bluuebunny/crossref_metadata_2025 huggingface dataset:
Storage of Human Fetal Retina in Optisol Prior to Subretinal Transplantation
Authors: Diloreto David A. , Del Cerro Coca , Lazar Eliot S. , Cox Christopher , Del Cerro Manuel
Published: June 2017

We have previously reported that Optisol, a commercially available medium (Chiron Vision, Irvine, CA), is a favorable fluid for the storage of neural retina. We now characterize the cell survival and histologic integrity of the tissue following storage and then transplantation. Neural retinas from human fetal eyes obtained within 1 h of pregnancy termination were stored in Optisol-GS medium (containing TC-199 and MEM medium, 2.5% chondroitin sulfate, 1% dextran, HEPES buffer, gentamicin sulfate, 0.1 mmol/L nonessential amino acids, sodium bicarbonate, 1 mmol/L sodium pyruvate, and additional antioxidants), at 4°C for the following time periods: 0, 1, 2, and 7 days, and then tested for viability. Retinas stored for 2 and 7 days were mechanically dissociated and grafted into the eyes of light-damaged Fischer 344 rats (17 animals, 28 eyes). Non-transplanted eyes received injections of vehicle only (5 animals, 6 eyes). All animals were immunosuppressed daily with cyclosporine (10 mg/kg). All experiments were conducted in strict accordance with institutional, federal, and ARVO guidelines. Cell viability averaged 94.8%, 90.2%, 83.2%, and 76.8% at 0, 1, 2, and 7 days storage, respectively. Light microscopy demonstrated that the fetal retina was preserved without evident changes for up to 48 h. Up to 7 days there was a good to very good preservation of the cells in the outer neuroblastic layer. One month after transplantation subretinal grafts of retinal tissue stored for 2 and 7 days in Optisol showed good integration with host retina and initial photoreceptor differentiation. Optisol is a widely used medium for the preservation of human corneas prior to transplantation. Our results show that this medium is also highly suitable for preserving human fetal retinas prior to transplantation.
Rescue of Motoneurons from Axotomy-Induced Cell Death by Polymer Encapsulated Cells Genetically Engineered to Release CNTF
Authors: Tan S.A. , Déglon N. , Zurn A.D. , Baetge E.E. , Bamber B. , Kato A.C. , Aebischer P.
Published: June 2017

The neurodegenerative disease amyotrophic lateral sclerosis (ALS) results from the progressive loss of motoneurons, leading to death in a few years. Ciliary neurotrophic factor (CNTF), which decreases naturally occurring and axotomy-induced cell death, may result in slowing of motoneuron loss and has been evaluated as a treatment for ALS. Effective administration of this protein to motoneurons may be hampered by the exceedingly short half-life of CNTF, and the inability to deliver effective concentration into the central nervous system after systemic administration in vivo. The constitutive release of CNTF from genetically engineered cells may represent a solution to this delivery problem. In this work, baby hamster kidney (BHK) cells stably tranfected with a chimeric plasmid construct containing the gene for human or mouse CNTF were encapsulated in polymer fibers, which prevents immune rejection and allow long-term survival of the transplanted cells. In vitro bioassays show that the encapsulated transfected cells release bioactive CNTF. In vivo, systemic delivery of human and mouse CNTF from encapsulated cells was observed to rescue 26 and 27% more facial motoneurons, respectively, as compared to capsules containing parent BHK cells 1 wk postaxotomy in neonatal rats. With local application of CNTF on the nerve stump and by systemic delivery through repeated subcutaneous injections, 15 and 13% more rescue effects were observed. These data illustrate the potential of using encapsulated genetically engineered cells to continuously release CNTF to slow down motoneuron degeneration following axotomy and suggest that encapsulated cell delivery of neurotrophic factors may provide a general method for effective administration of therapeutic proteins for the treatment of neurodegenerative diseases.
Successful Treatment of Diabetes in Nod Mice with Advanced Disease by Islet Isografts following Immunoregulation with Linomide (Quinoline-3-Carboxamide)
Authors: Slavin Shimon , Weiss Lola , Wenlang Xia , Gross David J.
Published: June 2017

We have recently documented that oral Linomide (quinoline-3-carboxamide) prevents autoimmune insulitis, islet destruction, and diabetes in NOD mice treated at an early stage (5 wk of age) of the disease. In this report, we show that treatment of female NOD mice with advanced disease (age 23-24 wk) by syngeneic islet transplantation and oral Linomide administration results in prevention of graft insulitis and diabetes in the Linomide group up to 40 wk (diabetes at 40 wk: isograft recipients with Linomide n = 0 of 6; isograft recipients alone n = 5 of 6; p < 0.0001). The extent of protection from glucose intolerance by the combination of transplantation with Linomide was superior to that of Linomide alone [blood glucose (mean ± SD) 60 min post-i.p. injection of 1 g/kg body weight glucose: Linomide plus isograft 6.7 ± 1 mmol/L; Linomide alone 18.7 ± 6.3 mmol/L; p < 0.0001]. Thus, Linomide should be considered a potential immunoregulatory modality in patients undergoing pancreatic islet or whole organ transplantation.
Cyclosporine Does not Inhibit the Process of Revascularization of Pancreatic Islet Transplantation
Authors: Mendola José F. , Goity Cristian , Esmatjes Enric , Saenz Alex , Fernandez-Cruz Laureano , Gomis Ramon
Published: June 2017

