Molecular support for morphology‐based family‐rank taxa: The contrasting cases of two families of Proseriata (Platyhelminthes)

Representatives of the Meidiamidae and Otomesostomidae (Platyhelminthes: Proseriata) are seldom encountered, and the monophyly and phylogenetic relationships of these families have never been assessed on molecular basis. Here, we present the first exhaustive molecular study of Proseriata at the family level, including species belonging to the genera Meidiama and Yorknia (Meidiamidae), and Otomesostoma auditivum (Otomesostomidae), using 18S and 28S genes as markers. We performed phylogenetic analyses (Maximum Likelihood [ML] and Bayesian Inference [BI] methods) and species delimitation methods (Single/Multiple Threshold‐Generalized Mixed Yule Coalescent [ST/MT‐GMYC] and Poisson Tree Processes [PTP/bPTP]). The taxon Meidiamidae was not supported, since the type species (Meidiama lutheri) and Meidiama etrusca sp. n. are nested within the Archimonocelididae, formerly restricted to specialized cnidarian feeders. Species belonging to the genus Yorknia resulted genetically well separated from species of Meidiama and from the rest of Archimonocelididae. The new family‐level taxon Yorkniidae fam. n. is thus here introduced, to include the type species of Yorknia (Yorknia aprostatica), and six new species, five of which are formally described here. Otomesostoma auditivum, representative of Otomesostomidae, the only exclusively freshwater taxon of the Proseriata, is the sister taxon of the predominantly marine Apingospermata. This result is not conflictual with the family level attributed to Otomesostomidae on morphological grounds, but it raises speculations on the marine versus freshwater origin of Apingospermata.     

Polymorphonuclear Cell Functional Impairment in Relapsing Remitting Multiple Sclerosis Patients: Preliminary Data

Multiple Sclerosis patients run an increased risk of microbial infections, which leads to high rates of hospitalization and infection-related mortality. Although immunotherapy may increase infection risk in some cases, data as to the relationship among microbial factors, immunotherapy and alterations in the innate immunity of these patients are still scanty. On these grounds, this interdisciplinary study aims at investigating the role the functional activity of polymorphonuclear cells (PMNs) play in relapsing remitting multiple sclerosis at different stages. The in vitro ability of PMNs from patients, either untreated or treated with immunosuppressant or immunomodulatory drugs to kill Klebsiella pneumonia or Candida albicans, were investigated and compared to PMNs from healthy subjects. The release of various cytokines was also assessed, as was the production of reactive oxygen species and their ability to regulate apoptosis after microbial stimulation. Our results indicate that although patients have a normal number of PMNs, they have a statistically significant (p<0.05) reduction in intracellular killing activity. Although variations are strongly related to the therapeutic management of patients, they are independent from their disease stage. As no statistically significant differences were observed between patients and controls in cytokine release values, reactive oxygen species production or apoptosis, we came to the conclusion that other factors may be involved. Supportive validation of these results from further studies might well help in identifying a subset of patients at high risk of infection who could benefit from a closer follow-up and/or antibiotic prophylaxis.     

Reproductive modes and genetic polymorphism in the tardigrade Richtersius coronifer (Eutardigrada, Macrobiotidae)

Abstract. Allozymes were assessed by starch gel electrophoresis in 3 populations of a eutardigrade, Richtersius coronifer , with different reproductive modes. One population from Italy (with 2 sub-populations) was amphimictic and 2 populations (1 from Italy and 1 from Sweden) were parthenogenetic. All populations, irrespective of their reproductive mode, were diploid with the same chromosome number (2n=12) and had bivalents in the oocytes. Of the 14 loci analyzed, only 3 were polymorphic. The amphimictic population had a higher degree of genetic variability (mean heterozygosity >0.25) than the parthenogenetic populations (mean heterozygosity of the 2 populations <0.01). In all female populations, allele frequencies at all 3 loci deviated from Hardy-Weinberg equilibria due to heterozygote deficiency. These results support a hypothesis of automictic parthenogenesis in R. coronifer .     

Symbiosis of Mycale (Mycale) vansoesti sp. nov. (Porifera, Demospongiae) with a coralline alga from North Sulawesi (Indonesia)

Abstract. The symbiotic association between the new sponge species Mycale vansoesti and the coralline alga Amphiroa sp. from the Bunaken Marine Park (North Sulawesi, Indonesia) is described. The alga completely pervades the sponge. The color of the sponge ectosome is white, both on the external surface and on the atrial wall, but where the alga is present the sponge takes on the light pink color of the alga. The sponge spicular complement is characterized by mycalostyles, anisochelae of two types, sigmas (often "C" shaped), and extremely abundant toxas organized in bundles forming toxadragma. In the association, the sponge shows very low silicate value, and consequently the alga represents the main skeleton of the sponge. On the other hand, the sponge affects the morphology of the alga, leading to a cylindrical shape, with thalli running parallel to the sponge surface. This association seems to be obligate for the sponge, as we found no sponges of this species living in isolation.     

