The invasive and metastatic properties of hormone-independent but hormone-responsive variants of MCF-7 human breast cancer cells

We have previously isolated a series of MCF-7 human breast cancer cell variants which no longer require estrogen-supplementation for tumor growth in nude mice (Clarke et al. Proc Natl Acad Sci USA 86: 3649-3653, 1989). We now report that these hormone-independent and hormone-responsive variants (MIII, MCF7/LCC1) can invade locally from solid mammary fat pad tumors, and produce primary extensions on the surface of intraperitoneal structures including liver, pancreas, and diaphragm. Both lymphatic and hematogenous dissemination are observed, resulting in the establishing of pulmonary, bone, and renal metastases. The pattern of metastasis by MIII and MCF7/LCC1 cells closely resembles that frequently observed in breast cancer patients, and provides the first evidence of metastasis from MCF-7 cells growing in vivo without supplementary estrogen. The interexperimental incidence of metastases, and the time from cell inoculation to the appearance of metastatic disease are variable. The increased metastatic potential is not associated with an increase in either the level of laminin attachment, laminin receptor mRNA expression, or secreted type IV collagenolytic activity. We also did not detect a significant decrease in the steady-state mRNA levels of the metastasis inhibitor nm23 gene. However, when growing without estrogen in vitro, MCF7/LCC1 cells produce elevated levels of the estrogen-inducible cathepsin D enzyme.

Transfection of MDA-MB-231 human breast carcinoma cells with bone sialoprotein (BSP) stimulates migration and invasion in vitro and growth of primary and secondary tumors in nude mice

We have investigated the role of bone sialoprotein (BSP), a secreted glycoprotein normally found in bone, in breast cancer progression. To explore functions for BSP in human breast cancer invasion and metastasis, the full-length BSP cDNA was transfected into the MDA-MB-231-BAG human breast cancer cell line under the control of the CMV promoter. Clones expressing BSP and vector control clones were isolated. BSP producing clones showed increased monolayer wound healing, a faster rate of stellate outgrowth in Matrigel and increased rate of invasion into a collagen matrix when compared to control clones. Clones were also examined in models of breast cancer growth and metastasis in vivo. BSP transfected clones showed an increased rate of primary tumor growth following mammary fat pad injection of nude mice. BSP transfected clones and vector control clones metastasized to soft organs and bone at a similar rate after intra-cardiac injection as determined by real-time PCR and X-ray analysis. Although these organs were targets for both BSP transfected and non-transfected cells, the size of the metastatic lesion was shown to be significantly larger for BSP expressing clones. This was determined by real-time PCR analysis for soft organs and by X-ray analysis of bone lesions. For bone this was confirmed by intra-tibial injections of cells in nude mice. We conclude that BSP acts to drive primary and secondary tumor growth of breast cancers in vivo.

An MMP13-selective inhibitor delays primary tumor growth and the onset of tumor-associated osteolytic lesions in experimental models of breast cancer

We investigated the effects of the matrix metalloproteinase 13 (MMP13)-selective inhibitor, 5-(4-{4-[4-(4-fluorophenyl)-1,3-oxazol-2-yl]phenoxy}phenoxy)-5-(2-methoxyethyl) pyrimidine-2,4,6(1H,3H,5H)-trione (Cmpd-1), on the primary tumor growth and breast cancer-associated bone remodeling using xenograft and syngeneic mouse models. We used human breast cancer MDA-MB-231 cells inoculated into the mammary fat pad and left ventricle of BALB/c Nu/Nu mice, respectively, and spontaneously metastasizing 4T1.2-Luc mouse mammary cells inoculated into mammary fat pad of BALB/c mice. In a prevention setting, treatment with Cmpd-1 markedly delayed the growth of primary tumors in both models, and reduced the onset and severity of osteolytic lesions in the MDA-MB-231 intracardiac model. Intervention treatment with Cmpd-1 on established MDA-MB-231 primary tumors also significantly inhibited subsequent growth. In contrast, no effects of Cmpd-1 were observed on soft organ metastatic burden following intracardiac or mammary fat pad inoculations of MDA-MB-231 and 4T1.2-Luc cells respectively. MMP13 immunostaining of clinical primary breast tumors and experimental mice tumors revealed intra-tumoral and stromal expression in most tumors, and vasculature expression in all. MMP13 was also detected in osteoblasts in clinical samples of breast-to-bone metastases. The data suggest that MMP13-selective inhibitors, which lack musculoskeletal side effects, may have therapeutic potential both in primary breast cancer and cancer-induced bone osteolysis.

