bluuebunny/crossref_metadata_2025 huggingface dataset:
Cystometrogram: the low-cost method for rural areas and its benefits
Authors: Gnanaraj J
Published: June 2017

Cystometrography (CMG) is a means of studying bladder pressure. It is a very useful diagnostic tool in patients with lower urinary tract symptoms for which a simple cystoscopy will not offer sufficient information to form a diagnosis. Of the 8893 patients who underwent screening for urological conditions in rural northeast India during 2010–2014, 280 with lower urinary tract symptoms were investigated with a combination of cystoscopy and CMG. By corresponding CMG diagnosis and treatment, we could examine patients’ overall satisfaction with both the procedure and the treatment. We describe a low-cost method of CMG and our results using this method in rural areas of India.
Relationships between structural and acoustic properties of maternal talk and children’s early word recognition
Authors: Suttora Chiara , Salerni Nicoletta , Zanchi Paola , Zampini Laura , Spinelli Maria , Fasolo Mirco
Published: June 2017

This study aimed to investigate specific associations between structural and acoustic characteristics of infant-directed (ID) speech and word recognition. Thirty Italian-acquiring children and their mothers were tested when the children were 1;3. Children’s word recognition was measured with the looking-while-listening task. Maternal ID speech was recorded during a mother–child interaction session and analyzed in terms of amount of speech, lexical and syntactic complexity, positional salience of nouns and verbs, high pitch and variation, and temporal characteristics. The analyses revealed that final syllable length positively predicts children’s accuracy in word recognition whereas the use of verbs in the utterance-final position has an adverse effect on children’s performance. Several of the expected associations between ID speech features and children’s word recognition skills, however, were not significant. Taken together, these findings suggest that only specific structural and acoustic properties of ID speech can facilitate word recognition in children, thereby fostering their ability to extrapolate sound patterns from the stream and map them with their referents.
Vocabulary growth in Armenian–English bilingual kindergarteners
Authors: Hovsepian Alice
Published: June 2017

Four-year-old ( n = 20) and five-year-old ( n = 22) bilingual children were tested twice in six months on Armenian (minority language) and English (majority language) picture identification and picture naming tasks to examine receptive and expressive vocabulary growth in both languages. Parental education, Armenian/English language exposure, and nonverbal cognitive ability were also measured as potential predictors of vocabulary. Children showed growth over time in all four vocabulary tasks. However, less growth was observed in Armenian expressive task than in others, which indicated a preference to use English. Armenian language exposure was the unique predictor of Armenian picture identification and picture naming, whereas English language exposure was the unique predictor of English picture identification.
The Use of an Autonomous Pedagogical Agent and Automatic Speech Recognition for Teaching Sight Words to Students With Autism Spectrum Disorder
Authors: Saadatzi Mohammad Nasser , Pennington Robert C. , Welch Karla C. , Graham James H. , Scott Renee E.
Published: June 2017

In the current study, we examined the effects of an instructional package comprised of an autonomous pedagogical agent, automatic speech recognition, and constant time delay during the instruction of reading sight words aloud to young adults with autism spectrum disorder. We used a concurrent multiple baseline across participants design to evaluate the efficacy of intervention and conducted post-treatment probes to assess maintenance and generalization. Our findings suggest that all three participants acquired and maintained new sight words and demonstrated generalized responding.
Compression treatment of pelvic congestion syndrome
Authors: Gavrilov SG , Karalkin AV , Turischeva OO
Published: June 2017

Aim

To study the influence of compression treatment on clinical manifestations and venous hemodynamics of the pelvis in patients with pelvic congestion syndrome.

Materials and methods

A prospective study of the various options and modes of compression treatment was carried out and included 74 patients with pelvic congestion syndrome in 2008–2015. The patients were divided into three groups. The first group consisted of 48 patients with symptoms of pelvic congestion syndrome and chronic pelvic pain. They used Class II compression shorts. In the second group, there were 14 patients with pelvic congestion syndrome, vulvar varicosities without pelvic pain. They used Class II compression shorts and stockings. In the third group, 12 women with pelvic congestion syndrome and chronic pelvic pain used only the Class II compression stockings. The treatment continued for 14 days. A clinical criterion was the change of severity of chronic pelvic pain. The evaluation of the treatments has been performed using radionuclide venography and emission computed tomography with labeled in vivo red blood cells.

