bluuebunny/crossref_metadata_2025 huggingface dataset:
Bilateral Fetal Striatal Grafts in the 3-Nitropropionic Acid-Induced Hypoactive Model of Huntington's Disease
Authors: Borlongan Cesario V. , Koutouzis Theodore K. , Poulos Stephen G. , Saporta Samuel , Sanberg Paul R.
Published: June 2017

We investigated the 3-nitropropionic acid (3-NP)–induced hypoactive model of Huntington's disease (HD) to demonstrate whether fetal tissue transplantation can ameliorate behavioral deficits associated with a more advanced stage of HD. Twelve-week-old Sprague–Dawley rats were introduced to the 3-NP dosing regimen (10 mg/kg, i.p., once every 4 days for 28 consecutive days), and were then tested for general spontaneous locomotor activity in the Digiscan locomotor apparatus. All rats displayed significant hypoactivity compared to their pre-3-NP injection locomotor activity. Randomly selected rats then received bilateral intrastriatal solid grafts of fetal striatal (lateral ganglionic eminence, LGE) tissues from embryonic day 14 rat fetuses. Approximately 1/3 of each LGE in hibernation medium was infused into each lesioned host striatum. In control rats, medium alone was infused intrastriatally. A 3-mo posttransplant maturation period was allowed prior to locomotor activity testing. Animals receiving fetal LGE grafts exhibited a significant increase in locomotor activity compared to their post-3-NP injection activity or to the controls’ posttransplant activity. Surviving striatal grafts were noted in functionally recovered animals. This observation supports the use of fetal striatal transplants to correct the akinetic stage of HD. To the best of our knowledge, this is the first study that has investigated the effects of fetal striatal transplantation in a hypoactive model of HD.
Neuronal Progenitor Cells of the Neonatal Subventricular Zone Differentiate and Disperse following Transplantation into the Adult Rat Striatum
Authors: Zigova Tanja , Pencea Viorica , Betarbet Ranjita , Wiegand Stanley J. , Alexander Charlie , Bakay Roy A. E. , Luskin Marla B.
Published: June 2017

We have investigated the suitability of a recently identified and characterized population of neuronal progenitor cells for their potential use in the replacement of degenerating or damaged neurons in the mammalian brain. The unique population of neuronal progenitor cells is situated in a well-delineated region of the anterior part of the neonatal subventricular zone (referred to as SVZa). This region can be separated from the remaining proliferative, gliogenic, subventricular zone encircling the lateral ventricles of the forebrain. Because the neurons arising from the highly enriched neurogenic progenitor cell population of the SVZa ordinarily migrate considerable distances and ultimately express the neurotransmitters GABA and dopamine, we have examined whether they could serve as an alternative source of tissue for neural transplantation. SVZa cells from postnatal day 0-2 rats, prelabeled by intraperitoneal injections of the cell proliferation marker BrdU, were implanted into the striatum of adult rats approximately 1 mo after unilateral denervation by 6-OHDA. To examine the spatio-temporal distribution and phenotype of the transplanted SVZa cells, the experimental recipients were perfused at short (less than 1 wk), intermediate (2-3 wk) and long (5 mo) postimplantation times. The host brains were sectioned and stained with an antibody to BrdU and one of several cell-type specific markers to determine the phenotypic characteristics of the transplanted SVZa cells. To identify neurons we used the neuron-specific antibody TuJ1, or antimembrane-associated protein 2 (MAP-2), and anti-GFAP was used to identify astrocytic glia. At all studied intervals the majority of the surviving SVZa cells exhibited a neuronal phenotype. Moreover, morphologically they could be distinguished from the cells of the host striatum because they resembled the intrinsic granule cells of the olfactory bulb, their usual fate. At longer times, a greater number of the transplanted SVZa cells had migrated from their site of implantation, often towards an outlying blood vessel, and the density of cells within the core of the transplant was reduced. Furthermore, there were rarely signs of transplant rejection or a glial scar surrounding the transplant. In the core of the transplant there were low numbers of GFAP-positive cells, indicating that the transplanted SVZa cells, predominantly TuJ1-positive/MAP2-positive, express a neuronal phenotype. Collectively, the propensity of the SVZa cells to express a neuronal phenotype and to survive and integrate in the striatal environment suggest that they may be useful in the reconstruction of the brain following CNS injury or disease.
Effect of Cryopreservation on the Survival and Function of Murine Islet Isografts and Allografts
Authors: Cattral Mark S. , Lakey Jonathan R.T. , Warnock Garth L. , Kneteman Norman M. , Rajotte Ray V.
Published: June 2017

