CD73 and CD29 concurrently mediate the mechanically induced decrease of migratory capacity of mesenchymal stromal cells

The assumption that mesenchymal stromal cell (MSC)-based therapies are capable of augmenting physiological regeneration processes has fostered intensive basic and clinical research activities. However, to achieve sustained therapeutic success in vivo, not only the biological, but also the mechanical microenvironment of MSCs during these regeneration processes needs to be taken into account. This is especially important for e.g., bone fracture repair, since MSCs present at the fracture site undergo significant biomechanical stimulation. This study has therefore investigated cellular characteristics and the functional behaviour of MSCs in response to mechanical loading. Our results demonstrated a reduced expression of MSC surface markers CD73 (ecto-5’-nucleotidase) and CD29 (integrin β1) after loading. On the functional level, loading led to a reduced migration of MSCs. Both effects persisted for a week after the removal of the loading stimulus. Specifi c inhibition of CD73/CD29 demonstrated their substrate dependent involvement in MSC migration after loading. These results were supported by scanning electron microscopy images and phalloidin staining of actin fi laments displaying less cell spreading, lamellipodia formation and actin accumulations. Moreover, focal adhesion kinase and Src-family kinases were identified as candidate downstream targets of CD73/CD29 that might contribute to the mechanically induced decrease in MSC migration. These results suggest that MSC migration is controlled by CD73 CD29, which in turn are regulated by mechanical stimulation of cells. We therefore speculate that MSCs migrate into the fracture site, become mechanically entrapped, and thereby accumulate to fulfil their regenerative functions.

PEGylation of lysine residues reduces the pro-migratory activity of IGF-I

Background: The insulin-like growth factor (IGF) system is composed of ligands and receptors which regulate cell proliferation, survival, differentiation and migration. Some functions are regulated via intracellular signaling cascades, others by involvement of the extracellular milieu, including binding proteins and other extracellular matrix proteins. However, understanding of their functions and the exact nature of these interactions remains incomplete. Methods: IGF-I was PEGylated at its lysine sites - K27, K65 and K68. Binding of PEG-IGF-I to the IGFBPs was analyzed using BIAcore and its ability to activate the IGF-IR was assessed using IGF-IR phosphorylation assay. Furthermore, functional consequences of PEGylating the lysine residues of IGF-I was investigated using cell viability and cell migration assays. In addition, particular downstream signaling pathways regularly implicated in these mechanisms were also dissected using phospho-AKT and phospho-ERK1/2 assays. Results: In this study, IGF-I specifically PEGylated at lysine 27 (PEG-K27), 65 (PEG-K65) or 68 (PEG-K68) were employed. Receptor phosphorylation was only reduced by 2-fold with PEG-K65 and PEG-K68 over all the time points tested, and as observed in two cell types, 3T3 fibroblasts and MCF-7 breast cancer cells. PEGylation at K27 resulted in a much larger effect, with more than 10-fold lower activation for 3T3 fibroblasts and a ~3 fold reduced IGF-IR activation for MCF-7 breast cancer cells over 15 minutes. In addition, all PEG-IGF-I variants demonstrated a ten-fold reduction in the association rate to IGF binding proteins (IGFBPs). Functionally, all PEG variants completely lost their ability to induce cell migration in the presence of IGFBP-3/vitronectin (VN) complexes as compared to IGF-I; in contrast, cell viability was fully preserved. Further investigations into the downstream signaling pathways revealed that the PI3-K/AKT pathway was preferentially affected upon treatment with the PEG-IGF-I variants compared to the MAPK/ERK pathway. Conclusion: PEGylation of IGF-I has an impact on cell migration but not cell viability. General significance: PEG-IGF-I may differentially modulate IGF-I mediated functions that are dependent on its interaction with its receptor as well as key extracellular proteins such as VN and IGFBPs.

In vitro assessment of migratory behavior of two cell populations in a simple multichannel microdevice

Recent literature suggests that mesenchymal stem/stromal cells (MSC) could be used as Trojan Horses to deliver “death-signals” to cancer cells. Herein, we describe the development of a novel multichannel cell migration device, and use it to investigate the relative migration rates of bone marrow-derived MSC and breast cancer cells (MCF-7) towards each other. Confluent monolayers of MSC and MCF-7 were established in adjacent chambers separated by an array of 14 microchannels. Initially, culture chambers were isolated by air bubbles (air-valves) contained within each microchannel, and then bubbles were displaced to initiate the assay. The MCF-7 cells migrated preferentially towards MSC, whilst the MSC did not migrate preferentially towards the MCF-7 cells. Our results corroborate previous literature that suggests MSC migration towards cancer cells in vivo is in response to the associated inflammation rather than directly to signals secreted by the cancer cells themselves.

