenalidomide, an immune-modulating drug, could overcome suppression.

Methods

Purified NK cells were cultured with IL-2, neuroblastoma/monocyte-conditioned culture medium (CM), IL-6, TGFβ1, and lenalidomide in various combinations and then characterized using cytotoxicity (direct and antibody-dependent cell-mediated cytotoxicity), cytokine, flow cytometry, and Western blotting assays. Anti-tumor activity of NK cells with lenalidomide, ch14.18, or both was evaluated with a xenograft model of neuroblastoma.

Results

CM from neuroblastoma/monocyte co-cultures contains IL-6 and TGFβ1 that suppress IL-2 activation of NK cell cytotoxicity and IFNγ secretion. IL-6 and TGFβ1 activate the STAT3 and SMAD2/3 pathways in NK cells and suppress IL-2 induction of cytotoxicity, granzymes A and B release, perforin expression, and IFNγ secretion. Lenalidomide blocks IL-6 and TGFβ1 activation of these signaling pathways and inhibits their suppression of NK cells. Neuroblastoma cells in NOD/SCID mice exhibit activated STAT3 and SMAD2/3 pathways. Their growth is most effectively inhibited by co-injected peripheral blood mononuclear cells (PBMC) containing NK cells when mice are treated with both ch14.18 and lenalidomide.

Conclusion

Immunotherapy with anti-tumor cell antibodies may be improved by lenalidomide, which enhances activation of NK cells and inhibits their suppression by IL-6 and TGFβ1.