The immunosuppressive drug cyclosporin-A (CsA) has been widely used to prevent pancreatic islet allograft rejection. Because it has been suggested that CsA may inhibit the process of revascularization of transplanted islets, the purpose of the study was to analyze by a double indirect immunofluorescence technique the revascularization process of isolated islets grafted in the liver and in the renal subcapsular space of rats treated with immunosuppressive doses of CsA. Lewis rats were grafted with either Lewis (isografts) or Wistar (allografts) pancreatic islets obtained by collagenase digestion. Rats were killed at different days after implantation and the liver and kidney bearing the grafted islets were snap frozen and immunohistochemically stained with a double immunofluorescence technique using a rabbit antifactor-VIII antiserum (which labels endothelial cells) and a guinea pig antiinsulin antibody. Islets implanted into nonimmunosuppressed hosts completed revascularization by days 3-7 after transplantation, as shown by the detection of endothelial cells within and surrounding the islets. The identical staining pattern of revascularization was observed in nonrejecting allografts as well as in isografts treated with CsA. We conclude that CsA did not inhibit the process of revascularization of rat islets after free transplantation. This finding is relevant for human islet transplantation, where CsA is currently employed to prevent kidney and islet allograft rejection. Copyright © 1997 Elsevier Science Inc.
Differential Dissection of the Rat E16 Ventral Mesencephalon and Survival and Reinnervation of the 6-Ohda-Lesioned Striatum by a Subset of Aldehyde Dehydrogenase-Positive th Neurons
Authors: Haque Nadia S.K. , Leblanc Celeste J. , Isacson Ole
Published: June 2017

The retinoic acid-generating enzyme, aldehyde dehydrogenase (AHD), is expressed in a subpopulation of dopaminergic neurons found in the substantia nigra. Using AHD and tyrosine hydroxylase (TH) as immunohistochemical markers, we determined whether differential dissection of the embryonic (E16) ventral mesencephalon (VM) into its lateral and medial portions contributed equally to the number of TH cells surviving transplantation, if grafted AHD/TH neurons reinnervate the host striatum according to their normal projection patterns, and examined the functional recovery caused by the implanted cells as assessed by amphetamine-induced rotation in a 6-OHDA-lesioned model of Parkinson's disease. The embryonic tissue was transplanted as solid pieces injected via a 20-gauge lumbar puncture needle into the center of the deafferented striatum. Groups received either one complete ventral mesencephalic piece (VM), two medial pieces of ventral mesencephalic tissue (MVM), or two lateral pieces of ventral mesencephalic tissue (LVM). Both VM and MVM groups showed a significant decrease in amphetamine-induced rotation over time and, there was no difference in the degree of reduction observed between the two groups. Histological evaluation of the transplants revealed a much larger total number of surviving TH cells in grafts from the VM and MVM groups compared to the LVM group. Surviving AHD/TH neurons were found in all groups. Whereas TH staining of the transplanted striatum displayed a halo of graft-derived fibers all around the transplant and integration of these fibers into the host neuropil, AHD staining showed a preferential reinnervation of the dorsolateral striatum corresponding to the normal projection pattern of AHD/TH neurons. In summary, selective dissection of the embryonic ventral mesencephalon is possible, functional recovery as assessed by amphetamineinduced rotation in animals transplanted with MVM is similar to that seen in animals grafted with VM, and AHD/TH neurons have a selective reinnervation pattern in the PD transplantation paradigm. These findings may have implications for the grafting of fetal mesencephalic tissue in PD patients.
Control of Insulin Secretion and Glucose Homeostasis in Exercising Diabetic Rats with Intrasplenic or Kidney Subcapsular Islet Grafts
Authors: Houwing Harmina , Hilbrands Luchiena G. , Van Suylichem Paul T.R. , Bruggink Jan E. , Steffens Anton B. , Strubbe Jan H.
Published: June 2017

This study was designed 1) to investigate mechanisms of insulin secretion during exercise after transplantation of islets in the spleen and under the kidney capsule, and 2) to compare these organs as transplantation site regarding an adequate portal or systemic delivery of insulin and glucose homeostasis during exercise. Diabetic rats were provided with 5 μL isogenic islet tissue in the spleen or under the kidney capsule, which results in normoglycemia, and were submitted to a swimming test Portal plasma insulin levels were higher than simultaneously sampled systemic insulin levels in the control and in the intrasplenic islet grafted group, but not in the kidney subcapsular islet-grafted group. Plasma portal and systemic insulin levels decreased, and glucose levels increased during exercise in all groups. The exercise-induced increase in levels of catecholamines was larger in systemic than in portal plasma, suggesting catecholamine extraction by the lungs or intestines. The experiments were repeated after removing of adrenal medulla, resulting in nondetectable or very low plasma adrenaline levels. Despite these low adrenaline levels, insulin levels decreased during exercise. The results indicate that 1) the exercise-induced reduction of insulin secretion is not mediated by circulating adrenaline, but is probably under control of the sympathetic nervous system, which could be the result of reinnervation of the transplanted islets. 2) Although a portal-systemic insulin gradient was absent in rats with kidney subcapsular islet grafts, the absence of a difference in glucose homeostasis during exercise between the sites revealed that all investigated sites are preferential to transplant islets.
Efficiency of Neural Differentiation of Mouse P19 Embryonal Carcinoma Cells is Dependent on the Seeding Density
Authors: Kitani Hiroshi , Ikeda Hidetoshi , Atsumi Tadao , Watanabe Rihito
Published: June 2017

