text:
- Research
- Open Access
The effect of bone marrow microenvironment on the functional properties of the therapeutic bone marrow-derived cells in patients with acute myocardial infarction
© Miettinen et al; licensee BioMed Central Ltd. 2012
- Received: 28 November 2011
- Accepted: 2 April 2012
- Published: 2 April 2012
Treatment of acute myocardial infarction with stem cell transplantation has achieved beneficial effects in many clinical trials. The bone marrow microenvironment of ST-elevation myocardial infarction (STEMI) patients has never been studied even though myocardial infarction is known to cause an imbalance in the acid-base status of these patients. The aim of this study was to assess if the blood gas levels in the bone marrow of STEMI patients affect the characteristics of the bone marrow cells (BMCs) and, furthermore, do they influence the change in cardiac function after autologous BMC transplantation. The arterial, venous and bone marrow blood gas concentrations were also compared.
Blood gas analysis of the bone marrow aspirate and peripheral blood was performed for 27 STEMI patients receiving autologous stem cell therapy after percutaneous coronary intervention. Cells from the bone marrow aspirate were further cultured and the bone marrow mesenchymal stem cell (MSC) proliferation rate was determined by MTT assay and the MSC osteogenic differentiation capacity by alkaline phosphatase (ALP) activity assay. All the patients underwent a 2D-echocardiography at baseline and 4 months after STEMI.
As expected, the levels of pO2, pCO2, base excess and HCO3 were similar in venous blood and bone marrow. Surprisingly, bone marrow showed significantly lower pH and Na+ and elevated K+ levels compared to arterial and venous blood. There was a positive correlation between the bone marrow pCO2 and HCO3 levels and MSC osteogenic differentiation capacity. In contrast, bone marrow pCO2 and HCO3 levels displayed a negative correlation with the proliferation rate of MSCs. Patients with the HCO3 level below the median value exhibited a more marked change in LVEF after BMC treatment than patients with HCO3 level above the median (11.13 ± 8.07% vs. 2.67 ± 11.89%, P = 0.014).
Low bone marrow pCO2 and HCO3 levels may represent the optimal environment for BMCs in terms of their efficacy in autologous stem cell therapy in STEMI patients.
- Blood gas analysis
- Bone marrow stem cells
- Cell therapy
- Mesenchymal stem cells
- Myocardial infarction
Acute myocardial infarction (AMI) is the major cause of congestive heart failure and subsequent mortality in the developed countries. Despite the major advances in treatment methods, myocardial infarction usually causes irreversible damage to heart muscle. Cell therapy based on autologous stem cell transplantation and potential myocardial regeneration has been the focus of many clinical trials for more than a decade. These trials have yielded contradictory results which have been proposed to be attributable to the heterogeneity of the study designs [1]. A recent experimental study showed that donor myocardial infarction impaired the therapeutic potential of bone marrow cells in mice and it was hypothesized that this might explain why human cell therapy trials have not matched the success achieved in rodent experiments [2]. Moreover, it has not been previously studied if the bone marrow microenvironment of patients with ST-elevation myocardial infarction (STEMI) can affect the functionality of the cells used in autologous cell therapy. Myocardial infarction has been shown to result in metabolic acidosis [3] and base excess in the venous blood has proved to be an independent predictor for intra-intensive care unit mortality of STEMI patients [4]. Since the blood gases measured from venous blood have been shown to correlate with bone marrow measurements [5], the acid-base status of STEMI patients likely affects also the microenvironment of bone marrow. After a careful literature review, we were not able to find any previous data on bone marrow physiological conditions in adults. Thus, the aim of the present study was to assess if the blood gas or electrolyte levels in the bone marrow of STEMI patients can influence the characteristics of the bone marrow cells and, subsequently, the success of autologous stem cell transplantation. We also compared arterial, venous and bone marrow blood gas and electrolyte concentrations in patients with STEMI. This study is part of a pilot study (FINCELL II), which followed the original FINCELL (FINnish stem CELL) trial [6].
Patients characteristics
Low dose
(5 × 107), n = 6
High dose
(7 × 108), n = 21
Age (years)
56 ± 10
60 ± 9
Male sex (%)
Body mass index
28 ± 5
28 ± 3
Hypertension [n(%)]
2 (33)
5 (24)
Diabetes [n(%)]
1 (5)
TnI max (μg/L)
52 ± 47
51 ± 56
Infarct-related vessel [n(%)]
3 (50)
6 (29)
3 (50)
14 (67)
1 (5)
LVEF at baseline (%)
47.4 ± 9.2
44.4 ± 6.3
LVEF at 4 months' follow-up (%)
50.5 ± 13.2
52.3 ± 11.6
TIMI flow before PCI [n(%)]
2 (33)
4 (19)
1 (17)
2 (10)
1 (17)
3 (14)
1 (17)
9 (43)
TIMI flow after PCI [n(%)]
1 (17)
2 (10)
4 (33)
17 (81)
%Stenosis of the infarct-related artery before PCI
95 ± 5
93 ± 7
Time from STEMI to BMC injection (h)
268 ± 162
214 ± 94
Number of injected BMCs
Number of mononuclear cells (× 106)
54 ± 22
701 ± 254
Number of CD34+ cells (× 106)
0.75 ± 0.46
8.9 ± 5.1
Adenosine infusion [n(%)]
9 (43)
Medication at discharge [n(%)]
6 (100)
20 (95)
6 (100)
21 (100)
6 (100)
21 (100)
ACE inhibitor/AT II receptor blocker
6 (100)
21 (100)
6 (100)
18 (86)
2 (33)
9 (43)
Study design
The day of acute STEMI was defined as day 0. On days 1-5, patients were randomly assigned in a double-blinded fashion, either to the group receiving the small cell dose (10 ml, mean cell count 5 × 107, n = 6) or to the group receiving the high cell dose (160 ml, mean cell count 7 × 108, n = 21). The patients and investigators performing the cell injection procedures and analysis of data were unaware of the randomization throughout the study. Bone marrow aspiration, collection and preparation of cells were performed in the morning preceding the intracoronary injection of BMCs, which was performed 4-14 days post-STEMI.
Cell preparation, administration and culturing
BMCs were collected in the morning of the cell administration day under local anesthesia and sterile conditions. 0.5-1 ml sample was aspirated from one-two iliac crest into a RAPIDLyte® heparinized syringe and the blood gas and electrolyte levels were analysed using an i-STAT® Point-of-Care System (Abbot Laboratories, Illinois, USA). 10 ml or 160 ml of bone marrow was aspirated and placed into tubes containing heparinized phosphate buffered saline (PBS; Gibco, Paisley, UK). The bone marrow cells were then transported to a culture laboratory dedicated to aseptic cell manipulation for autologous stem cell transplantation. The mononuclear cell fraction containing stem and progenitor cells was isolated by Ficoll density gradient centrifugation, washed twice with PBS and resuspended with 50% autologous serum in heparinized saline or with heparinized saline alone (total volume of 10 ml). The cells were kept at +4°C until use. The BMC separation procedure took approximately 3 hours and the cells were administered within 24 hours to the target coronary artery. One aliquot of the cell sample was subjected to quality-control procedures, i.e. microbial culture for sterility and flow cytometer analysis for CD34+ cell counting and determination of cell viability in the accredited laboratory of the Oulu University Hospital, which is subjected to both outside and inside quality control. The validity of the cell preparation system was assessed as described previously [6]. One aliquot of the BMC fraction was plated into 25 cm2 tissue culture flask and cultured in 5 ml medium containing alpha MEM (Gibco) buffered with 20 mM HEPES (Gibco) and containing 10% heat-inactivated fetal calf serum (FCS; Bioclear, Netherlands), 100 U/ml penicillin, 0.1 mg/ml streptomycin, 2 mM L-glutamine (Gibco). Cells were cultured at +37°C in 5% CO2 and 95% air. After one day, the medium was changed and non-attached cells were washed away. The attached cells (mesenchymal stem cells, MSCs) were cultured in a flask and the medium was replaced two times per week until near-confluence. The cells were passed two times before the analyses. In the cell counting analyses, the cells were washed with PBS and adherent cells detached using trypsin-EDTA solution (Gibco).
Flow cytometric analysis of cell surface antigens
Flow cytometric analysis of cell surface antigens was performed as previously described [7]. The minimum criteria panel of cell surface markers for hMSCs proposed by International Society for Cellular Therapy [8] was evaluated from all MSC lines analyzed, and each of the lines met the criteria. The percentage values of positive MSC markers were CD90 99.81 ± 0.23%, CD105 99.58 ± 0.37%, CD73 99.66 ± 0.28% and HLA-ABC 97.58 ± 3.06%. The percentage value of negative markers (CD14, CD34, CD45, CD19 and HLA-DR) was 1.80 ± 0.58%. Thus, the marker analysis showed that the cell cultures contained almost purely MSCs.
MTT proliferation assay
MSCs were cultured in 96-well plates in six replicates. 500 cells/well were plated and half of the medium was changed twice a week. Cell proliferation was measured by MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay after 1, 4, 7, and 14 days culture. The medium was removed and MTT reagent (Sigma-Aldrich, St. Louis, MO, USA) (0,5 mg/ml in medium) was added to the cells and incubated for 2 h at +37°C in 5% CO2 and 95% air. After incubation, the MTT solution was removed and 100 μl/well of dimethyl sulfoxide (DMSO; Sigma-Aldrich) was added. The absorbance of the reduced form of MTT was measured at 550 nm and 650 nm (background) in a plate reader (Victor 2, Wallac Oy, Turku, Finland).
Alkaline phosphatase activity assay
Human MSCs were seeded at 10000 cells/well into 24-well plates in four replicate wells and were cultured with the medium described above and another four replicates in a medium containing also 100 mM dexamethasone (Sigma, St. Louis, MO, USA), 10 mM β-glycerol (Sigma) and 0.05 mM ascorbic acid (Sigma). After 3 weeks, cultured cells were assayed in the following way: the assay buffer containing 0.1% Triton X-100, pH 7.6, was added to each well, and the plates were frozen. After thawing, alkaline phosphatase (ALP) activity was determined using 0.1 mM 4-p-nitrophenylphosphate (Sigma-Aldrich) as the substrate and absorbance was read at 405 nm in a plate reader (Victor 2, Wallac Oy, Turku, Finland). Each sample was measured in duplicate. The protein content of the wells was determined by the BIO-RAD Protein Assay (Bio-Rad Laboratories, Richmond, California, USA). The enzyme activities were expressed as units/mg protein.
Blood sampling, blood gas and biochemical laboratory analyses
All the patients underwent venous and arterial blood sampling on the day of cell transplantation. pH, pO2, pCO2, HCO3, BE, Na+ and K+ levels were measured in the central laboratory of Oulu University Hospital.
Measurement of left ventricular ejection fraction
A 2-dimensional echocardiogram was performed 2-5 days after STEMI and at 4 months after STEMI. The LVEF was measured by an experienced investigator in the core laboratory, unaware of the patient's treatment assignment, using the technique described previously [9].
Statistical analysis
Variables are expressed as means ± SD, or medians with interquartile range with skewed data. Paired samples t-test was used to determine the statistical significance of the difference in laboratory values between bone marrow and arterial/venous blood. Correlation analyses between bone marrow and arterial/venous blood laboratory values were conducted using paired samples correlation. In other analyses, Pearson's correlation or Spearman's correlation with skewed data was used. Analysis-of-variance (ANOVA) was used in the between-group comparisons. The variance analyses of the LVEF change in HCO3, BE and potassium subgroups (above/below median) were corrected with the cell dose group (low/high dose). All P values are two-tailed and statistical significance was set at P < 0.05. Analyses were performed with SPSS software, version 14.0.
Blood gas and electrolyte analyses of bone marrow, venous and arterial blood
Laboratory analyses of bone marrow, venous and arterial blood
Laboratory variable
Bone marrow
Venous blood
Arterial blood
P value for paired difference (BM vs. vein)
P value for paired difference (BM vs. artery)
7.38 ± 0.036
7.40 ± 0.031
7.42 ± 0.048
pO2 (kPa)
5.11 ± 0.68
5.63 ± 2.17
11.15 ± 2.79
< 0.001
pCO2 (kPa)
5.78 ± 0.81
5.62 ± 0.87
4.93 ± 0.84
< 0.001
HCO3 (mmol/L)
25.43 ± 2.83
25.25 ± 2.85
22.92 ± 1.86
< 0.001
BE (mmol/L)
-0.042 ± 2.46
0.20 ± 2.27
-1.09 ± 1.57
Na+ (mmol/L)
138.96 ± 1.83
140.29 ± 2.26
K+ (mmol/L)
4.96 ± 0.64
4.06 ± 0.37
Association of bone marrow blood gas and electrolyte levels with the functional properties of bone marrow cells
Association of bone marrow blood gas and electrolyte levels with functional properties of bone marrow cells
Laboratory variable
CD34+ cell viability
MSC proliferation rate
MSC osteoblast differentiation
Correlation coefficient
Correlation coefficient
Correlation coefficient
pO2 (kPa)
pCO2 (kPa)
HCO3 (mmol/L)
BE (mmol/L)
Na+ (mmol/L)
K+ (mmol/L)
Association of bone marrow blood gas and electrolyte levels with the functional recovery of the heart after cell therapy
Association of bone marrow blood gas and electrolyte levels with the change of LVEF after cell therapy
Laboratory variable
Change of LVEF (%-points)
Correlation coefficient
pO2 (kPa)
pCO2 (kPa)
HCO3 (mmol/L)
BE (mmol/L)
Na+ (mmol/L)
K+ (mmol/L)
In an attempt to clarify the individual differences in the levels of bone marrow blood gases, the associations of several other parameters (e.g. age, sex, body mass index, hypertension, hypercholesterolemia, diabetes, heart failure, severity of CAD, medication at discharge) with the bone marrow blood gas values were tested. The bone marrow BE and HCO3 levels were found to be higher in patients using diuretic drugs at the time of hospital discharge than in patients with no diuretic medication (BE 1.10 ± 2.81 vs. -0.87 ± 1.81 mmol/L, P = 0.043 and HCO3 26.69 ± 3.46 vs. 24.54 ± 1.89 mmol/L, P = 0.056, respectively). In addition, the patients using diuretics experienced a smaller improvement in the LVEF than patients without diuretic medication (1.81 ± 12.42 vs. 10.24 ± 7.41%-units, respectively, P = 0.036). No significant associations were found between any other patient characteristics and the bone marrow blood gases (P = NS in each analysis, data not shown).
