text:
Mesenchymal stem cell-based therapy: a new paradigm in regenerative medicine.
ABSTRACT Mesenchymal stem cells (MSCs), adherent fibroblastoid cells, present in bone marrow and many other tissues can be easily isolated and expanded in vitro. They are capable of differentiating into different cell types such as osteoblasts, chondrocytes, adipocytes, cardiomyocytes, hepatocytes, endothelial cells and neuronal cells. Such immense plasticity coupled with their ability to modulate the activity of immune cells makes them attractive for stem cell-based therapy aimed at treating previously incurable disorders. Preclinical studies have reported successful use of MSCs for delivering therapeutic proteins and repairing defects in a variety of disease models. These studies highlighted the in vivo potential of MSCs and their ability to home to injury sites and modify the microenvironment by secreting paracrine factors to augment tissue repair. Their therapeutic applicability has been widened by genetic modification to enhance differentiation and tissue targeting, and use in tissue engineering. Clinical trials for diseases such as osteogenesis imperfecta, graft-versus-host disease and myocardial infarction have shown some promise, demonstrating the safe use of both allogeneic and autologous cells. However, lack of knowledge of MSC behaviour and responses in vitro and in vivo force the need for basic and animal studies before heading to the clinic. Contrasting reports on immunomodulatory functions and tumorigenicity along with issues such as mode of cell delivery, lack of specific marker, low survival and engraftment require urgent attention to harness the potential of MSC-based therapy in the near future.
Article: Adipose tissue displays trophic properties on normal lung cellular components without promoting cancer cells growth.[Show abstract] [Hide abstract]
ABSTRACT: Surgical removal is the mainstay for early lung cancer treatment and persistent air leaks represent one of the most common clinical complications after lung surgery. Adipose tissue transplantation has been proposed as a new strategy for regenerative therapy after breast cancer surgery; however its efficacy and safety of lung tissue healing after lung resections are unknown. The purpose of this study was to test the biological activity of adipose tissue to facilitate lung tissue healing and evaluate its effect on cancer cells growth, thus providing insight for a possible clinical application. Different in vitro cellular models were used to prove the potential biologic effect of autologous fat tissue (AFT) in repairing injured lung tissue, and in vivo xenograft models were used to evaluate tumor promoting potential of AFT on putative residual cancer cells. Treatment of both embryonic (WI-38) and adult lung fibroblasts and of normal bronchial epithelial cells (HBEC-KT) with AFT samples, harvested from subcutaneous tissue layer of 20 patients undergoing pulmonary metastasectomy, improved wound healing and cell proliferation indicating a trophic effect on both mesenchymal and epithelial cell types. Conversely AFT-conditioned medium was unable to stimulate in vitro proliferation of a lung adenocarcinoma reporter cellular system (A549). Moreover, co-injection of AFT and A549 cells in nude mice did not promote engraftment and progression of A549 cells. These preclinical findings provide preliminary evidence on the potential efficacy of AFT to accelerate lung tissue repair without undesired tumor promoting effects on putative residual cancer cells. J. Cell. Physiol. © 2012 Wiley Periodicals, Inc.Journal of Cellular Physiology 11/2012; · 4.22 Impact Factor
Article: Establishment of a pancreatic β cell proliferation model in vitro and a platform for diabetes drug screening.[Show abstract] [Hide abstract]
ABSTRACT: Diabetes, a disease resulting from loss of functional β cells, is globally an increasingly important condition. Based on the islet-differentiation ability of ductal epithelial cells and stimulating β cell proliferation ability of the Reg Iα gene, we aimed to establish an in vitro pancreatic β cell proliferation model for screening therapeutic drugs of diabetes in the future. Pancreatic ductal epithelial cells were isolated from male Wistar rats, and induced to differentiate into pancreatic β cells. Immunofluorescence staining assay, western blot, RT-PCR analysis, and dithizone staining were used to characterize the cells. Rat Reg Iα protein was transiently expressed in vitro by transfection of HEK 293 cells with the PCMV6-entry-REG Ia plasmid, and expression was verified by RT-PCR analysis, proliferation assay, and apoptosis assay. The pancreatic β cell proliferation model was further validated by a proliferation assay using differentiated pancreatic β cells treated with transfection supernatant. Finally, we have successfully established an in vitro pancreatic β cells proliferation model using transiently expressed rat Reg Iα protein and differentiated pancreatic β cells from pancreatic ductal epithelial cells. This model could be used as a platform to screen new drugs for islet neogenesis to cure diabetes, especially Chinese herbal drugs in the future.Cytotechnology 08/2013; · 1.32 Impact Factor
Article: Central role for disabled-2 in mesenchymal stem cardiac protein expression and functional consequences after engraftment in acute myocardial infarction.[Show abstract] [Hide abstract]
ABSTRACT: Adult mesenchymal stem cells (MSCs) have been shown to spontaneously express cardiac proteins (CP) in vitro and to improve cardiac function after transplantation into experimentally induced acute myocardial infarction (AMI). However, if these effects are the result of MSC cardiac differentiation or a mere cooperative cellular interaction is a matter of active debate. Additionally, the molecular mechanisms involved in CP expression by adult stem cells in vitro and its possible benefit for cardiac regeneration and improved function remain unclear. Here we show that although MSCs effectively engraft in AMI tissue, this engraftment leads to downregulation of CP expression in the implanted MSCs. We also found that pretransplantation cardiac specification of MSCs by exposure of the cells to transforming growth factor beta 1 (TGF-β1) led to sustained MSC CP expression without altering engraftment efficiency. This increase in CP expression was associated with greater improvement in cardiac function 1 and 4 weeks after AMI with TGF-β1-pretreated MSCs. We discovered that the TGF-β1-enhanced cardiac potential of MSCs was mediated by downregulation of disabled-2 (Dab2) expression, suggesting an inverse correlation between Dab2 levels and CP expression/cardiac functional improvement after MSC engraftment. Our investigations further demonstrate that loss of Dab2 expression was sufficient to induce MSC CP expression and improve cardiac function after MSC engraftment after AMI. In summary, we define a novel role for the TGF-β1 receptor adaptor protein Dab2 as a regulator of CP expression in MSCs and its potential as a molecular target for the enhancement of stem cell cardiac specification for transplantation therapies.Stem cells and development 04/2011; 20(4):681-93. · 4.15 Impact Factor
Page 1
Mesenchymal Stem Cell-based Therapy: A New Paradigm in
Regenerative Medicine
Neeraj Kumar Satija1, Vimal Kishor Singh1, Yogesh Kumar Verma1, Pallavi Gupta1, Shilpa
Sharma1, Farhat Afrin2, Menka Sharma1, Pratibha Sharma1, R.P. Tripathi1 and G.U.
1Stem Cell & Gene Therapy Research Group, Institute of Nuclear Medicine & Allied
Sciences, Lucknow Road, Timarpur, Delhi-110054, India.