Dormant but migratory tumour cells in desmoplastic stroma of invasive ductal carcinomas

Mortality in breast cancer is linked to metastasis and recurrence yet there is no acceptable biological model for cancer relapse. We hypothesise that there might exist primary tumour cells capable of escaping surgery by migration and resisting radiotherapy and chemotherapy to cause cancer recurrence. We investigated this possibility in invasive ductal carcinoma (IDC) tissue and observed the presence of solitary primary tumour cells (SPCs) in the dense collagen stroma that encapsulates intratumoural cells (ICs). In IDC tissue sections, collagen was detected with either Masson's Trichrome or by second harmonics imaging. Cytokeratin-19 (CK-19) and vimentin (VIM) antibodies were, respectively, used to identify epithelial-derived tumour cells and to indicate epithelial to mesenchymal transition (EMT). Confocal/multiphoton microscopy showed that ICs from acini were mainly CK-19 +ve and were encapsulated by dense stromal collagen. Within the stroma, SPCs were detected by their staining for both CK-19 and VIM (confirming EMT). ICs and SPCs were subsequently isolated by laser capture microdissection followed by multiplex tandem-PCR studies. SPCs were found to be enriched for pro-migratory and anti-proliferative genes relative to ICs. In vitro experiments using collagen matrices at 20 mg/cm 3, similar in density to tumour matrices, demonstrated that SPC-like cells were highly migratory but dormant, phenotypes that recapitulated the genotypes of SPCs in clinical tissue. These data suggest that SPCs located at the breast cancer perimeter are invasive and dormant such that they may exceed surgical margins and resist local and adjuvant therapies. This study has important connotations for a role of SPCs in local recurrence.

Vimentin expression in cervical carcinomas : association with invasive and migratory potential

Vimentin is an intermediate filament protein normally expressed in mesenchymal cells, but evidence is accumulating in the literature which suggests that the aberrant expression of vimentin in epithelial cancer cells might be related to local invasiveness and metastatic potential. Vimentin expression has previously been associated with invasive properties in an in vitro model consisting of a set of HPV-33-transformed cervical keratinocyte cell lines. In the present study, in order to emphasize those in vitro findings, the expression of vimentin has been investigated in cervical neoplasms of different grades, using immunohistochemistry. A clear association is reported between vimentin expression and metastatic progression, since vimentin was detected in all invasive carcinomas and lymph node metastases, but not in CIN III lesions. These in vivo results are compared with present and previous data obtained in vitro on cervical keratinocyte cell lines, where vimentin expression also correlated with in vitro invasiveness.

High connectivity in rastrelliger kanagurta: Influence of historical signatures and migratory behaviour inferred from mtDNA Cytochrome b

Phylogeographic patterns and population structure of the pelagic Indian mackerel, Rastrelliger kanagurta were examined in 23 populations collected from the Indonesian-Malaysian Archipelago (IMA) and the West Indian Ocean (WIO). Despite the vast expanse of the IMA and neighbouring seas, no evidence for geographical structure was evident. An indication that R. kanagurta populations across this region are essentially panmictic. This study also revealed that historical isolation was insufficient for R. kanagurta to attain migration drift equilibrium. Two distinct subpopulations were detected between the WIO and the IMA (and adjacent populations); interpopulation genetic variation was high. A plausible explanation for the genetic differentiation observed between the IMA and WIO regions suggest historical isolation as a result of fluctuations in sea levels during the late Pleistocene. This occurrence resulted in the evolution of a phylogeographic break for this species to the north of the Andaman Sea.

