bluuebunny/crossref_metadata_2025 huggingface dataset:
Improved Survival of Macroencapsulated Islets of Langerhans by Preimplantation of the Immunoisolating Device: A Morphometric Study
Authors: Rafael E. , Wu G. S. , Hultenby K. , Tibell A. , Wernerson A.
Published: September 2014

Encapsulation of cells in a semipermeable membrane may in the future provide an opportunity to treat a variety of endocrine and neurological disorders, without the need for lifelong immunosuppression. The physiological conditions in the device are crucial factors for graft survival. Previously, we have shown that the exchange across the immunoisolating membrane and the microcirculation around the TheraCyte™ device increase around 3 months after implantation. The aim of this study was to determine whether preimplantation of the TheraCyte™ device would improve the survival of a later transplanted islet graft. A TheraCyte™ device was implanted SC on one side of the back of a nondiabetic SD rat. After 3 months, 1500 islets isolated from SD rats were transplanted via the device port. At the same time, another device, loaded with the same number of islets, was implanted on the other side of the back. Both devices were explanted 2 weeks after islet transplantation (i.e., 3.5 months and 0.5 month after device implantation, respectively). Six pairs of devices were evaluated by morphometery. The volume densities of viable islets were 0.22 ± 0.04 in the preimplanted device vs. 0.06 ± 0.03 in the nonpreimplanted one (p < 0.05). The corresponding volume densities of fibrosis and necrosis were 0.64 ± 0.13 vs. 0.85 ± 0.08 (p < 0.05) and 0.11 ± 0.14 vs. 0.09 ± 0.07 (ns), respectively. When the absolute volumes (mm3) were calculated, preimplanted devices contained 1.1 ± 0.7 endocrine cells while nonpreimplanted ones contained 0.4 ± 0.2 (p < 0.05). The percentages of insulin-positive β-cells in the preimplanted versus nonpreimplanted device were 80 ± 5% and 67 ± 6%, respectively (p < 0.01). The corresponding volumes of fibrotic tissue were 3.0 ±1.8 vs. 5.2 ± 1.2 (p < 0.05), while the amount of necrotic tissue did not differ significantly (0.42 ± 0.5 vs. 0.50 ± 0.3). Preimplantation of the TheraCyte™ device seems to improve the survival of an encapsulated islet graft and reduce fibroblast outgrowth in the device.
Maintenance of Cold-Preserved Porcine Hepatocyte Function with UW Solution and Ascorbic Acid-2 Glucoside
Authors: Takesue Michihiko , Maruyama Masanobu , Shibata Norikuni , Kunieda Takemi , Okitsu Teru , Sakaguchi Masakiyo , Totsugawa Toshinori , Kosaka Yoshikazu , Arata Akira , Ikeda Hideaki , Matsuoka Junji , Oyama Toshie , Kodama Makoto , Ohmoto Kenji , Yamamoto Shinichiro , Kurabayashi Yuzuru , Yamamoto Itaru , Tanaka Noriaki , Kobayashi Naoya
Published: September 2014

Normal human hepatocytes are an ideal source of liver-targeted cell therapies, such as hepatocyte transplantation and bioartificial livers, but availability of human donor livers for liver cell isolation is severely limited. To effectively utilize scarce donor organs for cell therapies, it is of extreme importance to establish an efficient isolation technique and an effective cold preservation solution for transportation of isolated cells. A lateral segment of the liver was surgically resected from pigs weighing 10 kg and a four-step collagenase and dispase digestion was conducted. Isolated hepatocytes were subjected to 8-h cold storage on ice. The following preservation solutions were tested: 1) University of Wisconsin (UW) solution, 2) UW with 100 μg/ml of ascorbic acid-2 glucoside (AA2G), 3) 100% fetal bovine serum (FBS), and 4) Dulbecco's modified Eagle's medium (DMEM) supplemented with 100% FBS. The mean viability of porcine hepatocytes was 95.5 ± 2.5% when isolated in three independent experiments. Viability, plating efficiency, membrane stability, and ammonia metabolic capacity of cold-preserved hepatocytes were significantly better maintained by the use of UW solution. When AA2G (100 μg/ml) was combined with UW solution, such parameters were further improved. It was explained by inhibition of caspase-3 activation and retention of ATP at high levels of hepatocytes preserved with UW solution containing AA2G. The present work demonstrates that a combination of UW solution with AA2G (100 μg/ml) would be a useful cold preservation means for the development of cell therapies.
Autologous Transplantation of Gingival Fibroblast-Like Cells and a Hydroxylapatite Complex Graft in the Treatment of Periodontal Osseous Defects: Cell Cultivation and Long-Term Report of Cases
Authors: Hou L.-T , Tsai Alex Yi-Min , Liu C.-M , Feng F.
Published: September 2014

