bluuebunny/crossref_metadata_2025 huggingface dataset:
Stem Cell Transplantation in Amyotrophic Lateral Sclerosis Patients: Methodological Approach, Safety, and Feasibility
Authors: Martínez Héctor R. , Molina-Lopez Juan Francisco , González-Garza María Teresa , Moreno-Cuevas Jorge E. , Caro-Osorio Enrique , Gil-Valadez Alfonso , Gutierrez-Jimenez Eugenio , Zazueta-Fierro Oscar E. , Meza J. Alfonso , Couret-Alcaraz Patricio , Hernandez-Torre Martin
Published: December 2011

Amyotrophic lateral sclerosis is characterized by the selective death of motor neurons. Stem cells have been proposed as a potential therapeutic strategy. The safety of stem cell transplantation into the frontal motor cortex to improve upper motor neuron function is described. Sixty-seven patients with definite amyotrophic lateral sclerosis were included. After giving their informed consent, the patients underwent magnetic resonance imaging, functional rating, pulmonary function test, and laboratory tests. Their bone marrow was stimulated with daily filgrastim (300 μg) given subcutaneously for 3 days. Peripheral blood mononuclear cells were obtained by leukapheresis. Isolated CD133+ stem cells were suspended in 300 μl of the patient's cerebrospinal fluid and implanted into the motor cortex. Adverse events were recorded at each step of the procedure and were classified according to the Common Terminology Criteria for Adverse Events v3.0. The survival at 1 year was 90% after transplantation. with a mean long-term survival rate of 40.17 months from diagnosis. The most common adverse events were in grades I–II and involved transient skin pain (19.5% of patients) attributed to the insertion of the Mahurkar catheter into the subclavian vein, minor scalp pain (15.9%), and headache (12.2%) from the surgical procedure. Several patients (1.5 – 4.5%) reported diverse grade I adverse events. There were two deaths, one considered to be associated with the procedure (1.5%) and the other associated with the disease. Autologous stem cell transplantation into the frontal motor cortex is safe and tolerated well by patients. Further controlled studies are required to define the efficacy of this procedure.
Quantitative in Situ Analysis of FoxP3+ T Regulatory Cells on Transplant Tissue Using Laser Scanning Cytometry
Authors: Takahashi Hidenori , Ruiz Phillip , Ricordi Camillo , Delacruz Victor , Miki Atsushi , Mita Atsuyoshi , Misawa Ryosuke , Barker Scott , Burke George W. , Tzakis Andreas G. , Ichii Hirohito
Published: December 2011

There is abundant evidence that immune cells infiltrating into a transplanted organ play a critical role for destructive inflammatory or regulatory immune reactions. Quantitative in situ analysis (i.e., in tissue sections) of immune cells remains challenging due to a lack of objective methodology. Laser scanning cytometry (LSC) is an imaging-based methodology that performs quantitative measurements on fluorescently and/or chromatically stained tissue or cellular specimens at a single-cell level. In this study, we have developed a novel objective method for analysis of immune cells, including Foxp3+ T regulatory cells (Tregs), on formalin-fixed/paraffin-embedded (FFPE) transplant biopsy sections using iCys® Research Imaging Cytometer. The development of multiple immunofluorescent staining was established using FFPE human tonsil sample. The CD4/CD8 ratio and the population of Tregs among CD4+ cells were analyzed using iCys and compared with the results from conventional flow cytometry analysis (FCM). Our multiple immunofluorescent staining techniques allow obtaining clear staining on FFPE sections. The CD4/CD8 ratio analyzed by iCys was concordant with those obtained by FCM. This method was also applicable for liver, small intestine, kidney, pancreas, and heart transplant biopsy sections and provide an objective quantification of Tregs within the grafts.
Banking Human Umbilical Cord-Derived Mesenchymal Stromal Cells for Clinical Use
Authors: Gong Wei , Han Zhibo , Zhao Hui , Wang Youwei , Wang Jiming , Zhong Jian , Wang Bin , Wang Shanshan , Wang Yongjuan , Sun Lingyun , Han Zhongchao
Published: December 2011

A great deal of interest has arisen recently with respect to human mesenchymal stem cells (MSCs), due to their broad therapeutic potential. However, the safety and efficacy of MSCs expanded ex vivo for clinical applications remain a concern. In this article, we establish a standardized process for manufacture of human umbilical cord-derived MSCs (UC-MSCs), which encompasses donor screening and testing, recovery, two-stage expansion, and administration. The biological properties and safety of UC-MSCs were then characterized and tested. The safety data from use in human patients have also been reported. After clinical-scale expansion, a yield of 1.03–3.78 × 108 MSCs was achieved in 10 batch manufacturing runs. The biological properties, such as plastic adherence, morphology, specific surface antigen (CD105, CD73, CD90, positive ≥ 95%; CD45, CD34, CD31, CD11b, CD19, HLA-DR, negative ≤2%), and multipotent differentiation potential (osteogenesis and adipogenesis) were retained. Bacterial and mycoplasma tests were negative and endotoxin levels were lower than 2 EU/ml. No adverse events were noted in two patients treated with intravenously and/or intrathecally administered MSCs. The data obtained indicate that banking UC-MSCs for clinical use is feasible.
Human Neural Stem Cells Overexpressing Choline Acetyltransferase Restore Cognitive Function of Kainic Acid-Induced Learning and Memory Deficit Animals
Authors: Park Dongsun , Joo Seong Soo , Kim Tae Kyun , Lee Sun Hee , Kang Hyomin , Lee Hong Jun , Lim Inja , Matsuo Akinori , Tooyama Ikuo , Kim Yun-Bae , Kim Seung U.
Published: December 2011