2Department of Biotechnology, Hamdard University, Hamdard Nagar, New Delhi-110062,
Received date: 12-Aug-08, Revised date: 27-Mar-09, Accepted date: 7-Jul-09
*Corresponding author: Dr. G. U. Gurudutta
Stem Cell & Gene Therapy Research Group,
Institute of Nuclear Medicine & Allied Sciences,
Lucknow Road, Timarpur,
Phone: 91-11-23905144
Fax: 91-11-25737049
Email: email@example.com
Please cite this article as a "Postprint";10.1111/j.1582-4934.2009.00857.x
has yet to undergo copy-editing and proof correction. See
http://www.blackwell-synergy.com/loi/jcmm for details.
approved for publication in the Journal of Cellular and Molecular Medicine, but
This is an Accepted Work that has been peer-reviewed and
Page 2
Mesenchymal stem cells (MSCs), adherent fibroblastoid cells, present in bone marrow and
many other tissues can be easily isolated and expanded in vitro. They are capable of
differentiating into different cell types such as osteoblasts, chondrocytes, adipocytes,
cardiomyocytes, hepatocytes, endothelial cells and neuronal cells. Such immense plasticity
coupled with their ability to modulate the activity of immune cells makes them attractive for
stem cell-based therapy aimed at treating previously incurable disorders. Preclinical studies
have reported successful use of MSCs for delivering therapeutic proteins and repairing
defects in a variety of disease models. These studies highlighted the in vivo potential of
MSCs and their ability to home to injury sites and modify the microenvironment by secreting
paracrine factors to augment tissue repair. Their therapeutic applicability has been widened
by genetic modification to enhance differentiation and tissue targeting, and use in tissue
engineering. Clinical trials for diseases like osteogenesis imperfecta, graft-versus-host disease
and myocardial infarction have shown some promise demonstrating the safe use of both
allogeneic and autologous cells. However, lack of knowledge of MSC behaviour and
responses in vitro and in vivo forces the need for basic and animal studies before heading to
the clinic. Contrasting reports on immunomodulatory functions and tumorigenicity along
with issues like mode of cell delivery, lack of specific marker, low survival and engraftment
require urgent attention to harness the potential of MSC-based therapy in the near future.
Keywords: mesenchymal stem cells, stem cell therapy, genetic modification, protein therapy,
tissue engineering
Page 3
List of main topics:
• Introduction
• Mesenchymal stem cells and its characteristics
• Experimental/Preclinical MSC-based studies
o MSC transplantation
o Genetically modified-MSC-based therapy
o MSC-based protein therapy
o Tissue engineering using MSCs
• Clinical studies
• Challenges and future prospects
Page 4
Bone marrow harbours cells of hematopoietic and non-hematopoietic lineages and their
precursors, known as stem/progenitor cells. The non-hematopoietic stem/progenitor cell
compartment contains mesenchymal stem cells (MSCs), which are involved in remodeling of
the mesenchymal tissues throughout adult life. These multipotent cells are easily isolated
from bone marrow and are capable of expansion and differentiation into mesodermal lineage
cells including osteoblasts, chondrocytes and adipocytes, under appropriate conditions, in
culture [1, 2]. This led to the evaluation of their potential for treating diseases, and the birth
of MSC-based therapy.
Recent clinical trials with MSCs for treating debilitating disorders like osteogenesis
imperfecta, myocardial infarction, stroke and graft-versus-host disease have shown some
promise [3-6]. Numerous preclinical studies have established the therapeutic potential of
MSCs in tissue engineering and as cellular protein factory for delivery of cytokines and
anticancer agents [7-9]. Genetically modified-MSCs have also been successfully evaluated in
animal models for diabetes, skeletal defects and myocardial infarction [10-14].
Cotransplantation of MSCs with hematopoietic stem cells (HSCs) has been documented to
improve HSC engraftment in mice [15, 16]. Though there has been a surge in preclinical and
clinical trials using MSCs, caution must be taken in planning such studies since MSC biology
is only beginning to be understood. However, the question arises: what makes MSCs unique
and preferable for cell-based therapies?
In this review, we focus on the suitability of MSCs in the field of regenerative medicine. We
provide an overview of the current status of research on MSC-based therapies in
Page 5
experimental animals and humans. Different therapeutic designs along with preclinical cases
which also address the mechanisms of MSC action are discussed. Clinical trials with MSCs
are critically evaluated followed by a discussion on the controversies surrounding the use of
MSCs and the challenges that need to be overcome for translation of the therapy from the
bench to the clinic.
Mesenchymal stem cells and its characteristics
MSCs were first identified about 30 years ago by Friedenstein and colleagues as an adherent
fibroblast-like population in the bone marrow capable of differentiating into bone [17]. Since
then MSCs have been isolated from the human bone marrow based on their ability to adhere
to tissue culture plastic [1]. Though occurring at a very low frequency of 1 in 10,000 to
100,000 bone marrow mononuclear cells, these cells are capable of proliferating in vitro
without significant loss of differentiation potential during early passages [1, 2, 18].
Originally isolated from the bone marrow, similar populations have also been isolated from
peripheral blood [19], periosteum [20], umbilical cord blood [21], synovial membrane [22],
trabecular bone [23], adipose tissue [24], limbal stroma [25], amniotic fluid [26], lung [27],
dermis and muscle [28]. These populations have been functionally characterized based on
their ability to differentiate into osteoblasts, chondrocytes and adipocytes in culture upon
induction due to lack of specific markers for MSCs [29]. However, phenotypically they are
defined as positive for CD105, CD73 and CD90, and negative for hematopoietic markers
(CD34, CD45, CD11b and CD19) and HLA-DR [30]. Since these surface markers are used
for characterizing cultured MSCs, immense efforts are underway to identify markers for their
direct isolation from tissue. Positive selection approaches using antibodies against low-
Page 6
affinity nerve growth factor receptor [31], stage-specific embryonic antigen (SSEA)-1 [32]
and SSEA-4 [33] have been used for isolation of primitive MSCs. Transplantation of a single
cell-derived population of SSEA-1+ mesenchymal cells in mice is the first in vivo study
demonstrating their capability of differentiating into different mesenchymal cell types, thus
showing their true stem cell properties. However, these cell populations were similar
phenotypically but heterogeneous in their functionality since all clones did not demonstrate
same differentiation potential suggesting only enrichment of MSCs using these markers.
Antibodies have also been raised against MSCs for their prospective isolation such as STRO-
1, SH-2, SH-3 and SH-4, but none of them recognize an epitope exclusively present on MSCs
[34]. Though the use of non-homogenous MSCs in preclinical and clinical studies has proved
safe and effective (as will be discussed ahead), the search for surface markers exclusive to
MSCs for their isolation and characterization is extremely important.
Playing a role in the homeostasis of mesenchymal lineages, these cells differentiate into
osteoblasts, adipocytes, chondrocytes, tenocytes, myoblasts and stromal fibroblasts [1, 35,
36]. Recent identification of various MSC populations such as mesodermal progenitor cells
(MPCs) [35], marrow-isolated adult multilineage inducible (MIAMI) cells [37], very small
embryonic-like stem cells (VSELs) [38] and SSEA-1+ mesenchymal cells [32] have
demonstrated their differentiation into mesodermal, endodermal and neuroectodermal
lineages, such as cardiomyocytes, hepatocytes, neural cells and endothelial cells [32, 35, 39-
41]. Hematopoietic differentiation has also been observed upon transplantation of SSEA-1+
cells in mice, signifying their primitiveness compared to all other populations [32]. However,
the transdifferentiation potential of MSCs has been questioned due to differences in the MSC
populations, culture conditions, experimental models and evaluation methods [42]. Many of
the observed morphological changes could be a culture artifact or a result of fusion with
Page 7
somatic cell [43, 44]. Therefore, a thorough evaluation of the plasticity of MSCs in vivo is
essential since in vitro conditions might not represent the true in vivo milieu.