Inhibitory Effect of Aqueous Extracts from Marine Sponges on the Activity and Expression of Gelatinases A (MMP-2) and B (MMP-9) in Rat Astrocyte Cultures

The aim of this study was to evaluate whether water soluble compounds present in aqueous extracts from seven Mediterranean demosponges exert biological activity towards matrix metalloproteinases (MMPs), which represent important pathogenic factors of human diseases. Aqueous extracts were tested on LPS-activated cultured rat astrocytes, and levels and expression of MMP-2 and MMP-9 were assessed by zymography and RT-PCR, respectively. Our results demonstrated that the studied extracts contain water soluble compounds able to inhibit MMP-2 and MMP-9 activity and expression. We also compared the anti-MMP activities present in aqueous extracts from wild and reared specimens of Tethya aurantium and T. citrina. The results obtained revealed that the reared sponges maintain the production of bioactive compounds with inhibitory effect on MMP-2 and MMP-9 for all the duration of the rearing period. Taken together, our results indicate that the aqueous extracts from the selected Mediterranean demosponges possess a variety of water-soluble bioactive compounds, which are able to inhibit MMPs at different levels. The presence of biological activity in aqueous extracts from reared specimens of T. aurantium and T. citrina strongly encourage sponge aquaculture as a valid option to supply sponge biomass for drug development on a large scale.     

Tubulin glycylases and glutamylases have distinct functions in stabilization and motility of ependymal cilia

Microtubules are subject to a variety of posttranslational modifications that potentially regulate cytoskeletal functions. Two modifications, glutamylation and glycylation, are highly enriched in the axonemes of most eukaryotes, and might therefore play particularly important roles in cilia and flagella. Here we systematically analyze the dynamics of glutamylation and glycylation in developing mouse ependymal cilia and the expression of the corresponding enzymes in the brain. By systematically screening enzymes of the TTLL family for specific functions in ependymal cilia, we demonstrate that the glycylating enzymes TTLL3 and TTLL8 were required for stability and maintenance of ependymal cilia, whereas the polyglutamylase TTLL6 was necessary for coordinated beating behavior. Our work provides evidence for a functional separation of glutamylating and glycylating enzymes in mammalian ependymal cilia. It further advances the elucidation of the functions of tubulin posttranslational modifications in motile cilia of the mammalian brain and their potential importance in brain development and disease.     

Inherited Brugada and long QT-3 syndrome mutations of a single residue of the cardiac sodium channel confer distinct channel and clinical phenotypes.

Defects of the SCN5A gene encoding the cardiac sodium channel alpha-subunit are associated with both the long QT-3 (LQT-3) subtype of long-QT syndrome and Brugada syndrome (BrS). One previously described SCN5A mutation (1795insD) in the C terminus results in a clinical phenotype combining QT prolongation and ST segment elevation, indicating a close interrelationship between the two disorders. Here we provide additional evidence that these two disorders are closely related. We report the analysis of two novel mutations on the same codon, Y1795C (LQT-3) and Y1795H (BrS), expressed in HEK 293 cells and characterized using whole-cell patch clamp procedures. We find marked and opposing effects on channel gating consistent with activity associated with the cellular basis of each clinical disorder. Y1795H speeds and Y1795C slows the onset of inactivation. The Y1795H, but not the Y1795C, mutation causes a marked negative shift in the voltage dependence of inactivation, and neither mutation affects the kinetics of the recovery from inactivation. Interestingly, both mutations increase the expression of sustained Na+ channel activity compared with wild type (WT) channels, although this effect is most pronounced for the Y1795C mutation, and both mutations promote entrance into an intermediate or a slowly developing inactivated state. These data confirm the key role of the C-terminal tail of the cardiac Na+ channel in the control of channel gating, illustrate how subtle changes in channel biophysics can have significant and distinct effects in human disease, and, additionally, provide further evidence of the close interrelationship between BrS and LQT-3 at the molecular level.     

The affinity of metergoline for 3H-Serotonin (5HT) binding sites is regulated by guanine nucleotide

100 μM guanine nucleotide Gpp (NH)p reduces the affinity of the serotonergic antagonist metergoline for ³H-5HT binding sites in rat cerebral cortex. This effect is present both in inhibition binding and in saturation experiments. The hypothesis that the interaction of some serotonergic antagonists with ³H-5HT binding sites is regulated by guanine nucleotides is discussed.     

Vitamin C Effect on Mitoxantrone-Induced Cytotoxicity in Human Breast Cancer Cell Lines

In recent years the use of natural dietary antioxidants to minimize the cytotoxicity and the damage induced in normal tissues by antitumor agents is gaining consideration. In literature, it is reported that vitamin C exhibits some degree of antineoplastic activity whereas Mitoxantrone (MTZ) is a synthetic anti-cancer drug with significant clinical effectiveness in the treatment of human malignancies but with severe side effects. Therefore, we have investigated the effect of vitamin C alone or combined with MTZ on MDA-MB231 and MCF7 human breast cancer cell lines to analyze their dose-effect on the tumor cellular growth, cellular death, cell cycle and cell signaling. Our results have evidenced that there is a dose-dependence on the inhibition of the breast carcinoma cell lines, MCF7 and MDA-MB231, treated with vitamin C and MTZ. Moreover, their combination induces: i) a cytotoxic effect by apoptotic death, ii) a mild G2/M elongation and iii) H2AX and mild PI3K activation. Hence, the formulation of vitamin C with MTZ induces a higher cytotoxicity level on tumor cells compared to a disjointed treatment. We have also found that the vitamin C enhances the MTZ effect allowing the utilization of lower chemotherapic concentrations in comparison to the single treatments.     

Developmental stage on day-5 and fragmentation rate on day-3 can influence the implantation potential of top-quality blastocysts in IVF cycles with single embryo transfer

Background  

In IVF-ICSI cycles with single embryo transfer (SET), embryo selection for transfer is of crucial importance. The present study aimed to define which embryo parameters might be related to the implantation potential of advanced blastocysts.