We compared the efficacy of fresh and frozen/thawed islets by determining the minimum number required to consistently reverse diabetes in mice. Defined numbers of islets, isolated from Balb/c (H-2d) and CBA/J (H-2k) mice, were transplanted into streptozotocin-induced diabetic Balb/c mice. Frozen/thawed grafts were cooled slowly to −40°C, stored at −196°C, and thawed rapidly. At 100 days after transplantation, isografts were recovered for measurement of insulin content. Mean (±SD) recovery of cryopreserved islets after thawing was 80 ± 3% (range 67–89%). For both fresh and frozen/thawed isografts and allografts, 200 islets were required to establish normoglycemia. The degree of metabolic function provided by equivalent quantities of fresh and frozen/thawed grafts was similar; and all normoglycemic isograft recipients remained so until graft nephrectomy. The insulin content of fresh and frozen/thawed isografts containing 200 and 300 islets were 151 ± 25 and 126 ± 8 mU and 259 ±36 and 278 ± 20 mU, respectively. Among allograft recipients, median survival ranged from 15 to 20 days, and was not influenced by cryopreservation or graft size. The results of this study demonstrate a high rate of recovery of viable islets following cryopreservation. The function of equivalent quantities of fresh and cryopreserved islet isografts and allografts in nonimmunosuppressed recipients is similar.
Implantation of Collagen Iv/Poly(2-Hydroxyethyl Methacrylate) Hydrogels Containing Schwann Cells into the Lesioned Rat Optic Tract
Authors: Plant Giles W. , Chirila Traian V. , Harvey Alan R.
Published: June 2017

Poly (2-hydroxyethylmethacrylate) (PolyHEMA) hydrogels, when combined with extracellular matrix molecules and infiltrated with cultured Schwann cells, have the capability to induce CNS axonal regrowth after injury. We have further investigated these PolyHEMA hydrogels and their potential to bridge CNS injury sites. Collagen IV-impregnated hydrogels containing Schwann cells were implanted into the lesioned optic tract in 14 rats. On examination 2–4 months later, there was good adherence between the implants and CNS tissue, and large numbers of viable Schwann cells (S100+, GFAP+, Laminin+, and LNGFR+) were seen within the hydrogel matrices. Immunohistochemical analysis showed that the collagen IV-impregnated PolyHEMA hydrogels preferentially supported the transplanted Schwann cells and not host glial cells such as astrocytes (GFAP+) or oligodendroglia (CAII+). Macrophages (ED1+) were also seen within the sponge structure. Eighty-three percent of the implanted hydrogels contained RT97+ axons within their trabecular networks. Regrowing axons were associated with the transplanted Schwann cells and not with the small number of infiltrating astrocytes. RT97+ axons were traced up to 510 μm from the nearest host neuropil. These axons were sometimes myelinated by the transplanted Schwann cells and expressed the peripheral myelin marker Po+. WGA/HRP-labeled retinal axons were seen within transplanted hydrogel sponges, with 40% of the cases growing for distances up to 350–450 μm within the polymer network. The data indicate that impregnating PolyHEMA sponges with collagen IV can modify the host glial reaction and support the survival of transplanted Schwann cells. This study thus provides new information on how biomaterials could be used to modify and bridge CNS injury sites.
Pretreatment of Crude Pancreatic Islets with Mitomycin C (Mmc) Prolongs Islet Graft Survival in a Xenogeneic Rat-To-Mouse Model
Authors: Grochowiecki Tadeusz , Gotoh Mitsukazu , Dono Keizo , Takeda Yutaka , Nishihara Masayoshi , Ohta Yoshihiko , Kimura Fumihiko , Ohzato Hiroki , Umeshita Koji , Sakon Masato , Monden Morito
Published: June 2017

In this study, we examined the effect of mitomycin C (MMC) treatment on graft survival and evaluated its efficacy in immunomodulation of islet graft for transplantation. Male WS rats were used as islet donors and streptozotocin-induced diabetic C57BL/6 mice as recipients. The isolated islets were treated with MMC at concentrations of 0, 0.1, 1, 3.2, 10, 32, 100, 320, and 1000 μg/mL for 30 min, and were cultured for 20 h. Then, 300–400 islets were transplanted into the renal subcapsular space of diabetic mice. Significant prolongation of graft survival was obtained when the islets were treated with MMC at a concentration of 10, 32, or 100 μg/mL (MST 23 ± 7.4, 17.5 ± 5.4, 29.6 ± 9.7 days: p < 0.003, p < 0.012, p < 0.001, respectively, vs. 12.3 ± 2.7 days for culturing alone). Islets treated with MMC at a concentration of 320 μg/mL or more failed to restore normoglycemia in the diabetic recipient mice after transplantation. Viability of islets incubated with doses up to 100 μg/mL, assessed under the confocal microscope after propidium iodide and Hoechst 33342 staining, was maintained well comparable to that of freshly isolated islets, while those treated at 320 μg/mL was significantly decreased. Thus, a therapeutic window for MMC efficacy was found at concentrations from 10 μg/mL to 100 μg/mL. This modality is simple and effective and underlying molecular mechanisms need to be determined in the future.
Higher Efficiency of Retrovirus Transduction in the Late Stage of Primary Culture of Hepatocytes from Nontreated than from Partially Hepatectomized Rat
Authors: Enosawa Shin , Suzuki Seiichi , Li Xiao-Kang , Okuyama Torayuki , Fujino Masayuki , Amemiya Hiroshi
Published: June 2017

