Two <Emphasis Type="Italic">Streptomyces</Emphasis> Species Producing Antibiotic,Antitumor, and Anti-Inflammatory Compounds Are Widespread Among Intertidal Macroalgae and Deep-Sea Coral Reef Invertebrates from the Central Cantabrian Sea

Streptomycetes are widely distributed in the marine environment, although only a few studies on their associations to algae and coral ecosystems have been reported. Using a culture-dependent approach, we have isolated antibiotic-active Streptomyces species associated to diverse intertidal marine macroalgae (Phyllum Heterokontophyta, Rhodophyta, and Chlorophyta), from the central Cantabrian Sea. Two strains, with diverse antibiotic and cytotoxic activities, were found to inhabit these coastal environments, being widespread and persistent over a 3-year observation time frame. Based on 16S rRNA sequence analysis, the strains were identified as Streptomyces cyaneofuscatus M-27 and Streptomyces carnosus M-40. Similar isolates to these two strains were also associated to corals and other invertebrates from deep-sea coral reef ecosystem (Phyllum Cnidaria, Echinodermata, Arthropoda, Sipuncula, and Anelida) living up to 4.700-m depth in the submarine Avilés Canyon, thus revealing their barotolerant feature. These two strains were also found to colonize terrestrial lichens and have been repeatedly isolated from precipitations from tropospheric clouds. Compounds with antibiotic and cytotoxic activities produced by these strains were identified by high-performance liquid chromatography (HPLC) and database comparison. Antitumor compounds with antibacterial activities and members of the anthracycline family (daunomycin, cosmomycin B, galtamycin B), antifungals (maltophilins), anti-inflamatory molecules also with antituberculosis properties (lobophorins) were identified in this work. Many other compounds produced by the studied strains still remain unidentified, suggesting that Streptomyces associated to algae and coral ecosystems might represent an underexplored promising source for pharmaceutical drug discovery.     

Complexation and Full Characterization of the Tretinoin and Dimethyl-βeta-Cyclodextrin Complex

The aim of this work is to prepare tretinoin/dimethyl-beta-cyclodextrin complexes and fully characterize them through various analytical techniques. According to the phase solubility studies performed, the equilibrium for maximum complexation is reached in about 8 days presenting an A_L-type diagram (soluble complexes) corresponding mainly to 1:1 stoichiometry (K _s = 13,600 M⁻¹), although the possibility of the presence of 1:2 complexes was mathematically proven. Differential scanning calorimetry, X-ray diffraction and all the other analytical techniques have proven the presence of true complex formation in all the preparation methods tested. H-NMR and FTIR spectra allowed the selection of the best complexation method. The comparison between Raman spectra revealed that the more relevant feature is the band at 1,573 cm⁻¹, which corresponds to the entire delocalization of the superconjugated system, and after inclusion is observed as a positive frequency shift. Based on these results and the data obtained by molecular modelling calculations, it is proposed that the structure of the drug included into the cyclodextrin corresponds to the side chain including the functional group COOH. The complex was also analysed by atomic force microscopy to determine its size distribution which was heterogeneous and polymodal. However, it could be observed that they all have the same phase constitution.     

Proline as a biochemical marker in relation to the ecology of two halophytic Juncus species

Aims Osmolytes, used for maintaining osmotic balance and as 'osmoprotectants', are synthesized in plants as a general, conserved response to abiotic stress, although their contribution to stress-tolerance mechanisms remains unclear. Proline, the most common osmolyte, accumulates in many plant species in parallel with increased external salinity and is considered a reliable biochemical marker of salt stress. We have measured proline levels in two halophytic, closely related Juncus species under laboratory and field conditions to assess the possible relevance of proline biosynthesis for salt tolerance and therefore for the ecology of these two taxa.Methods Proline was quantified in plants treated with increasing NaCl concentrations and in plants sampled in two salt marshes located in the provinces of Valencia and Alicante, respectively, in southeast Spain. Electrical conductivity, pH, Na + and Cl ? concentrations were measured in soil samples collected in parallel with the plant material.Important findings Treatment with NaCl inhibited growth of J. acutus plants in a concentration-dependent manner, but only under high salt conditions for J. maritimus. Salt treatments led to proline accumulation in both species, especially in the more salt-tolerant J. maritimus. The results, obtained under laboratory conditions, were confirmed in plants sampled in the field. In all the samplings, proline contents were significantly lower in J. acutus than in the more tolerant J. maritimus growing in the same area. No direct correlation between soil salinity and proline levels could be established, but seasonal variations were detected, with increased proline contents under accentuated water deficit conditions. Our results suggest that proline biosynthesis is not only an induced, general response to salt stress but also an important contributing factor in the physiological mechanisms of salt tolerance in Juncus, and that it therefore correlates with the ecology of both species.     