Serum-free culture conditions for retinoic acid-induced neural differentiation of mouse P19 embryonal carcinoma cells were determined for future ex vivo retroviral gene transfer and brain transplantation studies. Neural differentiation of P19 cells was dependent on the seeding densities, and both neurons and astroglia differentiated efficiently at high seeding densities (2 × 104 and 5 × 104 cells/cm2) but not at low seeding density (1 × 104 cells/cm2). In addition, P19 cells cultured at 5 × 104 cells/cm2 showed neural differentiation whether or not they were infected with Friend leukemia virus FrC6-V, which inhibited neural differentiation at 2 × 104 cells/cm2. These results indicate that FrC6-V-infected P19 embryonal carcinoma cells should be seeded at high density to achieve efficient neural differentiation in vitro for ex vivo gene transfer with a FrC6-V–derived retroviral vector system.
Immunoregulation via Adhesion Molecules in Allogenic and Xenogenic Hepatocyte Transplantation to Nagase's Analbuminemic Rats
Authors: Horimoto Hitoshi , Nozawa Masumi , Kokudo Norihiro , Nakao Masatomo , Takahashi Shigeki , Miyasaka Masayuki , Seino Kenichiro , Yagita Hideo , Okumura Ko
Published: June 2017

We investigated the effects of monoclonal antibodies (mAbs) against lymphocyte function-associated antigen-1 (LFA-1) and intercellular adhesion molecule-1 (ICAM-1) on intrasplenic allogenic and xenogenic hepatocyte transplantation (HCTx) to analbuminemic rats. Ten to 12-wk-old male Nagase's analbuminemic rats (RT1l) were used as recipients, Wistar/Shi rats (RTlk) were used as donors for allografts and BALB/C mice were used as donors for xenografts. The experimental groups were as follows: group 1, allo-HCTx (n = 7); group 2, allo-HCTx + antirat ICAM-1/antirat LFA-1 mAbs (1.0 mg/kg/day, for 7 days, respectively) (n = 6); group 3, xeno-HCTx (n = 5); group 4, xeno-HCTx + mAbs (antimouse LFA-1/antirat ICAM-1) (n = 5). group 5, xeno-HCTx + mAbs (antirat LFA-1/antimouse ICAM-1) (n = 5). Serum rat albumin levels were measured in groups 1 and 2, and serum mouse albumin levels were measured in groups 3, 4, and 5, as indicators of the function of grafted hepatocytes. In allotransplantation groups, the serum rat albumin levels in the mAbs-treated group were significantly higher than those in the control group for 6 wk after transplantation. In xenotransplantation groups, no increase in the serum mouse albumin levels was detected in any group.
Electrostatic Endothelial Cell Seeding Technique for Small-Diameter (<6 MM) Vascular Prostheses: Feasibility Testing
Authors: Bowlin Gary L. , Rittgers Stanley E.
Published: June 2017

Multiple studies have indicated the importance of surface charge in the adhesion of multiple cardiovascular cell lines including platelets and endothelial cells on the substrate materials (1,4,7-10,12-15). It is the purpose of this article to report a feasibility study conducted using an electrostatic endothelial cell seeding technique. The feasibility study was conducted using human umbilical vein endothelial cells (HUVEC), a static pool apparatus, a voltage source, and a parallel plate capacitor. The HUVEC concentration and seeding times were constant at 560,000 HUVEC/ml and 30 min, respectively. Scanning electron microscopy examination of the endothelial cell adhesion indicated that an induced temporary positive surface charge on e-PTFE graft material enhances the number and the maturation (flattening) of HUVECs adhered. The results indicated that the total number of endothelial cells adhered (70.9 mm2) was increased from 9198 ± 1194 HUVECs on the control (no induced surface charge) e-PTFE to 22,482 ± 4814 HUVECs (2.4 × control) on the maximum induced positive surface charge. The total number of cells in the flattened phase of adhesion increased from 837 ± 275 to 6785 ± 1012 HUVECs (8.1 ×) under identical conditions. Thus, the results of the feasibility study support the premise that electrostatic interaction is an important factor in both the endothelial cell adhesion and spreading processes and suggest that the electrostatic seeding technique may lead to an increased patency of small diameter (<6 mm) vascular prostheses.
Generation of Primary Peptide-Specific Cd8+Cytotoxic T-lymphocytes in Vitro using allogeneic dendritic cells
Authors: Peshwa Madhusudan V. , Bemke Claudia , Dupuis Marc , Kundu Smriti K. , Engleman Edgar G. , Merigan Thomas C. , Van Schooten Wim C. A.
Published: June 2017

Dendritic ceils (DC) are potent antigen-presenting cells (APC) capable of inducing strong T-cell–mediated immunity. Infusion of lymphoma-specific antigen-loaded autologous DC has been demonstrated to result in the generation of antigen-specific immunity and reduction in tumor burden in B-cell lymphoma patients. Cellular immunotherapy employing antigen-loaded DC could have a potential therapeutic impact in tumors and viral infections, including HIV infection. However, DC in HIV-infected individuals and breast cancer patients are believed to be functionally defective. Therefore, the potential of using allogeneic DC offers significant implications for DC immunotherapy in AIDS and immunocompromised cancer patients. To explore the potential of allogeneic DC therapy in vivo, we tested the ability of allogeneic DC to generate primary peptide-specific CD8+cytotoxic T-lymphocyte (CTL) responses in vitro. Our results indicate that DC from HLA class I-matched individuals elicit primary immune responses in vitro using viral peptides as naive antigens. A primary peptide-specific immune response could also be detected even when only one HLA allele (HLA-A*0201) was matched between the allogeneic DC and T-lymphocytes. The ability to generate primary peptide-specific responses in vitro is strongly indicative of the in vivo therapeutic potential of allogeneic DC.
Tissue Engineered Skeletal Muscle: Preparation of Highly Dense, Highly Oriented Hybrid Muscular Tissues
Authors: Okano Takahisa , Matsuda Takehisa
Published: June 2017