The aim of this investigation was to optimize the conditions for gene transduction by a retroviral vector into primary cultured hepatocytes. Because a retrovirus infection is dependent upon the proliferative activity of the target cells, we examined the thymidine incorporation of the primary culture, using cells from nontreated and partially hepatectomized donor rats. Partial hepatectomy 1 day before cell isolation greatly enhanced the thymidine incorporation, and similarly, the transduction of LacZ, the gene for E. coli β-galactosidase, although the percentage of transduced cells remained low. With cells from the normal rat, the thymidine incorporation increased gradually after the beginning of the culture and reached 16 times the initial activity on day 2, while hepatocytes from the partially hepatectomized donor showed no increase at that stage of the culture. Correspondingly, the number of gene-transduced hepatocytes was increased when the vector was added to the cells 1 or 2 days after isolation. Therefore, hepatocytes cultured for 1 or 2 days after isolation are suitable for retroviral transduction.
Article Commentary: Transplants of Neuronal Cells Bioengineered to Synthesize GABA Alleviate Chronic Neuropathic Pain
Authors: Eaton M. J. , Plunkett J. A. , Martinez M. A. , Lopez T. , Karmally S. , Cejas P. , Whittemore S. R.
Published: June 2017

The use of cell lines utilized as biologic “minipumps” to provide antinociceptive molecules, such as GABA, in animal models of pain is a newly developing area in transplantation biology. The neuronal cell line, RN33B, derived from E13 brain stem raphe and immortalized with the SV40 temperature-sensitive allele of large T antigen (tsTag), was transfected with rat GAD67 cDNA (glutamate decarboxylase, the synthetic enzyme for GABA), and the GABAergic cell line, 33G10.17, was isolated. The 33G10.17 cells transfected with the GAD67 gene expressed GAD67 protein and synthesized low levels of GABA at permissive temperature (33°C), when the cells were proliferating, and increased GAD67 and GABA during differentiation at nonpermissive temperature (39°C) in vitro, because GAD67 protein expression was upregulated with differentiation. A control cell line, 33V1, transfected with the vector alone, contained no GAD67 or GABA at either temperature. These cell lines were used as grafts in a model of chronic neuropathic pain induced by unilateral chronic constriction injury (CCI) of the sciatic nerve. Pain-related behaviors, including cold and tactile allodynia and thermal and tactile hyperalgesia, were evaluated after CCI in the affected hind paw. When 33G10.17 and 33V1 cells were transplanted in the lumbar subarachnoid space of the spinal cord 1 week after CCI, they survived greater than 7 weeks on the pia mater around the spinal cord. Furthermore, the tactile and cold allodynia and tactile and thermal hyperalgesia induced by CCI was significantly reduced during the 2–7-week period after grafts of 33G10.17 cells. The maximal effect on chronic pain behaviors with the GABAergic grafts occurred 2–3 weeks after transplantation. Transplants of 33V1 control cells had no effect on the allodynia and hyperalgesia induced by CCI. These data suggest that a chronically applied, low local dose of GABA presumably supplied by transplanted cells near the spinal dorsal horn was able to reverse the development of chronic neuropathic pain following CCI. The use of neural cell lines that are able to deliver inhibitory neurotransmitters, such as GABA, in a model of chronic pain offers a novel approach to pain management.
Effects of Osteogenic Protein-1 (OP-1) Treatment on Fetal Spinal Cord Transplants to the Anterior Chamber of the Eye
Authors: Granholm A.-Ch. , Sanders L. A. , Ickes B. , Albeck D. , Hoffer B. J. , Young D. A. , Kaplan P. L.
Published: June 2017

Spinal cord injury represents a serious medical problem, and leads to chronic conditions that cannot be reversed at present. It has been suggested that trophic factor treatment may reduce the extent of damage and restore damaged neurons following the injury. We have tested the effects of osteogenic protein-1 (OP-1, also known as BMP-7), a member of the transforming growth factor-β superfamily of growth factors, on developing spinal cord motor neurons in an intraocular transplantation model. Embryonic day 13 or 18 spinal cord tissue was dissected, incubated with OP-1 or vehicle, and injected into the anterior chamber of the eye of adult rats. Injections of additional doses of OP-1 were performed weekly, and the overall growth of the grafted tissue was assessed noninvasively. Four to 6 weeks postgrafting, animals were sacrificed and the tissue was processed for immunohistochemistry using antibodies directed against choline acetyltransferase, neurofilament, and the dendritic marker MAP-II. We found that OP-1 treatment stimulated overall growth of spinal cord tissue when dissected from embryonic day 18, but not from embryonic day 13. OP-1 treatment increased cell size and extent of cholinergic markers in motor neurons from both embryonic stages. The neurons also appeared to have a more extensive dendritic network in OP-1-treated grafts compared to controls. These findings indicate that OP-1 treatment may reduce the extent of axotomy-induced cell death of motor neurons, at least in the developing spinal cord.
Extracorporeal Circulation as a New Experimental Pathway for Myoblast Implantation in mdx Mice
Authors: Torrente Y. , D'angelo M. G. , Del Bo R. , Deliso A. , Casati R. , Benti R. , Corti S. , Comi G. P. , Gerundini P. , Anichini A. , Scarlato G. , Bresolin N.
Published: June 2017