Alzheimer disease (AD) is a progressive neurodegenerative disease, which is characterized by loss of memory and cognitive function. In AD patients dysfunction of the cholinergic system is the main cause of cognitive disorders, and decreased activity of choline acetyltransferase (ChAT), an enzyme responsible for acetylcholine (ACh) synthesis, is observed. In the present study we investigated if brain transplantation of human neural stem cells (NSCs) genetically modified to encode ChAT gene improves cognitive function of kainic acid (KA)-induced learning deficit rats. Intrahippocampal injection of KA to hippocampal CA3 region caused severe neuronal loss, resulting in profound learning and memory deficit. F3.ChAT human NSCs transplanted intracerebroventricularly improved fully the learning and memory function of KA-induced learning deficit animals, in parallel with the elevation of ACh levels in cerebrospinal fluid. F3.ChAT human NSCs migrated to the KA-induced injury site (CA3) and differentiated into neurons and astrocytes. The present study demonstrates that human NSCs expressing ChAT have lesion-tropic property and improve cognitive function of learning deficit model rats with hippocampal injury by increasing ACh level.
The Colayer Method as an Efficient Way to Genetically Modify Mesencephalic Progenitor Cells Transplanted into 6-OHDA Rat Model of Parkinson's Disease
Authors: Ratzka Andreas , Kalve Ieva , Özer Meltem , Nobre André , Wesemann Maike , Jungnickel Julia , Köster-Patzlaff Christiane , Baron Olga , Grothe Claudia
Published: December 2011

Exogenous cell replacement represents a potent treatment option for Parkinson's disease. However, the low survival rate of transplanted dopaminergic neurons (DA) calls for methodological improvements. Here we evaluated a method to combine transient genetic modification of neuronal progenitor cells with an optimized cell culture protocol prior to intrastriatal transplantation into 6-hydroxydopamine (6-OHDA) unilateral lesioned rats. Plasmid-based delivery of brain-derived neurotrophic factor (BDNF) increases the number of DA neurons, identified by tyrosine hydroxylase immunoreactivity (TH-ir), by 25% in vitro, compared to enhanced green fluorescence protein (EGFP)-transfected controls. However, the nucleofection itself, especially the cell detachment and reseeding procedure, decreases the TH-ir neuron number to 40% compared with nontransfected control cultures. To circumvent this drawback we established the colayer method, which contains a mix of nucleofected cells reseeded on top of an adherent sister culture in a ratio 1:3. In this setup TH-ir neuron number remains high and could be further increased by 25% after BDNF transfection. Comparison of both cell culture procedures (standard and colayer) after intrastriatal transplantation revealed a similar DA neuron survival as seen in vitro. Two weeks after grafting TH-ir neuron number was strongly reduced in animals receiving the standard EGFP-transfected cells (271 ± 62) compared to 1,723 ± 199 TH-ir neurons in the colayer group. In contrast to the in vitro results, no differences in the number of grafted TH-ir neurons were observed between BDNF, EGFP, and nontransfected colayer groups, neither 2 nor 13 weeks after transplantation. Likewise, amphetamine and apomorphine-induced rotational behavior improved similarly over time in all groups. Nevertheless, the colayer protocol provides an efficient way for neurotrophic factor release by transplanted progenitor cells and will help to study the effects of candidate factors on survival and integration of transplanted DA neurons.
Assessment of Neuroprotective Effects of Human Umbilical Cord Blood Mononuclear Cell Subpopulations In Vitro and in Vivo
Authors: Boltze Johannes , Reich Doreen M. , Hau Susann , Reymann Klaus G. , Strassburger Maria , Lobsien Donald , Wagner Daniel-Christoph , Kamprad Manja , Stahl Tobias
Published: December 2011

Experimental transplantation of human umbilical cord blood (hUCB) mononuclear cells (MNCs) in rodent stroke models revealed the therapeutic potential of these cells. However, effective cells within the heterogeneous MNC population and their modes of action are still under discussion. MNCs and MNC fractions enriched (CD34+) or depleted (CD34-) for CD34-expressing stem/progenitor cells were isolated from hUCB. Cells were transplanted intravenously following middle cerebral artery occlusion in spontaneously hypertensive rats and directly or indirectly cocultivated with hippocampal slices previously subjected to oxygen and glucose deprivation. Application of saline solution or a human T-cell line served as controls. In vivo, MNCs, CD34+and CD34-cells reduced neurofunctional deficits and diminished lesion volume as determined by magnetic resonance imaging. MNCs were superior to other fractions. However, human cells could not be identified in brain tissue 29 days after stroke induction. Following direct application on postischemic hippocampal slices, MNCs reduced neural damage throughout a 3-day observation period. CD34+cells provided transient protection for 2 days. The CD34-fraction, in contrast to in vivo results, failed to reduce neural damage. Direct cocultivation of MNCs was superior to indirect cocultivation of equal cell numbers. Indirect application of up to 10-fold MNC concentrations enhanced neuroprotection to a level comparable to direct cocultivation. After direct application, MNCs migrated into the slices. Flow cytometric analysis of migrated cells revealed that the CD34+cells within MNCs were preferably attracted by damaged hippocampal tissue. Our study suggests that MNCs provide the most prominent neuroprotective effect, with CD34+cells seeming to be particularly involved in the protective action of MNCs. CD34+cells preferentially home to neural tissue in vitro, but are not superior concerning the overall effect, implying that there is another, still undiscovered, protective cell population. Furthermore, MNCs did not survive in the ischemic brain for longer periods without immunosuppression.
Perfluorocarbon Emulsions Prevent Hypoxia of Pancreatic β-Cells
Authors: Maillard E. , Juszczak M. T. , Langlois A. , Kleiss C. , Sencier M. C. , Bietiger W. , Sanchez-Dominguez M. , Krafft M. P. , Johnson P. R. V. , Pinget M. , Sigrist S.
Published: December 2011