We prepared highly dense, highly oriented hybrid muscular tissues that are composed of C2C12 cells (skeletal muscle myoblast cell line) and type I collagen. A cold mixture of C2C12 cells suspended in DMEM and type I collagen solution was poured into capillary tube molds of two different sizes (inner diameters; 0.90 and 0.53 mm, respectively). One end of each mold was sealed. Upon centrifugation (1000 rpm, 5 min) and subsequent thermal gelation, a rod-shaped gel was obtained. It was cultured in an agarose gel-coated dish for 7 days (first for 3 days in a growth medium and then for 4 days in a differentiation medium), during which time it shrank to become a highly dense tissue. Small-diameter rod-shaped, highly dense cellular assemblages with multinucleated myotubes were formed and only few necrotic cells at the core of the tissue were observed. On the other hand, a ring-shaped tissue prepared using a specially devised agarose gel mold was subjected to cyclic stretching at 60 rpm, resulting in the formation of a highly dense, highly oriented hybrid muscular tissue in which both densely accumulated cells and collagen fiber bundles tended to be aligned in the direction of stretching. The hybrid muscular tissues that were prepared using via sequential procedures of a centrifugal cell packing method and a mechanical stress-loading method became closer to native muscular tissues in terms of cell density and orientation.
Intrasplenic Liver Parenchymal Cells in Conjunction with Low-Dose Rapamycin and Cyclosporine Induce a Unique and Specific Prolongation of Rat Cardiac and Small Bowel Allograft Survival
Authors: Boyle Michael J. , Baghdassarian Vano , Stepkowski Stanislaw M. , Dumble Lynette J. , Kahan Barry D.
Published: June 2017

These experiments investigated the immunosuppressive properties of liver tissue. Brown Norway (BN; RT1n) rat heart allografts survived in untreated control Wistar Furth (WFu; RTlu) rat recipients for 6.2 ± 1.5 days, while allografts in animals that received rapamycin (RAPA) 0.0075 mg/kg/day and cyclosporine (CsA) 0.375 mg/kg/day delivered for 14 days by continuous intravenous infusion (civi) using osmotic pumps in conjunction with intrasplenic (i.s.) saline survived to 18.4 ± 1.3 days. i.s. addition of 3 M-KCl extracted BN hepatic antigen or unpurified BN hepatocytes (liver parenchymal cells—5 × 107/kg), which exhibited a 4.8% class II antigen expression, and which alone failed to prolong allograft survival (MST = 6.0 ± 1.4 days), increased heart allograft survival to 25.3 ± 2.3 and 27.2 ± 1.9 days, respectively (p < 0.01). Hepatocyte purification using Dynabeads and Percoll reduced class II expression to 0.9% and increased allograft survival to 32.8 ± 1.6 days (p < 0.01). In contrast, the effect of 5 × 108/kg BN erythrocytes, exhibiting only 0.1% class II expression, was much less (23.8 ± 1.9 days). Administration i.s. of BN splenocytes or nonparenchymal liver cells, demonstrated by flow cytometry to exhibit a 47.3 or 55.1% expression of class II antigen, respectively, failed to induce any significant increase in allograft survival (18.4 ± 4.6 and 19.4 ± 0.5 days, respectively). Survival of BN rat small bowel allografts was increased in Lewis (LEW; RTl1) rat recipients treated with RAPA, CsA, and unfractionated BN hepatocytes from 10.2 ± 1.9 to 21.2 ± 1.5 days. Pretreatment with i.s. BN hepatocytes, 14 days prior to harvesting, reduced WFu lymphocyte responses to allogeneic stimulation with BN or ACI spleen cells by 75 and 70%, respectively. Addition of 1 × 105unpurified donor-specific BN or third-party Buffalo (BUF; RTlb) hepatocytes, but not supernatant, to the responder wells of MLCs resulted in a 61 and 40% suppression, respectively, of the WFu lymphocyte response induced by BN allogeneic stimulation. These findings suggest that while class I MHC expression has a significant role to play in exerting the immunosuppressive effects of hepatocytes, other influences more specific to liver may also prevail.
Response of Cultured Fetal and Adult Rat Hepatocytes to Growth Factors and Cyclosporine
Authors: Lilja Helene , Blanc Pierre , Demetriou Achilles A. , Rozga Jacek
Published: June 2017