Autogenous cell transplantation via hydroxylapatite (HA) vehicle has been reported to have beneficial effects on the treatment of human periodontal osseous defects. The aim of this study was to explore the possibility of using gingival fibroblast-like cells in the therapy of osseous defects caused by inflammatory periodontitis by reporting long-term results of gingival fibroblast-coated hydroxylapatite (GF–HA) grafting for healing these defects. Gingival fibroblasts were cultured from healthy gingivae of treated subjects. Growth of cells on HA particles was established in vitro, and then the GF–HA complex was transplanted into the periodontal osseous defects. Clinical parameters of gingival and plaque indices, probing depth, and periapical x-ray were monitored at baseline and at various periods from 50 months to 6 years after surgery. Grafting with only HA in the osseous defects of the same patient was used for comparison. The present study shows that GF–HA-treated sites could achieve marked pocket reduction and probing attachment gain at reentry and later recalls. Good clinical bone filling of osseous defects in GF–HA-treated sites was also demonstrated in periapical radiographs (increased bone height and reappearance of the crestal cortex) and in some reentry sites. One HA-treated site was filled with connective tissue only, and the absence of new bone formation was noted during a reentry operation. Another HA-treated site exhibited a comparable increase in radiographic density, while part of HA particles were gradually lost in longer recalls. These limited observations conclude that GF–HA grafting may provide a treatment modality leading to regeneration of periodontal tissues in periodontitis-affected osseous defects. Further studies including more cases and demonstration of the deposition of differentiated periodontal tissues are necessary before further application of this therapy.
Long-Term Hibernation of Human Fetal Striatal Tissue does Not Adversely Affect its Differentiation In Vitro or Graft Survival: Implications for Clinical Trials in Huntington's Disease
Authors: Hurelbrink Carrie B. , Tyers Pam , Armstrong Richard J. E. , Dunnett Stephen B. , Barker Roger A. , Rosser Anne E.
Published: September 2014

Transplantation of human fetal CNS tissue is a promising therapy for neurodegenerative conditions such as Huntington's disease (HD), but its widespread adoption is limited by restricted tissue availability. One method of overcoming this problem would be to store the tissue in hibernation medium, an approach that we reported previously for human fetal striatal tissue stored for up to 24 h. We now demonstrate the feasibility of storing such tissue for up to 8 days in hibernation medium. When either fresh or 8-day hibernated striatal cells were cultured under standard conditions for 4 days, the proportion of DARPP-32-positive neurons did not differ significantly, although the total number of cells was significantly less from tissue that had been hibernated. Six weeks after transplantation into cyclosporin A-immunosuppressed unilateral quinolinic acid-lesioned rats, there was no significant difference between fresh and hibernated grafts, both in terms of graft volume and extent of striatal phenotypic markers. This study therefore clearly demonstrates that hibernation of human fetal striatal tissue for up to 8 days is not deleterious to its differentiation in culture or survival following transplantation, and is therefore an appropriate method of storage for this tissue.
A Simple and Cost-Effective Method for the Isolation of Islets from Nonhuman Primates
Authors: O'neil John J. , Tchipashvili Vaja , Parent Richard J. , Ugochukwu Obinna , Chandra Gaurav , Koulmanda Maria , Ko Dicken , Kawai Tatsuo
Published: September 2014

Recent advances in islet cell transplantation have led to insulin independence in a majority of islet transplant recipients. However, there exists a need to overcome the shortage of donor tissue and the necessity for lifelong immunosuppression. Preclinical studies in large animal models are necessary to evaluate the safety and efficacy of alternative approaches for clinical islet transplantation. The nonhuman primate serves as an appropriate animal model for such investigations; however, a major impediment in performing such preclinical research has been the difficulty in isolating islets of sufficient quantity and quality. The current study describes a simple and cost-effective method to isolate nonhuman primate islets to support preclinical islet transplantation research. The results of islet isolations from 54 cynomolgus monkeys and 4 baboons are reported. The pancreas was infused with Liberase HI and subjected to static digestion. The digested tissue was shaken, filtered through a mesh screen, applied to a discontinuous gradient, and centrifuged in much the same manner as with conventional rodent islet isolations. Islets were collected from the two interfaces, washed, and transplanted. Following purification, cynomolgus monkey islet isolation yields were 50,100 ± 3120 IE total or 8760 ± 420 IE/g pancreas with the percent purity and viability of 90.8 ± 0.9 and 90.7 ± 0.7, respectively. Total insulin content of the isolated islets was 405 ± 53 μg insulin with DNA content being and 976 ± 117 μg DNA, corresponding to a ratio of 0.57 μg insulin/μg DNA. STZ-induced diabetes was reversed in both mouse and nonhuman primate recipients, which possessed significant levels of c-peptide following transplantation and well-granulated islet grafts. The technique yields sufficient numbers of pure and viable islets to support preclinical research to develop improved strategies to prevent the immune destruction of the transplanted islet graft.
Hyperthermic Preconditioning Protects Pig Islet Grafts from Early Inflammation but Enhances Rejection in Immunocompetent Mice
Authors: Brandhorst Daniel , Brandhorst Heide , Kumarasamy Vidya , Maataoui Adel , Alt Alexandra , Brendel Mathias D. , Bretzel Reinhard G.
Published: September 2014

The induction of heat shock proteins (HSP) protects isolated islet cells against the cytotoxicity of inflammatory mediators in vitro. Very little information is available about the effect of HSP overexpression on function of preconditioned islet grafts. The present study investigated the function of heat-exposed pig islets after transplantation into immunocompetent mice in comparison with in vitro resistance against inflammatory mediators. Pig islets were preconditioned at 43°C or sham treated prior to subcapsular transplantation into diabetic C57/Bl6j mice. Nondiabetic mice simultaneously receiving preconditioned and control islets were subjected to bilateral nephrectomy for determination of pig insulin. Resistance against H2O2, NO, human Il-1β, IFN-γ, or TNF-α was assessed by trypan blue exclusion and insulin determination. Heat-induced protein expression was confirmed by Western blot analysis. Graft preconditioning increased resistance against H2O2, NO, or cytokines (p < 0.05) but decreased survival in nondiabetic mice (p < 0.05) and function in diabetic mice (p < 0.01). Upregulation of caspase-3 activity as well as Bax, Fas, FasL, and DFF expression (p < 0.05) indicated simultaneous induction of apoptosis. The coexpression of HSP and proapoptotic proteins reveals the dual character of the stress response simultaneously starting mechanisms for protection and apoptosis. In vitro assays seem to reflect only insufficiently the situation of islets after transplantation.
Y Chromosome Detection of Three-Dimensional Tissue-Engineered Skeletal Muscle Constructs in a Syngeneic Rat Animal Model
Authors: Beier J. P. , Kneser U. , Stern-Sträter J. , Stark G. B. , Bach A. D.
Published: September 2014

