Hypoxia selects for androgen independent LNCaP cells with a more malignant geno- and phenotype.

Hypoxia confers resistance to common cancer therapies, however, it has also has been shown to result in genetic alterations which may allow a survival advantage and increase the tumorigenic properties of cancer cells. Additionally, it may exert a selection pressure, allowing expansion of tumor cells with a more aggressive phenotype. To further assess the role of hypoxia in malignant progression in prostate cancer we exposed human androgen dependent prostate cancer cells (LNCaP) to cycles of chronic hypoxia and isolated a subline, LNCaP-H1. This article describes the partial characterization of this cell line. The LNCaP-H1 subline showed altered growth characteristics and exhibited androgen independent growth both in vitro and in vivo. Furthermore, these cells were resistant to mitochondrial-mediated apoptosis, probably since the endogenous levels of Bax was lower and Bcl-2 higher than in the parental LNCaP cells. Microarray analysis revealed that a complex array of pathways had differential gene expression between the 2 cell lines, with LNCaP-H1 cells exhibiting a genetic profile which suggests that they may be more likely metastasize to distant organs, especially bone. This was supported by an in vitro invasion assay, and an in vivo metastasis study. This study shows that hypoxia can select for androgen independent prostate cancer cells which have a survival advantage and are more likely to invade and metastasize.

Cholinergic-induced Ca2+ signaling in interstitial cells of Cajal from the guinea pig bladder.

Acetylcholine released from parasympathetic excitatory nerves activates contraction in detrusor smooth muscle. Immunohistochemical labeling of guinea pig detrusor with anti-c-Kit and anti-VAChT demonstrated a close structural relationship between interstitial cells of Cajal (ICC) and cholinergic nerves. The ability of guinea pig bladder detrusor ICC to respond to the acetylcholine analog, carbachol, was investigated in enzymatically dissociated cells, loaded with the Ca(2+) indicator fluo 4AM. ICC fired Ca(2+) transients in response to stimulation by carbachol (1/10 microM). Their pharmacology was consistent with carbachol-induced contractions in strips of detrusor which were inhibited by 4-DAMP (1 microM), an M(3) receptor antagonist, but not by the M(2) receptor antagonist methoctramine (1 microM). The source of Ca(2+) underlying the carbachol transients in isolated ICC was investigated using agents to interfere with influx or release from intracellular stores. Nifedipine (1 microM) or Ni(2+) (30-100 microM) to block Ca(2+) channels or the removal of external Ca(2+) reduced the amplitude of the carbachol transients. Application of ryanodine (30 microM) or tetracaine (100 microM) abolished the transients. The phospholipase C inhibitor, U-73122 (2.5 microM), significantly reduced the responses. 2-Aminoethoxydiethylborate (30 microM) caused a significant reduction and Xestospongin C (1 microM) was more effective, almost abolishing the responses. Intact in situ preparations of guinea pig bladder loaded with a Ca(2+) indicator showed distinctively different patterns of spontaneous Ca(2+) events in smooth muscle cells and ICC. Both cell types responded to carbachol by an increase in frequency of these events. In conclusion, guinea pig bladder detrusor ICC, both as isolated cells and within whole tissue preparations, respond to cholinergic stimulation by firing Ca(2+) transients. PMID: 18171995 [PubMed - indexed for MEDLINE]

Rheological characterization of thermoresponsive semisolids for vaginal HIV vaccine delivery

Purpose. To manufacture and characterize, through oscillatory rheology, thermoresponsive rheologically structured vehicles
(RSV’s) capable of enhanced retention times within the vagina for the purposes of HIV vaccine delivery.
Methods. Pluronics F127, F108 and F68 were investigated and RSV’s were prepared by dissolving sorbic acid (0.1% w/w)
and mucoadhesive component (Gantrez SBF97 or Noveon AA1, 3% w/w) in the required amount of H2O and NaOH.
Pluronic (10% w/w) was added via mixing in an ice-bath followed by hydroxyethylcellulose (5%) and subsequently
poly(vinylpyrollidone) (4%w/w). Oscillatory temperature sweeps between 10-38°C were preformed within the linear
viscoelastic region of the formulations on an AR2000 rheometer (T.A. Instruments, Surrey, England) with a 2cm diameter
parallel plate geometry and a plate gap of 1000µm at 1Hz.

Dispersal mode and assessments of recovery on the shores of Gruinard, the 'anthrax island'

Formaldehyde run-off was an unintended impact of the anthrax decontamination procedure on the island of Gruinard. The death of intertidal organisms was observed where formaldehyde reached the shore during 1986. The extent to which shores on Gruinard have recovered was assessed with survey work in 2000. Recovery estimates were based on the hypothesis that the process of recolonization is partly dependent on species' dispersal capability. Underdevelopment of the assemblage of species lacking planktonic dispersal stages (direct developers) is therefore evidence that the process of recolonization is ongoing, rather than complete. A novel multivariate test showed that, when comparing quadrats from Gruinard and nearby mainland shores, assemblages of direct developing molluscs were significantly more distinct than assemblages of molluscs with planktonic dispersal stages. The average densities of species with direct development were generally lower on Gruinard than on mainland shores. While some species with direct development have similar densities on Gruinard and on the surrounding shores, the recovery of the overall assemblage was still incomplete after 14 years. In contrast, the harvested species, Littorina littorea, appeared to benefit from the absence of humans visiting Gruinard's shores. Populations of L. littorea on Gruinard contained significantly higher proportions of large individuals. Depending on the dispersal capabilities of different species, Gruinard is either still in recovery or acts as a reserve.