Another distinguishing feature of MSCs is their ability to expand in vitro under normal
culture conditions [2]. We have observed 88- to 560-fold expansion in a single passage (15-
20 days) upon culturing early passage MSCs at a density of 50-500 cells/cm2 [our
unpublished data]. Colter et al have reported extensive expansion of a subpopulation of
MSCs, designated recycling stem (RS) cells, to the order of 109-fold in 6 weeks by culturing
cells at low density of 1.5 or 3 cells/cm2 [45]. Clinical feasibility of culture expanded MSCs
has been validated by a number of studies [4, 46-49]. Thus, a small amount of bone marrow
aspirate is sufficient for generation of large number of cells needed for transplantation
following in vitro expansion.
Immunological characterization of human MSCs revealed intermediate expression levels of
human leukocyte antigen major histocompatibility complex (MHC) class I, and no expression
of MHC class II antigen and costimulatory molecules CD40, CD80 and CD86 [50-52]. The
expression of MHC class I prevents them from the action of natural killer cells while absence
of costimulatory molecules leaves T cells anergic [reviewed in 53, 54]. In addition, MSCs
have been demonstrated to suppress T lymphocyte proliferation and activation [50, 51]. As a
consequence, MSCs are able to modulate the immune response making them immune
privileged and suitable for allogeneic transplantation, as has been reported in numerous
clinical studies [3, 55, 56]. Further, MSCs have been reported to home to sites of injury and
disease following intravenous infusion and contribute to the repair process [5, 48, 57]. The
expression of chemokine receptors on MSCs might be responsible for their ability to sense
and respond to signals like chemokines expressed by injured tissues [58], causing them to
Page 8
extravasate from the blood vessels, like immune cells [59], via a coordinated rolling and
adhesion behaviour on endothelial cells in a P-selectin- and VCAM1-dependent manner [60].
Their contribution to tissue repair is also mediated by secretion of paracrine factors having
angiogenic and anti-apoptotic properties [61-63]. These paracrine factors not only attract
endothelial cells and macrophages but are also likely to stimulate the resident stem/progenitor
cells to aid in the process of tissue repair [64].
MSCs can be easily isolated from readily accessible blood and bone marrow compared to
other stem cells from tissues like brain, heart and liver [65, 66]. Additionally, ex vivo
expansion potential enables generation of a sufficient number of cells for transplantation [45].
Immunomodulatory functions, homing ability to injured sites and capability to modify the
microenvironment by paracrine factors makes intravenous delivery feasible in comparison to
site-specific delivery of neural [67], cardiac [68] and muscle stem cells [69]. Thus, making
MSCs a promising candidate for stem cell-based therapy (Figure 1).
Experimental/Preclinical MSC-based studies
Capitalizing the extraordinary properties of MSCs, several studies have been undertaken to
evaluate their potential for tissue repair in animal models. Depending on the type of
disease/injury, different strategies involving site-specific delivery, genetic modification and
use of scaffolds have been designed. Basic studies to identify the mode of action of MSCs
and their responses to damages have also been addressed highlighting the therapeutic
potential as well as safety and efficacy of using MSCs. However, certain issues remain to be
resolved before translation of MSC-based therapy to the clinic (Figure 2).
Page 9
MSC transplantation
To begin with, numerous studies using systemic administration of MSCs have been
performed at preclinical level to assess their in vivo behaviour and suitability for the
treatment of a number of injuries and diseases (Table 1). Ortiz and colleagues evaluated the
ability of intravenously infused MSCs in bleomycin exposed mice, which represents a lung
injury model, to engraft in the lung tissue [70]. Bleomycin treatment resulted in 23-fold
increase in engraftment levels of MSCs compared to mice not exposed to bleomycin. Further,
the engrafted cells adopted an epithelium-like morphology and reduced bleomycin-induced
inflammation and collagen deposition in the lung [70, 71]. Whether MSCs actually
underwent transdifferentiation into alveolar epithelial type II cells or fused with epithelial
cells was not evaluated. However, transplanted mice exhibited increased level of G-CSF and
GM-CSF which might have mobilized endogenous stem cells aiding in repair [71]. The anti-
inflammatory action of MSCs was mediated by secretion of IL-1 receptor antagonist (IL1RN)
which suppressed expression of pro-inflammatory cytokines TNF-α and IL-1α [72]. In vitro
migration assays demonstrated the release of, as yet unknown, chemotactic factors from
damaged lung cells which attracted MSCs to the injury site [71].
Shimizu's group standardized transdifferentiation of MSCs into keratinocytes in culture and
investigated whether MSCs could migrate and engraft into wounded skin in murine model.
They found that intravenously injected MSCs transdifferentiated into keratinocytes,
endothelial cells and pericytes at the wound site, thereby accelerating the repair process [73].
Evaluating the migratory mechanism using in vitro and in vivo migration assays, they
identified chemokine receptor CCR7 to play a major role since its ligand SLC/CCL21
induced MSC migration [73]. Expression of keratin by transplanted MSCs and formation of
glandular structures was reported by Wu and colleagues upon injection of MSCs around
Page 10
wound in an excisional wound splinting model in diabetic mice [74]. They observed
reduction in the number of donor-derived cells in the wound during the 4 week follow-up
suggesting that MSC effects are transient and do not provide long-term self-renewal stem
cells for keratinocytes. Since MSCs have also been observed to return to the bone marrow
after wound healing [75], the local concentration of the chemokine signals or the expression
of a particular chemokine in response to injury at the site might have a significant role in
retaining MSCs [76], which needs further evaluation. Apart from undergoing
transdifferentiation, MSCs are also likely to contribute to the repair process by secreting
paracrine factors including VEGF-α, EGF, keratinocyte growth factor, SDF-1, IGF-1 and
Angiopoietin-1 (Ang-1), which facilitate the recruitment of macrophages, keratinocytes and
endothelial cells to the wound site and enhance angiogenesis and wound healing [63, 74].
Thus, reduction in inflammatory responses and accelerated angiogenesis contribute to the
ongoing reparative process but functionality of MSC-generated tissue like sebaceous and
sweat glands, if any, is not known. Therefore, complete regeneration of the tissue is debatable
Similarly, transplantation of human MSCs in hyperglycemic NOD/SCID mice resulted in
homing to islets associated with an increase in pancreatic islets and mouse insulin production
[78]. No human insulin was detected in blood and the reduction in blood glucose levels was
mainly a result of stimulation of islets and β-cell [78-80], similar to that observed for neural
stem cells in mice [81], as well as inhibition of T cell responses against the new β-cells [82]
These studies bring to light the potential of MSCs to migrate to injury site and modify the
microenvironment, thereby modulating the immune response and facilitating tissue repair by
stimulating endogenous stem/progenitor cells. It is, therefore, necessary that studies
suggesting transdifferentiaion clearly define the experimental conditions and thoroughly
Page 11
evaluate the true nature of the differentiated cells by expression profiling and functional
assays. Genetic marking approach may be useful in assessing the differentiation potential of
putative MSCs upon transplantation in animal model systems [83]. Further, these animal
models represent excellent systems to elucidate the mechanism of action of MSCs in
mediating various therapeutic effects, in order to improve the present treatment regimens and
facilitate the development of new approaches.