Surgical reconstruction of muscle tissue lost by trauma or tumor ablation is limited by the lack of availability of functional native tissue substitution. Moreover, so far most inherited or acquired muscle diseases are lacking sufficient treatment, because only few alternatives exist to provide functional restoration of lost muscle tissues. Engineering those tissues and transplantation into sites of dysfunction may be an alternative approach and may allow replacement of such damaged or failing skeletal muscle tissues. Techniques attempting reconstruction of some human tissues and organs (tissue engineering) have been introduced into clinical practice recently. One major problem that previous transplantation studies were facing is the ability of detection of transplanted cells after integration. Using the Y chromosome in situ hybridization technique in a syngeneic rat model allows transplantation of cell constructs orthotopically, without manipulation of the cells, with no rejection or immunosuppression being implied, but providing a nondilutable genetic marker to identify transplanted cells. The purpose of our study was to create functional skeletal muscle tissue in vivo using the transplantation of primary myoblasts precultivated within a three-dimensional (3D) fibrin matrix and to determine the fate of the transplanted cells using the Y chromosome detection technique. 3D myoblast cultures were established derived from male donor rats and after 7 days of cultivation we performed an orthotopic transplantation of 3D cell constructs into a created muscle defect within the gracilis muscle of syngeneic female rats. Anti-desmin immunostaining and Y chromosome in situ hybridization indicated the survival and integration of transplanted male myoblasts into the female recipient animal, thus demonstrating the feasibility of this approach in tissue engineering and the research of cell transplantation in general.
CD86 Blockade in Genetically Modified Porcine Cells Delays Xenograft Rejection by Inhibiting T-Cell and NK-Cell Activation
Authors: Costa Cristina , Pizzolato Maryellen C. , Shen Yamin , Wang Yi , Fodor William L.
Published: September 2014

Porcine xenografts transplanted into primates are rejected in spite of immunosuppression. Identification of the triggering mechanisms and the strategies to overcome them is crucial to achieve long-term graft survival. We hypothesized that porcine CD86 (pCD86) contributes to xenograft rejection by direct activation of host T cells and NK cells. Formerly, we designed the human chimeric molecule hCD152-hCD59 to block pCD86 in cis. To test the efficacy in vivo, we have utilized a pig-to-mouse xenotransplant model. First, we showed that hCD152-hCD59 expression prevents the binding of murine CD28Ig to pCD86 on porcine aortic endothelial cells (PAEC) and dramatically reduces IL-2 secretion by Con A-stimulated mouse splenocytes in coculture. Moreover, IFN-γ secretion by IL-12-stimulated mouse NK cells was averted after coculture with hCD152-hCD59 PAEC. In vivo, control PAEC implanted under the kidney capsule were rapidly rejected (2–4 weeks) in BALB/c and BALB/c SCID mice. Rejection of hCD152-hCD59 PAEC was significantly delayed in both cases. Signs of immune modulation in the hCD152-hCD59-PAEC BALB/c recipients were identified such as early hyporesponsiveness and diminished antibody response. Thus, simply modifying the donor xenogeneic cell can diminish both T cell and NK cell immune responses. We specifically demonstrate that pCD86 contributes to rejection of porcine xenografts.
Galantamine Effects on Memory, Spatial Cue Utilization, and Neurotrophic Factors in Aged Female Rats
Authors: French K. L. , Bimonte-Nelson H. A. , Granholm A.-Ch.
Published: September 2014

Galantamine is an acetylcholine esterase inhibitor that has been approved for use in Alzheimer's disease. However, even though clinical studies indicate efficacy in attenuating some of the symptoms associated with the disease, there are a paucity of studies evaluating the effects of galantamine administration on cognitive performance and brain parameters in aged rats. Further, because all previous animal studies using galantamine have been performed in male rats, there is no information on how females respond to galantamine treatment. Therefore, we studied the effects of 0.3, 0.6, and 1.2 mg/kg/day galantamine in 20-month-old female rats in terms of performance on the working and reference memory water radial arm maze task. Galantamine did not influence maze performance. Furthermore, a probe trial procedure to determine extramaze cue utilization while solving the water radial arm maze established that aged female rats utilized extramaze cues, and that they did not rely on a nonspatial chaining strategy to locate hidden platforms. Galantamine treatment had no effect on use of extramaze cues or chaining. In addition, there were no significant changes in neurotrophin levels in the frontal cortex, entorhinal cortex, hippocampus, or basal forebrain after galantamine administration. Therefore, the data reported here suggest that aged animals do utilize spatial strategies for solving a working memory task, but galantamine has no appreciable effects on this task, at least not at the doses tested.
Acute Rejection of White Adipose Tissue Allograft
Authors: Ablamunits Vitaly , Goldstein Alan J. , Tovbina Marina H. , Gaetz Harold P. , Klebanov Simon
Published: September 2014