Interactions between adult migratory striped bass (Morone saxatilis) and their prey during winter off the Virginia and North Carolina Atlantic coast from 1994 through 2007

The migratory population of striped bass (Morone saxatilis) (>400 mm total length[TL]) spends winter in the Atlantic Ocean off the Virginia and North Carolina coasts of the United States. Information on trophic dynamics for these large adults during winter is limited. Feeding habits and prey were described from stomach contents of 1154 striped bass ranging from 373 to 1250 mm TL, collected from trawls during winters of 1994-96, 2000, and 2002-03, and from the recreational fishery during 2005-07. Nineteen prey species were present in the diet. Overall, Atlantic menhaden (Brevoortia tyrannus) and bay anchovy (Anchoa mitchilli) dominated the diet by boimass (67.9%) and numerically (68.6%). The percent biomass of Atlantic menhaden during 1994-2003 to 87.0% during 2005-07. Demersal fish species such as Atlantic croaker (Micropogonias undulatus) and spot (Leiostomus xanthurus) represented <15% of the diet biomass, whereas alosines (Alosa spp.) were rarely observed. Invertebrates were least important, contributing <1.0% by biomass and numerically. Striped bass are capable of feeding on a wide range of prey sizes (2% to 43% of their total length). This study outlines the importance of clupeoid fishes to striped bass winter production and also shows that predation may be exerting pressure on one of their dominant prey, the Atlantic menhaden.

The decline of migratory salmonid stocks: a new threat to pearl mussels in Scotland

The general decline of the endangered freshwater pearl mussel Margaritifera margaritifera (L.) throughout its holarctic range is well documented. Scotland is considered to be a stronghold of margaritifera, containing approximately half of the world's known remaining viable populations. However, even here the majority of populations have declined and many have disappeared completely. This article provides an overview of the freshwater pearl mussel life-cycle and the life-cycle of salmonids which are the host fish during the freshwater mussels short parasitic larval phase. The authors highlight the potential implications of the decline of salmonids for freshwater populations in Scotland.

Tracking migratory salmonids

The River Ribble drains into the Irish sea on the West coast of England. The estuary is approximately 20Km long, tunnel shaped, tapering from a 100m width at Preston dock to 5 km at Lytham where it enters the sea. This is a preliminary report on a study of oxygen requirements of Atlantic salmon (Salmo salar)and sea trout (Salmo trutta) in an estuary. Oxygen sending transmitters attached to fish were used to determine exposure of individuals to different dissolved oxygen concentrations as they moved in from the sea through the estuary of the River Ribble. This estuary is subject to extreme variations in dissolved oxygen concentrations. This report is based on the latest transcription and analysis of data completed in October 1983. The aim of this report is to give an overview of the work done and conclusions which are apparent at this stage.

An analysis of the historical catch data from the migratory salmonid fisheries of the River Ribble

The Ribble catchment is the largest and most diverse river system within National Rivers Authority (NRA), North West's Central Area. The river is approximately 100km in length and rises in a limestone area west of the Pennines. This report examines changes in the size and composition of the salmon and sea trout catches from the Ribble migratory salmonid fisheries during the years 1937 to 1991. Comparisons are made between the rod and net fisheries for both salmon and sea trout of the Ribble and Hodder. Patterns of catches shown by the Ribble fisheries are compared with those of other individual rivers and with patterns for the North West Region as a whole. An attempt is made to identify if any relationship exists between catch and stock abundance. Catch patterns shown by the Ribble and Hodder salmon fisheries are compared with electronic resistivity counter data from the two rivers. Annual salmon catch patterns and redd count data are compared both locally and regionally. Recommendations for future studies are made in the light of the report's findings.

Environmental quality standards to protect identified uses of controlled waters: salmonid fisheries, cyprinid fisheries, migratory fisheries, commercial harvesting of marine fish for public consumption, commercial harvesting of shellfish for for public consumption

This report considers the development of environmental quality standards (EQSs) for the salmonid fishery, cyprinid fishery, migratory fishery, commercial harvesting of marine fish for public consumption and commercial harvesting of shellfish for public consumption uses of controlled surface waters. Previous reports have been used to identify those parameters necessary for the maintenance of these five uses. Each water use is considered in a separate section within which identified parameters are discussed and standards proposed, a summary of the proposed standards is presented at the beginning of the relevant section. For salmonid, cyprinid and migratory fisheries, EQSs for substances in water have been proposed for the protection of these fisheries. For the commercial harvesting of marine fish and shellfish for public consumption uses 'Warning Levels' of substances in waters have been proposed. These 'Warning Levels' have been proposed by considering data on bioaccumulation and food standards and aim to prevent acceptable intake values and concentrations in fish/shellfish flesh exceeding statutory or recommended levels. For the commercial harvesting of marine fish for public consumption it has been concluded that the current EQSs for most List II substances for the protection of salt water life should be adequately stringent to protect this use, however for the commercial harvesting of shellfish for public consumption, these List II EQSs do not appear adequate to protect this use and more stringent 'Warning Levels' have been proposed. For all five uses considered in this report there has been found to be limited information on a number of the parameters considered and in general for indigenous species, this has been found to be especially so when considering migratory fisheries and the commercial harvesting of marine fish and shellfish.