The results of the present study demonstrate that there is a significant difference in blood gas and electrolyte levels between bone marrow and arterial blood in patients with STEMI. In addition, a significant difference was found between bone marrow aspirate and venous blood in pH, sodium and potassium levels. In contrast, the levels of pO2, pCO2, BE and HCO3 were similar in venous blood and bone marrow. In a further blood gas analysis, the proliferation rate of bone marrow-derived MSCs was associated with the pCO2 and HCO3 levels in the bone marrow. The MSC proliferation was more efficient when bone marrow pCO2 and HCO3 levels were low. It was also shown that a low HCO3 level in bone marrow was associated with better functional recovery of the heart after myocardial infarction treated with intracoronary injection of autologous BM cells. Patients with an HCO3 level below the median value experienced a more marked improvement of LVEF than patients with the HCO3 level above the median value. In contrast, the osteogenic differentiation potential of the BM-derived MSCs was shown to be better when bone marrow pCO2 and HCO3 levels were high.
One objective of this study was to compare arterial, venous and bone marrow blood gas and electrolyte concentrations in STEMI patients. As far as we are aware, only one previous study has assessed the levels of blood gases and electrolytes in bone marrow and venous blood in humans [5]. Grisham et al. studied children with acute leukemia or other hematological disorders. In those patients, a significant correlation was found between venous and bone marrow samples for pH, pCO2, HCO3, BE and sodium concentration, but no predictable relationship was observed for pO2 and potassium concentration. In the present study, pCO2, BE and HCO3 also correlated between bone marrow and venous blood as well as bone marrow and arterial blood. However, pH, sodium and potassium levels differed significantly between venous blood and bone marrow. In the study of Grisham et al. the actual differences between bone marrow and venous blood were not assessed. However, they also noticed that the potassium levels differed significantly between bone marrow and venous blood. They considered that this could be due to the traumatic nature of specimen collection and the high negative pressure generated in aspirating marrow evoking hemolysis [5]. A portion of the cells inevitably break during the bone marrow aspiration but we think that the cell breakage does not fully explain the difference in potassium concentration between bone marrow and venous blood because we found a difference also in the sodium concentration. Instead, it may be a true finding being related to the different activity of sodium-potassium pump.
Bicarbonate is alkaline, and it is one of the components that maintain acid-base homeostasis in the body. Most of the CO2 in the body is converted into carbonic acid (H2CO3), which rapidly forms bicarbonate (HCO3-). The pH in the body depends on the pCO2 and the level of HCO3-. In this study MSC proliferation was found to be more efficient when the bone marrow aspirate pCO2 and HCO3 levels were low. The pH in the bone marrow was not directly associated to the proliferation rate but this may be due to other compensatory buffering mechanisms, such as hemoglobin, which is known to stabilize the pH value. It is also known that active inflammation creates an acidic environment [10]. In the present study, the levels of pCO2 and HCO3 displayed a nonsignificant negative correlation with bone marrow interleukin (IL)-6 levels (data not shown). Thus, the levels of pCO2 and HCO3 may have declined in an attempt to buffer the acidosis caused by inflammatory processes. We have previously shown that inflammation (mediated by tumour necrosis factor alpha) enhances the proliferation of MSCs and activates many immunosuppressive pathways in vitro[7]. Thus, if inflammation is more extensive in the bone marrow of the patients with lower levels of pCO2 and HCO3, it can be postulated to have an influence on the resident MSCs and enhance also their in vitro proliferation. On the contrary, the osteogenic differentiation potential of the BM cells was found to be better when bone marrow pCO2 and HCO3 levels were high. This result is in line with previous studies which have shown that acidic environment impairs osteoblast function,[11, 12] reduces osteoblast alkaline phosphatase (ALP) activity [13, 14] and inhibits osteogenic differentiation of MSCs [14, 15].
Several previous trials have shown that the number of bone marrow CD34+ cells does not correlate with the change of LVEF after cell transplantation [16–19]. In the present study, the viability of CD34+ cells was not associated with the pCO2 and HCO3 levels. In addition, the observation that low bone marrow pCO2 and HCO3 is beneficial for BM-derived MSC proliferation but not for the osteoblast differentiation suggests that maintaining the stemness of MSCs or some other cell lineage is predominant in this kind of environment. It has been previously shown that hMSCs cultured in hypoxic conditions (2% oxygen) display significantly improved expansion characteristics while maintaining their multi-lineage potential [20]. On the other hand, hypoxia as such can lead to unfavorable effects in the cultured cells, such as chromosomal aberrations [21]. Obviously, one of the most important functions of the bone marrow microenvironment is to support the stemness of the resident BMCs and protect the infinitely dividing cell population from oxygen derived free radicals that could theoretically promote the cancer propagation of these cells. It has also been shown that bone marrow contains many different microenvironments in which MSCs are the prominent cell components; these cells play many important roles including controlling hematopoiesis [22, 23]. Based on our results, it seems that the optimal environment for BM cells in this respect is associated with a low bone marrow pCO2 value and reduced HCO3 levels which support the stemness of BM cells making them more efficient for cell therapy use and also more capable of undergoing intrinsic repair mechanisms. However, the results of this study are still preliminary findings and the importance of bone marrow microenvironment on MSC functionality after two MSC passages in culture media with 5% CO2 and 95% air, which are completely different conditions compared to the bone marrow of STEMI patients, remain to be elucidated. Further tests in cell cultures are currently ongoing and will later provide us more insights about the effect of different environmental factors of the bone marrow on the functionality of the BM cells.
As far as we are aware, this is the first study to assess the effects of the metabolic state of bone marrow on the functional capacity of the bone marrow-derived stem cells in STEMI patients. It has been previously shown that age, risk factors for coronary artery disease, diabetes and heart failure diminish the capacity of the blood-derived circulating cells to contribute to the functional repair compared to healthy controls [24–26]. In addition, when the number and functionality of bone marrow-derived cells have been analyzed, the presence of heart failure also has been shown to affect these cells. BMCs isolated from bone marrow aspirates of patients with ischemic heart failure have been less effective in achieving a recovery of blood flow after hind limb ischemia compared to cells from healthy controls [27]. The comparison of the composition and function of BMCs has demonstrated that the number of granulocyte-macrophage colony-forming units (GM-CFUs) is significantly lower and the migratory capacity of the cells is reduced in patients with ischemic heart failure as compared to healthy controls [27, 28]. Moreover, chronic heart failure has been shown to be an independent predictor of bone marrow cell impairment, whereas the cardiovascular risk factors have not been statistically predictive [28]. We have also earlier shown that the energy metabolism of bone marrow mesenchymal cells is different in aged (> 50 years old) patients compared to young (< 18 years) and umbilical cord blood mesenchymal stem cells [29]. In the present study, the age of the patients was not associated with the parameters describing the metabolic state of bone marrow or the viability of BMCs. Heart failure, hypercholesterolemia, diabetes or hypertension were not associated to these parameters either. However, this may be due to the small patient number in each disease group. Further studies with larger patient populations are needed to clarify the association between these diseases and the bone marrow metabolic state. There were also attempts to collect samples from healthy hip fracture patients and patients undergoing spinal surgery to compare their bone marrow blood gas levels with STEMI patients. However, there were serious technical difficulties in the measurement of the thick bone marrow sample taken from the open bone and also arterial blood 'contamination'. Thus, the comparison of blood gas values between healthy subjects and patients with myocardial infarction remains to be elucidated in future studies.
The LVEF change between baseline and follow-up visit has been the most widely used endpoint to evaluate the effectiveness of stem cell transplantation after myocardial infarction [1]. In the present study, patients with the bone marrow HCO3 level below the median value experienced a more marked improvement of LVEF after stem cell treatment than their counterparts with an HCO3 level above the median value. This clinical trial was designed to be a dose-response study but no difference in the LVEF recovery could be observed between the groups receiving low or high dose of BMCs in this pilot phase. Thus, it is not likely that the differences in the cell doses would have affected the EF results of the present substudy. However, because of the low patient number, the results of this study have to be interpreted as descriptive rather than confirmatory when considering the importance of the blood gas values of the bone marrow on the clinical outcome of BMC treated STEMI patients. More studies with higher patient number are needed to confirm these preliminary findings.
There is a significant difference in blood gas and electrolyte levels between bone marrow and arterial blood in patients with STEMI. There are also significant differences in the pH, sodium and potassium levels between bone marrow aspirate and venous blood. Even though bone marrow acid-base status and blood gas levels resemble more closely venous blood than its arterial counterpart, the bone marrow seems to form a unique compartment for maintenance and storage of stromal cells, similarly to that reported in the context of the hematopoietic stem cell niche. The proliferation rate of bone marrow-derived MSCs is more efficient when bone marrow pCO2 and HCO3 levels are low. In contrast, the osteogenic differentiation potential of the BM-derived MSCs is better when bone marrow pCO2 and HCO3 levels are high. The STEMI patients with lower HCO3 levels experience a more marked change in LVEF. Moreover, patients using diuretic drugs at the time of hospital discharge tended to have higher bone marrow pCO2 and HCO3 levels and these patients experienced a poorer improvement in the LVEF after stem cell therapy, most probably because of the underlying cardiac or venous congestion affecting also the bone marrow metabolic state. In this respect, low bone marrow pCO2 and HCO3 levels represent the optimal environment for BM cells in terms of their efficacy in autologous stem cell therapy in STEMI patients.
We would like to thank Ms. Kirsi Kvist-Mäkelä, Ms. Päivi Kastell, Ms. Pirkko Huikuri, Ms. Päivi Koski, and Ms. Sari Kaarlenkaski for their excellent assistance in this study.
This study was supported by grants from the Medical Council of the Academy of Finland, Helsinki, Finland and Finnish Foundation for Cardiovascular Research, Helsinki, Finland, and to Dr. Miettinen; grant from Ida Montin Foundation, Helsinki, Finland.
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- Askmyr M, Sims NA, Martin TJ, Purton LE: What is the true nature of the osteoblastic hematopoietic stem cell niche?. Trends Endocrinol Metab. 2009, 20: 303-309. 10.1016/j.tem.2009.03.004.View ArticlePubMedGoogle Scholar
- Vasa M, Fichtlscherer S, Aicher A, Adler K, Urbich C, Martin H, Zeiher AM, Dimmeler S: Number and migratory activity of circulating endothelial progenitor cells inversely correlate with risk factors for coronary artery disease. Circ Res. 2001, 89: E1-E7. 10.1161/hh1301.093953.View ArticlePubMedGoogle Scholar
- Tepper OM, Galiano RD, Capla JM, Kalka C, Gagne PJ, Jacobowitz GR, Levine JP, Gurtner GC: Human endothelial progenitor cells from type II diabetics exhibit impaired proliferation, adhesion, and incorporation into vascular structures. Circulation. 2002, 106: 2781-2786. 10.1161/01.CIR.0000039526.42991.93.View ArticlePubMedGoogle Scholar
- Dimmeler S, Leri A: Aging and disease as modifiers of efficacy of cell therapy. Circ Res. 2008, 102: 1319-1330. 10.1161/CIRCRESAHA.108.175943.View ArticlePubMedPubMed CentralGoogle Scholar
- Heeschen C, Lehmann R, Honold J, Assmus B, Aicher A, Walter DH, Martin H, Zeiher AM, Dimmeler S: Profoundly reduced neovascularization capacity of bone marrow mononuclear cells derived from patients with chronic ischemic heart disease. Circulation. 2004, 109: 1615-1622. 10.1161/01.CIR.0000124476.32871.E3.View ArticlePubMedGoogle Scholar
- Kissel CK, Lehmann R, Assmus B, Aicher A, Honold J, Fischer-Rasokat U, Heeschen C, Spyridopoulos I, Dimmeler S, Zeiher AM: Selective functional exhaustion of hematopoietic progenitor cells in the bone marrow of patients with postinfarction heart failure. J Am Coll Cardiol. 2007, 49: 2341-2349. 10.1016/j.jacc.2007.01.095.View ArticlePubMedGoogle Scholar
- Pietilä M, Palomäki S, Lehtonen S, Ritamo I, Valmu L, Nystedt J, Laitinen S, Leskelä HV, Sormunen R, Pesälä J, Nordstrom K, Vepsäläinen A, Lehenkari P: Mitochondrial Function and Energy Metabolism in Umbilical Cord Blood- and Bone Marrow-Derived Mesenchymal Stem Cells. Stem Cells Dev. 2012, 21: 575-588. 10.1089/scd.2011.0023.View ArticlePubMedPubMed CentralGoogle Scholar
This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.