As oxygen carriers, perfluorocarbon emulsions might be useful to decrease hypoxia of pancreatic islets before transplantation. However, their hydrophobicity prevents their homogenisation in culture medium. To increase the surface of contact between islets and Perfluorooctyl bromide (PFOB), and consequently oxygen delivery, we tested effect of a PFOB emulsion in culture medium on β-cell lines and rat pancreatic islets. RINm5F β-cell line or pancreatic rat islets were incubated for 3 days in the presence of PFOB emulsion in media (3.5% w/v). Preoxygenation of the medium was performed before culture. Cell viability was assessed by apoptotic markers (Bax and Bcl-2) and by staining (fluoresceine diacetate and propidium iodide). β-Cell functionality was determined by insulin release during a glucose stimulation test and. Hypoxia markers, HIF-1α and VEGF, were studied at days 1 and 3 using RT-PCR, Western blotting, and ELISA. PFOB emulsions preserved viability and functionality of RINm5F cells with a decrease of HIF-1α and VEGF expression. Islets viability was preserved during 3 days of culture. Secretion of VEGF was higher in untreated control (0.09 ± 0.041 μg VEGF/mg total protein) than in PFOB emulsion incubated islets (0.02 ± 0.19 μg VEGF/mg total protein, n = 4, p < 0.05) at day 1. At day 3, VEGF secretion was increased as compared to day 1 in control (0.23 ± 0.04 μg VEGF/mg total protein) but it was imbalance by the presence of PFOB emulsion (0.09 ± 0.03 μg VEGF/mg total protein, n = 5, p < 0.05). While insulin secretion was maintained in response to a glucose stimulation test until day 3 when islets were incubated in the presence of PFOB emulsion preoxygenated (0.81 ± 0.16 at day 1 vs. 0.75 ± 0.24 at day 3), the ability to secrete insulin in the presence of high glucose concentration was lost in islets controls (0.51 ± 0.18 at day 1 vs. 0.21 ± 0.13 at day 3). Atmospheric oxygen delivery by PFOB emulsion might be sufficient to decrease islets hypoxia. However, to improve islets functionality, overoxygenation is needed. Finally, maintenance of islet viability and functionality for several days after isolation could improve the outcome of islets transplantation.
Trichostatin a Promotes Cardiomyocyte Differentiation of Rat Mesenchymal Stem Cells after 5-Azacytidine Induction or during Coculture with Neonatal Cardiomyocytes via a Mechanism Independent of Histone Deacetylase Inhibition
Authors: Yang Ge , Tian Jie , Feng Chuan , Zhao Li-Li , Liu Zhenguo , Zhu Jing
Published: December 2011

This study was to investigate the effect of trichostatin A (TSA), a histone deacetylase (HDAC) inhibitor, on cardiac differentiation of bone marrow mesenchymal stem cells (MSCs) in vitro. Rat MSCs were isolated and divided into six groups: 1) control; 2) 5-azacytidine treatment (5-aza, 10 μM); 3) treatment with TSA (100, 300, and 500 nM); 4) treatment with 5-aza followed by incubation with TSA; 5) coculture with neonatal cardiomyocytes (CMs); and 6) treatment with TSA then coculture with CMs. HDAC activity was significantly inhibited in TSA-treated cells with the maximal inhibition after 24 h of exposure to TSA at 300 nM. No changes in HDAC activity were observed in control, 5-aza-treated, or coculture groups. Following 7 days of differentiation, the expression of early cardiac transcription factors GATA-4, NKx2.5, MEF2c, and cardiac troponin T (cTnT) was increased by 6—8 times in the cells in 5-aza-treated, coculture, or TSA-treated groups over control as determined using real-time PCR, immunofluorescence staining, and Western blotting. However, the percent cTnT-positive cells were dramatically different with 0.7% for control, 10% for 5-aza-treated, 25% for coculture, and 4% for TSA-treated group (500 nM). TSA treatment of the cells pretreated with 5-aza or cocultured with CMs dramatically increased the expression of GATA-4, NKx2.5, and MEF2c by 35—50 times over control. The cTnT protein expression was also significantly increased by over threefold by TSA treatment (500 nM) in both 5-aza-treated and coculture group over control. The percent cTnT-positive cells in both 5-aza-pre-treated and coculture groups were significantly increased by TSA treatment after 1 week of differentiation by up to 92.6% (from 10.3% to 19.8%) and 23.9% (from 24.5% to 30.2%), respectively. These data suggested that TSA enhanced the cardiac differentiation of MSCs after 5-aza induction or during coculture with CMs through a mechanism beyond the inhibition of HDAC activity.
Effects of Myeloablation, Peripheral Chimerism, and Whole-Body Irradiation on the Entry of Bone Marrow-Derived Cells into the Brain
Authors: Lampron Antoine , Lessard Martine , Rivest Serge
Published: December 2011

Understanding how bone marrow-derived cells (BMDCs) enter the central nervous system (CNS) is critical for the development of therapies for brain-related disorders using hematopoietic stem cells. We investigated the brain damages and blood–brain barrier (BBB) modification following either whole-body irradiation or a myeloablative chemotherapy regimen in mice, and the capacity for these treatments to induce the entry of BMDCs into the CNS. Neither treatment had a lasting effect on brain integrity and both were equally efficient at achieving myeloablation. Injection of bone marrow cells from green fluorescent protein (GFP) transgenic mice was able to completely repopulate the hematopoietic niche in the circulation and in hematopoietic organs (thymus and spleen). However, GFP+cells only entered the brain following whole-body irradiation. We conclude that myeloablation, damages to the brain integrity, or the BBB and peripheral chimerism are not responsible for the entry of BMDCs into the CNS following irradiation.
Cotransplantation of Human Embryonic Stem Cell-Derived Neural Progenitors and Schwann Cells in a Rat Spinal Cord Contusion Injury Model Elicits a Distinct Neurogenesis and Functional Recovery
Authors: Niapour Ali , Karamali Fereshteh , Nemati Shiva , Taghipour Zahra , Mardani Mohammad , Nasr-Esfahani Mohammad Hossein , Baharvand Hossein
Published: December 2011