White adipose tissue (WAT) transplantation, although widely used in humans, has been done for cosmetic and reconstructive purposes only. Accumulating evidence indicates, however, that WAT is an important endocrine organ and, therefore, WAT transplantation may become valuable as a replacement therapy for a number of hereditary human diseases. Because the most readily available source for such transplantations would be allogeneic tissue, the mechanisms involved in the rejection of WAT allograft should be explored. We have established a model in which leptin-producing allogeneic WAT is transplanted into leptin-deficient ob/ob mice. Because ob/ob mice are obese, hyperphagic, and hypothermic, WAT allograft function is monitored as the reversal of this leptin-deficient phenotype. Here we report that allografted WAT is primarily nonfunctional. However, when WAT is transplanted into immunodeficient (Rag1–/–) ob/ob mice, or into ob/ob mice depleted of T cells by anti-CD3 antibody, a long-term graft survival is achieved as indicated by the reversal of hyperphagia, weight loss, and normalization of body temperature. The symptoms of leptin deficiency rapidly recur when normal spleen cells of the recipient type are injected, or when the antibody treatment is terminated. In contrast, selective depletion of either CD4+ or CD8+ cells alone does not prevent WAT allograft rejection. Similarly, WAT allografts that do not express MHC class I or class II molecules are rapidly rejected, suggesting that both CD4+ and CD8+ T cells may independently mediate WAT allograft rejection.
Long-Term Graft Function after Allogeneic Islet Transplantation
Authors: Lakey Jonathan R. T. , Kin Tatsuya , Warnock Garth L. , Shapiro A. M. James , Tsapogas Panagiotis , Imes Sharleen , Korbutt Gregory S. , Kneteman Norman M. , Rajotte Ray V. , Ryan Edmond A.
Published: September 2014

Islet transplants are emerging as a viable option for the treatment of type 1 diabetes mellitus. From 1989 to 1995 we conducted a series of simultaneous islet–kidney transplants in six uremic type 1 diabetic patients. We report two of these patients who have shown persistent islet graft function over many years. Two female patients with duration of diabetes of 27 and 37 years underwent simultaneous islet–kidney transplant under steroid- and cyclosporine-based immunosuppression. Freshly isolated islets were supplemented with cryopreserved islets from our low-temperature bank of frozen islets. A total islet mass of 9,866 and 15,061 islet equivalents/kg body weight, respectively, was transplanted into the liver through portal vein. Reasonable blood glucose control has been achieved for up to 6 years posttransplant in one patient, but there was minimum clinical benefit from the islet graft at 10 years. In contrast, sustained insulin secretion with nearly normal HbA1c at 13 years follow-up was observed in another patient, providing hope for improving long-term graft outcomes for islet transplant recipient.
1,25-Dihydroxyvitamin D3Increases the Transplantation Success of Human Muscle Precursor Cells in SCID Mice
Authors: Stephan Lionel , Bouchentouf Manaf , Mills Philippe , Lafreniere Jean-François , Tremblay Jacques P.
Published: September 2014

Human muscle precursor cell (hMPC) transplantation is a potential therapy for severe muscle trauma or myopathies. Some previous studies demonstrated that 1,25-dihydroxyvitamin-D3(1,25-D3) acted directly on myoblasts, regulating their proliferation and fusion. 1,25-D3is also involved in apoptosis modulation of other cell types and may thus contribute to protect the transplanted hMPCs. We have therefore investigated whether 1,25-D3could improve the hMPC graft success. The 1,25-D3effects on hMPC proliferation, fusion, and survival were initially monitored in vitro. hMPCs were also grafted in the tibialis anterior of SCID mice treated or not with 1,25-D3to determine its in vivo effect. Graft success, proliferation, and viability of transplanted hMPCs were evaluated. 1,25-D3enhanced proliferation and fusion of hMPCs in vitro and in vivo. However, 1,25-D3did not protect hMPCs from various proapoptotic factors (in vitro) or during the early posttransplantation period. 1,25-D3enhanced hMPC graft success because the number of muscle fibers expressing human dystrophin was significantly increased in the TA sections of 1,25-D3-treated mice (166.75 ± 20.64) compared to the control mice (97.5 ± 16.58). This result could be partly attributed to the improvement of the proliferation and differentiation of hMPCs in the presence of 1,25-D3. Thus, 1,25-D3administration could improve the clinical potential of hMPC transplantation currently developed for muscle trauma or myopathies.
Intrasplenic Hepatocyte Transplantation Prolonged the Survival in Nagase Analbuminemic Rats with Liver Failure Induced by Common Bile Duct Ligation
Authors: Ito Masahiro , Nagata Hideo , Yamamoto Toshiyuki , Yoshihara Daisuke , Fox Ira J. , Miyakawa Shuichi
Published: September 2014