Upregulation of matrix metalloproteinases (MMPs) in breast cancer xenografts : a major induction of stromal MMP-13

In human breast cancer (HBC), as with many carcinoma systems, most matrix metalloproteinases (MMPs) are largely expressed by the stromal cells, whereas the tumour cells are relatively silent in MMP expression. To determine the tissue source of the most relevant MMPs, we xenografted HBC cell lines and HBC tissues into the mammary fat pad (MFP) or bone of immunocompromised mice and measured the expression of human and mouse MMP-2, -9, -11, -13, membrane-type-1 MMP (MT1-MMP), MT2-MMP and MT3-MMP by species-specific real-time quantitative RT-PCR. Our data confirm a stromal origin for most tumour-associated MMPs and indicate marked and consistent upregulation of stromal (mouse) MMP-13 and MT1-MMP in all xenografts studied, irrespective of implantation in the MFP or bone environments. In addition, we show increased expression of both human MMP-13 and human MT1-MMP by the MDA-MB-231 tumour cells grown in the MFP compared to in vitro production. MMP protein and activity data confirm the upregulation of MMP mRNA production and indicate an increase in the activated MMP-2 species as a result of tumour implantation. These data directly demonstrate tumour induction of MMP production by stromal cells in both the MFP and bone environments. These xenografts are a valuable means for examining in vivo production of MMPs and suggest that MMP-13 and MT1-MMP will be relevant targets for inhibiting breast cancer progression.

Benefits and risks of large hydrostatic recesses in hydroelectric turbine thrust bearings

Operators of hydroelectric power stations sometimes call upon engineers to modify existing hydroelectric turbines, usually several decades old, for improved maintainability and reliability. One common modification is the hybridisation of plain thrust pads to allow hydrostatic operation to reduce the risk of bearing wipe at low speed (virtually all new installations benefit from this feature). A modification such as this is not a difficult undertaking; however, there are numerous factors that need to be considered in order to maximize bearing performance. One factor that stands out above the others is whether the thrust bearing should be designed to lift the turbine immediately from the standing condition, which presents an interesting challenge: the recess has to have a sufficiently large area in order for the supply pressure to be able to overcome the dead weight of the turbine. If the combination of groove area and pressure is insufficient, then lifting is neither immediate nor guaranteed. This need not be a significant problem, as the bearings have exhibited adequate performance even in the absence of a hydrostatic lubricant supply. A case study is presented whereby relatively large hydrostatic recesses are added to the pads of thrust bearing. It is demonstrated with the aid of simple numerical modelling that the impact of the recess relative to the original pad is small under normal operating conditions. Most surprising, however, is that significant reductions in average oil film temperature and power dissipation are predicted.

Ramipril - improving exercise tolerance in intermittent claudication?

Peripheral artery disease (PAD) is one of the most common manifestations of systemic atherosclerosis. It is estimated that 10-15% of the general population is affected by PAD, whereby the narrowed arteries lead to reduced blood flow to the extremeties - particularly the legs. While many people have mild or no systems with PAD, approximately one-third of people experience intermittent claudication (IC).

Image-based midsole insert design and the material effects on heel plantar pressure distribution during simulated walking loads

The primary objective of this paper is to study the use of medical image-based finite element (FE) modelling in subjectspecific midsole design and optimisation for heel pressure reduction using a midsole plug under the calcaneus area (UCA). Plugs with different relative dimensions to the size of the calcaneus of the subject have been incorporated in the heel region of the midsole. The FE foot model was validated by comparing the numerically predicted plantar pressure with biomechanical tests conducted on the same subject. For each UCA midsole plug design, the effect of material properties and plug thicknesses on the plantar pressure distribution and peak pressure level during the heel strike phase of normal walking was systematically studied. The results showed that the UCA midsole insert could effectively modify the pressure distribution, and its effect is directly associated with the ratio of the plug dimension to the size of the calcaneus bone of the subject. A medium hardness plug with a size of 95% of the calcaneus has achieved the best performance for relieving the peak pressure in comparison with the pressure level for a solid midsole without a plug, whereas a smaller plug with a size of 65% of the calcaneus insert with a very soft material showed minimum beneficial effect for the pressure relief.