Fish pass design - criteria for the design and approval of fish passes and other structures to facilitate the passage of migratory fish in rivers

Many of British rivers hold stocks of salmon (Salmo salar L.) and sea trout (Salmo trutta L.) and during most of the year some of the adult fish migrate upstream to the head waters where, with the advent of winter, they will eventually spawn. For a variety of reasons, including the generation of power for milling, improving navigation and measuring water flow, man has put obstacles in the way of migratory fish which have added to those already provided by nature in the shape of rapids and waterfalls. While both salmon and sea trout, particularly the former, are capable of spectacular leaps the movement of fish over man-made and natural obstacles can be helped, or even made possible, by the judicious use of fish passes. These are designed to give the fish an easier route over or round an obstacle by allowing it to overcome the water head difference in a series of stages ('pool and traverse' fish pass) or by reducing the water velocity in a sloping channel (Denil fish pass). Salmon and sea trout make their spawning runs at different flow conditions, salmon preferring much higher water flows than sea trout. Hence the design of fish passes requires an understanding of the swimming ability of fish (speed and endurance) and the effect of water temperature on this ability. Also the unique features of each site must be appreciated to enable the pass to be positioned so that its entrance is readily located. As well as salmon and sea trout, rivers often have stocks of coarse fish and eels. Coarse fish migrations are generally local in character and although some obstructions such as weirs may allow downstream passages only, they do not cause a significant problem. Eels, like salmon and sea trout, travel both up and down river during the course of their life histories. However, the climbing power of elvers is legendary and it is not normally necessary to offer them help, while adult silver eels migrate at times of high water flow when downstream movement is comparatively easy: for these reasons neither coarse fish nor eels are considered further. The provision of fish passes is, in many instances, mandatory under the Salmon and Freshwater Fisheries Act 1975. This report is intended for those involved in the planning, siting, construction and operation of fish passes and is written to clarify the hydraulic problems for the biologist and the biological problems for the engineer. It is also intended to explain the criteria by which the design of an individual pass is assessed for Ministerial Approval.

A survey of estuarine oxygen concentrations in relation to the passage of migratory salmonids

This report presents the first attempt at a national assessment of an Environmental Quality Standard (EQS) for dissolved oxygen (DO) in estuaries with the objective of allowing the passage of migratory salmonids. Under the Control of Pollution Act, Water Authorities and River Purification Boards have powers to control discharges to estuaries and need to define an EQS for the calculation of consent conditions. The object of any such standards is to permit the existence of good quality salmonid fisheries with only very occasional restrictions to the passage of fish. The report gives brief summaries of the DO regime in estuaries, the oxygen requirements of salmonids, and of tentative standards proposed by various authorities. These standards are then compared with DO and fishery data from UK estuaries, provided by the appropriate regulatory authorities. It concludes that a minimum annual lower 95-percentile of 5.0 mg/1 will meet the objective in most estuaries, and that a lower value of 3.0 mg/1 will permit the establishment of a more restricted fishery. However, more stringent standards may be needed in estuaries containing high concentrations of toxic pollutants. containing high concentrations of toxic pollutants.

Investigations into migratory fish propagation in the area of the Lancashire River Board

This report explores salmon propagation in the Lancashire River Board area (North West of England), looking at the Rivers Lune, Wyre, Ribble, and Hodder, showing salmon catch statistics, methods of propagation and habitat conditions investigations.

Summary document of the work being done in 1992-1993 regarding the migratory salmonid fisheries of the River Ribble catchment

This document lists the work on salmonid fisheries in the River Ribble catchment in the north west of England which was done in 1992-93. The document includes the following: analysis of historical catch data from the River Ribble covering 1937-1991; Ribble and Hodder stock assessment surveys 1992-93 including Swanside Beck; Ribble catch data from game angling log books 1993; results of salmon scale sampling 1993; results of microtagging programme in Hodder and Ribble 1993; Waddow Weir dish counter data 1993; Winkley Hall (Hodder), Locks Weir and Waddow Hall (Ribble) fish counter data 1993; number of fish and ova from Witcherwell Hatchery 1993.