The deficiency of dystrophin, a sarcolemmal associated protein, is responsible for Duchenne muscular dystrophy (DMD). Gene replacement is attractive as a potential therapy. In this article, we describe a new method for myoblast transplantation and expression of dystrophin in skeletal muscle tissue of dystrophin-deficient mdx mouse through iliac vessels extracorporeal circulation. We evaluated the extracorporeal circulation as an alternative route of delivering myoblasts to the target tissue. Two series of experiments were performed with the extracorporeal circulation. In a first series, L6 rat myoblasts, transfected with LacZ reporter gene, were perfused in limbs of 15 rats. In the second series, the muscle limbs of three 6–8-week-old mdx were perfused with myoblasts of donor C57BL10J mice. Before these perfusions, the right tibialis anterior (TA) muscle of the rats and mdx was injected three times at several sites with bupivacaine (BPVC) and hyaluronidase. The ability of injected cells to migrate in the host tissue was assessed in rats by technetium-99m cell labeling. No radioactivity was detected in the lungs, bowels, and liver of animals treated with extracorporeal circulation. The tissue integration and viability of the myoblasts were ultimately confirmed by genetic and histochemical analysis of LacZ reporter gene. Following a single extracorporeal perfusion of myoblasts from donor C57BL10J, sarcolemmal expression of dystrophin was observed in clusters of myofibers in tibialis anterior muscles previously treated with BPVC and hyaluronidase. Furthermore, large clusters of dystrophin-positive fibers were observed in muscles up to 21 days after repeated treatments. These clusters represented an average of 4.2% of the total muscle fibers. These results demonstrate that the extracorporeal circulation allows selective myoblast-mediated gene transfer to muscles, opening new perspectives in muscular dystrophy gene therapy.
Direct Effects of Cyclosporin a on Proliferation of Hematopoietic Stem and Progenitor Cells
Authors: Perry S. Scott , Kim Mijung , Spangrude Gerald J.
Published: June 2017

Cyclosporin A (Cy A) has been reported to both stimulate and inhibit bone marrow colony assays in a dose-dependent manner. The observation that anti-γ-IFN antibodies stimulate hematopoiesis to the same degree as Cy A has led several groups to propose that the stimulatory effects of Cy A are due to inhibition of γ-IFN production by T cells. In this study we observed that cultures of highly enriched hematopoietic stem/progenitor cells (HSPC), devoid of CD3/5/8+ T cells, also exhibit enhanced cloning efficiency when cultured in the presence of Cy A. Normal bone marrow cells or Thy-1.1lowSca-1+Linneg HSPC were incubated in methylcellulose cultures and stimulated with various combinations of steel factor (SF), interleukin (IL)-3, IL-6, granulocyte colony stimulating factor (G-CSF), and erythropoietin (EPO) in the presence of increasing concentrations of Cy A. HSPC cultures with SF, IL-3, and IL-6 stimulation and low Cy A concentrations had from 24% to 78% higher cloning efficiencies than did parallel cultures without Cy A, and did not fall below control levels until the Cy A concentration was increased to more than 1.25 μg/ml. The addition of EPO and G-CSF abrogated the Cy A stimulation observed with SF, IL-3, and IL-6. These results were reflected in whole bone marrow, but with a higher range of variability. Cultures in which FK-506 replaced Cy A showed no consistent stimulation or inhibition of colony formation. These studies show that Cy A can stimulate hematopoietic stem cell growth independent of mediation by T cells. Consequently, these results argue for a direct positive effect of Cy A on the signal transduction pathways in HSPC.
Novel Method for Isolation of Adult Porcine Pancreatic Islets with Two-Stage Digestion Procedure
Authors: Cui Wanxing , Gu Yuanjun , Miyamoto Masaaki , Tanaka Masaki , Xu Baoyou , Imamura Masayuki , Iwata Hiroo , Ikada Yoshihioto , Inoue Kazutomo
Published: June 2017

It is particularly difficult to isolate porcine islets (PI). Experience suggests that the success rate of porcine islet isolation (PII) is probably considerably influenced by the distension and digestion of the pancreas. In this study, we divided the digestion procedure into two stages and developed a new enzyme solution to improve both the distension and digestion procedures. As a result, we established a novel and stable method of large-scale adult porcine islet isolation (APII). The harvested pancreata of 2-year-old pigs weighing over 200 kg (n = 18) were distended by introducing our new enzyme solution gently and slowly through the pancreatic ducts. Two-stage digestion (cold, then warm) was then performed by first placing the distended pancreata on ice for 2 h to cause diffusion of the enzyme solution around the islets, and then by incubating the pancreata in a water bath at 37°C for 45 min without shaking. The islets were purified by a COBE 2991 cell processor on dextran T70 discontinuous density gradients. Histological study was performed on porcine pancreata sampled after 0, 15, 30, and 45 min of the second stage, and stained with H&E stain. Next, islet equivalent was calculated. Static incubation study was performed by stimulating the islets with 3.3 and 16.7 mM glucose in Krebs' Ringer bicarbonate buffer (KRBB) solution at 37°C for 1 h, and finally the insulin released was measured. The dilated acinar cells septa around the islets were observed at time 0. Destruction of the acinar cells around the islets by warm digestion was recognized at 15 and 30 min, and destroyed and separated acinar cells present around the islets at 45 min. During the entire course of the warm digestion, the islets remained intact. The number of isolated islets was 291,667 ± 240,452 IEQ/pancreas (n = 14) and 3,294 ± 2199 IEQ/g of pancreatic tissue. The purity of recovered porcine islets was over 90%. The concentration of the insulin secreted by 10,000 IEQ islets selected at random was 83.9 ± 13.4 μU/dish/h in response to 3.3 mM glucose and 104.1 ± 12.9 μU/dish/h in response to 16.7 mM glucose (n = 20). A success rate of approximately 80% was attained with APII. We demonstrated that this increase in the success rate was due to the improved distension and digestion provided by this method. This two-stage APII method with its new enzyme solution may facilitate the future use of porcine islets in clinical xenotransplantation trials.
Protective Effect of Nafamostat Mesilate on Injury of Porcine Hepatocytes by Human Plasma
Authors: Kawazoe Yasushi , Eguchi Susumu , Sugiyama Nozomu , Yuzawa Hiroyuki , Kawashita Yujo , Fujioka Hikaru , Kanematsu Takashi
Published: June 2017

