Mitochondrial DNA phylogeography reveals the existence of an Evolutionarily Significant Unit of the sand goby Pomatoschistus minutus in the Adriatic (Eastern Mediterranean)

The sand goby Pomatoschistus minutus is a major component of marine shelf and estuarine food webs and an important study organism in behavioural research. Yet, despite the sand goby’s significance, its past and present patterns of migration and gene flow are poorly understood. Here we use the mtDNA control region and parts of the flanking tRNA genes of 63 fish from six localities in the Adriatic (Eastern Mediterranean), Western Mediterranean, Atlantic, and North Sea to investigate the phylogeography of this gobiid. Phylogenetic analyses and population genetics statistics reveal the existence of an Evolutionarily Significant Unit, sensu Moritz (1994), in the Adriatic and another in the Western Mediterranean, Atlantic, and North Sea. A possible biogeographical scenario for the separation of the ancestral population is that sand gobies in the Adriatic and Western Mediterranean split between 10,000 and 5000 years ago when due to the rise in sea temperature they migrated northwards and were bisected by the Italian peninsula. A testable prediction of this scenario is that sand gobies from the Western Mediterranean, Adriatic, and Aegean form three reciprocally monophyletic groups which are the descendants of a three-way diversification event.     

Interaction of water-soluble porphyrins with single- and double-stranded polyribonucleotides

CD and uv-visible absorption studies with several tetracationic water-soluble porphyrin derivatives show that some of these species can serve as probes to discriminate between A- and B-conformational forms of single-stranded polynucleotides. It is also observed that these porphyrins can participate in the formation of double helices by forming transient intermediate complexes enroute to duplex formation. © 1994 John Wiley & Sons, Inc.     

Ergoline Derivatives as a Probe for Featuring the 5-HT1A Receptor Pharmacophore

A 5-HT_1A pharmacophore has been obtained employing a set of rigid templates encompassing the 5-HT1A structure. The use of rigid templates allowed us to overcome the discrepancy found when flexible structures where the energy of the active conformers are sometimes higher than the global minimum energy are used. On the basis of the results herein reported the three-dimensional requirements necessary for the binding interaction have been defined within this set of molecules. In this study forbidden zones of the receptor have been characterised. The pharmacophore model derived places some agonist/antagonist pharmacophore models appeared in the literature.     

Adult–Adult Play in Primates: Comparative Analyses of its Origin, Distribution and Evolution

Comparative analyses were conducted on a data set derived from the literature so as to test several hypotheses which were developed to explain the distribution of adult–adult play fighting within the order primates. Ratings for play occurring in sexual and non‐sexual contexts were developed. Three hypotheses were evaluated: (i) that play occurring in non‐sexual social contexts is a byproduct of its use in sex; (ii) that the occurrence of play is related to its use for social assessment and manipulation, and so is more likely to be present in species with reduced familiarity between individuals; and (iii) that phylogenetic affiliation influences the likelihood that species within clades engage in play. We used independent contrasts to test the first two hypotheses, and both were significant, with the presence of play in sexual contexts accounting for 14–16% of the variance of play in non‐sexual contexts, and reduced social familiarity accounting for 30–40% of the variance in the occurrence of play in non‐sexual contexts. To test the third hypothesis, we mapped the occurrence of both types of play onto known phylogenies. The overlap was not congruent, indicating that phylogenetic relationships did not account for the distribution of play. Given that play in both sexual and non‐sexual contexts was more likely to occur in species with a social organization involving reduced frequency of contact between the sexes and other social group members, we suggest that the likely adaptive value of play fighting is as a tool for social assessment and manipulation. The possible factors that mitigate the use of play fighting for these purposes, such as the availability of other forms of communication that could serve similar functions, are discussed.     