Recently, MSCs have also been shown to improve hematopoietic transplantation [15, 16, 47,
84, 85]. Transplantation of HSCs is used for the treatment of oncohematological disorders,
but marrow ablative therapy (involving high dose chemotherapy and radiotherapy) destroys
not only hematopoietic cells but also damages the stroma [86, 87]. This is likely to cause
reduction in the engraftment of HSCs in the hostile environment as has been demonstrated in
mice [88], thereby, decreasing the success of transplant. Koc et al reported rapid
hematopoietic recovery in 28 breast cancer patients undergone high-dose chemotherapy
following coinfusion of HSCs and MSCs [47]. Enhanced hematopoietic engraftment was also
reported upon infusion of limiting number of umbilical cord blood stem cells with unrelated
MSCs in mice [84]. Co-transplanting MSCs with HSCs (CD34+ cells) has been shown to
improve engraftment in the bone marrow in mice, though the underlying mechanism needs to
be elucidated [15, 16]. This will not only help in improving the present regimens to enhance
HSC engraftment, but represents a useful strategy that can be employed to enhance success of
transplantation of other adult stem cells, as documented by increased survival of MHC-
mismatched skin grafts in immunocompetent baboons [89].
Migration of MSCs to the sites of injury and disease has also been well documented in animal
models for myocardial infarction and cerebral ischemia [90, 91]. Also, culture expanded
Page 12
human MSCs have been shown to home to radiation-injured tissues in NOD/SCID mouse
model [92]. This portrays their ability to sense and respond to damage signals, thereby
avoiding the need for targeted delivery (such as intramyocardial, intrahepatic) to damaged
tissues. However, intravenous infusion would cause distribution of cells throughout the body
reducing the fraction of cells homing to the damaged site [93, 94]. Another issue is
entrapment of a large fraction of cells in the lung [90] resulting in very low engraftment
levels of the order of 0.1% to 2.7% in the tissues [3, 93]. In vitro expansion of MSCs is also
likely to result in low homing as demonstrated in murine study [95], but whether human
MSCs also exhibit similar effect remains to be determined. Another contributing factor is low
cell survival rate after transplantation [96, 97]. Thus, preconditioning of MSCs prior to
transplant by culturing in presence of SDF-1 [98] or under hypoxic conditions [99, 100] are
useful strategies which enhance cell survival in the hostile environment in vivo. Such
preconditioning leads to the activation of Akt survival pathway as well as increased
expression of pro-survival and pro-angiogenic factors like hypoxia-inducible factor 1, VEGF,
erythropoietin, Ang-1 and Bcl-2. Also increased expression of c-met leads to higher
migration rates to ischemic tissue in response to secreted hepatocyte growth factor as
demonstrated in rat hind limb ischemia model [100].
Further studies using disease models need to be carried out to elucidate the molecular
mechanism involved in MSC homing for the improvement of current therapies. For instance,
studies have revealed the involvement of integrin β1 in MSC migration and engraftment in
ischemic myocardium in mice [101], whereas CD44 has been implicated in migration and
localization of MSCs to kidneys in mouse model of acute renal failure [102]. Cytokine-
mediated up-regulation of CXCR4 expression in Flk1+ MSCs improved their engraftment in
bone marrow of sublethally irradiated NOD/SCID mice [103] while ectopic expression of α4
Page 13
integrin on mouse MSCs resulted in significant increase in bone specific retention of
transplanted MSCs in mouse [104]. These studies offer molecular targets for genetic
engineering of MSCs to enhance their homing and engraftment to injury sites and accelerate
recovery. Alternatively, cytokine treatment of MSCs to enhance expression of tissue-specific
adhesion molecule or tissue-specific administration of chemotactic factor like SDF-1α [105],
CCL12 [73] and MCP-3 [76] is likely to facilitate targeting to a particular tissue.
Genetically modified-MSC-based therapy
Integrating the strengths of genetic engineering and stem cell biology holds tremendous
potential for designing treatments for critical injuries and diseases by inducing differentiation
into a specific lineage and improving adhesion potential. Following transplantation, the fate
of MSCs would be determined stochastically in vivo depending on the niches they home and
therefore, not all transplanted cells might contribute to the repair of the damage. As recently
demonstrated in mice, transplanted MSCs differentiated into osteoblasts in the heart [106].
Thus, site-specific transplantation of functional, differentiated cells would be advantageous
under certain conditions. Though differentiated cells can be generated by chemical stimulants
or differentiation factors in vitro, the differentiation state might not be stable upon
transplantation. Such reversal of differentiation (i.e. dedifferentiation) has been shown in
vitro for MSCs upon the withdrawal of stimulants [107]. Therefore, genetically modifying
stem cell by a key differentiation factor would help to achieve directed and complete
differentiation into the desired lineage.
Studies on the therapeutic applicability of genetically modified - MSCs (GM-MSCs) have
been carried out in animal models (Table 2). MSCs transduced with BMP2 and BMP4 have
been shown to successfully repair a variety of musculoskeletal defects in animal models as
Page 14
BMPs are potent inducers of osteogenic differentiation [11, 12, 108]. The cells not only
themselves undergo differentiation but also stimulate the neighbouring cells to participate in
the repair process. It has also been reported that short-term expression (for 6 days) of BMP-2
in MSCs was sufficient to irreversibly induce osteochondral bone formation upon
implantation into tibialis anterior muscle or joints of SCID mice [109].
Differentiation being a process coordinately regulated by number of factors, expression of
combination of genes has proved more fruitful for orthopedic gene therapy. BMP-2/7 and
BMP-4/7 heterodimers exhibit higher activity than homodimers, therefore, simultaneous
transduction with BMP-2 and BMP-4 or BMP-7 in mesenchymal cells resulted in 2- to 3-fold
more bone formation in mice [110, 111]. However, BMPs are secreted factors and their
constitutive overexpression is likely to cause abnormal bone formation in vivo. Therefore,
regulated overexpression of osteogenic transcription factor Runx2 (using tetracycline-
regulated Tet-Off expression system) has been demonstrated to offer control over osteoblast
differentiation of engineered MSCs in mice [112]. These engineered cells provide a novel
approach for treatment of osteochondral disorders and use of regulateable expression systems
to prevent undesirable effects, but studies aimed at mapping the fate of GM-MSCs following
repair of the defect are required before leaping at the prospect of using them for human
clinical trials.
Use of GM-MSCs has been investigated in culture as a choice for the treatment of genetic
disorders. Genetic modification of MSCs with dominant-negative collagen type I protein
successfully repaired bones derived from osteogenesis imperfecta patients [113], whereas
dystrophin-transfected MSCs participated in myogenesis through cellular fusion and
complemented the genetic defect of muscular dystrophy myotubes in vitro [114].