Nafamostat mesilate (NM), a protease inhibitor, possesses a cytoprotective effect and inhibits the activation of complement. The present study investigated whether NM has any protective effect against injury of porcine hepatocytes by human plasma in a bioartificial liver support system. Porcine hepatocytes were harvested and seeded at a density of 2 × 105 cells on a 35-mm collagen-coated plate in Dulbecco's modified Eagle's medium (DMEM) with 10% fetal calf serum. Twenty-four hours later, the medium was replaced with human plasma with three concentrations of NM between 3.8 × 10–5 and 3.8 × 10–4 M and then cultured for 6 h. The viability of porcine hepatocytes, lactate dehydrogenase (LDH) levels, lidocaine clearance, porcine albumin production, and changes in complement (C3) levels were measured. The viability of porcine hepatocytes in human plasma decreased significantly to 37.7 ± 11.4% of that in DMEM. NM improved the viability of the hepatocytes, lowered the levels of LDH, and increased lidocaine clearance and albumin production in a concentration-dependent manner. The concentrations of C3, the marker of xenogeneic reactions, did not change significantly, indicating that no hyperacute xenogeneic reaction occurred in our series. Together, our results suggested that NM exerts favorable effects on porcine hepatocytes in human plasma through direct effect such as prevention of protease activity in the plasma membrane of porcine hepatocytes rather than inhibition of complement-dependent immunoreactions.
Long-Term Functional Assessment of Encapsulated Cells Transfected with Tet-On System
Authors: Hagihara Yasushi , Saitoh Youichi , Arita Norio , Eguchi Yutaka , Tsujimoto Yoshihide , Yoshimine Toshiki , Hayakawa Toru
Published: June 2017

In our previous study, xenogeneic mouse neuroblastoma cells bearing the POMC gene, the precursor of ACTH and β-endorphin, were implanted within polymer capsules into the CSF space of rats. Although ACTH and β-endorphin were secreted, we were not able to control the amounts or times of hormone release. A promoter that is inducible by administration of tetracycline derivatives (Tet) was linked to the POMC gene to control its gene expression (Neuro2A-Tet-On-POMC; NTP). The results showed that POMC gene expression in the implanted encapsulated NTP cells could be regulated in a dose-dependent manner by Tet administration to the hosts. However, no analysis of gene control with the Tet-On system over a long period has been performed. In this study, encapsulated NTP cells were treated in vitro with doxycycline (Dox) (1.0, 10, 100, 1000 ng/ml) continuously for a month. On day 4, the amount of ACTH secretion was dependent on the Dox dose. But in the course of the experiment, the difference of ACTH secretion among those treated with Dox 10, 100, and 1000 ng/ml was eliminated. On the other hand, NTP cells, which were treated with Dox (1000 ng/ml) just on days 7, 14, 21, and 28, secreted almost the same amount of ACTH in 24 h. From these results, for clinical use, an NTP cell line that secretes enough opiate to reduce pain sensitivity without Dox should be established, and Dox could then be administered if necessary.
Efficient Gene Transfer and Expression in Islets by an Adenoviral Vector that Lacks All Viral Genes
Authors: Guo Zhiguang , Shen Jikun , Mital Deepak , Hong Yuanze , Alemany Ramon , Zhong Wei-Wei , Jensik Stephen C. , Williams James W.
Published: June 2017

Although adenoviral vector-mediated gene transfer has significant potential for gene therapy, host immune responses to virally expressed proteins and small insert capacity may limit its clinical application. In order to overcome these disadvantages, a new adenoviral vector that lacks all viral genes has been developed. Using the green fluorescent (GFP) gene as a reporter gene, we investigated the efficiency of gene transfer by this all-viral-genes-deleted and minimal cis-element remaining adenoviral vector (miniAd-GFP) in islets in vitro and ex vivo, and compared it with the E1-deleted adenoviral vector (E1-GFP). One day after in vitro infection, GFP was expressed in both miniAd-GFP- and E1-GFP-infected islets. The percentage of GFP-positive single cells was not significantly different between miniAd-GFP-infected islets and E1-GFP-infected islets. When these islets were transplanted into syngeneic diabetic mice, both miniAd-GFP- and E1-GFP-infected islet grafts reversed diabetes, and normal blood glucose levels were maintained for over 20 weeks posttransplantation. Mild lymphocyte infiltration was found in all E1-GFP-infected islet grafts at all time points. However, this was not seen in most miniAd-GFP-infected islet grafts. Our results indicate that gene transfer by an adenoviral vector that lacks all viral genes is as efficient as E1-deleted adenoviral vector-mediated gene transfer in islets. Furthermore, this adenoviral vector might be less immunogeneic than the E1-deleted adenoviral vector.
MicroRNA-138 enhances TRAIL-induced apoptosis through interferon-stimulated gene 15 downregulation in hepatocellular carcinoma cells
Authors: Zuo Chaohui , Sheng Xinyi , Liu Zhuo , Ma Min , Xiong Shuhan , Deng Hongyu , Li Sha , Yang Darong , Wang Xiaohong , Xiao Hua , Quan Hu , Xia Man
Published: June 2017

