Hydrogel microwell arrays allow the assessment of protease-associated enhancement of cancer cell aggregation and survival

Current routine cell culture techniques are only poorly suited to capture the physiological complexity of tumor microenvironments, wherein tumor cell function is affected by intricate three-dimensional (3D), integrin-dependent cell-cell and cell-extracellular matrix (ECM) interactions. 3D cell cultures allow the investigation of cancer-associated proteases like kallikreins as they degrade ECM proteins and alter integrin signaling, promoting malignant cell behaviors. Here, we employed a hydrogel microwell array platform to probe using a high-throughput mode how ovarian cancer cell aggregates of defined size form and survive in response to the expression of kallikreins and treatment with paclitaxel, by performing microscopic, quantitative image, gene and protein analyses dependent on the varying microwell and aggregate sizes. Paclitaxel treatment increased aggregate formation and survival of kallikrein-expressing cancer cells and levels of integrins and integrin-related factors. Cancer cell aggregate formation was improved with increasing aggregate size, thereby reducing cell death and enhancing integrin expression upon paclitaxel treatment. Therefore, hydrogel microwell arrays are a powerful tool to screen the viability of cancer cell aggregates upon modulation of protease expression, integrin engagement and anti-cancer treatment providing a micro-scaled yet high-throughput technique to assess malignant progression and drug-resistance

Redshift clustering in the Hubble Deep Field

We present initial results from a redshift survey carried out with the Low Resolution Imaging Spectrograph on the 10~m W. M. Keck Telescope in the Hubble Deep Field. In the redshift distribution of the 140 extragalactic objects in this sample we find 6 strong peaks, with velocity dispersions of ${\sim}400${\kms}. The areal density of objects within a particular peak, while it may be non-uniform, does not show evidence for strong central concentration. These peaks have characteristics (velocity dispersions, density enhancements, spacing, and spatial extent) similar to those seen in a comparable redshift survey in a different high galactic latitude field (Cohen et al 1996), confirming that the structures are generic. They are probably the high redshift counterparts of huge galaxy structures (``walls'') observed locally.Comment: 14 pages, including 2 figures, to appear in ApJ Letter

Prostatic trypsin-like kallikrein-related peptidases (KLKs) and other prostate-expressed tryptic proteinases as regulators of signalling via proteinase-activated receptors (PARs)

The prostate is a site of high expression of serine proteinases including members of the kallikrein-related peptidase (KLK) family, as well as other secreted and membrane-anchored serine proteinases. It has been known for some time that members of this enzyme family elicit cellular responses by acting directly on cells. More recently, it has been recognised that for serine proteinases with specificity for cleavage after arginine and lysine residues (trypsin-like or tryptic enzymes) these cellular responses are often mediated by cleavage of members of the proteinase-activated receptor (PAR) family - a four member sub-family of G protein-coupled receptors. Here, we review the expression of PARs in prostate, the ability of prostatic trypsin-like KLKs and other prostate-expressed tryptic enzymes to cleave PARs, as well as the prostate cancer-associated consequences of PAR activation. In addition, we explore the dysregulation of trypsin-like serine proteinase activity through the loss of normal inhibitory mechanisms and potential interactions between these dysregulated enzymes leading to aberrant PAR activation, intracellular signalling and cancer-promoting cellular changes

Variable and invariable liaison in a corpus of spoken French

Using texts selected from the C-Oral-Rom corpus, this study considers how linguistic and sociolinguistic variables affect liaison. In the majority of cases, liaison appears on monosyllabic function words. Individual lexemes differ greatly in rate of liaison. With regard to sociolinguistic variation, female speakers realize liaison consonants more often than male speakers, younger speakers realize it more often than older speakers, and liaison rates for speakers without university degree are higher than for speakers with university degree. Results are discussed in the light of models of prosodic structure and with respect to their implications for models of socio- linguistic variation

A Kallikrein 15 (KLK15) single nucleotide polymorphism located close to a novel exon shows evidence of association with poor ovarian cancer survival.

BACKGROUND: KLK15 over-expression is reported to be a significant predictor of reduced progression-free survival and overall survival in ovarian cancer. Our aim was to analyse the KLK15 gene for putative functional single nucleotide polymorphisms (SNPs) and assess the association of these and KLK15 HapMap tag SNPs with ovarian cancer survival. RESULTS: In silico analysis was performed to identify KLK15 regulatory elements and to classify potentially functional SNPs in these regions. After SNP validation and identification by DNA sequencing of ovarian cancer cell lines and aggressive ovarian cancer patients, 9 SNPs were shortlisted and genotyped using the Sequenom iPLEX Mass Array platform in a cohort of Australian ovarian cancer patients (N = 319). In the Australian dataset we observed significantly worse survival for the KLK15 rs266851 SNP in a dominant model (Hazard Ratio (HR) 1.42, 95% CI 1.02-1.96). This association was observed in the same direction in two independent datasets, with a combined HR for the three studies of 1.16 (1.00-1.34). This SNP lies 15 bp downstream of a novel exon and is predicted to be involved in mRNA splicing. The mutant allele is also predicted to abrogate an HSF-2 binding site. CONCLUSIONS: We provide evidence of association for the SNP rs266851 with ovarian cancer survival. Our results provide the impetus for downstream functional assays and additional independent validation studies to assess the role of KLK15 regulatory SNPs and KLK15 isoforms with alternative intracellular functional roles in ovarian cancer survival.RIGHTS : This article is licensed under the BioMed Central licence at http://www.biomedcentral.com/about/license which is similar to the 'Creative Commons Attribution Licence'. In brief you may : copy, distribute, and display the work; make derivative works; or make commercial use of the work - under the following conditions: the original author must be given credit; for any reuse or distribution, it must be made clear to others what the license terms of this work are