Cotransplantation of neural progenitors (NPs) with Schwann cells (SCs) might be a way to overcome low rate of neuronal differentiation of NPs following transplantation in spinal cord injury (SCI) and the improvement of locomotor recovery. In this study, we initially generated NPs from human embryonic stem cells (hESCs) and investigated their potential for neuronal differentiation and functional recovery when cocultured with SCs in vitro and cotransplanted in a rat acute model of contused SCI. Cocultivation results revealed that the presence of SCs provided a consistent status for hESC-NPs and recharged their neural differentiation toward a predominantly neuronal fate. Following transplantation, a significant functional recovery was observed in all engrafted groups (NPs, SCs, NPs + SCs) relative to the vehicle and control groups. We also observed that animals receiving cotransplants established a better state as assessed with the BBB functional test. Immunohistofluorescence evaluation 5 weeks after transplantation showed invigorated neuronal differentiation and limited proliferation in the cotransplanted group when compared to the individual hESC-NP-grafted group. These findings have demonstrated that the cotransplantation of SCs with hESC-NPs could offer a synergistic effect, promoting neuronal differentiation and functional recovery.
Development and Application of Purified Tissue Dissociation Enzyme Mixtures for Human Hepatocyte Isolation
Authors: Gramignoli Roberto , Green Michael L. , Tahan Veysel , Dorko Kenneth , Skvorak Kristen J. , Marongiu Fabio , Zao Wenchen , Venkataramanan Raman , Ellis Ewa C. S. , Geller David , Breite Andrew G. , Dwulet Francis E. , McCarthy Robert C. , Strom Stephen C.
Published: December 2011

Human hepatocyte transplantation is gaining acceptance for the treatment of liver diseases. However, the reagents used to isolate hepatocytes from liver tissue are not standardized and show lot-to-lot variability in enzyme activity and endotoxin contamination. For clinical application, highly purified reagents are preferable to crude digest preparations. A purified tissue dissociating enzyme (TDE) preparation (CIzyme™ purified enzymes) was developed based on the enzyme compositions found in a superior lot of collagenase previously used by our group for human hepatocyte isolation. The performance of this enzyme preparation was compared to collagenase type XI on 110 liver cases by assessing hepatocyte yield, viability, and seven other functional assays that included plating efficiency, basal and induced CYP450 activities, phase II conjugation activity, and ammonia metabolism. No statistically significant difference was observed between these TDEs when they were used to isolate hepatocytes from liver resections or organ donor tissue on 54 hepatocyte isolations with type XI enzyme and 56 isolations using CIzyme™. These results show that a highly purified and defined TDE preparation can be formulated that provides excellent performance with respect to viability, yield, and functional activity of the isolated cells. In addition to reproducible formulation, these purified enzyme products have only 2–3% of the endotoxin of crude enzyme preparations. These results show that purified enzymes such as CIzyme™ will be a safe and effective for the isolation of human hepatocytes for clinical transplants.
CDNF Protects the Nigrostriatal Dopamine System and Promotes Recovery after MPTP Treatment in Mice
Authors: Airavaara Mikko , Harvey Brandon K. , Voutilainen Merja H. , Shen Hui , Chou Jenny , Lindholm Päivi , Lindahl Maria , Tuominen Raimo K. , Saarma Mart , Hoffer Barry , Wang Yun
Published: December 2011

Cerebral dopamine neurotrophic factor (CDNF) is a recently discovered protein, which belongs to the evolutionarily conserved CDNF/MANF family of neurotrophic factors. The degeneration of dopamine neurons following 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) treatment is well characterized, and efficacy in this model is considered a standard criterion for development of parkinsonian therapies. MPTP is a neurotoxin, which produces parkinsonian symptoms in humans and in C57/Bl6 mice. To date, there are no reports about the effects of CDNF on dopamine neuron survival or function in the MPTP rodent model, a critical gap. Therefore, we studied whether CDNF has neuroprotective and neurorestorative properties for the nigrostriatal dopamine system after MPTP injections in C57/Bl6 mice. We found that bilateral striatal CDNF injections, given 20 h before MPTP, improved horizontal and vertical motor behavior. CDNF pretreatment increased tyrosine hydroxylase (TH) immunoreactivity in the striatum and in the substantia nigra pars reticulata (SNpr), as well as the number of TH-positive cells in substantia nigra pars compacta (SNpc). Posttreatment with CDNF, given 1 week after MPTP injections, increased horizontal and vertical motor behavior of mice, as well as dopamine fiber densities in the striatum and the number of TH-positive cells in SNpc. CDNF did not alter any of the analyzed dopaminergic biomarkers or locomotor behavior in MPTP-untreated animals. We conclude that striatal CDNF administration is both neuroprotective and neurorestorative for the TH-positive cells in the nigrostriatal dopamine system in the MPTP model, which supports the development of CDNF-based treatment for Parkinson's disease.
Remote Processing of Pancreas can Restore Normal Glucose Homeostasis in Autologous Islet Transplantation after Traumatic Whipple Pancreatectomy: Technical Considerations
Authors: Khan Aisha , Jindal Rahul M. , Shriver Craig , Guy Stephen R. , Vertrees Amy E. , Wang Xiaojing , Xu Xiumin , Szust Joel , Ricordi Camillo
Published: December 2011