Hepatocellular carcinoma is a leading cause of cancer-related mortality worldwide. TRAIL (tumor necrosis factor–related apoptosis–inducing ligand) is a potential target for cancer therapy. However, many cancer cells are resistant to TRAIL-induced apoptosis and its mechanism is not well understood. In this study, to identify potential therapeutic targets for TRAIL-resistant cancer cells, we compared the expression levels of interferon-stimulated gene 15 in TRAIL-sensitive and TRAIL-resistant hepatocellular carcinoma cell lines. Western blot analysis showed that interferon-stimulated gene 15 expression levels were significantly higher in resistant HLCZ01and Huh7 cells than in sensitive LH86 and SMMC-7721 cells. Interferon-stimulated gene 15 knockdown in resistance cells led to TRAIL sensitivity. Conversely, interferon-stimulated gene 15 overexpression in sensitive cells resulted in TRAIL resistance. Our bioinformatics search detected a putative target sequence for microRNA miR-138 in the 3′ untranslated region of the interferon-stimulated gene 15. Real-time quantitative polymerase chain reaction analysis demonstrated that miR-138 was significantly downregulated in TRAIL-resistant cells compared to TRAIL-sensitive cells. Forced expression of miR-138 in resistant cells decreased both messenger RNA and protein levels of interferon-stimulated gene 15, and when exposed to TRAIL, activated poly(adenosine diphosphate-ribose) polymerase, indicating sensitization to TRAIL. The results suggested that miR-138 regulates the interferon-stimulated gene 15 expression by directly targeting the 3′ untranslated region of interferon-stimulated gene 15 and modulates the sensitivity to TRAIL-induced apoptosis. MiR-138 may be a target for therapeutic intervention in TRAIL-based drug treatments of resistant hepatocellular carcinoma or could be a biomarker to select patients who may benefit from the treatment.
Apatinib for metastatic breast cancer in non-clinical trial setting: Satisfying efficacy regardless of previous anti-angiogenic treatment
Authors: Lin Ying , Wu Zheng , Zhang Jian , Hu Xichun , Wang Zhonghua , Wang Biyun , Cao Jun , Wang Leiping
Published: June 2017

Apatinib is a novel tyrosine kinase inhibitor targeting vascular endothelial growth factor receptor 2. This study aimed to evaluate the efficacy and safety of apatinib in metastatic breast cancer (MBC) under non-clinical trial setting, and to study the impact of previous antiangiogenic treatment to the efficacy of apatinib. 52 MBC patients treated with apatinib under non-clinical trial setting in Fudan University Shanghai Cancer Center between January 1st 2015 and October 1st 2016 were included. All patients were included in time-to-treatment failure (TTF) analysis, while 45 patients were enrolled for progression-free survival (PFS) and overall survival (OS) analysis because 7 of the patients with treatment discontinuation due to intolerable toxicities had too short time for efficacy assessment. Impact of previous exposure to antiangiogenic treatment and other factors to patients’ survival were analyzed by Log-rank analysis and Cox multivariate analysis. The median PFS, median OS, and median TTF were 4.90 (95% confidence interval [CI] 3.44 – 6.36), 10.3 (unable to calculate 95% CI), and 3.93 (95% CI 1.96 – 5.90) months, respectively. Previous treatment of bevacizumab did not affect the efficacy of apatinib. Previous exposure to anthracycline, age of 60 years or older and palmar-plantar erythrodysesthesia syndrome were independent predictors for prolonged PFS. Discontinuation of treatment was more common in age group of 60 years or older than that in younger group, although the difference was not significant. Although toxicities were generally managable, a previously unrecorded grade 3~4 adverse event of dyspnea has been observed. This study confirmed the encouraging efficacy and manageable safety of apatinib on pretreated MBC patients in non-clinical trial setting. For the first time to our knowledge, this study found that previous treatment of bevacizumab did not affect the efficacy of apatinib, and reported an undocumented severe adverse effect of dyspnea.
State Export Promotion and Firm-level Employment
Authors: Cassey Andrew J. , Cohen Spencer
Published: June 2017

Most US states have export promotion programs, but it is unknown if these programs create long-term employment, which is often the policy’s stated goal. We merge administrative export promotion and employment data from Washington State to test the effect of firm-level export promotion on firm-level employment using the differences-in-differences estimator. We believe we are the first to have US state data at this level of detail. We find firm participation in an export assistance program increases firm-level employment fleetingly, but not in subsequent periods. Thus, we do not find a statistically significant impact to long-term employment from program participation.
Rethinking key–value store for parallel I/O optimization
Authors: Kougkas Anthony , Eslami Hassan , Sun Xian-He , Thakur Rajeev , Gropp William
Published: June 2017