Page 15
Generation of cells of different tissues for the purpose of transplantation can also be achieved
by genetic modification. Pancreatic transplantation is the only cure for type 1 diabetes
patients. However, shortage of pancreas donors calls for the development of alternative cell-
replacement therapy. Transdifferentiation of human bone marrow-MSCs into insulin-
producing cells by overexpression of pancreatic duodenal factor 1 (PDX1) has been achieved
in vitro [10, 115]. Only 50% of the cells expressed insulin and secreted it in response to
glucose in culture while other islet hormones were expressed by all cells. Since these cells did
not differentiate completely in vitro, as determined by microarray, transplantation under the
renal capsule in streptozotocin-diabetic immunodeficient mice induced further differentiation
and resulted in the reduction of hyperglycemia and stabilization of blood glucose levels
during the 5 week follow-up [10]. None of the transplanted cells were observed to migrate to
the pancreas, signifying the advantage of site-specific transplantation and avoidance of
unwanted effects due to homing to undamaged organs following systemic infusion. However,
to assess the maintenance of differentiated state, the cells can be transfected with vector
containing GFP or YFP cloned under the control of cell type-specific transcription factor
prior to transplantation to evaluate their fate in vivo, specifically when they are transplanted
in another tissue/organ since the microenvironment can alter their fate.
The cells must also be labeled properly (dyes like PHK26 or genetically like GFP) to track
them following transplantation in animal models. For instance, using GFP-labeled Akt-
overexpressing murine MSCs, Noiseux et al tracked MSC fate following intramyocardial
injection in mouse model of myocardial infarction [116]. They observed transient
engraftment of MSCs in the infarct zone and fusion of MSCs with recipient cardiomyocytes
as early as 3 days post-injection, raising concern regarding safety and long-term outcome of


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The president has signed a proclamation calling for flags to be flown at half-staff. The White House has postponed Obama's trip on Tuesday to visit a General Electric energy division in Schenectady, N.Y.
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Posts tagged adult-only
Sleeping Dogs receives adults-only rating in Japan
Sleeping Dogs, the upcoming Hong Kong-based gritty action title from Square Enix received a Z rating from the Japanese rating organization CERO (the equivalent of an AO rating from the ESRB). As a result, the game will be modified for Japanese audiences, particularly to penalize players for...
Silicon Valley company creating AO-only 'SexBox' console
It's been a while since we'd last heard the term "SexBox." Too long, in fact. Thankfully, Silicon Xtal Corporation is bringing it back, recently registering a trademark for the word first brought up surrounding the great Mass Effect sex debacle of 2008. According to the filing, SXC intends on...
Breaking News
Dying Light cancelled on Xbox 360, PS3
Featured Stories
Get a closer look at final retail Amiibos
Posted on Nov 22nd 2014 9:00AM
Barely Related: Star-Lord's dance-off, comet sounds
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Games we love and hate
Posted on Nov 21st 2014 3:00PM


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One Minute With: Deborah Moggach, novelist
Friday 29 March 2013
Where are you now and what can you see?
I'm in my study and watching two rats playing in the compost heap, then climbing up the bird feeder and hanging there upside down. Another hour's work gone.
What are you currently reading?
Chris Cleave's 'The Other Hand'. Utterly brilliant, and I long to adapt it as a film, but I suspect someone else has got there first.
Choose a favourite author and say why you admire him/her
Beryl Bainbridge because she understood the absurdity of life and how humour lurks in even the most tragic situation.
Describe the room where you usually write
My study, which overlooks the gambolling rats but also Keats's House, which happens to be where my grandmother was born in 1890. Behind it looms the Royal Free Hospital, where my parents died.
Which fictional character most resembles you?
Like a squillion other people, it's Elizabeth Bennet. She cut through the vapidity with her wit.
Who is your hero/heroine from outside literature?
My new hero is George Bellows, not only a great painter but a passionate socialist and larkey to boot. He even played the drums in a band.
Deborah Moggach's new novel is 'Heartbreak Hotel' (Chatto & Windus)
'At times I thought he was me'film
Review: One Direction, Fourmusic
Review: The World of Ice and Firebooks
Film More romcom than S&M
Review: The Imitation Gamefilm
Comedy...to show her mastectomy scars
TVNetflix gets cryptic
TV review A moving film showing kids too busy to enjoy their youth
Books Not even a Man Booker prize could save Richard Flanagan from a nomination
Arts & Ents blogs
- 1 To help fuel their propaganda machine against the poor, our government has now decided to redefine the word 'welfare'
- 2 Anti-gay hate preacher accidentally tweets 4,000 followers cartoon clip of him 'confessing' to be a 'homosexual sodomite'
- 3 Woman opens professional cuddling shop – gets 10,000 customers in first week
- 4 Grayson Perry: London needs affordable housing because 'rich people don't create culture'
- 5 Kenya bus attack: Al-Shabaab militants kill 28 non-Muslims who failed to recite Koran
Lee Evans announces his retirement from comedy on The Jonathan Ross Show
Iggy Azalea responds to Eminem rape lyrics: 'I'm bored of old men threatening young women'
Beyoncé '7/11' music video: Star bounces on bed in low-fi homage to viral video
Angelina Jolie confirms retirement from acting: 'I've never been comfortable on-screen'
Lana Del Rey rape video: Marilyn Manson denies involvement in shocking leaked footage
Rochester by-election: Ukip gains second MP as Tory defector Mark Reckless holds seat
'Beast of Bolsover' Dennis Skinner takes Ukip MP Mark Reckless to task moments after he is sworn in
Rochester by-election: Labour MP Emily Thornberry resigns after posting white van and England flags tweet
France 'blocks' Russian sailors from boarding a warship
Revealed: How the world gets rich – from privatising British public services
Myleene Klass: Ed Miliband 'strikes back' by comparing UK's need for Labour's mansion tax to Hear'Say track
WIN a trip for two to Cuba
Follow Simon Calder's journey in Cuba, and enter our competition to win a trip for two to Cuba
Rainbow list 2014: The LBGT people who make a real difference
Introducing our annual celebration of the lesbian, bisexual, gay, and transgender people who make a real difference
2014 WISE Summit
For the past four years, Qatar Foundation has been awarding an international prize for education each year, the first award of its kind in the world. Find out more...
Build yourself a brighter future
Chartered surveyors will be critical to shaping our new world
Why HD audio will lead to smarter calling
Telecom giant Vodafone is looking to improve voice quality, becoming the latest company to introduce new HD technology.
i100: A single mother on benefits who became one of the world's most successful authors
11 rags-to-riches underdog success stories
Made of London Series
From Britpop royalty to champion cheesemaker. Alex James has said that he would like to make more music with Blur.
From medieval to ultramodern
Mons is embracing the future as it prepares for its role as next year's European Capital of Culture, but it's also steeped in intriguing history. Philip Sweeney explores its two sides
Expertly tailored
From sharp cuts to discerning details, an expertly tailored look requires careful curation. Along with a few well-chosen accessories...
Enter the latest Independent competitions
Win anything from gadgets to five-star holidays on our competitions and offers page.
Business videos from commercial thought leaders
Watch the best in the business world give their insights into the world of business.