An emergency autologous islet transplant after a traumatic Whipple operation and subsequent total pancreatectomy was performed for a 21-year-old patient who was wounded with multiple abdominal gunshot wounds. After Whipple pancreatectomy, the remnant pancreas (63.5 g), along with other damaged organs, was removed by the surgeons at Walter Reed Army Medical Center (WRAMC) and shipped to Diabetes Research Institute (DRI) for islet isolation. The pancreas was preserved in UW solution for 9.25 h prior to islet isolation. Upon arrival, the organ was visually inspected; the pancreatic head was missing, the rest of the pancreas was damaged and full of blood; the tail looked normal. A 16-gauge catheter was inserted into the main duct and directed towards tail of the pancreas after the dissection of main duct in the midbody of the pancreas. The pancreas was distended with collagenase solution (Roche MTF) through the catheter. During 10 min of intraductal delivery of enzyme, the gland was distended uniformly. No leakage of the solution was observed. The pancreas was transferred to a Ricordi chamber for automated mechanical and enzymatic digestion. Islets were purified using a COBE 2991 cell processor. Islet equivalents (IEQ; 221,250) of 40% purity and 90% viability were recovered during the isolation, which were shipped back to WRAMC and infused by intraportal injection into the patient. Immediate islet function was demonstrated by the rapid elevation of serum C peptide followed by insulin independence with near normal oral glucose tolerance test (OGTT) 1 and 2 months later. It is possible to restore near normal glucose tolerance with autologous islet transplantation after total pancreatectomy even with suboptimal number of islets while confirming that islets processed at a remote site are suitable for transplantation.
Stem Cells in Burn Eschar
Authors: Van Der Veen Vincent C. , Vlig Marcel , Van Milligen Florine J. , De Vries Sharon I. , Middelkoop Esther , Ulrich Magda M. W.
Published: December 2011

This study compares mesenchymal cells isolated from excised burn wound eschar with adipose-derived stem cells (ASCs) and dermal fibroblasts in their ability to conform to the requirements for multipotent mesenchymal stem cells (MSCs). A population of multipotent stem cells in burn eschar could be an interesting resource for tissue engineering approaches to heal burn wounds. Cells from burn eschar, dermis, and adipose tissue were assessed for relevant CD marker profiles using flow cytometry and for their trilineage differentiation ability in adipogenic, osteogenic, and chondrogenic conditions. Although the different cell types did not differ significantly in their CD marker expression, the eschar-derived cells and ASCs readily differentiated into adipocytes, osteoblasts, and chondrocytes, while dermal fibroblasts only exhibited some chondrogenic potential. We conclude that the eschar-derived mesenchymal cells represent a population of multipotent stem cells. The origin of the cells from burn eschar remains unclear, but it is likely they represent a population of adult stem cells mobilized from other parts of the body in response to the burn injury. Their resemblance to ASCs could also be cause for speculation that in deep burns the subcutaneous adipose tissue might be an important stem cell source for the healing wound.
Inhibiting Matrix Metalloproteinase by Cell-Based Timp-3 Gene Transfer Effectively Treats Acute and Chronic Ischemic Cardiomyopathy
Authors: Tian Hai , Huang Ming-Li , Liu Kai-Yu , Jia Zhi-Bo , Sun Lu , Jiang Shu-Lin , Liu Wei , Kinkaid Heather Y. Mcdonald , Wu Jun , Li Ren-Ke
Published: December 2011

After a myocardial infarction (MI), an increase in the cardiac ratio of matrix metalloproteinases (MMPs) relative to their inhibitors (TIMPs) causes extracellular matrix modulation that leads to ventricular dilatation and congestive heart failure. Cell therapy can mitigate these effects. In this study, we tested whether increasing MMP inhibition via cell-based gene transfer of Timp-3 further preserved ventricular morphometry and cardiac function in a rat model of MI. We also measured the effect of treatment timing. We generated MI (coronary artery ligation) in adult rats. Three or 14 days later, we implanted medium (control) or vascular smooth muscle cells transfected with empty vector (VSMCs) or Timp-3 (C-TIMP-3) into the peri-infarct region ( n = 15—24/group). We assessed MMP-2 and −9 expression and activity, TIMP-3, and TNF-α expression, cell apoptosis, infarct size and thickness, ventricular morphometry, and cardiac function (by echocardiography). Relative to medium, VSMCs delivered at either time point significantly reduced cardiac expression and activity of MMP-2 and −9, reduced expression of TNF-α, and increased expression of TIMP-3. Cell therapy also reduced apoptosis and scar area, increased infarct thickness, preserved ventricular structure, and reduced functional loss. All these effects were augmented by C-TIMP-3 treatment. Survival and cardiac function were significantly greater when VSMCs or C-TIMP-3 were delivered at 3 (vs. 14) days after MI. Upregulating post-MI cardiac TIMP-3 expression via cell-based gene therapy contributed additional regulation of MMP, TIMP, and TNF-α levels, thereby boosting the structural and functional effects of VSMCs transplanted at 3 or 14 days after an MI in rats. Early treatment may be superior to late, though both are effective.
Multiple Intravenous Transplantations of Mesenchymal Stem Cells Effectively Restore Long-Term Blood Glucose Homeostasis by Hepatic Engraftment and β-Cell Differentiation in Streptozocin-Induced Diabetic Mice
Authors: Ho Jennifer H. , Tseng Tzu-Ching , Ma Wei-Hsien , Ong Wei-Kee , Chen Yu-Fan , Chen Ming-Hsiang , Lin Ming-Wei , Hong Chuang-Ye , Lee Oscar K.
Published: December 2011