Key–value stores are being widely used as the storage system for large-scale internet services and cloud storage systems. However, they are rarely used in HPC systems, where parallel file systems are the dominant storage solution. In this study, we examine the architecture differences and performance characteristics of parallel file systems and key–value stores. We propose using key–value stores to optimize overall Input/Output (I/O) performance, especially for workloads that parallel file systems cannot handle well, such as the cases with intense data synchronization or heavy metadata operations. We conducted experiments with several synthetic benchmarks, an I/O benchmark, and a real application. We modeled the performance of these two systems using collected data from our experiments, and we provide a predictive method to identify which system offers better I/O performance given a specific workload. The results show that we can optimize the I/O performance in HPC systems by utilizing key–value stores.
Use of computed tomography to define a sacral safe corridor for placement of 2.7 mm cortical screws in feline sacroiliac luxation
Authors: Philp Helen , Durand Alexane , De Vicente Felipe
Published: June 2017

Objectives

This study aimed to define a safe corridor for 2.7 mm cortical sacroiliac screw insertion in the dorsal plane (craniocaudal direction) using radiography and CT, and in the transverse plane (dorsoventral direction) using CT in feline cadavers. A further aim was to compare the values obtained by CT with those previously reported by radiography in the transverse plane.

Methods

Thirteen pelvises were retrieved from feline cadavers and dissected to expose one of the articular surfaces of the sacrum. A 2.7 mm screw was placed in the sacrum to a depth of approximately 1 cm in each exposed articular surface. Dorsoventral radiography and CT scanning of each specimen were performed. Multiplanar reconstructions were performed to allow CT evaluation in both the dorsal and transverse planes. Calculations were made to find the maximum, minimum and optimum angles for screw placement in craniocaudal (radiography and CT) and dorsoventral (CT) directions when using a 2.7 mm cortical screw.

Results

Radiographic measurement showed a mean optimum craniocaudal angle of 106° (range 97–112°). The mean minimum angle was 95° (range 87–107°), whereas the mean maximum angle was 117° (108–124°). Measurement of the dorsal CT scan images showed a mean optimum craniocaudal angle of 101° (range 94–110°). The mean minimum angle was 90° (range 83–99°), whereas the mean maximum angle was 113° (104–125°). The transverse CT scan images showed a mean dorsoventral minimum angle of 103° (range 95–113°), mean maximum angle of 115° (104–125°) and mean optimum dorsoventral angle of 111° (102–119°).

Conclusions and relevance

An optimum craniocaudal angle of 101° is recommended for 2.7 mm cortical screw placement in the feline sacral body, with a safety margin between 99° and 104°. No single angle can be recommended in the dorsoventral direction and therefore preoperative measuring on individual cats using CT images is recommended to establish the ideal individual angle in the transverse plane.
Adverse Drug Reactions and Cutaneous Manifestations Associated With Anticoagulation
Authors: Vu Trang T. , Gooderham Melinda
Published: June 2017

Anticoagulants are amongst the most commonly prescribed medications worldwide. Although rare, localised and systemic drug reactions have been reported with anticoagulants that can lead to significant morbidity and mortality. Some of the first signs of drug reactions to anticoagulants are cutaneous changes that, when recognised early, can prevent significant complications. Dermatologists should be aware of these changes to make an early and accurate diagnosis. This is particularly important in instances of skin-induced necrosis caused by systemic toxicity to anticoagulants. This review discusses adverse drug reactions to the traditional anticoagulants, warfarin and heparin, and the newer direct oral anticoagulants (DOACs) such as the thrombin inhibitor, dabigatran, and the factor Xa inhibitors, rivaroxaban, apixaban, and edoxaban. In particular, this review provides dermatologists with a framework for early diagnosis and management of patients with drug reactions to anticoagulants and alerts them to potential bleeding complications associated with minor procedures.
Austerity, luxury and uncertainty in the Indian emerald trade
Authors: Brazeal Brian
Published: June 2017

This article examines the bodily practices of austerity in a Jain community that dominates a large portion of the international emerald trade. The material characteristics of emeralds make them refractory towards formal markets. These gemstones are impossible to value with consistency. Prices fluctuate wildly as the stones move from hand to hand and from continent to continent. Among the Jains, bonds forged by kinship and religious observance guarantee international, multi-million dollar contracts in the absence of any legal enforcement mechanism. Religious ethics ensure that people deal with each other in good faith and make good on their debts. These ethics are inculcated in part through bodily practices. The bodily practices of Jain traders also include jewelers’ sensual relationships with the stones themselves. The pursuit of salvation through austerity and the pursuit of profit through luxury are conjoined in a community dedicated to making sense of the inscrutable mineral characteristics of emeralds.
The use of presumptions and duties in sustainable development equations
Authors: Ross Andrea , Zasinaite Agne
Published: June 2017

Sustainable development provides a forum for sometimes complementary, but often conflicting, factors to be raised and the best solution found. As new challenges present themselves, they need to fit into our decision making processes. This article examines the use of presumptions and duties in what it refers to as ‘sustainable development equations’. More specifically, it asks whether the regulatory regime for managing change in listed buildings is promoting and delivering the sustainable management of listed buildings. It does so by examining how various presumptions and duties are used and prioritized by the courts, by Historic Environment Scotland (HES) and local authorities in their policies on managing change and then in practice, in listed building consent decisions relating to solar panels. The paper concludes presumptions and duties are useful tools for ensuring certain factors are brought into sustainable development equations and given the appropriate status. However, in the context of listed buildings, the current balance is not capable of delivering or encouraging their sustainable management. Presumptions and duties are most useful when expressly part of particular regulatory regimes and where the policy that supports these sustainable development equations is sufficiently detailed to provide both regulators and regulated with reassurance and certainty.
Data collection, counterterrorism and the right to privacy
Authors: Taylor Isaac
Published: June 2017