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Title: The Infinite Sea (The 5th Wave #2)
Author: Rick Yancey
Rating: ★ ★ ★ 1/2
Genre: Science-Fiction
Format Read: Paperback, 336 pages
Publication Date: 16th September 2014 by Penguin Australia
Synopsis: "The hugely anticipated follow-up to the New York Times bestselling The 5th Wave, hailed by Justin Cronin as "wildly entertaining."
For Cassie Sullivan and the rest of Earth's remaining human survivors, the situation was already desperate when the 5th Wave hit. It's about to get worse.
No one yet knows the depths to which Earth's conquerors—the Others—will sink in order to rid the Earth of the human infestation, nor have they guessed the heights to which the human spirit can reach. Characters introduced in Book One will come to the fore—and others will face the ultimate test.
Readers will watch in awe as the Others give their answer to Cassie's defiance." (Taken from Goodreads)
[label type="alert"]This review contains spoilers of the first book.[/label]
This book is without a doubt one of my most anticipated books of the year. It is another crazy rollercoaster ride of emotions that I just…I just can't. I don't really know how to put my feelings into words.
The 5th Wave left us readers on a cliffhanger of 'What happened to Evan Walker?!?!?!' While I still found him quite odd and creepy, I couldn't help but like him either. He was so sweet and nice to Cassie … pretty fly for an alien guy (see what I did there?)
While the first book was about solitude, forming alliances and saving humanity, The Infinite Sea seems a lot more dire as it becomes each man for themselves. Cassie has joined Zombie's group of runaways and they're all at heads with each other on what to do, where to go and how to proceed as each day brings one of them closer to death. Cassie wants to stay at the hotel they've camped out at in case Evan Walker shows up, but the rest of the group thinks she's insane. No one truly believes that an alien could have fallen in love with her, let alone want to go against his own race. Or you know, have survived that deadly explosion from the first book.
The Infinite Sea is about survival at its core and we are treated with a range of different perspectives as Cassie gets sidelined while other characters take the stage. Through flashbacks, we are given a more thorough understanding of each character's background, especially the ever silent Poundcake, who refuses to speak, and the stoic Ringer. But more than just survival, this book is also about sacrifice. The things each character as had to give up from their old lives. Nugget, or Sams, has given up his right to a childhood. Cassie puts it perfectly when she says she doesn't recognise her brother anymore.
"I didn't recognize him. I'd never seen this kid before. Face ugly and twisted by rage and hate." (p. 39)
This change is fully realised when Cassie leaves him with a gun to protect himself. It truly symbolised the end of his childhood and what was soon to come.
Now if you're wondering whether we actually get some answers in this book, then I'm happy to say that we do. Do they make sense? Well, that's for you to judge. As for me, well …
Haha just kidding, but the explanation is not exactly straightforward. As Ringer journeys out by herself to find the group a new hideout, she ends up being captured by Vosch as she sacrifices herself to save someone. She's taken back to a different camp and is manipulated and used as a puppet in another of Vosch's games. All to capture the one person that could cause this whole operation to fail.
Vosch continually asks her why she thinks they're doing this to Earth. Why use manipulation to kill when a simple rock would have done the trick and wiped out the entire population. Well it's all about 'VINCIT QUI PATITUR'. I will not tell you what it means because it'll take away from the story, but if there's one central theme revolving around this book it's those three latin words.
This book definitely killed me slowly. Everything you wish would happen definitely doesn't as Yancey twists and steers the reader towards the ultimate ending. Don't get too happy because he's evil and will squish the hope that's blossomed in your heart. Seriously. I will show you my stack of used tissues.
While I didn't like this book as much as the first, it is a worthwhile sequel. You will get your answers, but you will also get your heart ripped out. Proceed with caution!
Thank you to Penguin Australia for providing a copy in exchange for an honest review.