It has already been established that hepatocyte transplantation (HTx) in animal models, such as both chemically and surgically induced acute liver failure, liver-based metabolic disease, and cirrhosis, resulted in significant improvement of liver function and survival. However, the efficacy of hepatocyte transplantation in secondary cholestatic liver disease is not well known. In this study, we transplanted hepatocytes into the spleen of Nagase analbuminemic rats (NARs) with common bile duct ligation (CBDL) to evaluate the function of transplanted hepatocytes by both of serum albumin levels and total bilirubin levels. CBDL was carried out on NARs to induce liver failure. Lewis rat hepatocytes were transplanted in NARs 7 days after CBDL. Animals, in groups of four, underwent the following interventions: group 1—intrasplenic transplantation of 30 × 106 primary Lewis rat hepatocytes in NARs with CBDL (n = 4), group 2—intrasplenic injection of 0.5 ml DMEM in NARs with CBDL (n = 4); group 3—CBDL only (n = 4); group 4—intrasplenic transplantation of 30 × 106 primary Lewis rat hepatocytes in NARs (n = 4). Both bilirubin levels and albumin levels in NARs with CBDL were significantly improved post-HTx. Animals receiving hepatocyte transplantation survived longer than animals in nontransplant control groups. This study indicates that hepatocytes can be transplanted to temporarily provide life-supporting liver-specific metabolic function and prolong the survival in recipient rats with liver failure induced by CBDL.
Role of Blood Glucose in Cytokine Gene Expression in Early Syngeneic Islet Transplantation
Authors: Montolio Marta , Téllez Noèlia , Soler Joan , Montanya Eduard
Published: September 2014

In islet transplantation, local production of cytokines at the grafted site may contribute to the initial nonspecific inflammation response. We have determined whether the metabolic condition of the recipient modulates the cytokine expression in islet grafts in the initial days after transplantation. Normoglycemic and hyperglycemic streptozotocin-diabetic Lewis rats were transplanted with 500 syngeneic islets, an insufficient beta cell mass to restore normoglycemia in hyperglycemic recipients. The expression of IL-1β, TNF-α, IFN-γ, IL-6, IL-10, and IL-4 genes was determined by real-time PCR in freshly isolated islets, in 24-h cultured islets and in islet grafts on days 1,3, and 7 after transplantation. IL-1β mRNA was strongly and similarly increased in normoglycemic and hyperglycemic groups on days 1, 3, and 7 after transplantation compared with freshly isolated and cultured islets. -α mRNA was also strongly increased on day 1, and it remained increased on days 3 and 7. IL-6 and IL-10 were not detected in freshly isolated islets, but their expression was clearly enhanced in 24-h cultured islets and islet grafts. IL-6 was further increased in hyperglycemic grafts. IL-10 expression was increased in both normoglycemic and hyperglycemic grafts on day 1 after transplantation, and remained increased in hyperglycemic grafts compared to 24-h cultured islets. IFN-γ mRNA was barely detected in a few grafts, and IL-4 mRNA was never detected. Thus, the inflammatory response in islet grafts was maximal on day 1 after transplantation, it was sustained, although at lower levels, on days 3 and 7, and it was partly enhanced by hyperglycemia.
Improved Quantity and In Vivo Function of Islets Isolated by Reduced Pressure-Controlled Injection of Collagenase in a Rat Model
Authors: Li Shiri , Sakai Tetsuya , Suzuki Yasuyuki , Goto Tadahiro , Tanaka Tomohiro , Yoshikawa Takuro , Kakinoki Keitaro , Tanioka Yasuki , Matsumoto Ippei , Fujino Yasuhiro , Kuroda Yoshikazu
Published: September 2014

In islet transplantation, insufficient yield is a major obstacle to one-donor/one-recipient transplant. Collagenase, which is injected via a pancreatic duct to separate islets from acini, can so easily distribute into the islet core that it may result in disruption of islets. The purpose of this study was to evaluate the superiority of reduced pressure-controlled collagenase injection (RPCI) at 80 mmHg on islet isolation to injection at 180 mmHg by examining in vivo transplant experiments besides the yield and the glucose stimulation test in a rat model. Lewis rat pancreases were distended with collagenase solution at 80 mmHg pressure as the RPCI group (group 1) and at 180 mmHg (group 2), followed by isolation. The yield in group 1 (1100 ± 160 islets with 2750 ± 530 IEQ) was significantly higher than that in group 2 (900 ± 130 islets with 1570 ± 350 IEQ, p < 0.01) due to the significant difference of the number of islets sized >150 μm in diameter, although the purity was not significantly different between the two groups. Stimulation indices in the glucose stimulation tests were 2.88 ± 1.12 in group 1 and 1.93 ± 0.62 in group 2 (p < 0.05). The cure rate by transplantation of 100 islets to diabetic nude mice in group 1 (8/10) was significantly higher than that in group 2 (3/10, p < 0.05). In a syngenic transplant model of 90% of islets isolated from one donor, the cure rates were 100% and 67% in groups 1 and 2, respectively (NS). The area under the curve on the graph of IPGTT on postoperative day 28 in group 1 was significantly smaller than that in group 2 (p < 0.05). In conclusion, our data show that RPCI at 80 mmHg could contribute to consistently high islet yield and in vivo function in a rat model. It was suggested that the current human protocol should be reviewed from this viewpoint.
Nitric Oxide-Containing Neurons in Long-Term Grafts in a Rat Model of Parkinson's Disease
Authors: Rajakumar B. , Flumerfelt B. A. , Hrycyshyn A. W. , Rajakumar N.
Published: September 2014