Depletion of pancreatic β-cells results in insulin insufficiency and diabetes mellitus (DM). Single transplantation of mesenchymal stem cells exhibits short-term effects in some preclinical studies. Here, we further investigated the long-term therapeutic effects of multiple intravenous MSC transplantations. In this study, multiple human MSC transplantations (4.2 × 107cells/kg each time) were performed intravenously at 2-week intervals into streptozocin (STZ)-induced diabetic mice for 6 months. Blood sugar, insulin, renal function, cholesterol, and triglyceride levels were monitored. We demonstrated that compared to single intravenous transplantation, which only transiently decreased hyperglycemia, multiple MSC transplantations effectively restored blood glucose homeostasis. Systemic oxidative stress levels were reduced from the seventh week of treatment. From the 11th week, production of human insulin was markedly increased. When MSC transplantation was skipped after blood sugar level returned to normal at the end of 15th week, a sharp rebound of blood sugar occurred, and was then controlled by subsequent transplantations. At the end of 6 months, histopathology examination revealed MSCs specifically engrafted into liver tissues of the recipients. Fifty-one percent of human cells in the recipient liver coexpressed human insulin, especially those surrounding the central veins. Taken together, intravenous MSC delivery was safe and effective for blood glucose stabilization in this preclinical DM model. Multiple transplantations were essential to restore and maintain glucose homeostasis through decreasing systemic oxidative stress in the early stage and insulin production in the late stage. Liver engraftment and differentiation into insulin-producing cells account for the long-term therapeutic effects of MSCs.
Metabolic Function of a Suboptimal Transplanted Islet Mass in Nonhuman Primates on Rapamycin Monotherapy
Authors: Soleimanpour Scott A. , Hirshberg Boaz , Bunnell David J. , Sumner Anne E. , Ader Marilyn , Remaley Alan T. , Rother Kristina I. , Rickels Michael R. , Harlan David M.
Published: December 2011

Although islet transplantation may restore insulin independence to individuals with type 1 diabetes mellitus, most have abnormal glucose tolerance. We asked whether the defective glucose tolerance is due to inadequate β-cell mass or to impaired insulin sensitivity. We performed metabolic studies on four cynomolgus primates before inducing diabetes with streptozotocin (STZ), then again 2–3 weeks after restoring insulin independence via intrahepatic islet transplantation utilizing a calcineurin inhibitor-free immunosuppressive regimen (induction with rabbit antithymocyte globulin and maintenance therapy with rapamycin). Engrafted β-cell mass was assessed by acute insulin and C-peptide responses to glucose (AIRglu and ACRglu) and arginine (AIRarg and ACRarg). Insulin sensitivity (SI) was determined in naive and transplanted primates from an intravenous glucose tolerance test using the minimal model. α-Cell function was determined by the acute glucagon response to arginine (AGRarg). Glucose tolerance ( K g) decreased from 4.1 ± 0.5%/min in naive primates to 1.8 ± 0.3%/min in transplanted primates ( p < 0.01). Following transplantation, AIRglu was 28.7 ± 13.1 μU/ml compared to 169.9 ± 43.1 μU/ml ( p < 0.03) in the naive condition, ACRglu was 14.5 ± 6.0 ng/ ml compared to 96.5 ± 17.0 ng/ml naive ( p < 0.01), AIRarg was 29.1 ± 13.1 μU/ml compared to 91.4 ± 28.2 μU/ml naive ( p < 0.05), and ACRarg was 1.11 ± 0.51 ng/ml compared to 2.79 ± 0.77 ng/ml naive ( p < 0.05). S I did not differ from naive to posttransplant states. AGRarg was reduced in transplanted primates (349 ± 118 pg/ml) when compared to both naive (827 ± 354 pg/ml) and post-STZ diabetic primates (1020 ± 440 pg/ml) ( p < 0.01 for both comparisons). These data suggest that impaired glucose tolerance observed in islet transplant recipients is secondary to low functional β-cell mass and not to insulin resistance shortly after transplant. Furthermore, improved glycemic control achieved via islet transplantation over the diabetic state might be attained, in part, via reduced glucagon secretion.
Lessons from Genetically Altered Mesenchymal Stem Cells (MSCs): Candidates for Improved MSC-Directed Myocardial Repair
Authors: Alfaro Maria P. , Young Pampee P.
Published: December 2011

The regenerative and reparative potential of mesenchymal stem cells (MSCs) make them attractive candidates for numerous cell-directed therapies. The variant degree of tissue repair by transplanted MSCs has been assessed in several published reports. There are many gaps in the knowledge of MSC biology and the underlying reasons for their disparate effectiveness in tissue repair. This review examines successful preclinical models of MSC-directed repair, particularly of myocardial repair, in an attempt to shed light into the events dictating MSC therapeutic efficacy. The reparative advantage of genetically altered MSCs will be described. This overview will elucidate possible molecular mechanisms that can influence MSC engraftment, differentiation, self-renewal, and ultimately increase wound repair.
De la pluralité des présidences dans l'Union européenne
Authors: Constantinesco Vlad
Published: December 2011

Résumé L’article étudie le nouveau système institutionnel de l’Union européenne issu du traité de Lisbonne. Il montre que le grand nombre de présidences qui découle du nouveau traité peut nuire à l’efficacité et à l’image de l’Union européenne dans l’opinion publique. En effet, ces diverses présidences fonctionnent selon des logiques, des légitimités et des rythmes différents, ce qui ne peut manquer de brouiller une image de l’Union européenne, déjà bien mise en cause par la crise qu’elle et ses États traversent, écho de la crise que diagnostiquaient, au début des années 1930, ceux qui allaient se réclamer du fédéralisme intégral.
Pour une Union européenne qui ne renonce pas
Authors: Lequesne Christian
Published: December 2011