Studies on characteristics of lightning generated currents in an interconnected lightning protection system

An in-depth knowledge about the characteristics of lightning generated currents will facilitate evaluation of the interception efficacy of lightning protection systems. In addition, it would aid in extraction of valuable statistics (from measured current data) on local lightning parameters. Incidentally, present day knowledge on characteristics of lightning induced current in typical lightning protection systems is rather limited. This is particularly true with closely interconnected protection systems, like the one employed in Indian Satellite Launch Pad-II. This system is taken as a specific example in the present study. Various aspects suggest that theoretical modelling would be the best possible approach for the intended work. From the survey of pertinent literature, it is concluded that electromagnetic modelling of lightning return-stroke with current source at the channel base is best suited for this study. Numerical electromagnetic code was used for the required electromagnetic field solution and Fourier transform techniques were employed for computing time-domain results. A validation for the numerical modelling is provided by laborator experiments on a reduced scale model of the system. Apart from ascertaining the influence of various parameters, salient characteristics of tower base currents for different kinds of events are deduced. This knowledge can be used in identifying the type of event, as well as its approximate location. A method for estimation of injected stroke current has also been proposed.

Linnunkiikut ja kallen kaulukset : Kotitekoiset kuukautissiteet 1900-luvun alkupuolen Suomessa

The aim of this thesis was to study what kind of home-made menstrual pads were used in the early 20th century in Finland, how the home-made pads were made and which techniques and materials were used. The use and taking care of menstrual pads were also explored. The history of menstrual pads has been studied in Sweden, Germany and United States but none of those studies has concentrated on home-made pads. Instead, there are many studies about womanhood and menstruation. In many studies home-made menstrual pads are only briefly mentioned. Menstrual pads were not commonly used in Finland at the beginning of the 20th century, but already in the 1940s the use of menstrual pads had become common in every stratum of society. Home-made menstrual pads were used even until the 1960s. In Finland, factory-made disposable menstrual pads became common only in the 1930s and they were only slowly accepted. The study material consisted of nine interviews, three archival inquiries, health care guidebooks from 1893 to 1943 and authentic menstrual pads, menstrual belts and other objects related to them. The interviewed women were born between 1915 and 1939. The narrative approach was used in the study and it also guided the analysis. The interview and archival data were studied according to the basic rules of oral history studies. Literature consisted of publications from several disciplines. The extensive primary material played the most important role in this study. The reconstructions of the menstrual pads were made according to the interviewed women s advice. In Finland there were innumerable variations of home-made menstrual pads. The pads were most commonly crocheted and knitted either by hand or by knitting machine. Pads were also sewn of cloth, old bed linen or old underwear. The menstrual pads were self-made or made by a female relative. Word of mouth was important in spreading information on how to make pads, because there were hardly any instructions available. The biggest pads were 54 cm long and 13 cm wide. The most widely used pad model was a rectangle, which had triangle-shaped ends with a buttonhole or a loop. The pad was attached to the menstrual belt or to the buttons of the suspender belt. Knitted and crocheted pads had one, two or three layers. In sewn pads, there could be even more layers. Cellulose wadding or pieces of cloth could be placed inside the pad to increase the absorption ability. The experiences of the comfort of self-made pads varied. The crocheted and sewn pads were found chafing, knitted ones were found soft and comfortable. The menstrual pads were laborious to wash and boil in lye water. Therefore disposable pads made everyday life easier. The home-made menstrual pads were part of a unique tradition of handicrafts and folk culture. Hand-made pads were one of the most common handicraft products and were a part of every woman s life. Even so, the menstrual pads were unnoticeable. The large number of variations was probably caused by the silence around menstrual topics and by the lack of instructions for making pads. Variations are also explained by the uniqueness of every handicraft product. In Finland the home-made pads were used until relatively recent times. This was caused by the conditions of wartime and the following years and the rarity of commercial pads. Furthermore, until the late 20th century Finland was an agricultural society where all innovations spread slowly. Home-made menstrual pad was a secret handicraft of women and every woman needed to know how to make it by herself.

Preparation, structural analysis and prebiotic potential of arabinoxylo-oligosaccharides