The role that nitric oxide may play in modulating graft function in long-term fetal ventral mesencephalic grafts in an animal model of Parkinson's disease was investigated. Mature grafts harvested from the entire fetal ventral mesencephalon possessed a large number of neuronal nitric oxide synthase (nNOS)/NADPH-diaphorase-containing neurons throughout the graft intermingled with dopaminergic neurons. The morphological and neurochemical characteristics of these NADPH-diaphorase neurons resembled those in centers adjacent to the substantia nigra of adult brain but not that of the striatum. Pretreatment with the nNOS blocker, 7-nitroindazole, resulted in contralateral rotations following methamphetamine challenge in long-term grafted animals that previously showed normalized rotational behavior. In contrast, mature grafts derived from fetal ventral mesencephalon without the midline areas possessed only a few nNOS-containing neurons within the grafts, and a similar methamphetamine challenge following 7-nitroindazole pretreatment in long-term grafted rats that previously showed normalized rotational behavior resulted in random movements. Our results indicate that nitric oxide-containing neurons inadvertently included during grafting may affect graft function, and excluding the midline areas of the ventral mesencephalon during tissue harvesting may minimize this effect.
Ex Vivo Expansion and Transplantation of Hematopoietic Stem/Progenitor Cells Supported by Mesenchymal Stem Cells from Human Umbilical Cord Blood
Authors: Huang Guo-Ping , Pan Zhi-Jun , Jia Bing-Bing , Zheng Qiang , Xie Chun-Gang , Gu Jiang-Hong , Mcniece Ian K. , Wang Jin-Fu
Published: September 2014

Human mesenchymal stem cells (MSCs) are multipotential and are detected in bone marrow (BM), adipose tissue, placenta, and umbilical cord blood (UCB). In this study, we examined the ability of UCB-derived MSCs (UCB-MSCs) to support ex vivo expansion of hematopoietic stem/progenitor cells (HSPCs) from UCB and the engraftment of expanded HSPCs in NOD/SCID mice. The result showed that UCB-MSCs supported the proliferation and differentiation of CD34+ cells in vitro. The number of expanded total nucleated cells (TNCs) in MSC-based culture was twofold higher than cultures without MSC (control cultures). UCB-MSCs increased the expansion capabilities of CD34+ cells, long-term culture-initiating cells (LTC-ICs), granulocyte-macrophage colony-forming cells (GM-CFCs), and high proliferative potential colony-forming cells (HPP-CFCs) compared to control cultures. The expanded HSPCs were transplanted into lethally irradiated NOD/SCID mice to assess the effects of expanded cells on hematopoietic recovery. The number of white blood cells (WBCs) in the peripheral blood of mice transplanted with expanded cells from both the MSC-based and control cultures returned to pretreatment levels at day 25 posttransplant and then decreased. The WBC levels returned to pretreatment levels again at days 45–55 posttransplant. The level of human CD45+ cell engraftment in primary recipients transplanted with expanded cells from the MSC-based cultures was significantly higher than recipients transplanted with cells from the control cultures. Serial transplantation demonstrated that the expanded cells could establish long-term engraftment of hematopoietic cells. UCB-MSCs similar to those derived from adult bone marrow may provide novel targets for cellular and gene therapy.
Effect of Pretransplant Preconditioning by Whole Body Hyperthermia on Islet Graft Survival
Authors: Brandhorst H. , Olbrich M. , Neumann A. , Jahr H. , Brandhorst D.
Published: September 2014

Previous observations in heat-shocked pig islets revealed the ambivalent character of the stress response simultaneously inducing processes of protection and apoptosis. To clarify whether the proapoptotic character of the stress response is reduced in heat-exposed islets still embedded in their native environment, hyperthermia was performed in the present study either as whole body hyperthermia (WBH) prior to pancreas resection or as in vitro heat shock (HS) after isolation. HS (42°C/45 min) was induced in donors 12 h before isolation (WBH, n = 32) or in freshly isolated islets prior to 12 h of culture at 37°C (in vitro HS, n = 25). Islets continuously incubated at 37°C served as controls (n = 34). Proinflammatory treatment was performed with H2O2, DETA-NO, or a combination of IL-1β, TNF-α, and IFN-γ. Quality assessment included islet yield, viability staining, static glucose incubation, and nude mouse transplantation. WBH was significantly less effective than in vitro HS to induce HSP70 overexpression and to increase islet resistance against inflammatory mediators. Although characterized by an unaltered Bax to Bcl-2 ratio, islets subjected to WBH partially failed to restore sustained normoglycemia in diabetic nude mice. The inflammatory response observed in the pancreas of WBH-treated rats was associated with significantly reduced viability that seems to have a higher predictive value for posttransplant outcome compared to islet in vitro function or mitochondrial activity. In contrast, in vitro HS significantly decreased transcript levels of Bcl-2, but did not affect posttransplant function compared to sham-treated islets. These findings suggest that WBH is primarily associated with increased necrosis as a secondary tissue type-specific effect of pancreas damage while in vitro HS mainly induces apoptosis.
Good Manufacturing Practice-Compliant Expansion of Marrow-Derived Stem and Progenitor Cells for Cell Therapy
Authors: Gastens Martin H. , Goltry Kristin , Prohaska Wolfgang , Tschöpe Diethelm , Stratmann Bernd , Lammers Dirk , Kirana Stanley , Götting Christian , Kleesiek Knut
Published: September 2014

