Protistan microbial observatory in the Cariaco Basin,Caribbean. I. Pyrosequencing vs Sanger insights into species richness

Microbial diversity and distribution are topics of intensive research. In two companion papers in this issue, we describe the results of the Cariaco Microbial Observatory (Caribbean Sea, Venezuela). The Basin contains the largest body of marine anoxic water, and presents an opportunity to study protistan communities across biogeochemical gradients. In the first paper, we survey 18S ribosomal RNA (rRNA) gene sequence diversity using both Sanger- and pyrosequencing-based approaches, employing multiple PCR primers, and state-of-the-art statistical analyses to estimate microbial richness missed by the survey. Sampling the Basin at three stations, in two seasons, and at four depths with distinct biogeochemical regimes, we obtained the largest, and arguably the least biased collection of over 6000 nearly full-length protistan rRNA gene sequences from a given oceanographic regime to date, and over 80 000 pyrosequencing tags. These represent all major and many minor protistan taxa, at frequencies globally similar between the two sequence collections. This large data set provided, via the recently developed parametric modeling, the first statistically sound prediction of the total size of protistan richness in a large and varied environment, such as the Cariaco Basin: over 36 000 species, defined as almost full-length 18S rRNA gene sequence clusters sharing over 99% sequence homology. This richness is a small fraction of the grand total of known protists (over 100 000–500 000 species), suggesting a degree of protistan endemism.     

Interferon-γ effect on growth regulation of cells with different levels of EGF receptor expression

Interferon gamma (IFNγ) is known to inhibit the proliferation of some transformed cell lines. Recently, we demonstrated the transactivation of the epidermal growth factor receptor (EGFR) in response to IFNγ (Burova et al., 2007) and provided direct evidence for the dependence of IFNγ-induced EGFR transactivation on the EGFR expression level in epithelial cells (Gonchar et al., 2008). This study examines an antiproliferative effect of IFNγ on human epithelial cell lines—A431 and HeLa that express high levels of EGFR, as well as HEK293 that expresses low levels of EGFR. To characterize the IFNγ-induced changes in these cells, we studied cell growth, the cell cycle, and induction of apoptosis. The response to IFNγ differed in the compared cell lines; cell growth was inhibited in both A431 and HeLa cells, but not in HEK293 cells, as was shown by the cell count and MTT. The cell-cycle phases analyzed by flow cytometry were disturbed in A431 and HeLa cells in response to IFNγ. On the contrary, in HEK293 cells, the IFNγ treatment did not alter distribution by cell cycle phases. Our results indicate that IFNγ produces an antiproliferative effect that depends on the increased expression of EGFR in A431 and HeLa cells. Furthermore, it was demonstrated that IFNγ induced the caspase 3 activation in A431 cells, which suggests the involvement of active caspase 3 in the IFNγ-induced apoptosis.     

Immune relations between mother and foetus in the viviparous poeciliid fish Xiphophorus hellerii Haeckel

Advanced embryos of the viviparous poeciliid fish Xiphophorus hellerii (swordtail) were removed from the site of gestation within the ovary and re-implanted into the peritoneal cavities of unrelated immunologically competent adults of the same species. Most of those transplanted with intact fertilization membranes were normal in appearance, and many were clearly still alive. Those transplanted without a fertilization membrane were all dead, with lymphocyte infiltration and other evidence of rejection as allografts. The integrity of the membrane in late pregnancy was indicated by swelling of follicles in hypoosmotic liquid. It was concluded that the fertilization membrane is capable of protecting an antigenically foreign embryo in an immunologically hostile environment, and that it probably serves this function during normal pregnancy.     

Preliminary clinical experience with the antiarrhythmic effect of PGF2a

A total dosage up to 1 mg PGF_2a as i.v. infusions of 10–40 μg/min. was investigated on patients with arrhythmias of several kinds. We found therapeutic effects in 5 of 6 patients with constant extrasystoles and in one patient with digitalis - induced partial AV-block respectively. In 3 of 4 patients with acute tachyarrhythmias the results were not convincing, probably due to a dosage not high enough. An increase of the diastolic stimulation threshold usually seen with other antiarrhythmics was not to be observed in 3 patients. The mechanism of action of PGF_2a has not yet been clarified.