Hepatocyte transplantation is a promising alternative to orthotopic liver transplantation in experimental animal models with genetic disorders of liver metabolism and liver failure. Fetal hepatocytes have several characteristics that make them potentially suitable as donor cells. In contrast to adult hepatocytes, fetal hepatocytes are thought to be highly proliferative, which may facilitate engraftment, expansion of transplanted cell population, and gene transfer requiring active DNA synthesis. The present study was undertaken to evaluate the proliferative capacity of fetal and adult rat hepatocytes under standardized culture conditions. Fetal (20 days of gestation) and adult hepatocytes were cultured in serum-free media at low densities and treated with growth factors. Proliferation was assessed by [3H]-thymidine incorporation and cell cycle analysis by flow cytometry. In nonstimulated cells, DNA synthesis at 4 h was about × 100 higher and after 10 days in culture ×20 higher in fetal compared to adult hepatocytes. When epidermal growth factor (EGF) was added, maximal DNA synthesis in fetal hepatocytes was seen at 48 h, whereas in adult hepatocytes at 72 h. For adult hepatocytes, the average increase compared to untreated cells was × 13.8 with EGF, ×18.5 with transforming growth factor alpha (TGF-α), and ×7.6 with hepatocyte growth factor (HGF). For fetal hepatocytes, the increase was twofold with either EGF, TGF-α or HGF. EGF-, TGF-α- and HGF-dependent DNA synthesis was inhibited by transfroming growth factor beta-1 (TGF-β1) in both fetal and adult hepatocyte cultures; this antiproliferative effect was significantly stronger in adult hepatocyte cultures. With cyclosporine, EGF-, TGF-α- and HGF-dependent DNA synthesis in fetal hepatocyte cultures decreased by 36–46%, whereas in adult hepatocytes by 19–27%. These results show that in contrast to adult hepatocytes, fetal hepatocytes have high spontaneous proliferative activity independently of growth factors and are relatively resistant to the inhibitory effect of TGF-β1. It was also found that cyclosporine suppresses proliferation of cultured fetal hepatocytes.
Hypothermic Storage of Periportal and Perivenous Rat Hepatocytes
Authors: Guibert Edgardo Elvio , Mediavilla María Gabriela , Mamprin María Eugenia , Rodríguez Joaquín Valentín
Published: June 2017

High yields of intact parenchymal cells are produced by the two-step Digitonin-collagenase perfusion of whole liver, and it has gained wide acceptance for biochemical and cellular analyses of zonal hepatocytes. The development reached by this methodology is in contrast to the time-limited use of the isolated cells unless those other methods, such as primary cultures, are employed. An alternative option to have cells ready to be used for several days, is the cold storage in University of Wisconsin solution as a preservation solution. This procedure is easy, not too expensive, and does not require specialized equipment. We study the competence of this system to maintain liver cells: mixed or total cells and cell-enriched fractions. We affirm viability of hepatocytes during hypothermic storage (UW-96h-4°C) by Trypan Blue exclusion, the capacity to retain cytoplasmic enzymes, metabolic competence to maintain total Glutathione content, and immunocytochemistry (gene detection). After 96 h of cold storage, mixed cells and cell-enriched fractions, were submitted to normothermic incubation (120 min, 37°C) and we check Trypan Blue exclusion, cytoplasmic enzyme release, and the capacity of cell populations to synthesize urea. The results show that it is possible to use, after several days of storage, mixed liver cells and cell-enriched fractions in metabolic and gene expression studies. This procedure allows us to reduce the number of experimental animals needed, to save experimental time and costs, and to facilitate further studies in vitro about the basis and consequences of metabolic heterogeneity of the liver cell plate.
In Vitro and in Vivo Grafting of Xeno Pig Fetal Liver Fragments Using Ultrafiltration Membrane
Authors: Kanai Nobuyuki , Morita Naokatsu , Munkhbat Batmunkh , Gansuvd Balgansuren , Hagihara Masao , Nagamachi Yukio , Tsuji Kimiyoshi
Published: June 2017

Transplantation of xeno fetal liver fragments (FLF) could be an alternative or supplementary therapy for acute and chronic liver failure not resolved by routine medical therapies. However, the xenografts themselves are rejected by the host immune system. To overcome these problems, immunoisolate capsules with various cutoff points, from 50,000 (YM30) to 500,000 (ZM500) were tested for their protective effects on FLF graft survival. In an in vitro study, the capsule with the smallest cutoff size (YM30) had an excellent protective effect on the grafts it contained, and showed the lowest GOT values in the culture supernatant and the normal histological structure. In an in vivo study using rats, the same capsule enabled a FLF graft to survive as long as 21 days, even with severe IgG deposition on and within the graft. In another in vivo study, which used beagle dog, however, it did not improve the natural course of survival of the graft, which had totally degenerated by day 7. In conclusion, 1) Immunocapsules, especially those with the smallest cutoff values, impeded the infiltration of the (xeno) humoral attacking factor, but the blocking effect was not complete, as shown by the immunoglobulin (IgG) deposit on the grafts they contained. 2) The FLFs with capsules survived longer than those without capsules—only in rats, not in beagles. This difference may be attributable to the difference of the extent of humoral or nutritional response to the xenografts.
Intraocular Transplantation of E1A-Immortalized Retinal Precursor Cells
Authors: Seigel Gail M. , Takahashi Masayo , Adamus Grazyna , Mcdaniel Thurma
Published: June 2017