subset: commoncrawl


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- Poster presentation
- Open Access
The PRINTO provisional definition of remission in juvenile dermatomyositis
© Lazarevic et al; licensee BioMed Central Ltd. 2011
- Published: 14 September 2011
- Active Phase
- Myositis
- Clinical Remission
- Dermatomyositis
- Creatine Phosphokinase
To define a definition of remission in juvenile dermatomyositis (JDM) that is stringent but achievable and could be applied uniformly as an outcome measure in clinical trials.
275 patients in active phase of JDM <18 years, median disease duration 7.7 months, were evaluated at baseline and 24 months. Out of 275 patients, all patients (39/275) who were off treatment at 24 months were defined as being in clinical remission and were included as the "gold standard" group. A random sample of patients (n=78) who were in an active phase of JDM at baseline and on medications at 24 months, were selected as the "comparison group". Literature was reviewed for definitions of remission and in addition other definitions were tested which included PRINTO core set variables, muscle enzymes, and other JDM activity measures. Accuracy measurements were calculated: sensitivity, specificity, Youden index, Cohen's kappa (≥0.8 almost perfect).
Four best single variables with kappa≥0.8 were Manual Muscle Testing (MMT)≥78 (sensitivity: 0.92, specificity 0.97, kappa 0.9), followed by Physician Global Assessment of Muscle Activity (PhyMAVAS) and Physician Global Visual Analogue Scale (PhyGloVAS)≤0.2 and Childhood Myositis Assessment Scale (CMAS)≥48. The most accurate combination of variables with kappa≥0.8 included 4 variables with 3 out of 4 required to be present: creatine phosphokinase ≤150, CMAS≥48, MMT≥78, PhyGloVAS≤0.2 (sensitivity 0.94, specificity 1.00, kappa 0.96).
The combination definition performed the best overall in defining remission in JDM and could be used for the evaluation of global response to therapy in future clinical trials.
Authors' Affiliations
This article is published under license to BioMed Central Ltd. This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.


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These days many students are looking forward to studying abroad because of various reasons. Some popular countries that student opt for are New Zealand, UK, USA and Australia. One of the most visited places by students is Australia. Many students choose Australia because it suits their requirements in various ways.
To get an Australian students visa, a student needs to know the following procedures and the requirements. A student should be aware of it to avoid any problems in the future.
For a student to get a student visa, the student must satisfy the basic requirements and go by the visa conditions. This is required for an academic performance and attendance which is considered after the student is in Australia.
Every student's visa has a list of conditions that are specified for every individual. It thus becomes very important for every student to abide by the visa conditions so that the person can prevent any visa cancellation. Every student's application is assessed on a few criteria's:
The important thing for a student is that the enrolment should be accepted by the Government registered Australian education institution for the desired course. This is then further checked by the Australian institution. This is given to a student who later gets an enrolment for a full time course. No tuition fee should be paid before the students get the PVA.
Financial Status
The person who is sponsoring should give the proof of cash assets which should be at least 6 months before the visa application date. Any person can sponsor the student.
The person who is sponsoring should have a proof of A$12000 p.a worth of cash assets as well as additional funds so that he/she can pay for the rest of the expenses for the student.
The IELTS will rate the student's proficiency in English and the overall score should be 6.0 other than the students who are opting for a Diploma course which has an overall score of 5.5. TOEFL score is not taken into consideration.
Every student's current circumstances are assessed by a visa officer. These officers check the condition in the student's home country. They check the student's financial and personal commitments that may force a student to go back to his/her home country.
Important Matters
If the student is adviced to meet the requirements of the PVA, then a further processing is required. The students have to go through a medical examination and pay the course tuition fee to the Australian educational institution. The dependent and student has to meet the requirements of the Australian Health before he/she gets the visa. Every individual has to go through an x-ray and a medical checkup. These results may be forwarded to Australia which would take about 3-4 weeks.
An Australian student visa application generally takes about 4-6 weeks. It may also get delayed because of verification of documents, medical tests or an incomplete document that requires the help of a visa officer.
It is a mandatory thing for all international student's to have an Overseas Student Health Cover while the visa is still valid. Once the PVA requirements are fulfilled, that when the premium for OSHC should be paid.