Ex vivo expansion is being used to increase the number of stem and progenitor cells for autologous cell therapy. Initiation of pivotal clinical trials testing the efficacy of these cells for tissue repair has been hampered by the challenge of assuring safe and high-quality cell production. A strategy is described here for clinical-scale expansion of bone marrow (BM)-derived stem cells within a mixed cell population in a completely closed process from cell collection through postculture processing using sterile connectable devices. Human BM mononuclear cells (BMMNC) were isolated, cultured for 12 days, and washed postharvest using either standard open procedures in laminar flow hoods or using automated closed systems. Conditions for these studies were similar to long-term BM cultures in which hematopoietic and stromal components are cultured together. Expansion of marrow-derived stem and progenitor cells was then assessed. Cell yield, number of colony forming units (CFU), phenotype, stability, and multilineage differentiation capacity were compared from the single pass perfusion bioreactor and standard flask cultures. Purification of BMMNC using a closed Ficoll gradient process led to depletion of 98% erythrocytes and 87% granulocytes, compared to 100% and 70%, respectively, for manual processing. After closed system culture, mesenchymal progenitors, measured as CD105+CD166+CD14–CD45– and fibroblastic CFU, expanded 317- and 364-fold, respectively, while CD34+ hematopoietic progenitors were depleted 10-fold compared to starting BMMNC. Cultured cells exhibited multilineage differentiation by displaying adipogenic, osteogenic, and endothelial characteristics in vitro. No significant difference was observed between manual and bioreactor cultures. Automated culture and washing of the cell product resulted in 181 × 106 total cells that were viable and contained fibroblastic CFU for at least 24 h of storage. A combination of closed, automated technologies enabled production of good manufacturing practice (GMP)-compliant cell therapeutics, ready for use within a clinical setting, with minimal risk of microbial contamination.
The Effects of Cell Density and Device Arrangement on the Behavior of Macroencapsulated β-Cells
Authors: La Flamme Kristen E. , Latempa Thomas J. , Grimes Craig A. , Desai Tejal A.
Published: September 2014

Over the last several decades, considerable research has focused on the development of cell encapsulation technology to treat a number of diseases, especially type 1 diabetes. One of the key advantages of cell encapsulation is that it permits the use of xenogenic tissue, particularly animal-derived cell lines. This is an attractive idea, because it circumvents the issue of a limited human organ supply. Furthermore, as opposed to whole islets, cell lines have a better proliferative capacity and can easily be amplified in culture to provide an endless supply of uniform cells. We have previously described a macroencapsulation device for the immunoisolation of insulin-secreting β-cells. The aim of this work was to optimize the viability and insulin secretion of cells encapsulated within this device. Specifically, the effects of cell packing density and device membrane configuration were investigated. The results indicated that cell density plays an important role in the secretory capacity of the cells, with higher cell density leading to increased insulin secretion. Increasing the transport area of the capsule by modifying the membrane configuration also led to an improvement in the insulin output of the device.
Clonal growth: Invasion or stability? A comparative study of clonal architecture and diversity in native and introduced lineages of Phragmites australis (Poaceae)
Authors: Douhovnikoff Vladimir , Hazelton Eric L. G.
Published: September 2014

• Premise of the study: The characteristics of clonal growth that are advantageous in invasive plants can also result in native plants’ ability to resist invasion. In Maine, we compared the clonal architecture and diversity of an invasive lineage (introduced Phragmites) and a noninvasive lineage (native Phragmites) present in much of North America. This study is the first on stand‐scale diversity using a sample size and systematic spatial‐sampling scheme adequate for characterizing clonal structure in Phragmites. Our questions included: (1) Does the structure and extent of clonal growth suggest that the potential for clonal growth contributes to the invasiveness of the introduced lineage? (2) Is clonal growth common in the native lineage, acting as a possible source of ecological resistance and resilience?

• Methods: Microsatellite markers were used to measure clonal sizes, architecture, and diversity within each lineage in stands within four marshes in Maine.

• Key results: Clonal diversity measures indicated that clonal growth was significantly greater in stands of the native lineage than in the introduced. While lineage was a consistent predictor of clonal diversity relative ranking, the marsh location was a much stronger predictor of the absolute range of these values.

• Conclusions: Our results indicate an important role for clonal growth in the space consolidation of native Phragmites and could explain why the introduced lineage, with stronger competitive traits, has not replaced the native where they co‐occur. These results with regard to clone size, size distributions, singleton occurrence, and clonal architecture provide some evidence for stand development that follows a genotypic initial floristics model.
REAL ESTATE STRATEGIC MANAGEMENT MODEL FOR LITHUANIAN MUNICIPALITIES
Authors: Klumbyte Egle , Apanaviciene Rasa
Published: September 2014

Within the municipal boundaries, municipalities themselves are usually the largest real estate owners and managers. Such significant amount of real estate property could be expected to be professionally managed; however, the situation is different. According to the latest publications, only about 25% of major European cities are able to follow the quantity and value of their real estate portfolios. The Lithuanian Free Market Institute has recently introduced its first Index of Municipalities in Lithuania and states that none of the Lithuanian municipalities has developed its real estate management strategy. This paper reviews the scientific research on municipal real estate management and analyses the system of real estate management in Lithuanian municipalities. The authors of the paper collaborated with the Association of Local Authorities in Lithuania as well as the representatives of municipalities, and together they identified the main problems of real estate management. On the basis of real estate management research and practice within Lithuanian municipalities the authors of the paper present a brand new model which would help to manage municipal real estate effectively by taking into account the priorities of strategic economic and social development tendencies of the region.
Effect of the strontium aluminate and hemihydrate contents on the properties of a calcium sulphoaluminate based cement
Authors: Velazco G. , Almanza J. M. , Cortés D. A. , Escobedo J. C. , Escalante-Garcia J. I.
Published: September 2014