Arabinoxylo-oligosaccharides (AXOS) can be prepared enzymatically from arabinoxylans (AX) and AXOS are known to possess prebiotic potential. Here the structural features of 10 cereal AX were examined. AX were hydrolysed by Shearzyme® to prepare AXOS, and their structures were fully analysed. The prebiotic potential of the purified AXOS was studied in the fermentation experiments with bifidobacteria and faecal microbiota. In AX extracted from flours and bran, high amounts of a-L-Araf units are attached to the b-D-Xylp main chain, whereas moderate or low degree of substitution was found from husks, cob and straw. Nuclear magnetic resonance (NMR) spectroscopy showed that flour and bran AX contain high amounts of a-L-Araf units bound to the O-3 of b-D-Xylp residues and doubly substituted b-D-Xylp units with a-L-Araf substituents at O-2 and O-3. Barley husk and corn cob AX contain high amounts of b-D-Xylp(1→2)-a-L-Araf(1→3) side chains, which can also be found in AX from oat spelts and rice husks, and in lesser amounts in wheat straw AX. Rye and wheat flour AX and oat spelt AX were hydrolysed by Shearzyme® (with Aspergillus aculeatus GH10 endo-1,4-b-D-xylanase as the main enzyme) for the production of AXOS on a milligram scale. The AXOS were purified and their structures fully analysed, using mass spectrometry (MS) and 1D and 2D NMR spectroscopy. Monosubstituted xylobiose and xylotriose with a-L-Araf attached to the O-3 or O-2 of the nonreducing end b-D-Xylp unit and disubstituted AXOS with two a-L-Araf units at the nonreducing end b-D-Xylp unit of xylobiose or xylotriose were produced. Xylobiose with b-D-Xylp(1→2)-a-L-Araf(1→3) side chain was also purified. These AXOS were used as standards in further identification and quantification of corresponding AXOS from the hydrolysates in high-performance anion-exchange chromatography with pulsed amperometric detection (HPAEC-PAD) analysis. The prebiotic potential of AXOS was tested in in vitro fermentation experiments. Bifidobacterium adolescentis ATCC 15703 and B. longum ATCC 15707 utilized AXOS from the AX hydrolysates. Both species released L-arabinose from AXOS, but B. adolescentis consumed the XOS formed, whereas B. longum fermented the L-arabinose released. The third species tested, B. breve ATCC 15700, grew poorly on these substrates. When cultivated on pure AXOS, the bifidobacterial mixture utilized pure singly substituted AXOS almost completely, but no growth was detected with pure doubly substituted AXOS as substrates. However, doubly substituted AXOS were utilized from the mixture of xylose, XOS and AXOS. Faecal microbiota utilized both pure singly and doubly substituted AXOS. Thus, a mixture of singly and doubly substituted AXOS could function as a suitable, slowly fermenting prebiotic substance. This thesis contributes to the structural information on cereal AX and preparation of mono and doubly substituted AXOS from AX. Understanding the utilization strategies is fundamental in evaluating the prebiotic potential of AXOS. Further research is still required before AXOS can be used in applications for human consumption.

Project Energise: Using participatory approaches and real time computer prompts to reduce occupational sitting and increase work time physical activity in office workers

Objectives This efficacy study assessed the added impact real time computer prompts had on a participatory approach to reduce occupational sedentary exposure and increase physical activity. Design Quasi-experimental. Methods 57 Australian office workers (mean [SD]; age = 47 [11] years; BMI = 28 [5] kg/m2; 46 men) generated a menu of 20 occupational ‘sit less and move more’ strategies through participatory workshops, and were then tasked with implementing strategies for five months (July–November 2014). During implementation, a sub-sample of workers (n = 24) used a chair sensor/software package (Sitting Pad) that gave real time prompts to interrupt desk sitting. Baseline and intervention sedentary behaviour and physical activity (GENEActiv accelerometer; mean work time percentages), and minutes spent sitting at desks (Sitting Pad; mean total time and longest bout) were compared between non-prompt and prompt workers using a two-way ANOVA. Results Workers spent close to three quarters of their work time sedentary, mostly sitting at desks (mean [SD]; total desk sitting time = 371 [71] min/day; longest bout spent desk sitting = 104 [43] min/day). Intervention effects were four times greater in workers who used real time computer prompts (8% decrease in work time sedentary behaviour and increase in light intensity physical activity; p < 0.01). Respective mean differences between baseline and intervention total time spent sitting at desks, and the longest bout spent desk sitting, were 23 and 32 min/day lower in prompt than in non-prompt workers (p < 0.01). Conclusions In this sample of office workers, real time computer prompts facilitated the impact of a participatory approach on reductions in occupational sedentary exposure, and increases in physical activity.

Intensive versus conventional glycaemic control for treating diabetic foot ulcers