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Click Here for Articles About THE BIG BANG THEORY
VIDEO: First Look - Next Week's THE BIG BANG THEORY
|Related Links
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November 21, 2014
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November 21, 2014
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November 21, 2014
|Scoop: THE BIG BANG THEORY on CBS - Thursday, December 4, 2014
November 20, 2014
|Related: THE BIG BANG THEORY, CBS
On next week's episode of THE BIG BANT THEORY, entitled "The Cooper/Kripke Inversion," Sheldon is forced to work with Barry Kripke and faces a crisis of confidence. Meanwhile, Howard and Raj spend $1,000 on action figures of themselves, Thursday, Jan. 31 (8:00 - 8:31 PM, ET/PT) on the CBS Television Network. Check out a first look below!
Leonard and Sheldon are brilliant physicists - wizards in the laboratory, but socially inept outside of it. Enter beautiful, free-spirited neighbor, Penny, who aims to teach them a thing or two about life. While Leonard spends his days looking for love, Sheldon is quite content video-chatting with platonic partner Amy Farrah Fowler, or playing Star Trek 3D Chess with their ever-expanding universe of friends, including fellow scientists Koothrappali and Wolowitz, and Wolowitz' new wife, the adorable microbiologist Bernadette.
About Author
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Echo channels Siri-like intelligence to power your home. Also, it might spy on you.
So shiny. So smart.
As someone with Asperger's Syndrome, I know how important it is to find support, even from an iPhone program.
Siri has some competition.
The company has come a long way since its "Think Different" campaign. But that's not all good news.
When Apple shows off its newest killer feature, it will come with a few restrictions


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Sunday, 09 Sep 2012 03:22 PM
The pledge, made to the nation in a prime time television interview, comes four months into his presidency as tumbling ratings and talk of inertia have forced him to become more proactive on the economy.
"I am in a battle and will not look back," he said. "I am setting up a calendar ... two years to create a policy for work and competitiveness. I am accelerating," he said.
Hollande's speech was clearly aimed at turning around perceptions that he is not moving fast enough.
"I know where I am going. I will assume all the responsibility and I will talk regularly to the French people," he said.
Hollande, whose immediate challenge will be to find 30 billion euros ($38.40 billion) in savings in the 2013 budget, said the government would lower its growth forecast to 0.8 percent from 1.2 percent for next year.
"Growth has been shrinking for several months. We will be just over zero percent in 2012 ... so I have asked the government to base the predictions on realistic forecasts. It will be inferior to 1 percent, probably 0.8 percent," he said.
The budget is expected to be the most austere in 30 years as France tries to hit a deficit target of 3 percent of gross domestic product next year or risk losing investors' trust.
It will be presented at a Sept. 28 Cabinet meeting, which has been pushed back by two days to allow for Hollande's trip to the United Nations' General Assembly in New York.
Hollande said last week that by holding state spending steady next year in nominal terms, excluding debt servicing and pension payments, his government would save 10 billion euros in inflation-adjusted terms.
However, that would amount to just one third of the more than 30 billion euros in savings which Hollande says are needed to hit next year's deficit target and stay on course to balance the budget by the end of his five-year mandate.
With his government refusing to cut staffing levels, the bulk of the adjustment will have to come from tax rises.
He said the remaining 20 billion euros would come from tax hikes on companies and wealthier households.
"It's a considerable effort, it's never been done in the history of the Vth Republic, but it's my responsibility," he said.
Having won a May election with 51.6 percent of the vote, Hollande's ratings have slid below 50 percent in less than half the time it took Sarkozy to fall from favour.
A BVA poll published for Le Parisien on Sunday suggested that almost 60 percent of French people are "relatively unhappy" with the president's start compared with 34 percent on May 31. A second survey from pollsters Opinionway showed his popularity rating falling 14 points to 46 percent since June.
Hollande said he expected unemployment, which is at 13 year highs, to begin to fall within a year as his proposals to create 80,000 subsidised jobs and to hire 60,000 people in the education sector as well as a so-called "generation contract" to encourage companies to hire young workers kicks in.
The new Socialist head of state has tried to break from his predecessor Nicholas Sarkozy's pushy manner, promising to consult with social partners but gave unions until the end of the year to reach a "historic compromise" on labour reform or said the government would act unilaterally.
"He seemed to be a totally helpless president, expressing hopes for growth, some minor measures that practically came from the (President Francois) Mitterrand era," Marine Le Pen, president of the far-right National Front, said. "It was almost prehistoric, but what he did confirm was that he would punch a hole of 33 billion euros in the middle classes." ($1 = 0.7812 euros)
© 2014 Thomson/Reuters. All rights reserved.
Tuesday, 25 Nov 2014 15:45 PM
From the Chinese point of view, wars of the future are looking more like Star Wars every day.
. . .
Tuesday, 25 Nov 2014 13:15 PM
Americans disagree on President Barack Obama taking unilateral executive action on amnesty for illegal aliens, but they . . .
Tuesday, 25 Nov 2014 12:55 PM
Iranian leaders are celebrating the seven-month extension of nuclear talks with the United States and other world powers . . .
Sincere thanks to all members of the grand jury who deliberated under extreme...
Well...,it looks like we can anticipate the Ferguson Grand Jury decision this...
Not a smidgeon of evidence . 30,000 emails from lois alone ion that time period...
Take a minute to ask yourself.
What would you do? When you have a a sworn enemy...
1. Ok we know the problem, now what is Congress going to DO about it? Stop...
Newsmax, Moneynews, Newsmax Health, and Independent. American. are registered trademarks of Newsmax Media, Inc. Newsmax TV, and Newsmax World are trademarks of Newsmax Media, Inc.