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Examiner.com is fully powered by Examiners, thousands of writers who are self-motivated independent contributors. Each Examiner is able to express through words and photos a deep expertise in a topic. Their knowledge is enhanced by a viewpoint unique to their experiences and oftentimes, their location.
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Have you ever compressed watermelon? You won't believe what a difference it makes! The flavor of the watermelon becomes sweeter and more intense, and in this recipe it's balanced perfectly with a bit of spice, fresh tomatoes and a tangy, creamy avocado dressing. I have so much to talk about that I don't even know where to begin.
(If my husband were editing this, he would have just told me I've already told you more that you need to know.)
Since the editor is out, I'll carry on. 😉
The thing is, I have a lot on my mind — about this recipe, and otherwise.
The Compressed Watermelon Salad Recipe
If you've never tried compressed watermelon, you must. Sometimes adding one more step, a detail, if you will, to a recipe, is all it takes to bring it from very good, to over the top. And in this case, compressing the watermelon does it.
And guess what? It's quite easy. (See recipe instructions below.)
Compressing the watermelon intensifies the flavor and the texture becomes more dense and the color more vibrant. (Here's why.)
This Compressed Watermelon Salad is a most delightful appetizer course, and would be lovely for a summer lunch.
Now, for the "otherwise" —
Among other things (which my husband might edit out), tomorrow my boys, Mason and Zack, go back to school. They'll be in 4th and 6th grade, and for the first time they'll be at two different schools. How did this even happen!? How are they not still in preschool!?
How will I be able to take Mason to his new school and go the the new parent meeting and be able to meet Zack's new teacher at the same time?
Of course everyone will survive and be just fine — except for me, that is. Survive, yes. Be just fine, I'm not so sure. I might just show up at the preschool with a box of tissues.
In the meantime, I'll be distracting myself with this this super delicious and utterly stunning Compressed Watermelon Salad.
I hope you enjoy it as much as I do!
Oh, and here are a few more of my favorite watermelon recipes:
- Watermelon Pizza
- Watermelon Cucumber Salsa
- Balsamic Glazed Fresh Melon Cake
- Balsamic Watermelon Avocado Chopped Salad
*Please note that the Creamy Lemon-Lime Avocado Dressing should be made in advance -- it takes about 10 minutes.
- about 1/4 of a large seedless watermelon
- about 1/2 cup Creamy Lemon-Lime Avocado Dressing (click here for the recipe)
- 1 cup tiny, multi-color heirloom tomatoes, sliced in half or quarters
- 1 small to medium-sized jalapeño pepper, super thinly sliced
- a few sprigs of fresh thyme
Use a long slicing knife to cut the watermelon in half, horizontally. Place one half flat side down on a clean, dry surface. Then slice a portion off the round top and then each side, making it a square or rectangle.
Now decide on your serving plates. Cut the watermelon flesh to match the shape of the plate -- could be circular, square, rectangular, or into cubes if you'd like --- it's up to you! Whatever you choose, the slices should be about ½-thick, and you should have 4 of them, or 4 generous handful of the cubes.
Place the slices (or cubes) in a large, heavy-duty zip-lock bag -- be sure the watermelon pieces are laying flat and as close together as possible. Remove as much air possible form the bag as you seal it. You may need more than one, if not a few bags. Now lay the bags flat on a baking sheet or tray that will fit into your freezer. Place something flat on top -- I use another baking sheet, but if you that doesn't work for you, you can even use a piece of cardboard. Now place heavy items on top to weigh it down -- could be cans of food, packages of frozen meat or even ice packs. Whatever works. Leave it in the freezer for 2 hours.
When you're ready to plate the watermelon, after the 2 hours, use the bottom of a spoon to thickly spread about 2 tablespoon of the Creamy Lemon-Lime Avocado Dressing on the plate -- mostly in the center. Add a watermelon slice on top and gently press it into the dressing so that it spreads beneath it.
Add about ¼ of the tomatoes to each plate, and a few slices of the jalapeño pepper (use as many as you'd like, depending on how much heat you want). Add a spring or two of thyme and serve!
Most recipes for compressed watermelon will call for vacuum sealed bags, but it seems this works just as well.


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THE FASH PACK. get all your fashion info fast, fresh, and free! every day! plus new feature: outfit posts! view my print-a-day challenge. no repeats! (until i say so.) you can also follow me on instagram @ilardeliza
Friday, November 25, 2011
style weekend - nov 25
pls buy manila bulletin today (nov 25) for your free copy of style weekend for men (out every last friday of the month)! on the cover: the face shop celebrity endorser and k-pop phenomenon, kim hyun joong! don't hate me because i got to interview him one-on-one in seoul, korea!
also in this issue: driving the yet-to-be-released BMW M5 at ascari race resort in spain; fragrances, skin care, and business accessories for men; red and green clothes & stuff; ancestral homes converted in restos; LV's architect & interiors book; and red-hot anne curtis in the centerfold!
Posted by the fash pack at 7:27 AM
Labels: style weekend
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Drama One-Act Shows held at MIHS this weekend
On March 26 and 27, the Drama 2 cast will be performing their original one-act short plays. For the past two months, the students have been working closely with professional writer and performer Andrea Karin Nelson on scripting and performing their own one-person shows. "We had to write like, 100 ideas, and then pick one and be like, 'Alright I'm gonna do it!' And now … Continue reading Drama One-Act Shows held at MIHS this weekend


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We believe everyone can find their place at Community Church. We understand that exploring Jesus or visiting a new church for the first time can be intimidating and uncomfortable experience. Community Church has a rich history spanning over 25 years of helping people just like you come to know Jesus. Today, Community Church is home to thousands of people. We're a dynamic, family church for everyone with people of all ages and from all walks of life who make up the community of Community Church.
Whether you're completely new to church or just haven't been for awhile, we'd love to see you. Join us on Sundays. At Community Church you can expect a friendly welcome, a dynamic service, and relevant teaching.
We're glad you're here.
Losing It - Week 6
Losing It - Week 5
Losing It - Week 4
Losing It - Week 3


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Artist: Band: Two One Nine
CD Review:
Barenaked Ladies better watch their back. A talented newcomer is right on their tails. In fact they might want to put in a call to their lawyers as Antipasti Insomniac is dangerously similar in guitar licks and melody to Pinch Me. Dont misunderstand me, Two One Nine is fun and intelligent music and very enjoyable. Two One Nines main writer is J. Williams, who does have some very clever and humorous lyrics, also like Barenaked Ladies. It is perfectly fine to have influences and use them as your foundation. But as Williams continues to write I hope he continues to flesh out his own voice. Everything people love about Barenaked Ladies they will love about Two One Nine. These songs are funny, sweet, clever and expertly played. In fact the production and guest players are all quite solid and impressive. This is definitely an entertaining CD and I look forward to the music written by J. Williams coming into its own singular voice. http://www.jeremywilliams.us


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Author Archives: Harrielle
Smyth Jewelers East Coast's Largest Jewelry Store
Smyth Jewelers located in Timonium, MD offers free shipping on orders of $200 or more. They have a 100% Satisfaction Guarantee: You must be thrilled with any item purchased from Smyth… They guarantee it! 100% Satisfaction guarantee Since our establishment … Continue reading
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Chamilia Quality & Unique Designs
Chamilia is one of the fastest-growing personalized accessory brands in the world creating extraordinary, innovative jewelry for the woman with an eye for fashion. Chamilia's premium quality and unique designs distinguish it from all other jewelry brands. Our collection of … Continue reading
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Great review by Matej Istak from Mono and Stereo magazine.
"High-End Audio, high performance, highest level of the aesthetics? These three should all go hand in hand without any second thought. Especially at present times when our niche industry is morphing into a newer, better self."
"The Block amplifier creation is intimately connected with G LAB Design Fidelity passion for beauty"
"BLOCK creates an open sounding experience, that never loose a focus or close connection of the music being sourced."
Read full review here.


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A (attempted) debate with a McCain shill who also blogged about what caused the fall of the Roman Empire, resulted in this astonishing revelation:
It was unrestrained immigration that finally caused the Empire to fall.
It's pretty simple stuff.
You can see the discussion on HotAir for yourself here.
This is an important discussion to be addressed in more detail later.
Update: 12:00 P.M.
It's looks like just what I figured.
It's the battle over immigration that is at stake here, with the establishment GOP, supporting McCain who is a shameless supporter of amnesty by default.
Read Daily Pundit's blog that highlights this very issue in more detail (as linked by HotAir commentator Connie.
Sunday, February 3, 2008
The cause of the fall of The Roman Empire.
Labels: Dobson, Huckabee, immigration, Keyes, Romney
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Courtney Marie Taylor
Pen and ink drawing on decoupage medium
Size: 12 x 12"
Year: 2018
My body of work is strongly influenced by the natural world. Water is a common theme throughout my paintings. I have always been drawn to the ocean, and frequently use my work to call attention to it's care. Oil painting allows me to explore the shapes and colors reflected in an ever-changing body of water. Other themes that show up in my work include: the exploration of humanity, diversity, astrology and the practice of living a mindful life. In art school, I was exposed to artist's works like Chuck Close and Kehinde Wiley. I was strongly influenced by their ability to paint highly naturalistic imagery in a more contemporary way. After art school, I became an art educator, this continued my journey learning about how art can awaken complex issues and get important conversations going. Now, I am creating art full time, living in downtown Raleigh, and continuing to practice living a mindful life.


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"Scrappy T comes away with the lead! But Afleet Alex is–oh Scrappy T blew the turn!"
We all remember that terrifying moment at the top of the stretch in the 2005 Preakness when Afleet Alex clipped heels, fell to his knees and almost went down. No one remembers it better than Scrappy T.
"It still haunts me to this day," said Scrappy T while pacing along the fence line in his pasture, "what if he actually fell? What if he died? After all the hype and feel good story with Alex and her lemonade… My god I almost ruined everything."
11 years later it is clear he is still shaken from the experience.
"I still have nightmares about it. Afleet Alex has said its fine, no one got hurt so its all good. That's not the point, it was almost a tragedy and it was all my fault. How was I supposed to know Ramon would go left handed at the top of the stretch? Who does that?! Of course I'll blow the turn!" Visibly distraught, he needed a break to go roll in some mud.
After a solid racing career, Scrappy T was originally retired to a farm in Virginia where he competed in shows. Like he always does he completely blew that opportunity, so now he is living down in Florida like most grumpy old men. His owner bought that new place as a pinhooking operation and turns Scrappy out with the yearlings to teach them a thing or two about life. "Don't panic when you get hit on your left side," Scrappy told the yearlings. "It will scare you, it will come out of no where, you wont know how to react. But whatever you do, do not bolt!"The yearlings took note of Scrappy's wise words.
Today Scrappy and Alex are good friends, they snapchat almost every day. But sometimes Alex will make low blow jokes, "LOL almost tripped!" and it really hits home for Scrappy.