The purpose of this study was to examine the effect of the ocular environment on the survival, tumorigenicity, and phenotypic marker expression of immortalized retinal precursor cells transplanted into immunocompetent adult and neonatal Sprague–Dawley rats. E1A-NR.3, a rat immortalized retinal precursor cell culture, was used as an inexhaustible source of experimental graft material. These cells were prelabeled with the fluorescent marker diI (1,1′-dioctadecyl-3,3,3′,3′-tetramethylindocarbocyanine perchlorate) and transplanted intravitreally (50,000 cells per μL) into 11 adult and 31 neonatal Sprague–Dawley rat eyes. At 1 mo posttransplant, animals were sacrificed and retinal tissue sections examined histologically for the presence of grafted cells, signs of tumor formation, and retinal phenotypic marker expression. No obvious signs of tumor formation or rejection were seen in a total of 42 eyes in the immunocompetent hosts. Our results indicate that E1A-NR.3 cells survive at least 1 month in vivo, and can migrate from the vitreous into neuroretinal cell layers. Subpopulations of surviving grafted cells were seen to express photoreceptor markers rhodopsin and recoverin comparably between in vitro and in vivo conditions. However, the number of cells immunoreactive for vimentin and E1A decreased significantly under in vivo conditions. This report represents the first experimental intravitreal transplantation of E1A-immortalized retinal precursor cells into adult and neonatal rats. The intraocular location and environment appears to affect phenotypic expression of surviving grafted cells, especially with respect to vimentin and E1A expression. The fact that E1A-NR.3 cells survived intraocularly at least 1 mo without tumor formation suggests that the cells may continue to be useful for further in vivo studies of experimental retinal transplantation, and effects of histological location on retinal cell phenotype and histogenesis in immunocompetent hosts. © 1998 Elsevier Science Inc.
The Potential Contribution of Rejection to Survival of Transplanted Human Islets
Authors: Swift S.M. , Clayton H.A. , London N.J.M. , James R.F.L.
Published: June 2017

Clinical islet transplantation is potentially the treatment of choice for people with type I diabetes. Rates of insulin independence in islet transplant recipients are disappointingly low, and the relative contribution of the rejection response compared with the loss of islet function is still unclear. We have compared the mixed lymphocyte islet coculture (MLIC) with the mixed lymphocyte acinar cell coculture (MLAC) and the mixed lymphocyte response (MLR) as in vitro models of allograft rejection to MHC and tissue-specific antigens expressed by human islets and acinar cells. The reduced number of MHC class II antigen-positive cells in islets and acinar tissue compared to those in the stimulator lymphocyte population of the MLR, correlated with a reduced proliferative response in the MLIC and MLAC. Enhancement of MHC class II antigen expression by islets using TNFα and IFNγ did not increase their stimulatory capacity in the islet cocultures, which may have been due to a corresponding absence of B7 expression. The lack of T cell proliferation to acinar cells despite cytokine-induced enhancement of MHC class II expression and detectable B7 expression appeared to be due to the inhibitory effect of exocrine enzymes on lymphocyte proliferation. In conclusion, we suggest that a rejection response to islets and acinar tissue is possible due to the accompanying MHC class II-positive cells and that, in this model, islet and acinar-specific antigens do not significantly contribute to that response. Acinar cells may have the potential to stimulate lymphocytes directly, but this was not evident by proliferation in the MLAC. Rejection appears to contribute to the low survival rate of human islet allografts, but it is unlikely that this is the sole explanation, and other factors should be considered. © 1998 Elsevier Science Inc.
Intranigral Transplantation of Fetal Substantia Nigra Allograft in the Hemiparkinsonian Rhesus Monkey
Authors: Starr Philip A. , Wichmann Thomas , Van Horne Craig , Bakay Roy A. E.
Published: June 2017

Current clinical protocols for fetal cell transplantation for Parkinson's disease (PD) have focused on restoring dopamine in the striatum. However, there are now a number of human transplant recipients who have had robust innervation of the striatum by dopaminergic grafts (documented by positron emission tomography or by autopsy), but only a partial improvement in parkinsonian motor signs. Thus, there is a need for improved transplant strategies. In animal models of PD, there is recent evidence that restoring dopamine in the substantia nigra, instead of or in addition to the striatum, may be important to correct abnormal motor behavior. This pilot study examined the morphological features and behavioral effects of fetal dopaminergic neuronal allografts placed into the substantia nigra of three 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-treated hemiparkinsonian rhesus monkeys. We show that grafts can survive in host substantia nigra. Characteristics of the graft–host interface were variable. In one animal, reinnervation of host substantia nigra was observed, and this animal showed behavioral improvement in a reach-and-retrieval task.
Neuron-Enriched Second Trimester Human Cultures: Growth Factor Response and in Vivo Graft Survival
Authors: White Michael G. , Hammond Robert R. , Sanders Virginia J. , Bonaroti Eugene A. , Mehta Anil P. , Wang Guoji , Wiley Clayton A. , Achim Cristian L.
Published: June 2017

Grafts of first trimester fetal tissue show limited survival and integration in the adult CNS. Alternative grafting strategies have been sought for treatment of neurodegenerative disease. We have developed cultures of human second trimester fetal tissues to study neuronal differentiation. Grafted into the SCID mouse striatum, aggregates of these cultures formed neuron-rich xenografts for at least 8 months. We examined the influence of various neurotrophic factors, including basic fibroblast growth factor (bFGF), brain-derived neurotrophic factor (BDNF), transforming growth factor-beta 1 (TGF-β1), and hepatocyte growth factor (HGF), on the growth and differentiation of neuronal and glial cell populations. BDNF promoted the survival and differentiation of second trimester neurons whereas bFGF exhibited a strong proliferative effect on precursors and the astroglial population. Our data suggest that second trimester human fetal cultures contain neuroprogenitor cells that can be directed to the neuronal lineage. This process may be amplified by treatment with BDNF, which we hypothesize could improve the long-term in vivo survival of neuron-enriched grafts.
A New Bioartificial Liver Using Porcine Hepatocyte Spheroids in High-Cell-Density Suspension Perfusion Culture: In Vitro Performance in Synthesized Culture Medium and in 100% Human Plasma
Authors: Sakai Yasuyuki , Naruse Katsutoshi , Nagashima Ikuo , Muto Tetsuichiro , Suzuki Motoyuki
Published: June 2017