Results

Group 1: The compression shorts had a positive effect on the disease in 81.3% of patients. Chronic pelvic pain decreased from 6.4 ± 1.6 to 1.2 ± 0.7 points. The coefficient of pelvic congestion syndrome (Cpcs) decreased from 1.73 ± 0.32 to 1.12 ± 0.27 (p < 0.05). In 18.8% of patients, no positive effect was observed. Group 2: The results of radionuclide venographyshowed accelerating outflow of blood from the lower limbs and reduction of insufficiency of perforating veins. Mean radionuclide transit time decreased in all patients in the tendon, muscle pump parts, popliteal vein and was respectively: 23.6 ± 2.2 s, 29.6 ± 3.4 s, 32.3 ± 4.2 s and after treatment 16.4 ± 3.1 s, 22.1 ± 2.5 s, 25.7 ± 1.9 s (p < 0.05). Group 3: The use of compression stockings class II on the clinical manifestations of pelvic congestion syndrome in the patients. Cpcs also remained unchanged.

Conclusion

Research has shown the efficiency of class II compression shorts in the treatment of patients with isolated extension of intrapelvic venous plexuses. Class II compression stockings do not have any impact on the clinical manifestations of pelvic congestion syndrome.
Are Polydrug Users More Physically and Verbally Aggressive? An Assessment of Aggression Among Mono- Versus Polydrug Users in a University Sample
Authors: Steele Jennifer L. , Peralta Robert L.
Published: June 2017

Early research has revealed that patterns of aggression and antisocial behavior are present among polydrug users. Often missing from this discourse is the examination of whether polydrug users are quantitatively different from monodrug users in their use of aggression. Theoretical perspectives are often centered on the psychopharmacological effects of substance use on behavior. Consideration of possible poly- versus monodrug use differences and their impact on aggression has not been investigated. Data from this study were derived from a sample of Midwestern university students ( N = 793). The relationship between violence, aggression, and concurrent polydrug use in the last year is assessed with a series of multivariate ordinary least squares (OLS) regression models. Results demonstrate that higher incidents of physical and verbal aggression are reported among polydrug users compared to monodrug users and abstainers. When analyses were broken down by polydrug users (those who engaged in alcohol/marijuana and alcohol/NMUPD [nonmedical use of prescription drugs] stimulants), polydrug users reported higher levels of physical aggression compared to monodrug users. Similarly, monodrug users reported higher levels of physical aggression compared to nonusers. This research extends our understanding of aggression among users from two different subcategories: polydrug users in comparison to those who only engage in one form of substance use. Scholars and practitioners who work with violent offenders should consider patterns of drug use behavior when addressing substance use–related aggression.
Comparing Grids With Vertical and Horizontal Item-by-Item Formats for PCs and Smartphones
Authors: Revilla Melanie , Couper Mick P.
Published: June 2017

Much research has been done comparing grids and item-by-item formats. However, the results are mixed, and more research is needed especially when a significant proportion of respondents answer using smartphones. In this study, we implemented an experiment with seven groups ( n = 1,476), varying the device used (PC or smartphone), the presentation of the questions (grids, item-by-item vertical, item-by-item horizontal), and, in the case of smartphones only, the visibility of the “next” button (always visible or only visible at the end of the page, after scrolling down). The survey was conducted by the Netquest online fieldwork company in Spain in 2016. We examined several outcomes for three sets of questions, which are related to respondent behavior (completion time, lost focus, answer changes, and screen orientation) and data quality (item missing data, nonsubstantive responses, instructional manipulation check failure, and nondifferentiation). The most striking difference found is for the placement of the next button in the smartphone item-by-item conditions: When the button is always visible, item missing data are substantially higher.
The Heterogeneity of Competitive Forces: The Impact of Competition for Resources on United Way Fundraising
Authors: Paarlberg Laurie E. , Hwang Hyunseok
Published: June 2017