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As an extracurricular place of learning, the museum is a destination for excursions, day hiking and school trips, as well as a platform for national and international school meetings. In accordance with its guiding principle, it offers different forms of education that are introduced in stories about the Peenemünde research stations and the Nazi armament programme for the creation of new, innovative long-range weapons, expand on important aspects and put topical expansion up for discussion.
The pedagogical offers include all age groups, various subjects and different levels of detail. Tours and learning offers in the form of workshops can be booked for an introduction to the key topic of the museum, which can also be expanded upon by more in-depth learning offers.
Topics covered in the museum are presented in a 90-minute tour. The tours take into consideration the respective prior knowledge from grade level 2 to secondary II level.
Insights into specific parts of exhibitions, the depot and the restoration workshop along with a material counter you can touch can be optionally provided.
For more than 30 participants, we recommend booking two parallel tours.
Pupils find out about the museum independently, but under instruction from the museum's pedagogical staff in around 3 hours, thus gaining knowledge relevant to their syllabus.
The offers include a joint tour, work assignments in individual, partner or small group work, and a joint session where the results are gone over and presented with a subsequent assignment-based concluding discussion. Each learning opportunity has subgoals according to the capacity model.
During the concluding discussion, topics dealt with in the exhibitions are deconstructed from their specific historical context and their validity for our current society is discussed. Relation to the present also emerges in the additional opportunity of examining the example of historical discoveries in the area of the contaminated munition and armament sites in Peenemünde. At a material counter in the seminar room or museum depot, pupils can touch selected artefacts.
To supplement the introductory offers or during a repeated visit, the museum has various in-depth learning offers, each of which lasts up to two hours and can be selected parallel to one another or after one another:
- Creative analysis (collage, memory game, puzzle, fliers, reconstruction)
- Transfer of learning content into other media (storyboard, clip)
- Examination of learning content with performative methods (reconstructing images, historical game)
In the sense of explorative learning, pupils can work with written, image or item collections from the museum, solve problems presented to them themselves and deliver presentations by designing a small exhibition.
Pupils work individually or in small groups and then present their findings together. The material is provided on a learning counter.
During tours of the monument landscape, pupils can discover the authentic grounds of the Peenemünde research facilities and therefore experience history actively and without media. In the museum there is a limited number of rental bicycles available.
Because of the heavy demand for education offers from the museum, early registration beforehand is advised. The pedagogical staff at the museum will be happy to help provide advice or answer any content-related queries.
School trips to the museum in the form of extracurricular learning are promoted by the State of Mecklenburg-Vorpommern. You can find more information here:


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Friends of ours gave us a Daisy Gardenia as a living and loving memory to Byron. New growth started appearing early spring and then without any indication, the first bloom appeared and I was awestruck. Not because it finally bloomed, but because it bloomed this morning, on his birthday. Further affirmation that he hasn't left us, he's everywhere, at all times, with us always.
Category Archives: memorial
Day of the Dead
Dia de los Muertos ~ Honoring, remembering and celebrating those who have crossed before us. Praying in support of their spiritual journey –
j.a.terry 11.1.15
Created by my sister-in-law, Gretta –
www.gofundme.com/byronterry –
This is a memorial set up by the family of Byron Terry to benefit his wife and son at home as they are left to deal with many unexpecteds. We lost a most caring, compassionate, and loving person and the hole in our hearts is huge. BT, as most called him, had two grown children plus a son who will be graduating from high school this year. Money doesn't take away the hurt, but if we can help alleviate the burden on them some, we want to have our part in doing so. Please share this page to raise more support, thank you.


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9/9/99 – the date that changed gaming forever. The birth of the Dreamcast and the birth of Generation 6!
This was when shit started getting real; and by real I mean casulties, this console war gave us our first fatality…poor SEGA ceased to be a hardware creator and became a machine for pumping out average titles!
As a SEGA kid this was a blow I thought i would never recover from, but the Dreamcast is one of the greatest consoles of all time. SEGA went out and left us with a wealth of gaming gold that will never be replicated…the gaming landscape was inexorably changed during this generation.
What classics are oft-forgotten I hear you scream at the top of your lungs… simmer down kids…i'm 'bout to lay down that knowledge.
6) Zombie Revenge (Dreamcast; 2000) – the least known entry into the House of the Dead series; Zombie revenge took the formula and turned it on its head! Gone was the Dreamcast Gun (yes that was it's actual name!) and in its place was a scrolling beat'em'up in the mold of lesser known Saturn game 'Die Hard Arcade' (and it's Dreamcast sequel 'Dynamite Cop') you assume the role of Stick Breitling, Linda Rotta and Rikiya Busujima and join their adventures in punching, kicking and shooting zombies in the face! A very fun game that is very hard to find, but worth it for arcade style high-jinx!
5) Freek Style (PS2, Xbox, GCN; 2002) – SSX is oft-remembered as the best stunt/racing game hybrid of its generation (and rightly so as SSX is a bloody brilliant series) however there was another EA 'BIG' game that ran it bloody close! The formula is the same; a racing game in which you need to crack out the stunts in order to fill a turbo meter and gain an advantage; just replace a snowboard with a motocross bike and you've got the idea here. Freekstyle had some great tracks in which to facilitate the action and in my view it surpassed SSX in pure racing stakes – the only downside is the abrasive 'Nu-Metal' soundtrack that was 'a la mode' in 2002. Stick her on mute, kickstart the bike and get read to FREEK OUT! Give us a deadly descent style reboot EA! DO YOU HEAR ME?!
4) Transformers (PS2; 2004) – I must admit i did not play this game on release, i had that honour in 2007 during University, but what a game! Better that the High Moon games on many levels (especially the aircraft carrier level – which turns out to be it's own final boss…epic!) this game is based on the Generation 1 series of transformers so it is instant win. But its a very competent third person shooter that makes great use of its licence! If you like Transformers; play it. If you life third person shooters; play it. Hell if you like video games at all, play it! Just play it!
3) Freedom Fighters (PS2; 2003) – This is another game i didn't sample until University and again, what a game! The plot revolves around a Soviet invasion of the USA and the small scale fight back by citizens; basically it's Homefront, before Homefront ruined the idea for everyone. Take control of plumber-turned-revolutionary Christopher Stone; use your charisma points and assemble a squad (that's right it's a squad based shooter) of like-minded people to take down the Commie scum. Freedom Fighters was excellent, hell it's still excellent. Although age has not been good to its looks (it was running on a modified version of the Hitman 2 engine) it still plays well today.
2) Outtrigger (Dreamcast; 2001) – WAAAAaaaaay before fan-boys were quoting fps to one-up each other on Xbox One and PS4, SEGA unleashed a 60fps, online first-person shooter helmed by the great Yu Suzuki. Outtrigger was frenetic, tight and an absolute joy to play. Online (powered by the DC's ridiculous 33.6K modem) was seamless and lag free. Outtrigger is THE great, forgotten console FPS. Although the arenas are compact, there is more than enough room for 4 players to murder each other with gay-abandon. Forget Quake 3 and Unreal Tournament (both ports of these games are outstandingly good though) Outtrigger is the place to get split screen shooting on the Dreamcast!
1) Black (PS2/Xbox; 2006) – People will argue over what was the best FPS this generation and many will cite Halo 2 as the champion; i disagree with this, Black is the greatest FPS of the 6th Generation. It had the unfortunate honour of being released AFTER the Xbox 360 and was thusly relegated to the annuls of history, doomed to obscurity. However it deserves to be plucked from the purgatory it finds itself in and played by as many people as possible. Made by Criterian Games it certainly has pedigree, and it shows in every aspect of the game, the gun play is tight, level design is very interesting (and allows for deformation of textures), when you're close to death the screen will fade to black (and you though COD started that?). Black was also gorgeous to look at, heavily influenced by cinema and just a joy to play. Find it. Play it.
Hopefully we've opened your eyes to some great games, if you do decide to play any of these games please leave your thoughts on our twitter page, or indeed, in the comment section below! Remember we still have part five to come, Generation 7!
If you hated these games, then flame us! We're big enough to take your hate!
Basically, if you have any thoughts on what we're doing here at MrLuvvaLuvva let us know!