The effect of strontium aluminate (SrAl2O4) on the hydration process of a calcium sulphoaluminate (C4A3Ŝ) cement was investigated. Cement pastes were prepared by mixing C4A3Ŝ , hemihydrate (CaSO4· ½H2O, CŜH0.5) and 0, 10 or 20wt% of SrAl2O4 (SrA). The amount of CŜH0.5 was 15, 20 or 25wt% based on the C4A3Ŝ quantity. The cement pastes were hydrated using water to cement ratios (w/c) of 0.4 and 0.5. Samples were cured from 1 to 28 d. The compressive strength and setting time were evaluated and the hydration products were characterized. It was found that the setting time was delayed up to 42 min for the samples containing SrAl2O4 compared to samples without addition. The samples with 25wt% hemihydrate containing 20wt% SrAl2O4 developed the highest compressive strength (60 MPa) after 28 d of curing. The main product after hydration was ettringite (C6AŜ3H32). The morphology of this phase consisted of thin needle-shaped crystals.
Infrastructural Innovation and Generative Information Infrastructures
Authors: Hanseth Ole , Nielsen Petter
Published: September 2014

This chapter addresses issues related to how to enable the broadest possible innovative activities by infrastructural technology design. The authors focus on the development of high-level services based on mobile telecommunication technologies that for matters of simplicity are termed the development of a Mobile Internet. The focus of the analysis is how features of the technology itself enable or constrain innovations. The authors do this by looking on a few embryos of the Mobile Internet (primarily the Norwegian CPA platform, but also two pre-CPA platforms in Norway and Japan's i-mode) through the concepts of end-to-end architecture, programmability of terminals, and generativity. This analysis illustrates that the change from closed infrastructures like MobilInfo and SMSinfo to more open ones like CPA and i-mode increased the speed and range of innovation substantially. At the same time, the differences between CPA and i-mode regarding programmability of terminals and the billing service provided by the CPA network enabling the billing of individual transactions also contributed to basically the same speed and range of innovations around CPA as i-mode in spite of the huge differences in investments into the networks made by the owners. However, the analysis also points out important differences between the Internet and the existing Mobile Internet regarding technological constraints on innovations. It points out important ways in which powerful actors' strategies inhibit innovations, and how they embed their strategies into the technology and, accordingly, create technological barriers for innovation.
Responsible Innovation and Standard Selection
Authors: van de Kaa Geerten
Published: September 2014

This chapter discusses the notion of “responsible innovation” and “value sensitive design”. It applies these notions to standardization and more specifically to standard selection. Based on earlier research (Van de Kaa, 2013; Van den Ende, Van de Kaa, Den Uyl, & De Vries, 2012), it is proposed that standards should be flexible to facilitate changes related to ethical and societal values. An acceptable standard can be achieved by involving users in the standard development process. The understanding of standardization and standard selection in particular can be improved by incorporating concepts and theories from the discipline of philosophy. This chapter discusses three conceptualizations of standard selection: market dominance, socio-political acceptance, and acceptability.
Assessing IPR Disclosure within Standard Setting
Authors: Layne-Farrar Anne
Published: September 2014

As part of its “policy project to examine the legal and policy issues surrounding the problem of potential patent ‘hold-up' when patented technologies are included in collaborative standards,” the Federal Trade Commission held an all-day workshop on June 21, 2011. The first panel of the day focused on patent disclosure rules intended to encourage full knowledge of patents “essential” for a standard and therefore to prevent patent ambush. When patents are disclosed after a standard is defined, the patent holder may have enhanced bargaining power that it can exploit to charge excessive royalties (e.g., greater than the value the patented technology contributes to the product complying with the standard). In this chapter, the authors present a case study on patent disclosure within the ICT sector. Specifically, they take an empirical look at the timing of patent disclosures within the European Telecommunications Standards Institute, the body responsible for some of the world's most prevalent mobile telephony standards. They find that most members officially disclose their potentially relevant patents after the standard is published, and sometimes considerably so. On the other hand, the authors also find that the delay in declaring patents to ETSI standards has been shrinking over time, with disclosures occurring closer to (although for the most part still after) the standard publication date for more recent standard generations as compared to earlier ones. This latter finding coincides with ETSI policy changes, suggesting that standards bodies may be able to improve patent disclosure with more precise rules.
Privacy Preserving Distributed K-Means Clustering in Malicious Model Using Verifiable Secret Sharing Scheme
Authors: Patel Sankita , Sonar Mitali , Jinwala Devesh C.
Published: September 2014

In this article, the authors propose an approach for privacy preserving distributed clustering that assumes malicious model. In the literature, there do exist, numerous approaches that assume a semi honest model. However, such an assumption is, at best, reasonable in experimentations; rarely true in real world. Hence, it is essential to investigate approaches for privacy preservation using a malicious model. The authors use the Pederson's Verifiable Secret Sharing scheme ensuring the privacy using additively homomorphic secret sharing scheme. The trustworthiness of the data is assured using homomorphic commitments in Pederson's scheme. In addition, the authors propose two variants of the proposed approach - one for horizontally partitioned dataset and the other for vertically partitioned dataset. The experimental results show that the proposed approach is scalable in terms of dataset size. The authors also carry out experimentations to highlight the effectiveness of Verifiable Secret Sharing scheme against Zero Knowledge Proof scheme.