Résumé Dans cet article, l’auteur considère que le projet européen des pères fondateurs a disparu avec la fin de la guerre froide, pour être remplacé par un projet européen à l’échelle d’un continent, vecteur de valeurs occidentales, de démocratie et de règles de marché. Dans son ensemble, le nouveau projet européen doit être considéré comme un succès, qu’il faut asseoir en dynamisant l’économie et défendant la démocratie.
Europe 2020 and the Future of the European Union
Authors: Van Iersel Joost P.
Published: December 2011

Résumé Dans cet article, l’auteur donne une vision de l’Europe 2020 et des possibilités de son impact bénéfique dans le contexte politique et économique actuel. En remplaçant la stratégie de Lisbonne qui a échoué, l’Europe 2020 – programmes nationaux de réforme, initiatives phares, marché unique – se conçoit comme une méthode et un programme globaux pour l’Union européenne de la prochaine décennie, renforçant les liens entre les États membres et l’Union, et approfondissant ainsi l’intégration. Dans le cadre de l’Europe 2020, de saines politiques fiscales et financières doivent être mises en corrélation avec une nécessaire stratégie de croissance, dans laquelle les domaines qui sont soumis à la méthode communautaire et ceux réservés aux États membres seraient coordonnés de manière efficace. Le rôle de la Commission devrait être renforcé. La gouvernance est donc une question centrale. Dans une perspective politique, la stratégie Europe 2020 peut être considérée comme une ouverture vers un gouvernement économique. Sous les strictes conditions de sa réalisation selon les décisions du Conseil en 2010, l’Europe 2020 peut favoriser de façon décisive la solidité de l’économie et un environnement plus stable orienté vers l’avenir. Les acteurs non gouvernementaux comme les entreprises, les organisations et la société civile devraient être considérés comme acteurs engagés dans ce processus.
L'Europe malmenée
Authors: Barthalay Bernard
Published: December 2011

Résumé Bernard Barthalay, dans une Union européenne aux prises avec une crise sans précédent et en proie à une idéologie de démission de la puissance publique et l’intergouvernementalisme laissez-fairiste, analyse les difficultés de la zone euro en un triple oubli, celui de la finalité (les États-Unis d’Europe), de la méthode (un objectif, des étapes, un calendrier) et du peuple (à qui l’Europe est devenue étrangère tandis qu’elle conditionne plus que jamais la vie quotidienne). Il en résulte, non seulement un risque majeur de récession, qui précipiterait la paupérisation relative de l’Europe dans le monde, mais aussi une crise de la démocratie, dans les institutions communes comme dans les États eux-mêmes, qui réduirait l’Europe à l’insignifiance. L’auteur propose une sortie de crise par le haut, avec un plan en trois étapes de fédération budgétaire et politique de la zone euro, mobilisant au service de l’emploi la capacité de financement d’un Trésor fédéral et ouvrant un nouveau chantier constitutionnel, mettant en œuvre les ressources des quatre démocraties : directe, représentative, sociale et participative.
A Pilot Study Testing a Fall Prevention Intervention for Older Adults: Determining the Feasibility of a Five-Sensor Motion Detection System
Authors: Ferrari Marisa , Harrison Barbara , Rawashdeh Osamah , Rawashdeh Muawea , Hammond Robert , Maddens Michael
Published: December 2011

Interventions to prevent hospital-based falls in older adults are critically important to reduce morbidity, mortality, and health care costs. The purpose of this pilot study was to test the accuracy and acceptability of a wireless five-sensor motion detection system (5S-MDS) for detecting falls. Wearable motion sensors, which measure and integrate movement in space, may identify human movement patterns that immediately precede falls, thus allowing prevention. However, sensors must be accurate, and older adults must find wearable sensors acceptable. This descriptive feasibility study recruited 5 healthy older adults (mean age = 69.6) who wore the 5S-MDS while performing 35 movement scenarios. All participants agreed the sensors were acceptable, and skin integrity was maintained for all. The 5S-MDS accurately reflected the patients’ movements and was found acceptable to the older adults; thus, the 5S-MDS has potential as an early warning system for falls.
Revising a Medication Education Program on an Inpatient Child and Adolescent Psychiatric Unit
Authors: Eisenmann Carmen M.
Published: December 2011

Adherence to a medication regimen can be challenging for children and adolescents with mental health disorders. Medication education can be a beneficial tool for nurses to help promote adherence to psychotropic medications. This article describes an initiative to improve medication education offered to children and adolescents and their families on an acute child and adolescent inpatient unit in a mental health facility. Strategies included adding a game to medication education groups, creating and distributing medication education handouts, and developing and implementing medication education sessions for parents. When used by the patients and families, the interventions were appreciated. Having successful interventions in place may help meet the diverse educational needs of this population as nurses seek to improve medication adherence.
PTP1B Deficiency Exacerbates Inflammation and Accelerates Leukocyte Trafficking In Vivo
Authors: Berdnikovs Sergejs , Pavlov Vladimir I. , Abdala-Valencia Hiam , McCary Christine A. , Klumpp David J. , Tremblay Michel L. , Cook-Mills Joan M.
Published: December 2011