How does competition for resources affect the fundraising performance of local United Way (UW) affiliates? Drawing upon population ecology, we hypothesize a nonlinear relationship between competition and organizational performance. Using a 21-year panel data set that includes UW campaign data, contributions to specialized fundraising organizations, and contributions to the general population of nonprofit organizations, we estimate a fixed effects regression model. We find that the effect of competition differs depending upon the degree of niche overlap. Contributions to organizations with greatest niche overlap have a negative effect on contributions to local UWs. Consistent with population ecology, other types of fundraising organizations that have less niche overlap show beneficial relationships at certain levels of giving. However, few communities reach the philanthropic level at which benefits of competition occur.
Development of an arthroscopically compatible polymer additive layer manufacture technique
Authors: Partridge Simon W , Benning Matthew J , German Matthew J , Dalgarno Kenneth W
Published: June 2017

This article describes a proof of concept study designed to evaluate the potential of an in vivo three-dimensional printing route to support minimally invasive repair of the musculoskeletal system. The study uses a photocurable material to additively manufacture in situ a model implant and demonstrates that this can be achieved effectively within a clinically relevant timescale. The approach has the potential to be applied with a wide range of light-curable materials and with development could be applied to create functionally gradient structures in vivo.
When weak governments confront inclusive trade unions: The politics of protecting labour market outsiders in the age of dualization
Authors: Rathgeb Philip
Published: June 2017

Some European welfare states protect outsiders from economic uncertainty better than others. Conventional theories attribute this either to the material interests of producer groups or the vote-seeking actions of political parties. By contrast, this article focuses on the interaction between trade unions and governments, drawing on empirical experience in Austria and Sweden. High levels of inclusiveness provide trade unions with a powerful interest in protecting outsiders. Governments, however, have moved towards dualistic policy choices, but when weakened by intra-coalitional divisions or a hung parliament, they retain a political incentive to negotiate political deals with trade unions, and this forces them to compensate outsiders for economic uncertainty.
Freshwater radiocarbon reservoir age in the lower Yellow River floodplain during the late Holocene
Authors: Yu Shi-Yong , Chen Xue-Xiang , Cheng Peng , Chen Shiyue , Hou Zhanfang
Published: June 2017

The radiocarbon concentration of the dissolved inorganic carbon (DIC) pool of most freshwater bodies such as lakes and rivers tends to be depleted with respect to that of the contemporaneous atmosphere, resulting in an apparent radiocarbon age of inorganic carbon therein known as freshwater reservoir effect. This phenomenon of radiocarbon age offset is usually site specific and varies over time. By radiocarbon dating pairs of twigs and gastropod shells preserved in an ancient lake, as well as gastropod shells in pre-bomb flooding sediments of known age, we are able to determine the freshwater radiocarbon reservoir effect in the lower Yellow River floodplain during the late Holocene. The regional freshwater radiocarbon reservoir age ranges from 1081 to 233 years with an average of 575 ± 317 years, slightly older than that obtained from Lake Dongping by radiocarbon dating of live submerged aquatic plants. Our results show that the freshwater radiocarbon reservoir age is much older than its marine counterpart of the Yellow Sea and Bohai Bay. A source inversion using dual carbon isotopes (i.e. 14C and 13C) and a linear mixing model revealed that the DIC in the surface water bodies was mainly from shallow groundwater and the Yellow River. Our results have broad relevance to the geological and archaeological chronologies based on the radiocarbon method for this area.
Increased Survival of Dopaminergic Neurons in Striatal Grafts of Fetal Ventral Mesencephalic Cells Exposed to Neurotrophin-3 or Glial Cell Line-Derived Neurotrophic Factor
Authors: Espejo Mónica , Cutillas Blanca , Arenas Ernest , Ambrosio Santiago
Published: June 2017