The potential role of lycopene for the prevention and therapy of prostate cancer: From molecular mechanisms to clinical evidence

Lycopene is a phytochemical that belongs to a group of pigments known as carotenoids. It is red, lipophilic and naturally occurring in many fruits and vegetables, with tomatoes and tomato-based products containing the highest concentrations of bioavailable lycopene. Several epidemiological studies have linked increased lycopene consumption with decreased prostate cancer risk. These findings are supported by in vitro and in vivo experiments showing that lycopene not only enhances the antioxidant response of prostate cells, but that it is even able to inhibit proliferation, induce apoptosis and decrease the metastatic capacity of prostate cancer cells. However, there is still no clearly proven clinical evidence supporting the use of lycopene in the prevention or treatment of prostate cancer, due to the only limited number of published randomized clinical trials and the varying quality of existing studies. The scope of this article is to discuss the potential impact of lycopene on prostate cancer by giving an overview about its molecular mechanisms and clinical effects. © 2013 by the authors; licensee MDPI, Basel, Switzerland

Effectiveness of private land conservation areas in maintaining natural land cover and biodiversity intactness

CITATION: Shumba, T. et al. 2020. Effectiveness of private land conservation areas in maintaining natural land cover and biodiversity intactness. Global Ecology and Conservation, 22:e00935, doi:10.1016/j.gecco.2020.e00935.The original publication is available at https://www.journals.elsevier.com/global-ecology-and-conservationPrivate land conservation areas (PLCAs) are increasingly looked to for meeting the deficit left by state-owned protected areas in reaching global conservation targets. However, despite the increasing extent and recognition of PLCAs as a complementary conservation strategy, little research has been done to quantify their effectiveness; a critical consideration if they are to be counted towards international biodiversity conservation targets. The long history of PLCAs in South Africa provides an interesting case study to address this knowledge gap. Here, we quantified the effectiveness of South African PLCAs by comparing losses in natural land cover and biodiversity intactness within PLCAs with different levels of protection to that of unprotected control points. Points within PLCAs were matched with unprotected control points to test the prediction that if PLCAs offer effective protection, losses in natural land cover and biodiversity intactness would be significantly lower within their boundaries in comparison to unprotected controls exposed to similar conditions. Consequences of natural land cover loss on biodiversity intactness were thus assessed, thus advancing standard approaches for quantifying effectiveness. Between 1990 and 2013, PLCAs lost significantly less natural land cover (3%) and biodiversity intactness (2%) than matched unprotected areas (6% and 4%, respectively). Of the natural land cover lost within PLCAs, most was converted to cultivated land. Farms can support more species than other land uses (e.g. mines), a likely explanation for why losses in biodiversity intactness were less than losses in natural land cover. Contrary to the predicted pattern, effectiveness did not increase with level of protection; informal PLCAs with no legal protection had comparable natural land cover and biodiversity intactness retention to strictly protected PLCAs, with most losses recorded among PLCAs with moderate protection. This study provides the first national-scale evidence that PLCAs can be an effective mechanism for conserving natural land cover and biodiversity intactness, which is highly relevant given current discussions around their likely long-term biodiversity conservation capacity.Publisher's versio

Enter the Dragon: The Dynamic and Multifunctional Evolution of Anguimorpha Lizard Venoms

While snake venoms have been the subject of intense study, comparatively little work has been done on lizard venoms. In this study, we have examined the structural and functional diversification of anguimorph lizard venoms and associated toxins, and related these results to dentition and predatory ecology. Venom composition was shown to be highly variable across the 20 species of Heloderma, Lanthanotus, and Varanus included in our study. While kallikrein enzymes were ubiquitous, they were also a particularly multifunctional toxin type, with differential activities on enzyme substrates and also ability to degrade alpha or beta chains of fibrinogen that reflects structural variability. Examination of other toxin types also revealed similar variability in their presence and activity levels. The high level of venom chemistry variation in varanid lizards compared to that of helodermatid lizards suggests that venom may be subject to different selection pressures in these two families. These results not only contribute to our understanding of venom evolution but also reveal anguimorph lizard venoms to be rich sources of novel bioactive molecules with potential as drug design and development lead compounds