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Search results for 'instinct'
Samsung DS to 3.5mm Headset Adapter - AARM071FBE
Regular Price: $9.99
Use the Samsung DS to 3.5mm Headset Adapter to connect your phone to any external headset with a 3.5mm audio input jack. Simply plug one end of the adapter into the Samsung DS port on your phone and plug your favorite headset or speaker into the 3.5mm audio jack (female), and you're set! The Samsung DS to 3.5mm Headset Adapter makes it easy to listen to the music on your phone, even if your phone doesn't have a 3.5mm audio jack.Learn More
Scosche Retractable Charge & Sync Cable - USB to Apple 30-Pin / Micro-USB (Bulk)
Regular Price: $20.00
Sick of having to pick through a giant pile of wires strewn about your office or home to find the right cable for charging your phone, camera, or gaming device? Why keep all those wires out when you just need one? That's right, the Scosche Retractable Charge & Sync Cable combines all the charging & syncing power of your Apple 30-Pin connector cable and your micro-USB cable into a single super-cable, capable of charging hundreds of devices. Plus, at a respectable 3.5 foot total length and retractable into a few mere inches of untangled wire, you can pocket this cable and take it anywhere.Learn More
Universal Micro-USB Charge & Sync Cable w/ LED Checker - Black
Regular Price: $25.00
The Universal Micro-USB Charge & Sync Cable w/ LED Checker provides high-speed USB 2.0 data transfer rates, while simultaneously charging your cellular device via your computer's USB port. Organize digital files (music, images, and contacts) on your PC, then transfer files directly to your cell phone using this cable (file types are limited to your cell phone model).This unique charge / sync cable has an LED light which turns blue when the device is fully charged or is idle, and red when it's charging. The Universal Micro-USB Cable with LED Checker is compatible with most Micro-USB devices.Learn More
Samsung ETA0U61JBE Wall Charger
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Key Features:
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- High-quality and ultra portable, great for travel use
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NOTE: This product contains the charger head only.Learn More
Bullet USB Car Charger
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- Comes in a variety of colors
LG STA-U15WS Micro-USB Wall Charger (Bulk)
Regular Price: $11.99
The LG STA-U15WR Micro-USB Wall Charger Features:
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- Input: 100-240V, 50/80 Hz 0.3A
- Output: 5V === 1A
- IC chip prevents overcharging
- Perfect as replacement or spare charger
- Includes a micro-USB data cable (4 ft. long)
- Energy Star complaint
- AC adapter is compatible with LG phones and many other USB enabled devices
NOTE: Ships in Eco-friendly bulk packaging.Learn More
T-Mobile USB and Wall Charging System
Regular Price: $9.99
The T-Mobile USB and Wall Charging System product includes a charging USB data cable and home charger adapter. The home charger adapter features a compact folding blade design, which plugs into any US outlet. It also has a USB port for the included data cable, which charges your device from your computer and allows you to sync your device with a computer, as well.Learn More
Medis 24 / 7 Xtreme Portable Charger
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The Medis 24/7 Xtreme Portable Charger is the world's first personal, portable, fuel cell power system. The Medis 24/7 Xtreme Portable Charger uses electrochemical energy to power your phone, eliminating the need for outlets, wall chargers and the limitations of batteries. Just squeeze it and give it a shake and your phone can be powered up wherever you are. Included are a variety of connectors, such as a standard USB, Mini USB, Micro USB, Palm Treo, and Nokia tips.Learn More
Samsung micro-USB Travel Charger - ETA0U20JBE
Regular Price: $14.99
High-quality and portable, the Samsung ETA0U20JBE USB Travel Charger is a perfect replacement or spare charger. Made with compatibility in mind, the Samsung DTA Charger Head connects via a standard USB 2.0 port, enabling you to charge any device that connects via USB. The Samsung ETA0U20JBE USB Travel Charger has a built-in IC chip, preventing overcharging and shorting, and automatically adapts to meet various voltage requirements throughout the world. It's perfect for travel use.Learn More
Samsung Micro-USB Data Cable - ECC1DU4BBE
Regular Price: $14.99
This Samsung ECC1DU4BBE Micro-USB Data Cable provides high-speed data transfer rates, while simultaneously charging your Galaxy S II or other Samsung Micro-USB compatible phone through your computer's USB port. Organize digital files (music, images, and contacts) on your PC, then transfer files directly to your cell phone using this cable (file types are limited to your cell phone model). The Samsung ECC1DU4BBE Micro-USB Data Cable is compatible with most Micro-USB devices.Learn More


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Joe's Opens Two Stores in So. California
Popular denim brand Joe's has opened two new stores in Southern California: Melrose Place and South Coast Plaza.
Sophia Bush played hostess alonside brand founder Joe Dahan at the Los Angeles opening saying, "I built my whole look around its punk schoolgirl vibe" wearing the the tartan bomber jacket from the Holiday collection.
About Joe's
Los Angeles-based Joe's was created in 2001 by founder and creative director, Joe Dahan. The emergence of premium denim made history in the fashion industry with Joe's at the forefront pioneering the concept of body-specific fits. Over the last decade Joe's has stayed true to it's dedication to timeless style by constantly modernizing wardrobe staples from one season to the next. Today Joe's extends beyond premium denim and includes a casual, chic lifestyle collection for men, women and kids.
Photo by enewsimage.com/Splash News
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May 28, 2014
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How far off shore are we talking? Are either one of these close enough to get to via kayak?
Both web sites mention boat and kayak rentals at Robbie's, but don't say whether you need a power boat or can kayak over.
Thanks again!
Lignumvitae Key Botanical State Park & Indian Key Historic State Park
How far off shore are we talking? Are either one of these close enough to get to via kayak?
3 Replies | Jump to last reply |
Report Abuse
Both are close enough to kayak to (in the same trip - -but one is on bayside and one is on ocean side).
Buy a little soft sided cooler and pack a lunch, drinks, snacks to enjoy on the trip.
Note that Lingnumvitae is much buggier than Indian Key. I would not go there in the summer. In the winter, put on bug spray before leaving the dock.
Report Abuse
Thank you so much for the most helpful reply! We're going end of February/Middle of March. Do you think those two places are good for kayaking? Much to see?
Thanks again!
Report Abuse
Yes - it's a fun (and easy) trip. You'll enjoy the tarpon at Robbie's too.
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Lowtax - Oh. Male. Why didn't you ask that in the first place?
Roy - May I ask where you live, sir?
Lowtax - Sure, go for it.
Roy - Where do you live?
Lowtax - California. Roy.
Roy - How old are you?
Lowtax - Let me ask you a question, Roy. I've seen the movies about people who buy life insurance and then their bosses and kids kill them because they want to collect the money. Is there any way to keep people from collecting the money if I die, Roy?
Roy - Excuse me?
Lowtax - I don't want anybody collecting my life insurance money when I die, Roy. That's just asking to be killed! Roy! Haven't you seen those movies before? I'm not making this up, Roy, it happens.
Roy - I understand your fears, but you can list the dependant as a spouse or other loved one in this situation.
Lowtax - You don't undestand, Roy. I don't want ANYBODY getting the cash if I'm killed, Roy. If I don't, it will seem like my death is just some crazy lotto casino payout, Roy, and I don't want those vulture bastard kids of mine dropping arsenic in my beef soup every night, Roy. Do you know what it's like to have kids, Roy?
Roy - Sorry sir, I dont.
Lowtax - Roy, every day it's like I'm fighting for my own life, Roy. They're playing music loud, driving around like crazy freaks, and just getting on my last damn nerve. I don't know how long I can take this, Roy, before I just snap and burn the place down.
Lowtax - Hey Roy, are you there?
Roy - Yes, sorry, I had to leave for a second.
Lowtax - So tell me more about your program. I want to make sure I'm safe when I die, Roy. I don't want to end up in the gutter like my uncle.
Roy - Well sir, I'll need some more information from you. How old are you?
Lowtax - 42.
Lowtax - Years old.
Roy - Are you a smoker?
Lowtax - Well Roy, I've been quitting for the past two years, so I'm gradually becoming less of a smoker. I don't do the hard stuff though, because my uncle once did some stuff laced with crystal, and he coughed up white crap every hour until the day he died, God bless his soul.
Roy - You are talking about cigeretts, right?
Lowtax - No, my uncle did crank.
Roy - But what about you?
Lowtax - I don't do crank, Roy. It's against my religion.
Speaking of religion, do I have to tell you what religion I am for your life insurance?
Roy - No we dont need that information.
Lowtax - Good. Because my religion gets me all worked up sometimes, and I just get really, really angry until I'm breaking things and my heart starts thumping like a goddamn bass drum. My wife has to lay me down and push my chest until my eyes roll down and I'm able to breathe without the machine, Roy.
By the way, I'm a Lothorarian. The whole "sinister diarama" way of thinking thing.
Roy - Do you have heart problems or medical conditions like those?
You Will Read This Headline. Then You'll Laugh. Then You'll Realize You Were Wrong All Along.
'We're going to be in trouble!' Little Sister wailed, clutching her favorite book to her chest and sobbing. 'This isn't fun like a story anymore!' But Big Sister was not listening, she was thinking. She grabbed Little Sister's book from her and ran into town, yelling 'Help! A book made me and my sister hurt someone!'