Governments around the world collect huge amounts of personal data from their citizens (as well as foreigners) for counterterrorist purposes. While mining this data has arguably increased the security of populations, the practices through which these data are currently collected in many countries have been criticised for violating individuals’ rights to privacy. Yet it is not clear what a permissible data collection regime (if one is possible) would look like and thus also how we could reform existing regimes to make them morally acceptable. This article explores a number of ways in which we might justify a data collection regime to those affected in spite of the setbacks to their privacy. In contrast to existing justifications, I argue that individuals can be asked to surrender their personal data as a requirement of reciprocity in a cooperative system in which they gain security from others doing likewise. Relying on this justification, though, has significant implications for how we should reform existing data collection regimes. In particular, more stringent limits will need to be placed on the forms which these regimes can legitimately take.
Effects of a mindfulness-based intervention on symptoms and signs in chronic heart failure: A feasibility study
Authors: Norman Jonna , Fu Michael , Ekman Inger , Björck Lena , Falk Kristin
Published: June 2017

Aims:

Despite treatment recommended by guidelines, many patients with chronic heart failure remain symptomatic. Evidence is accumulating that mindfulness-based interventions (MBIs) have beneficial psychological and physiological effects. The aim of this study was to explore the feasibility of MBI on symptoms and signs in patients with chronic heart failure in outpatient clinical settings.

Methods:

A prospective feasibility study. Fifty stable but symptomatic patients with chronic heart failure, despite optimized guideline-recommended treatment, were enrolled at baseline. In total, 40 participants (median age 76 years; New York Heart Association (NYHA) classification II−III) adhered to the study. Most patients ( n=17) were randomized into MBI, a structured eight-week mindfulness-based educational and training programme, or controls with usual care ( n=16). Primary outcome was self-reported fatigue on the Fatigue severity scale. Secondary outcomes were self-reported sleep quality, unsteadiness/dizziness, NYHA functional classification, walking distance in the six-minute walk test, and heart and respiratory rates. The Mann–Whitney U test was used to analyse median sum changes from baseline to follow-up (week 10±1).

Results:

Compared with usual care (zero change), MBI significantly reduced the self-reported impact of fatigue (effect size −8.0; p=0.0165), symptoms of unsteadiness/dizziness ( p=0.0390) and breathlessness/tiredness related to physical functioning (NYHA class) ( p=0.0087). No adverse effects were found.

Conclusions:

In stable but symptomatic outpatients with chronic heart failure, MBI alleviated self-reported symptoms in addition to conventional treatment. The sample size is small and further studies are needed, but findings support the role of MBI as a feasible complementary option, both clinically and as home-based treatment, which might contribute to reduction of the symptom burden in patients diagnosed with chronic heart failure.
Planting the Seeds of Change: Growing Lifestyle Medicine Interest Groups With the Donald A. Pegg Award
Authors: Stiegmann Regan A. , Abreu Alyssa , Gardner James E. , Hipple Jessie M. , Poling P. Elainee , Frates Elizabeth Pegg
Published: June 2017

The practice of Lifestyle medicine (LM) focuses on helping patients make healthy choices to prevent and treat disease. While such interventions are considered first-line treatment for many diseases, many medical schools have not yet been able to include lifestyle medicine classes in the core curriculum but most are able to offer a parallel curriculum that does not interfere with the schedule of core classes. Lifestyle Medicine Interest Groups (LMIGs) are being created around the country and around the globe. Many students and faculty members are interested in starting and sustaining a LMIG at their schools, but some do not have enough funding or they lack the framework that provides structure to their efforts. To address this situation, the American College of Lifestyle Medicine (ACLM) has encouraged the development of LMIGs, which are student-run organizations that provide a parallel curriculum in LM. To support and strengthen this effort, the ACLM initiated the Donald A. Pegg award to fund four allied health students in founding and augmenting their institution’s LMIGs.

The 2016 inaugural winners were James Gardner, P. Elainee Poling, Alyssa Abreu, and Jessie M. Hipple. Their LMIG activities have included events such as nutrition and cooking classes, exercise prescription seminars, group fitness sessions, and patient lifestyle counseling in various clinical settings. Pearls of wisdom for building successful LMIGs include cultivating strong faculty mentorship, marketing the personal benefits to students who attend activities, and collaborating with other student groups.
The role of special education teachers in facilitating peer relationships among students with mild intellectual disabilities in lower secondary school
Authors: Sigstad Hanne Marie Høybråten
Published: June 2017

Background:

Students with intellectual disabilities may lack sufficiently developed skills to initiate qualitatively good social interactions; thus, they might be in need of assistance. This study examined special education teachers’ role in facilitating peer relationships among students with mild intellectual disabilities in a mainstream school context.

Materials and Methods:

The study was based on qualitative semi-structured interviews with nine special education teachers who belong to special education groups in lower secondary schools. A thematic structural analysis was used to identify themes.

Results:

The teachers made substantial efforts to promote social competence and ensure optimal conditions to foster peer interactions. An “academic-oriented” education, divergent attitudes, challenges in teacher collaboration, and organizational constraints may be barriers.

Conclusions:

In a mainstream school, the role of special education teachers appears to be dependent on the basic values of the school management in terms of real opportunities to foster peer relationships among students with mild intellectual disabilities.