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We are pleased to announce that the STUDIA Education Fair 2012 will be held in Helsinki, Finland on December 4-5. Studia 2012 is an international further education fair for upper secondary school students and graduates.Reasons to AttendThe Finland Education Fair 2012 is an international further education fair for upper secondary school students and graduates. The visitors are also more and more interested in educational and working possibilities abroad. The 2011 edition of the Finland Education Fair received over 14,900 visitors and 142 exhibitors.Market OverviewEducation has always been a high priority for Finland. At the moment, Finland has 16 university level institutions and 25 polytechnics. The latest statistics show that over 13,000 students from Finland were studying internationally in 2009.
Finland has developed its world class education system which offers students wonderful higher education opportunities in almost all fields of study. The reputation of its education system is high not only because it provides high standard of education but because also because it is continuously working and investing for increment in level of educational standards. As such, the national educational strategy is to invest substantial amount on research based education to develop the country as the information society. The higher educational wings of Finland constitutes of Universities and Polytechnics which provide most advanced state-of-art facilities to its students.Fair OutlineMost off the Finnish universities and polytechnic institutes participate in the fair. Participation of the Finnish further education field make Studia a vital part of the Finnish student counselling.
Exhibitor feedback


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Adwords is great, but its built-in tools don't always allow you to solve problems unique to your business. I was recently inspired by this post on Search Engine Land to build something similar off the Google Ads API.
There were a few problems on our account that couldn't be solved by the Brainlabs solution.
- Because of the way our checkout process is structured, Adwords doesn't know about our transactions. Until recently, this hadn't been a problem; we have an approximation for transactions set up in Google Analytics, and the main KPIs for most of our campaigns are app downloads and email collection (things Adwords is aware of). But I wanted to use real-time transactional information to inform our bidding.
- I didn't want to set up different scripts or bid modifier charts for each of our campaigns. People behave differently when buying Broadway tickets than they do when buying NBA tickets, and I wanted something that would calculate modifiers on the fly for each campaign.
- Finally, the major sports teams are spread out across three time zones. The "right time" to bid up a campaign for the Los Angeles Dodgers is different than for the New York Yankees, so I needed something that could handle those differences as well.
A few months back, we had our quarterly Hackathon and I decided to explore the Adwords API, which is pretty easy to sign up for.
There are several Google Ads API client libraries, like PHP, Ruby, Java, etc. I settled on the Python Libary because I hadn't ever built anything in Python. This will become abundantly clear when you dive into my code.
You can find the script and instructions on how to set it up here. There are three major components of the script.
- Pulling transactions or whatever KPI you're interested in from your database
- We use Amazon Redshift, so my script uses the Psycopg2 package to connect to and query our database. If you're running off MySQL, you can follow the instructions here in order to connect to your DB.
- Make sure that any reporting you do is in the same time zone as your Adwords account. For example, our database records timestamps in UTC, but our Adwords account runs everything on EST.
- Working with the Adwords API
- This file, adwords_api.py, contains every Adwords API-related call, from getting spend reports to modifying keyword bids.
- This whole operation only runs on campaigns with a specific label. There are instructions here on how to find your label IDs, or how to modify the script so it runs on every campaign on your account.
- Modifying bids
- This file, create_bids.py, actually calculates the bid modifier for each hour based on an ROI or CPA goal defined by you.
What will future versions do?
- I'd like to write versions that work on the adgroup and keyword level. Currently, the script only calculates modifiers on the campaign level and then attaches them to every keyword in said campaign.
- Right now, this script doesn't take day of week into account. While conversion rate from SEM doesn't change much day to day, I understand that day of week could be meaningful for some businesses. For SeatGeek's purposes in particular, a future version that integrates time-to-next-event for each team would be a big improvement.
While this is certainly a work in progress, I hope this script can act as a jumping-off point for others to build something that works for their needs.


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Worth the watch - 15:211786
30491 1 1
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Boomer Initiative.
Keep in mind this is ONLY US Boomers. And we are not the only 50+ somethings in the world.
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Allergan, Inc. Announces FDA Approval Of JUVEDERM R XC Dermal Filler Formulated With Lidocaine
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Allergan, Inc. (NYSE:AGN) announced the U.S. Food and Drug Administration's (FDA) approval of JUVEDERM(R) XC, a new formulation of the currently FDA-approved JUVEDERM(R) dermal filler and the latest advancement1 in hyaluronic acid (HA) dermal fillers. Allergan's new JUVEDERM(R) formulation contains the local anesthetic lidocaine to provide patients with enhanced comfort during treatment of moderate to severe facial wrinkles and folds, such as the nasolabial folds (or "parentheses") that appear around the nose and mouth. JUVEDERM(R) is the first and only hyaluronic acid dermal filler approved by the FDA to last up to one year from initial treatment2 and number-one selling hyaluronic acid dermal filler.3
"As the global leader in medical aesthetics, Allergan is committed to providing the latest scientific advancements in facial aesthetic products to meet patients' demands and further optimize their experiences," said Robert Grant, Allergan's Corporate Vice President and President, Allergan Medical. "We lead innovation in the dermal filler category with the first and only smooth-consistency gel hyaluronic acid dermal filler approved by the FDA to last up to a year. Now we have added lidocaine to JUVĂ DERM(R) to provide the same smooth, long-lasting result, but with additional comfort for patients."
The JUVEDERM(R) XC formulation with 0.3% preservative-free lidocaine numbs the treatment area within seconds, potentially reducing the need for an additional anesthetic. JUVEDERM(R) XC provides the same smooth, long-lasting results as demonstrated with existing formulations of JUVEDERM(R), and now offers a more comfortable injection experience and potentially less time spent in the physician's office when compared to the non-lidocaine JUVEDERM(R) formulation.
"Patients want a smooth and natural-looking result from a dermal filler treatment, but as a physician I am also interested in managing my patient's discomfort during the injection. Before the introduction of JUVEDERM(R) XC it often took up to 30 minutes for an anesthetic block to take effect. In the clinical trial leading up to the FDA approval, patients reported they experienced less pain with JUVEDERM(R) XC, compared to their previous dermal filler treatment without lidocaine. So with the new formulation, patients can receive the same smooth results as demonstrated with JUVEDERM(R) but enjoy a more comfortable injection experience," said Charles Boyd, M.D., Boyd Cosmetic Surgical Institute and JUVEDERM(R) XC clinical investigator.
The FDA approval of JUVEDERM(R) XC was based on data from a multicenter, double-blind, randomized clinical trial. A total of 72 subjects were followed for two weeks after treatment with one of two JUVEDERM(R) formulations (JUVĂ DERM(R) XC with lidocaine or JUVEDERM(R) without lidocaine) in each nasolabial fold. In the clinical study (n=72), 93 percent of patients reported less pain when treated with the new formulation of JUVEDERM(R) compared to those treated with the non-lidocaine formulation of JUVEDERM(R).4 JUVEDERM(R) XC was found to be more effective in reducing procedural pain during correction of facial wrinkles and folds while maintaining a similar safety and effectiveness profile to the non-lidocaine formulation of JUVEDERM(R).
Following FDA's approval, the new formulation of JUVEDERM(R) with lidocaine is available for ordering nationwide. JUVEDERM(R) is a prescription-only treatment and should be administered by a qualified medical practitioner who has been trained in JUVEDERM(R) injection techniques.
Allergan encourages individuals interested in being treated with the JUVEDERM(R) family of products to visit www.Juvederm.com and access the JUVEDERM(R) Online Treatment Visualizer to upload a picture and help visualize how JUVEDERM(R) can potentially smooth away moderate to severe facial wrinkles and folds. This tool is for visualization and illustrative purposes only and is not a substitute for a consultation with a qualified medical practitioner.
JUVEDERM(R) Family of Products
Allergan's JUVEDERM(R) dermal filler product line includes JUVEDERM(R) Ultra and JUVEDERM(R) Ultra Plus in the United States, providing physicians with the flexibility to tailor each treatment to the particular needs of the patient. Both of these formulations will now be offered with lidocaine under the brand names, JUVEDERM(R) Ultra XC and JUVEDERM(R) Ultra Plus XC. JUVEDERM(R) XC is contraindicated in patients with a history of allergies to lidocaine.
A Brief Description of Indications for Use, Contraindications, Warnings, Precautions, and Adverse Events for JUVEDERM(R) Injectable Gel
Indication: In the United States, JUVEDERM(R) injectable gel (including JUVEDERM(R) Ultra, JUVEDERM(R) Ultra Plus, JUVEDERM(R) Ultra XC, and JUVEDERM(R) Ultra Plus XC) is indicated for correction of moderate to severe facial wrinkles and folds (such as nasolabial folds).
Contraindications: JUVEDERM(R) injectable gel should not be used in patients who have severe allergies marked by a history of anaphylaxis or history or presence of multiple severe allergies. JUVEDERM(R) should not be used in patients with a history of allergies to Gram-positive bacterial proteins. JUVEDERM(R) Ultra XC and JUVEDERM(R) Ultra Plus XC should not be used in patients with a history of allergies to lidocaine.
Warnings: JUVEDERM(R) injectable gel should not be injected into blood vessels. If there is an active inflammatory process or infection at specific injection sites, treatment should be deferred until the underlying process is controlled.
Precautions: The safety of JUVEDERM(R) for use during pregnancy, in breastfeeding females, or in patients under 18 years has not been established. The safety and effectiveness of JUVĂ DERM(R) injectable gel for the treatment of areas other than facial wrinkles and folds (such as lips) have not been established in controlled clinical studies. Patients who are using substances that can prolong bleeding, such as aspirin or ibuprofen, as with any injection, may experience increased bruising or bleeding at injection site. Patients should inform their physician before treatment if they are using these types of substances. As with all skin-injection procedures, there is a risk of infection. JUVĂ DERM(R) should be used with caution in patients on immunosuppressive therapy, or therapy used to decrease the body's immune response, as there may be an increased risk of infection. The safety of JUVĂ DERM(R) in patients with a history of excessive scarring (eg, hypertrophic scarring and keloid formations) and pigmentation disorders has not been studied. If laser treatment, chemical peel, or any other procedure based on active dermal response is considered after treatment with JUVĂ DERM(R) injectable gel, or if JUVĂ DERM(R) is administered before the skin has healed completely after such a procedure, there is a possible risk of an inflammatory reaction at the treatment site.
Adverse events: The most commonly reported side effects are temporary injection-site redness, swelling, pain/tenderness, firmness, lumps/bumps, and bruising. Most side effects are mild or moderate in nature, and their duration is short lasting (7 days or less).
CAUTION: Rx only.
Forward-Looking Statements
This press release contains "forward-looking statements," including the statements by Dr. Boyd and Mr. Grant, and other statements regarding the safety, effectiveness, adverse reactions, product availability, approval and market potential of JUVEDERM(R) injectable gel and JUVEDERM(R) injectable gel with lidocaine. These statements are based on current expectations of future events. If underlying assumptions prove inaccurate or unknown risks or uncertainties materialize, actual results could vary materially from Allergan's expectations and projections. Risks and uncertainties include, among other things, general industry and medical device market conditions; technological advances and patents attained by competitors; challenges inherent in the research and development and regulatory processes; challenges related to new product marketing, such as the unpredictability of market acceptance for new products and/or the acceptance of new indications for such products; inconsistency of treatment results among patients; potential difficulties in manufacturing a new product formulation; general economic conditions; and governmental laws and regulations affecting domestic and foreign operations. Allergan expressly disclaims any intent or obligation to update these forward-looking statements except as required by law. Additional information concerning these and other risk factors can be found in press releases issued by Allergan, as well as Allergan's public periodic filings with the Securities and Exchange Commission, including the discussion under the heading "Risk Factors" in Allergan's 2008 Form 10-K and Allergan's Form 10-Q for the quarter ended September 30, 2009.
About Allergan, Inc.
Founded in 1950, Allergan, Inc., with headquarters in Irvine, California, is a multi-specialty health care company that discovers, develops and commercializes innovative pharmaceuticals, biologics and medical devices that enable people to live life to its greatest potential - to see more clearly, move more freely, express themselves more fully. The Company employs approximately 8,500 people worldwide and operates state-of-the-art R&D facilities and world-class manufacturing plants. In addition to its discovery-to-development research organization, Allergan has global marketing and sales capabilities with a presence in more than 100 countries.
© 2010 Allergan, Inc. Irvine, CA 92612. (R) and (TM) Marks owned by Allergan, Inc. JUVEDERM(R) is a mark owned by Allergan Industries SAS
(1) Weinkle SH, Bank DE, Boyd CM, Gold MH, Thomas JA, Murphy DK. A multi-center, double-blind, randomized controlled study of the safety and effectiveness of JuvĂ derm injectable gel with and without lidocaine. J Cosmet Dermatol. 2009 Sep;8(3):205-10.
(2) Pinsky MA, Thomas JA, Murphy DK, Walker PS; for the JuvĂ derm vs Zyplast Nasolabial Fold Study Group. JuvĂ derm Injectable gel: A multicenter, double-blind, randomized study of safety and effectiveness. Aesthetic Surg J. 2008; 28(1):17-23.
(3) Guidepoint Global Tracking for US Dermal Fillers, January - October 2009
(4) Weinkle SH, Bank DE, Boyd CM, Gold MH, Thomas JA, Murphy DK. A multi-center, double-blind, randomized controlled study of the safety and effectiveness of JuvĂ derm injectable gel with and without lidocaine. J Cosmet Dermatol. 2009 Sep;8(3):205-10.
Source: Allergan, Inc
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xc, xc dermal, juvederm xc, xc ultra, plus xc, ultra xc, approval xc, xc formulation, xc provides, xc based