Val Kilmer lures 6 morons into the ultimate death schvitz to prove global warming is real.
The Goons have already started their preparations, because that pitch isn't going to carpet itself!
When it comes to gaming laptops, you can never have too many LEDs or too much bad taste.
Our Something Awful ICQ pranks target the worst and most idiotic folks on the Internet. Believe it or not, these ICQ pranks are all - unfortunately - real.
Copyright ©2014 Rich "Lowtax" Kyanka & Something Awful LLC.


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Jamil Ghalayini, London-based foreign exchange structurer at UBS, has left the bank.Ghalayini joined as a director in the forex structuring group from Commerzbank in the spring of last year. He worked alongside Zurich-based Benjamin Anderegg and reported to Marc Honegger, who heads global structuring. Ghalayini reported to Honegger, also in Zurich.
The bank said no replacement had been named. The departure of Ghalayini from the London office follows centralisation of the bank's operation in Zurich with Delia Wang, senior exotics options trader moving to Zurich from Singapore.
Topics: People, UBS, Commerzbank
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Home > Sport > Golf
Friday March 28, 2014 MYT 10:32:04 AM
Friday March 28, 2014 MYT 10:32:59 AM
(Reuters) - Pat Perez and Danny Lee grabbed the first-round clubhouse lead at the Texas Open on Thursday while Phil Mickelson endured a putting nightmare and plunged to nearly last on five-over-par 77.
American Perez, who moved to a cross-handed putting method two months ago, finished in style with birdies at the final three holes for a four-under 68 at the TPC San Antonio.
New Zealander Lee also plundered the closing stretch with four birdies in the final five holes to join Perez.
PGA Tour rookie Andrew Loupe was also four under after 11 holes when play was halted by darkness.
Loupe was among 45 players who did not complete the round due to morning fog that delayed the start of play until 10am.
Perez, 38, whose sole PGA Tour win came in 2009, went cross-handed at the San Diego tour stop in January and promptly finished tied second that week.
"I want to putt conventional, but I don't putt well doing that," he told Golf Channel. "So I went back to cross-handed and started to putt well. I got some old feelings back that I had when I won, and I've just taken it from there."
South Korean-born Lee also changed his putting grip recently, going to the "claw" grip, so-called because the player rests his bottom hand lightly on the club and takes most of the wrist action out of the stroke.
"I was hitting it really good but I wasn't making that many putts early in the round but at the end I was making quite a lot of putts," said Lee, whose runner-up finish at the Puerto Rico Open three weeks ago did wonders for his confidence.
Three-times Masters champion Mickelson, meanwhile, hit only nine greens in regulation to fall nine strokes from the lead.
He also struggled on the greens, running up 32 putts and missing several short attempts, including one from barely two feet for a double-bogey at his final hole.
Mickelson will need a sizzling second round merely to make the halfway cut as he works towards Augusta.
While some players, notably Tiger Woods and Adam Scott, generally prefer to take time off before the majors and prepare at home, Mickelson likes to play his way in.
He is making his first appearance at the Texas Open in more than two decades, and is also planning to contest next week's Houston Open.
Four-time major winner Ernie Els temporarily joined Mickelson at five over, before clawing back three late birdies for a 74.
The Texas Open, which dates to 1922, is the oldest PGA Tour event held in the same city every year.
(Reporting By Andrew Both in Cary, North Carolina; Editing by Frank Pingue)
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United Nations Security Council Demands Action In Disaster
Story By: Larry Stine
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Original Published: 7/21/2014
The United Nations Security Council is calling for a swift inquiry into the Malaysian Airlines disaster in Ukraine. Ambassadors met into the early morning hours Monday, as fighting continues between pro-Russian separatists and Ukrainian forces in the area. The Security Council met late Monday morning discussing a proposed resolution that would call for an independent, international investigation into the downing of that Malaysia jet. It also calls on all states to refrain from, in effect, interfering with any investigation, and calls for a halt in fighting in the area, so that investigators can have free reign with what remains of the Malaysian plane. The British ambassador predicted a vote late Monday afternoon. However, Russia was the main holdout, proposing several amendments, concerned that Ukraine would have a hand in the investigation. The Russian Ambassador, Vitaly Churkin, departed the Security Council session, saying, "We'll see how it works out." Earlier, the British ambassador had said, "Typical Russian stalling tactics."
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Barbara, a New York con-woman in debt to a mobster, finds an easy mark in Bob, a humble, small-town bachelor who, unbeknownst to him, is about to inherit $100,000. Barbara assumes a different identity and moves to the small town of Pascagoula, Mississippi, in order to romance and marry Bob. Barbara, now calling herself Nancy, gets a job teaching school and eventually Bobby and she marry, but her plans are foiled when she actually falls in love and can no longer steal his money. She leaves town, on the run from the mob, but they find her and a gunfight erupts, in which it appears that Barbara/Nancy is shot and killed. Bob is devastated, and leaves town with his inheritance. Soon Nancy is with him, having conspired together to fake her death.
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Iraq still seeking U.S. trainers, PM Maliki says
BAGHDAD (Reuters) - Iraq will continue talks with Washington on how U.S. trainers can work with Iraqi forces after a complete withdrawal of American troops at the end of the year, Prime Minister Nuri al-Maliki said on Saturday.
U.S. President Barack Obama announced on Friday that all U.S. troops would leave Iraq as scheduled by the end of this year after the two governments failed to reach an agreement over giving American soldiers legal immunity.
"Now that we have put this behind us, this will let us settle the issue of training," Maliki said. "Iraqis will ask to resume talks over the number of trainers, the duration (of their stay in Iraq) and how those trainers will be used."
More than eight years after the invasion to topple Saddam Hussein, violence has fallen since the sectarian slaughter of 2006-2007. But Iraq still faces a stubborn Sunni Islamist insurgency tied to al Qaeda and rival Shi'ite militias.
U.S. and Iraqi officials say the local armed forces can contain violence but they need trainers to help build up air defense, maritime capabilities, intelligence gathering and moving the military to conventional warfare tactics.
Iraq will get some military training from a U.S. embassy programme and from contractors who are part of a package with the U.S.-made military hardware Baghdad is purchasing, such as F-16 fighter jets and tanks.
Military advisers working at the embassy are covered by diplomatic immunity afforded to the State Department.
U.S. Secretary of State Hillary Clinton said on Saturday that U.S. commitment to the democratic future of Iraq remains strong despite the decision to go ahead with the withdrawal of American troops as scheduled.
"Even as our troops come home, the United States's commitment to Iraq's future as a secure, stable, democratic nation remains as strong as ever," she said at a news conference in the Tajik capital. "This will end the war and it will open a new chapter in our relationship."
Clinton said Washington expected to have a "significant" security training presence at the Baghdad embassy.
Washington had hoped a few thousand troops in Iraq would help buttress its stability and offset the influence of neighbouring Iran. U.S. officials initially had sought Baghdad's approval for around 3,400 troops.
Many Iraqis are worried about their country's stability and security without the buffer of U.S. military presence, and fear Iraq may slip back into the sectarian tensions that pushed the country to the brink of civil conflict.
(Writing by Patrick Markey; Editing by Michael Roddy)
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