Characterizing the marine Natura 2000 network for the Atlantic region

1. One of the goals for Natura 2000, a key European Community programme of nature conservation, is to produce a network of protected areas. An analysis of the Natura 2000 marine sites proposed in the most recently agreed list for the Atlantic region (northern Portugal to Denmark, n = 298) was used to characterize the network in terms of site areas and inter-site distances. Sites were considered as part of the network when they included any of the marine Natura 2000 Annex I habitat types found in the Atlantic region (excluding lagoons).

Coastline configuration as a determinant of structure in larval assemblages

The richness and turnover of coastal larval pools set upper limits for biodiversity in coastal systems. For particular local systems, such as embayments, the characteristics of the local larval pool are determined by the relative contributions of locally produced and external larvae. The balance between these sources partially reflects the extent of tidal exchange and is hence related to system size and flushing time. Larvae of benthic marine invertebrates were sampled from 8 bays along the Irish coast to investigate the effect of coastline configuration on the characteristics of the larval pool. Flushing time explained 34.5% of the variability in species richness from a series of daily samples. Many of the potentially relevant environmental variables are correlated, limiting the potential for individual variables to be examined in isolation. We therefore used a principal components analysis to describe the major patterns in environmental variability across bays. The second principal component separated bays along a gradient of increasing depth, salinity, tidal range and flushing time. Scores along this component were generally better predictors of the larval pool than single variables, explaining as much as 61.2% of the variation in species richness, diversity and similarity between dates. Deeper bays, with more saline water and longer flushing times, tended to have richer and more diverse larval pools, with a greater consistency in species composition between sample dates. No relationship was found between environmental variables and larval abundance. Our results suggest that flushing time, particularly when in combination with topographic variables, chlorophyll, tidal range and salinity, may be a useful predictor for the richness and turnover of local larval pools.

A new species of Pareledone (Cephalopoda : Octopodidae) from Antarctic Peninsula Waters

During recent cruises aboard RV Polarstern in the Antarctic Peninsula region, a new species of benthic octopodid was discovered whose generic affinities based on morphological characteristics were uncertain. Molecular sequence analysis of six mitochondrial and nuclear genes allows this species to be placed with confidence within the genus Pareledone. The species is described herein and morphological diagnostic characters are provided for its identification.

Redescription of the deep-sea octopod Benthoctopus normani (Massy 1907) and a description of a new species from the Northeast Atlantic

A long-synonymized species Benthoctopus normani (Massy 1907) (Cephalopoda: Octopodidae) is redescribed from material collected over 30 years by the National Oceanography Centre, Southampton and the National Museums of Scotland. It can be distinguished from other octopodid specimens found in deep waters of the Northeast Atlantic by its biserial suckers, lack of ink sac, and simple ligula, which lacks transverse ridges. Examination of the collections led to the identification of a new species of Benthoctopus from the Northeast Atlantic, which is described herein.

Cephalopods of the South Georgia slope

During January 2003 the bathymetric distribution of the cephalopod fauna of the South Georgia and Shag Rocks slope (100-900 m) was investigated using a commercial bottom trawl. Forty-four trawl stations caught 193 cephalopod specimens including six species of octopod and seven of squid. The benthic octopods Pareledone turqueti and Adelieledone polymorpha were abundant in shallow water at South Georgia, being replaced by Thaumeledone gunteri in greater depths. However, neither A. polymorpha nor T gunteri were caught on the adjacent Shag Rocks area. Two specimens of the deep-sea genus Graneledone were caught on the South Georgia slope. The most abundant squid species caught were Moroteuthis knipovitchi, Psychroteuthis glacialis and Slosarczykovia circumantarctica, which are primarily pelagic and may have been taken when their vertical migrations impinged on the slope.

Re-evaluation of Graneledone setebos (Cephalopoda : Octopodidae) and allocation to the genus Megaleledone

The holotype of the Antarctic octopodid Graneledone setebos was re-examined and found to lack the epidermal warts characteristic of the genus Graneledone. It is similar in its large size to another Southern Ocean species, Megaleledone senoi. A comparative study of G. setebos and specimens attributed to M. senoi led us to conclude that M. senoi is a junior synonym of G. setebos. Although M. senoi is not valid, the genus Megaleledone can be separated from other genera by the structure of the radula (which lacks marginal plates) and we therefore consider the genus to be valid. We propose the new combination of Megaleledone setebos and have refigured the beaks and radula of the holotype herein and expanded the description. A search of museum specimens and the literature shows that Megaleledone setebos is more common in Antarctic waters than previously supposed.

A redescription of Graneledone verrucosa (Verrill, 1881) (Octopoda : Octopodidae)

Graneledone verrucosa (Verrill 1881), the type species of the genus Graneledone, is redescribed based on historical material and previously unreported specimens that have resulted from an increase in deep-sea fishing in the North East Atlantic. Graneledone verrucosa var. media (Joubin 1918) was found to be invalid and is herein synonymized with G. verrucosa. Graneledone verrucosa is shown to inhabit deep water throughout the North Atlantic; its distribution extends from 20degrees to 65degrees N and from 9degrees to 75degrees W. A revised diagnosis is given for the genus Graneledone Joubin, 1918.