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Cost of Invisible Braces in Houston, TX
This post is also available in: Español (Spanish)
Similar to Invisalign®, the ClearCorrect™ system may improve the way you bite with your back teeth, the alignment of your front teeth, the appearance of your smile, and the overall health of your teeth and mouth. This ultimately can save you money by preventing future oral disease.
Keep in mind that treatment costs will vary depending on your specific needs, which will be discussed during your one-on-one consultation.
We are committed to providing quality, affordable care for you and your family. Summit Dental Center™ offers financing plans and payment options that will break the total cost of treatment into a series of monthly payments. We also accept most insurance plans and government assistance programs. Please visit our financing page to find the best option for your situation.
Why Choose Summit Dental Center™?
Known as a "great dentist for kids," Summit Dental Center™ has a reputation for providing a family friendly atmosphere and positive dental experience
- Offers ClearCorrect™ as an alternative to Invisalign®
- Quality care at affordable rates
- 7 convenient locations with flexible evening and weekend hours
What to Expect During Your Consultation
- All new patients receive a $25 initial consultation
- Your initial visit consists of x-rays, examination, and consultation
- Treatment options are discussed before any service is performed
- We make quality dental care easy and affordable
With seven convenient locations throughout the Houston area, we offer both extended and weekend hours to our patients. We are dedicated to providing quality dental service, so please contact us today to schedule your appointment and ask about our free consultations!


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The Commodore Amiga has become a classic target for nostalgic gamers to relive their early gaming experiences. iOS has seen its fair share of releases from the retro computing platform, including Battle Squadron One, Alien Breed, Babylonian Twins, and Defender of the Crown, just to name a few. There are several more in the works, as well.
Today, however, developer Cope-Com announced the release of classic two-player shoot-em-up, Battle Squadron, for OS X and PC. It follows up a successful iOS release in 2011 as well as one for Android in 2012.
Not only that, but Battle Squadron is returning to Amiga, with a version available for Amiga OS 4.X, which sort of brings the whole thing full circle.
Battle Squadron is available via download for about $14, or as a special collectors' edition (pre-order for $34), which has the signatures of the original developers on it. This special edition contains versions for all desktop versions, which is an exact replica of the original game, with some extra new features thrown in for the fans.
Whichever edition you choose, you'll get a sweet two-player spaceship shooter game with all the trimmings that you can play right on your Mac. Not too shabby for a game that's been around since 1989.
You can also get Battle Squadron for iOS on the App Store for $3.99, though it won't include two-player action.
Source: Cope-Com


text: Wilson Combat Ultralight Carry For Sale from Jays Gun | Positive feedback: 97% View | Verified Seller | 1709 Completed Sales View Sellers Items /UserImages/2672/982238487 6748437.jpg /UserImages/2672/982238487 6748436.jpg /UserImages/2672/982238487 6748435.jpg /UserImages/2672/982238487 6748434.jpg /UserImages/2672/982238487 6748433.jpg /UserImages/2672/982238487 6748432.jpg /UserImages/2672/982238487 6748431.jpg Description: he Ultralight Carry Compact has all the cutting-edge features of the Ultralight Carry in a lighter smaller package. A 1" shorter slide and ½ inch smaller gripframe than a full-size 1911 ensure that you will always have your Ultralight Carry Compact when needed. The fluted heavy bull barrel is not only attractive, but also functional-it traps dirt and debris that can otherwise cause a malfunction. Try an Ultralight Carry Compact and see why it is quickly becoming a top concealed carry choice of seasoned tactical shooters. Read More Condition: New In Box Caliber: 9mm Luger Location: NY Trades Accepted: NO Read More Shipping Notes: See Description Read More Share Item Item #: 982238487 Stock No.: 84834 Jays Gun Seller Since: Jun. 2000 Seller Level: Trusted Seller This seller is an FFL City: Uniondale State: NY Phone: 516-385-5812 Price: $4,150.00 Buy It Now Add To Cart Please Login/Register to Order Login or Register to buy Shipping: $35.00 (or FREE in-store pick up!) Accepted Payment Methods: Returns: No Returns Send Message to Jays Gun You must login or register to send a message to this seller. Contact Seller as Guest Message To Seller Email * That Email Address is already in use. If you typed your email correctly, you are already registered and you should login or click here for the password recovery system. First Name * Last Name * Phone * Address 1 * Address 2 City * State * -State- AK AL AR AZ CA CO CT DC DE FL GA GU HI IA ID IL IN KS KY LA MA MD ME MI MN MO MS MT NC ND NE NH NJ NM NV NY OH OK OR PA PR RI SC SD TN TX UT VA VI VT WA WI WV WY Zip * Already have an account? Login Your Recently Viewed Items