Background The estimated likelihood of lower limb amputation is 10 to 30 times higher amongst people with diabetes compared to those without diabetes. Of all non-traumatic amputations in people with diabetes, 85% are preceded by a foot ulcer. Foot ulceration associated with diabetes (diabetic foot ulcers) is caused by the interplay of several factors, most notably diabetic peripheral neuropathy (DPN), peripheral arterial disease (PAD) and changes in foot structure. These factors have been linked to chronic hyperglycaemia (high levels of glucose in the blood) and the altered metabolic state of diabetes. Control of hyperglycaemia may be important in the healing of ulcers. Objectives To assess the effects of intensive glycaemic control compared to conventional control on the outcome of foot ulcers in people with type 1 and type 2 diabetes. Search methods In December 2015 we searched: The Cochrane Wounds Specialised Register; The Cochrane Central Register of Controlled Trials (CENTRAL) (The Cochrane Library); Ovid MEDLINE; Ovid MEDLINE (In-Process & Other Non-Indexed Citations); Ovid EMBASE; EBSCO CINAHL; Elsevier SCOPUS; ISI Web of Knowledge Web of Science; BioMed Central and LILACS. We also searched clinical trial databases, pharmaceutical trial databases and current international and national clinical guidelines on diabetes foot management for relevant published, non-published, ongoing and terminated clinical trials. There were no restrictions based on language or date of publication or study setting. Selection criteria Published, unpublished and ongoing randomised controlled trials (RCTs) were considered for inclusion where they investigated the effects of intensive glycaemic control on the outcome of active foot ulcers in people with diabetes. Non randomised and quasi-randomised trials were excluded. In order to be included the trial had to have: 1) attempted to maintain or control blood glucose levels and measured changes in markers of glycaemic control (HbA1c or fasting, random, mean, home capillary or urine glucose), and 2) documented the effect of these interventions on active foot ulcer outcomes. Glycaemic interventions included subcutaneous insulin administration, continuous insulin infusion, oral anti-diabetes agents, lifestyle interventions or a combination of these interventions. The definition of the interventional (intensive) group was that it should have a lower glycaemic target than the comparison (conventional) group. Data collection and analysis All review authors independently evaluated the papers identified by the search strategy against the inclusion criteria. Two review authors then independently reviewed all potential full-text articles and trials registry results for inclusion. Main results We only identified one trial that met the inclusion criteria but this trial did not have any results so we could not perform the planned subgroup and sensitivity analyses in the absence of data. Two ongoing trials were identified which may provide data for analyses in a later version of this review. The completion date of these trials is currently unknown. Authors' conclusions The current review failed to find any completed randomised clinical trials with results. Therefore we are unable to conclude whether intensive glycaemic control when compared to conventional glycaemic control has a positive or detrimental effect on the treatment of foot ulcers in people with diabetes. Previous evidence has however highlighted a reduction in risk of limb amputation (from various causes) in people with type 2 diabetes with intensive glycaemic control. Whether this applies to people with foot ulcers in particular is unknown. The exact role that intensive glycaemic control has in treating foot ulcers in multidisciplinary care (alongside other interventions targeted at treating foot ulcers) requires further investigation.

Prevalence of foot disease and risk factors in general inpatient populations: A systematic review and meta-analysis

Objective: To systematically review studies reporting the prevalence in general adult inpatient populations of foot disease disorders (foot wounds, foot infections, collective ‘foot disease’) and risk factors (peripheral arterial disease (PAD), peripheral neuropathy (PN), foot deformity). Methods: A systematic review of studies published between 1980 and 2013 was undertaken using electronic databases (MEDLINE, EMBASE and CINAHL). Keywords and synonyms relating to prevalence, inpatients, foot disease disorders and risk factors were used. Studies reporting foot disease or risk factor prevalence data in general inpatient populations were included. Included study's reference lists and citations were searched and experts consulted to identify additional relevant studies. 2 authors, blinded to each other, assessed the methodological quality of included studies. Applicable data were extracted by 1 author and checked by a second author. Prevalence proportions and SEs were calculated for all included studies. Pooled prevalence estimates were calculated using random-effects models where 3 eligible studies were available. Results: Of the 4972 studies initially identified, 78 studies reporting 84 different cohorts (total 60 231 517 participants) were included. Foot disease prevalence included: foot wounds 0.01–13.5% (70 cohorts), foot infections 0.05–6.4% (7 cohorts), collective foot disease 0.2–11.9% (12 cohorts). Risk factor prevalence included: PAD 0.01–36.0% (10 cohorts), PN 0.003–2.8% (6 cohorts), foot deformity was not reported. Pooled prevalence estimates were only able to be calculated for pressure ulcer-related foot wounds 4.6% (95% CI 3.7% to 5.4%)), diabetes-related foot wounds 2.4% (1.5% to 3.4%), diabetes-related foot infections 3.4% (0.2% to 6.5%), diabetes-related foot disease 4.7% (0.3% to 9.2%). Heterogeneity was high in all pooled estimates (I2=94.2–97.8%, p<0.001). Conclusions: This review found high heterogeneity, yet suggests foot disease was present in 1 in every 20 inpatients and a major risk factor in 1 in 3 inpatients. These findings are likely an underestimate and more robust studies are required to provide more precise estimates.