A prototype of a bioartificial liver (BAL) based on suspension perfusion culture of porcine hepatocyte spheroids was developed at 150 ml scale. About 2% (4 × 109 cells) of whole human liver cells was immobilized. The cell density in the bioreactor was 2.7 × 107 cells/ml, which was almost comparable to that of presently developed packed-bed-type BALs. The bioreactor was perfused with culture medium while retaining spheroids. This was done using a rotating stainless filter (pore size 50 μm). In vitro 8-h perfusion experiments utilizing both synthesized culture medium and 100% human plasma demonstrated the spheroids in the bioreactor had almost the same functions on a unit/cell basis as those in small-scale rotational culture. This indicated that the functional deterioration often associated with scaling up had been minimized. Rapid spheroid aggregation and dysfunction in specific human plasma pool must be eliminated before clinical application, although this phenomenon seemed to be inherent to porcine hepatocyte-based BALs. This prototype shows promise in meeting present clinical demands by achieving maximal metabolic activities even in the short term.
Long-Term Treatment of Streptozotocin-Induced Diabetes by Continuous Insulin Minipump in the Syrian Hamster
Authors: Fisher William E.
Published: June 2017

It is difficult to normalize plasma glucose for a prolonged period of time by SC injection in experimental animals. The goal of this study was to determine the feasibility and the dosage of insulin needed to maintain 24-h normoglycemia in streptozotocin-diabetic Syrian hamsters with a SC-implanted osmotic minipump. The pumps, which release insulin at a constant rate, were replaced every 14 days with fresh pumps for as long as 52 days. A high insulin dose (1 U/kg/h) was required to normalize plasma glucose and fatty acid concentrations, water and food consumption, urine output, and body weight.
Xenografting Human T2 Sympathetic Ganglion from Hyperhidrotic Patients Provides Short-Term Restoration of Catecholaminergic Functions in Hemiparkinsonian Athymic Rats
Authors: Liu D. M. , Lin S. Z. , Wang S. D. , Wu M. Y. , Wang Yun
Published: June 2017

Previous studies have suggested that allografting peripheral sympathetic ganglia, such as superior cervical ganglia, partially relieves clinical or behavioral deficits in parkinsonian patients and animals. However, removal of these ganglia can cause Horner's syndrome, which limits the utilization of this approach. Hyperhidrosis, a disease of excessive sweating, is commonly seen in young Orientals. Treatment of hyperhidrosis often involves surgical removal of the second thoracic sympathetic ganglia (T2G), which contain catecholaminergic neurons. The purpose of our study was to investigate behavioral responses and tyrosine hydroxylase (TH) immunoreactivity in hemiparkinsonian rats at different time points after transplantation of human T2G from hyperhidrotic patients. Athymic Fisher 344 rats were injected unilaterally with 6-hydroxydopamine into the medial forebrain bundle to destroy the nigrostriatal dopaminergic (DA) pathway. The effectiveness of lesions was tested by measuring methamphetamine (MA)-induced rotations. These unilaterally lesioned rats were later transplanted with T2G or T2 fiber tract (T2F) obtained from adult hyperhidrotic patients. Animals grafted with T2G showed a reduction in MA-induced rotation by 2 weeks; however, rotation returned to the pregrafting levels by 3 months. Animals receiving T2F grafts did not show any reduction of rotation over a 3-month period. Animals were later sacrificed for TH immunostaining at different time points. Tyrosine hydroxylase-positive [TH(+)] cell bodies and fibers were found in the lesioned striatum 2–4 weeks after T2G grafting, suggesting the survival of transplants. Two to 3 months after grafting, TH(+) fibers were still found in almost all the recipients. However, TH(+) cell bodies were found in only three of seven rats studied. Animals receiving T2F grafting did not show any TH immunoreactivity in the lesioned striatum over the 3-month period. These data indicate that T2G transplants from adult hyperhidrotic patients can survive and provide transient normalization of the motor behavior in the hemiparkinsonian athymic rats. Because of the short-term improvement in behavior after grafting, the use of T2G in human trials should be cautious at the present time. Further laboratory research is required.
Subcutaneous Transplantation of Bovine and Human Adrenocortical Cells in Collagen Gel in scid Mice
Authors: Popnikolov Nikolay K. , Hornsby Peter J.
Published: June 2017

Adrenocortical cells of bovine origin and of adult and fetal human origin were transplanted subcutaneously (SC) in scid mice after being embedded in collagen gel. In this site the cells survived, became vascularized by invasion of host endothelial cells, and secreted steroids into the circulation. The animals' own adrenal glands were removed at the time of cell transplantation. Steroids secreted by the transplants replaced the essential functions of the animals' own adrenal glands. Adrenalectomized animals without transplanted cells died after several days, but most animals with transplanted bovine or adult human adrenocortical cells survived; fewer animals survived with transplanted fetal human adrenocortical cells. The histology of the tissues formed from transplanted cells resembled that of the normal adrenal cortex. A few proliferating cells were observed in tissue from bovine or adult human cells; there was a greater percentage of dividing cells in tissue derived from fetal cells. Subcutaneous transplantation of bovine or human primary adrenocortical cells in collagen provides a model for the study of the physiology, cell biology, and molecular biology of adrenocortical cells in a three-dimensional vascularized tissue structure in a host animal.
On demons and dreamers: Violence, silence and the politics of impunity in the Brazilian Truth Commission
Authors: Furtado Henrique
Published: June 2017