It is reported that PTP1B limits cytokine signaling in vitro. However, PTP1B’s function during inflammation in vivo is not known. In this report, we determined whether PTP1B deficiency affects allergic inflammation in vivo. Briefly, lungs of OVA-challenged PTP1B−/− mice had elevated numbers of eosinophils and eosinophil progenitors at 6 h after one OVA challenge and at 24 h after a third OVA challenge as compared with OVA-challenged wild-type mice. There was also an increase in numbers of CD11b+SiglecF+CD34+IL-5Rα+ eosinophil progenitors in the bone marrow, peripheral blood, and spleens of OVA-challenged PTP1B−/− mice. Intravital microscopy revealed that, in OVA-challenged PTP1B−/− mice, blood leukocytes rapidly bound to endothelium (5–30 min), whereas, in wild-type mice, blood leukocytes bound to endothelium at the expected 6–18 h. Consistent with early recruitment of leukocytes, lung eotaxin and Th2 cytokine levels were elevated early in the PTP1B−/− mice. Interestingly, spleen leukocytes from PTP1B−/− mice exhibited an increased chemotaxis, chemokinesis, and transendothelial migration in vitro. In summary, PTP1B functions as a critical negative regulator to limit allergic responses.
TLR and Nucleotide-Binding Oligomerization Domain-like Receptor Signals Differentially Regulate Exogenous Antigen Presentation
Authors: Wagner Claudia S. , Cresswell Peter
Published: December 2011

The effect of dendritic cell (DC) maturation on MHC class II-restricted Ag presentation is well studied, but less is known about the effects of DC maturation on MHC class I-restricted cross-presentation. We investigated the ability of mature DCs to present Ags from cells infected with HSV-1. Pretreatment with pure LPS increased cross-presentation in a manner dependent on both MyD88 and Toll/IL-1R domain-containing adaptor inducing IFN-β, whereas a similar dose of a less pure LPS preparation inhibited cross-presentation. The difference could not be attributed to differences in uptake or phenotypic maturation. The likely contaminant responsible for shutting down cross-presentation is peptidoglycan (PGN). Addition of PGN to pure LPS abrogated its ability to enhance cross-presentation. Direct activation of DCs with PGN inhibited cross-presentation through nucleotide-binding oligomerization domain-like receptor signaling. These results demonstrate that different maturation stimuli can have opposite impacts on the ability of DCs to cross-present viral Ags.
Regulatory T Cells Enhance Mast Cell Production of IL-6 via Surface-Bound TGF-β
Authors: Ganeshan Kirthana , Bryce Paul J.
Published: December 2011

Mast cell degranulation is a hallmark of allergic reactions, but mast cells can also produce many cytokines that modulate immunity. Recently, CD25+ regulatory T cells (Tregs) have been shown to inhibit mast cell degranulation and anaphylaxis, but their influence on cytokine production remained unknown. In this study, we show that, rather than inhibit, Tregs actually enhance mast cell production of IL-6. We demonstrate that, whereas inhibition of degranulation was OX40/OX40 ligand dependent, enhancement of IL-6 was due to TGF-β. Interestingly, our data demonstrate that the Treg-derived TGF-β was surface-bound, because the interaction was contact dependent, and no TGF-β was detectable in the supernatant. Soluble TGF-β1 alone was sufficient to enhance mast cell IL-6 production, and these supernatants were sufficient to promote Th17 skewing, but those from Treg–mast cell cultures were not, supporting this being surface-bound TGF-β from the Tregs. Interestingly, the augmentation of IL-6 production occurred basally or in response to innate stimuli (LPS or peptidoglycan), adaptive stimuli (IgE cross-linking by specific Ag), and cytokine activation (IL-33). We demonstrate that TGF-β led to enhanced transcription and de novo synthesis of IL-6 upon activation without affecting IL-6 storage or mRNA stability. In vivo, the adoptive transfer of Tregs inhibited mast cell-dependent anaphylaxis in a model of food allergy but promoted intestinal IL-6 and IL-17 production. Consequently, our findings establish that Tregs can exert divergent influences upon mast cells, inhibiting degranulation via OX40/OX40 ligand interactions while promoting IL-6 via TGF-β.
B Cell-Derived IL-10 Does Not Regulate Spontaneous Systemic Autoimmunity in MRL.FaslprMice
Authors: Teichmann Lino L. , Kashgarian Michael , Weaver Casey T. , Roers Axel , Müller Werner , Shlomchik Mark J.
Published: December 2011

B cells contribute to the pathogenesis of chronic autoimmune disorders, like systemic lupus erythematosus (SLE), via multiple effector functions. However, B cells are also implicated in regulating SLE and other autoimmune syndromes via release of IL-10. B cells secreting IL-10 were termed “Bregs” and were proposed as a separate subset of cells, a concept that remains controversial. The balance between pro- and anti-inflammatory effects could determine the success of B cell-targeted therapies for autoimmune disorders; therefore, it is pivotal to understand the significance of B cell-secreted IL-10 in spontaneous autoimmunity. By lineage-specific deletion of Il10 from B cells, we demonstrated that B cell-derived IL-10 is ineffective in suppressing the spontaneous activation of self-reactive B and T cells during lupus. Correspondingly, severity of organ disease and survival rates in mice harboring Il10-deficient B cells are unaltered. Genetic marking of cells that transcribe Il10 illustrated that the pool of IL-10–competent cells is dominated by CD4 T cells and macrophages. IL-10–competent cells of the B lineage are rare in vivo and, among them, short-lived plasmablasts have the highest frequency, suggesting an activation-driven, rather than lineage-driven, phenotype. Putative Breg phenotypic subsets, such as CD1dhiCD5+ and CD21hiCD23hi B cells, are not enriched in Il10 transcription. These genetic studies demonstrated that, in a spontaneous model of murine lupus, IL-10–dependent B cell regulation does not restrain disease and, thus, the pathogenic effects of B cells are not detectably counterbalanced by their IL-10–dependent regulatory functions.