On Biological Functions Mapping to the Heterochromatin of DROSOPHILA MELANOGASTER

We examined the behavior of an autosomal recessive maternal-effect mutation, abnormal-oocyte (abo), that is located in the euchromatin of the left arm of chromosome 2. When homozygous in females, abo results in a marked reduction in the probability that an egg produced by a mutant mother will develop into an adult. However, this probability is increased if the fertilizing sperm delivers to the egg either a normal allele of the maternal-effect gene or a specific type of heterochromatin (called ABO) that is located in small regions of the X and Y chromosome constitutive heterochromatin as well as in some autosomal heterochromatin. These regions, moreover, all react to Hoechst 33258 fluorescent dye identically and specifically. The amelioration of the maternal effect produced by this heterochromatin differs temporally from that caused by the normal allele of the euchromatic gene: the heterochromatin reduces only precellular blastoderm mortality, whereas the normal allele of the euchromatic gene reduces only postblastoderm mortality. Thus, although the genome of the preblastoderm Drosophila embryo is apparently mostly silent, the ABO-containing heterochromatin functions at this early time. Finally, preliminary data indicate that abo is but one member of a cluster of linked genes, each of which interacts with its own normal allele and with a different, locus-specific, heterochromatic factor. From these observations, it appears that Drosophila heterochromatin contains developmentally important genetic elements, and that a functional concomitant of heterochromatic location is gene action at a developmental stage during which the activity of the euchromatic genome is as yet undetectable. Some general implications of these inferences are considered.     

Active specific immunotherapy with extracted tumor-specific transplantation antigen, cyclophosphamide, and adoptive transfer of tumor-specific cytotoxic T-cell clones

An L3T4-, Lyt2+ tumor-specific, cloned T-lymphocyte cell line (RTT-2) was isolated from a spleen cell population harvested from C3H/HeJ mice, following in vivo immunization against a syngeneic MCA-induced fibrosarcoma (MCA-F) and in vitro restimulation with 1-butanol-extracted, isoelectrophoretically purified MCA-F tumor-specific transplantation antigen (TSTA). RTT-2 required exposure to homotypic-extracted MCA-F TSTA in combination with low-dose IL-2 to maintain its specific cytotoxic activity in vitro. In vivo local adoptive transfer of RTT-2 caused specific neutralization of homotypic MCA-F, but not heterotypic MCA-D, tumor cells. Systemic in vivo transfer of RTT-2 alone augmented host resistance. In combination with a triple regimen of weekly doses of purified TSTA (1 microgram SC) and a single ip injection of CY (20 mg/kg), adoptive transfer of RTT-2 cells (1 X 10(7)) retarded the neoplastic outgrowth in and prolonged the survival of primary hosts bearing 3-, 7-, and 14-day established MCA-F tumors. In a spontaneous pulmonary metastasis model following amputation of a tumor-bearing limb, the triple regimen of TSTA/CY/RTT-2 markedly reduced the number of lung colonies. Thus RTT-2, which displays specific tumoricidal activity in vitro and in vivo, may afford a suitable tool to dissect T-cell receptors recognizing tumor markers on 1-butanol-extracted, MCA-F TSTA.     

Effect of ovarian cancer ascites on SKOV-3 cells proteome: new proteins associated with aggressive phenotype in epithelial ovarian cancer

Background

Epithelial ovarian cancer is the second most lethal gynecological cancer worldwide. Ascites can be found in all clinical stages, however in advanced disease stages IIIC and IV it is more frequent and could be massive, associated with worse prognosis. Due to the above, it was our interest to understanding how the ascites of ovarian cancer patients induces the mechanisms by which the cells present in it acquire a more aggressive phenotype and to know new proteins associated to this process.

Methods

A proteomic analysis of SKOV-3 cells treated with five different EOC ascites was performed by two-dimensional electrophoresis coupled to MALDI-TOF. The level of expression of the proteins of interest was validated by RT-PCR because several of these proteins have only been reported at the messenger level.

Results

Among the proteins identified that increased their expression in ascites-treated SKOV-3 cells, were Ran GTPase, ZNF268, and Synaptotagmin like-3. On the other hand, proteins that were negatively regulated by ascites were HLA-I, HSPB1, ARF1, Synaptotagmin 1, and hnRNPH1, among others. Furthermore, an interactome for every one of these proteins was done in order to identify biological processes, molecular actions, and cellular components in which they may participate.

Conclusions

Identified proteins participate in cellular processes highly relevant to the aggressive phenotype such as nuclear transport, regulation of gene expression, vesicular trafficking, evasion of the immune response, invasion, metastasis, and in resistance to chemotherapy. These proteins may represent a source of information which has the potential to be evaluated for the design of therapies directed against these malignant cells that reside on ovarian cancer ascites.