Measures towards post-conflict or post-authoritarian justice have historically relied on the merging of the concepts of silence, violence and impunity in order to create a single promise of justice. Scholars and practitioners in the field usually defend a trifold agenda of breaking the silence about violations of human rights, denouncing systematic violence in the past and fighting impunity as the only way of ensuring that violence never happens again. This trope was mobilized in Brazil in 2014, when the report of the country’s National Truth Commission (CNV) was released. However, in the Brazilian case, truth-seeking also produced its own form of ‘silence’. Whereas the CNV commendably denounced 377 perpetrators as the ‘demons’ responsible for implementing a state of terror during the last dictatorship (1964–1985), it also created a depoliticized and victimized idea of leftist militants as mere dreamers who fought for liberty and democracy in the past. By representing leftist militants as freedom fighters, the CNV silenced their fundamental ideas (and actions) regarding the concept of revolutionary violence and its radical programme of structural change. In this article, I provide an explanation that connects the CNV’s ‘silencing’ of this political project to the unreflective merging between the concepts of silence, violence and impunity in the literature. Via a narrative analysis of the CNV’s report and a critique of transitional justice debates, I argue that the silence on the political project of the radical left in Brazil echoes transitional justice’s silence about the complexities of violence in general.
‘I don’t think they realise how good we are’: Innovation, inclusion and exclusion in women’s Olympic boxing
Authors: Tjønndal Anne
Published: June 2017

Marginalisation and exclusion of women in elite boxing has emerged as a substantial international problem, threatening women’s democratic right to equal participation in sport. Since the London 2012 Olympic Games women’s boxing has been an Olympic event. However, only 3 of the 10 weight categories were included in the Olympic programme for women. Today, male boxers compete in 10 out of 10 possible weight categories. While female boxers have gained accessed to a previously closed realm, discrimination, exclusion and unequal treatment of female athletes remains a widespread problem in international elite boxing. This article explores how the introduction of women’s boxing into the Olympic Games involves practices of inclusion and exclusion in elite boxing. In particular, this analysis focuses on the following two research questions: (a) how practices of social exclusion and inclusion affect the involvement and engagement of women in boxing; and (b) how, and in what ways, female boxers have experienced recognition and acceptance in elite boxing after the introduction of women’s boxing into the Olympic Games. Using a qualitative approach including document analysis of official press releases from the International Olympic Committee (IOC) and the International Boxing Association (AIBA), as well as interviews with athletes and coaches, this study reveals that although women’s boxing has gained access to the Olympics, the structural and cultural changes that comprise the social inclusion of women in boxing are yet to be implemented.
Acute-phase proteins in pregnant goats: a longitudinal study
Authors: Czopowicz Michał , Szaluś-Jordanow Olga , Mickiewicz Marcin , Witkowski Lucjan , Markowska-Daniel Iwona , Stefaniak Tadeusz , Reczyńska Daria , Bagnicka Emilia , Kaba Jarosław
Published: June 2017

We documented changes in serum amyloid A (SAA) and haptoglobin (Hp) concentration in goats during pregnancy, as measured by competitive ELISAs. Fifteen does (pregnant group) and 20 castrated males (control group) were enrolled in the study. Blood samples were collected on the same day from all 35 goats, 7 times throughout the study period: at mating, then once every month, during the week preceding kidding, and 1 mo after kidding. Baseline SAA and Hp concentrations at mating were identical in the 2 groups. In the pregnant group, SAA concentration rose significantly in the second month and remained elevated until the end of pregnancy, with the peak concentration at kidding. In the control group, SAA concentration remained unchanged compared to the baseline concentration throughout the study. SAA concentration was significantly higher in the pregnant than control group only at the end of the fourth month of pregnancy and at kidding. Hp concentration did not change during pregnancy or between groups. SAA concentration at kidding was affected only by parity of does – it was highest in does in the third and fourth pregnancy and gradually lower in older does.
The Development of Leader Character Through Crucible Moments
Authors: Byrne Alyson , Crossan Mary , Seijts Gerard
Published: June 2017

Business schools strive to develop leadership excellence in their students. In this essay, we suggest that educators should find ways to help students develop and deepen leader character, a fundamental component of exemplary leadership. Frequently, business school students have preconceived ideas of leadership, often neglecting leader character. We argue that educators can and should teach students that leader character is pivotal to leadership excellence and that they should actively develop students’ leader character. The foundational learning theories of Piaget and Kolb provide a useful framework to help achieve the development of leader character. We propose that leader character development arises from using accommodation learning strategies of crucible experiences, paired with assimilation learning methods of critical reflection, and further developed through equilibrium learning strategies where students can incorporate new information and work toward their personal character growth. While numerous teaching approaches can be used, we describe an experiential course, codesigned with members of the Canadian Forces, using these learning strategies to foster leader character development.
State Transportation Planning: Linking Quantifiable Performance Measures and Infrastructure Condition Outcomes
Authors: Yusuf Juita-Elena (Wie) , Jordan Meagan M.
Published: June 2017

This study focuses on whether state Departments of Transportations (DOTs) with quantifiable performance targets have better road repair and maintenance outcomes than DOTs without quantifiable performance targets. We also examine state DOT use of quantifiable pavement condition targets to examine differences across states in terms of agency and governance/legislative factors. While the results are mixed, they suggest that state agency capacity may be related to the use of concrete and specific performance measures. Furthermore, these concrete and specific performance measures may be related to the use of external accountability bodies and citizen participation. This suggests that quantifiable performance targets may be a useful tool for decision making, planning, and resource allocation in pursuit of desired performance outcomes.