Fiddler crab (Crustacea: Ocypodidae) distribution and the relationship between habitat occupancy and mouth appendages

Sediment characteristics, especially grain size, are usually considered the most important variables affecting Uca distribution, mainly due to its close relationship with mouth appendage morphology. The aim of this study was to verify, from an assemblage perspective, if sediment is the most important variable affecting Uca species distribution, and if mouth appendage morphology (setae type and curvature) would be related to habitat occupancy. Niche metrics and null model approaches were used to assess and test the hypothesis. The relevance of spoon-tipped setae curvature to Uca distribution was verified for the first time. A fragmented mangrove area was divided into seven subareas, and sampling of crabs and environmental variables took place in June and November 2010. Of 10 species recorded for Brazil, seven were found in the study area: U. burgersi, U. cumulanta, U. leptodactyla, U. maracoani, U. rapax, U. thayeri and U. uruguayensis. Multivariate analysis showed that sediment grain size and the presence of vegetation were the most important variables explaining distribution, reinforcing results commonly obtained by univariate approaches. The overlap of habitat occupancy was generally low and no relationship between mouth appendages was found with breadth and overlap measures. Contrary to predictions, most non-random overlap values were lower than expected by chance, suggesting that interspecific competition might influence species distribution. Also, variables such as the presence of vegetation are important and influence crab distribution, limiting the potential distribution that would be predicted by mouth adaptations alone. Thus, the use of these adaptations as surrogates of fiddler crab distribution is not recommended.     

Functional variants in NOS1 and NOS2A are not associated with progressive hearing loss in Ménière's disease in a European Caucasian population

Hearing loss in Ménière's disease (MD) is associated with loss of spiral ganglion neurons and hair cells. In a guinea pig model of endolymphatic hydrops, nitric oxide synthases (NOS) and oxidative stress mediate loss of spiral ganglion neurons. To test the hypothesis that functional variants of NOS1 and NOS2A are associated with MD, we genotyped three functional variants of NOS1 (rs41279104, rs2682826, and a cytosine-adenosine microsatellite repeat in exon 1f) and the CCTTT repeat in the promoter of NOS2A gene (rs3833912) in two independent MD sets (273 patients in total) and 550 controls. A third cohort of American patients was genotyped as replication cohort for the CCTTT repeat. Neither allele nor genotype frequencies of rs41279104 and rs2682826 were associated with MD, although longer alleles of the cytosine-adenosine microsatellite repeat were marginally significant (corrected p?=?0.05) in the Mediterranean cohort but not in a second Galicia cohort. Shorter numbers of the CCTTT repeat in NOS2A were significantly more frequent in Galicia controls (OR?=?0.37 [CI, 0.18-0.76], corrected p?=?0.04), but this finding could not be replicated in Mediterranean or American case-control populations. Meta-analysis did not support an association between CCTTT repeats and risk for MD. Severe hearing loss (>75?dB) was also not associated with any functional variants studied. Functional variants of NOS1 and NOS2A do not confer susceptibility for MD.     

Influence of phenological barriers and habitat differentiation on the population genetic structure of the balearic endemic Rhamnus ludovici-salvatoris Chodat and R. alaternus L.

Rhamnus ludovici-salvatoris, endemic to the Gymnesian Islands, coexists with the related and widespread R. alaternus in Mallorca and Menorca. In both species, the population genetic structure using RAPD, and flowering during a 3-year period to check for possible phenological barriers, were analyzed. Rhamnus ludovici-salvatoris showed lower genetic diversity and stronger population structure than R. alaternus, the Cabrera population being less diverse and the most differentiated. Rhamnus ludovici-salvatoris flowered one month later, although flowering of both species coincided sporadically. These congeners seem to have diverged through isolation by time and differentiation in habitat. The population genetic structure of R. ludovici-salvatoris could mainly be due to the existence of small populations on the one hand, and a gene flow caused by rare hybridization events on the other, which may also explain the presence of morphologically intermediate individuals in Menorca. The conservation of R. ludovici-salvatoris populations may include population reinforcements and other in situ interventions.