The transplantation of fetal mesencephalic cell suspensions into the brain striatal system is an emerging treatment for Parkinson's disease. However, one objection to this procedure is the relatively poor survival of implanted cells. The ability of neurotrophic factors to regulate developmental neuron survival and differentiation suggests they could be used to enhance the success of cerebral grafts. We studied the effects of neurotrophin-3 (NT-3) or glial cell line-derived neurotrophic factor (GDNF) on the survival of dopaminergic neurons from rat fetal ventral mesencephalic cells (FMCs) implanted into the rat striatum. Two conditions were tested: (a) incubation of FMCs in media containing NT-3 and GDNF, prior to grafting, and (b) co-grafting of FMCs with cells engineered to overexpress high levels of NT-3 or GDNF. One week after grafting into the rat striatum, the survival of TH+ neurons was significantly increased by pretreatment of ventral mesencephalic cells with NT-3 or GDNF. Similarly, co-graft of ventral mesencephalic cells with NT-3- or GDNF-overexpressing cells, but not the mock-transfected control cell line, increased the survival of graft-derived dopaminergic neurons. Interestingly, we also found that co-grafting of GDNF-overexpressing cells was less effective than NT-3 at improving the survival of fetal dopaminergic neurons in the grafts, and that only GDNF induced intense TH immunostaining in fibers and nerve endings of the host tissue surrounding the implant. Thus, our results suggest that NT-3, by strongly enhancing survival, and GDNF, by promoting both survival and sprouting, may improve the efficiency of fetal transplants in the treatment of Parkinson's disease.
Volume and Differentiation of Striatal Grafts in Rats: Relationship to the Number of Cells Implanted
Authors: Watts Colin , McNamara Iain R. , Dunnett Stephen B.
Published: June 2017

A growing body of evidence suggests that graft-mediated functional recovery in animal models of Huntington's disease is influenced by the morphology of the striatal grafts. Various parameters, including embryonic dissection, tissue preparation, and surgical delivery into the brain, have been investigated with the aim of increasing the proportion of the grafts comprising striatum-like tissue. While growing evidence suggests that implants derived from the selective dissection of the lateral ganglionic eminence (LGE) contain more striatal tissue, the relationship between the quantity of LGE tissue implanted and the striatum-like proportion of the resultant grafts has not been formally investigated. In this study the volume of striatum-like tissue within the grafts did not increase in a linear manner with increasing numbers of cells implanted. The proportion of the grafts that comprised the striatum-like patch compartment or P-zone remained constant after an initial rapid increase as the number of LGE cells implanted was increased. These results have important practical implications in determining the optimum number of LGE cells to implant and hence in the design of any surgical protocol for the clinical application of this technique.
Intrastriatal Ventral Mesencephalic Xenografts of Porcine Tissue in Rats: Immune Responses and Functional Effects
Authors: Larsson Lena C. , Czech Kimberly A. , Brundin Patrik , Widner Håkan
Published: June 2017

Transplantation of neural tissue from other species has the potential to improve function in patients with neurodegenerative disorders. We investigated the functional effects of embryonic porcine dopaminergic neurons transplanted in a rat model of Parkinson's disease and the immune responses to the grafts in immunosuppressed and nonimmunosuppressed hosts. Twenty-three rats with unilateral 6-hydroxydopamine lesions received dissociated, 27-day-old embryonic porcine ventral mesencephalic tissue in the right striatum. Eighteen rats received cyclosporine (10 mg/kg, IP, daily) during the whole period of 14 weeks, in combination with prednisolone (20 mg/kg, IP, daily) the first 4 days. Five rats served as nonimmunosuppressed controls. All rats were tested for amphetamine-induced rotational behavior at 3-week intervals. Two immunosuppressed rats were excluded due to severe side effects of the treatment. Functional recovery was seen in 9 of 16 immunosuppressed rats at 12 weeks. Six animals remained functionally recovered at 14 weeks and contained an average of 5750 ± 1450 (SEM) dopaminergic neurons. Between 9 and 14 weeks, three immunosuppressed rats rejected their grafts, based on rotation scores and immunohistochemical demonstration of cell infiltrates. One additional immunosuppressed rat showed evidence of ongoing rejection at 14 weeks. The striata in animals with ongoing or recent rejection contained large numbers of CD4- and CD8-positive lymphocytes, NK cells, macrophages, and microglia cells, whereas scar tissue was found in rats with grafts rejected at earlier time points (n = 11). Embryonic porcine ventral mesencephalic tissue matures in the adult rat striatum, reinnervates the host brain, and restores behavioral defects. Immunosuppressive treatment was necessary for long-term graft survival and functional recovery, but did not sufficiently protect from rejection mechanisms. Porcine neural tissue is an interesting alternative to embryonic human tissue for intracerebral transplantation in neurodegenerative diseases. However, to achieve stable graft survival in discordant xenogeneic combinations, an appropriate immunosuppressive treatment or donor tissue modifications are needed.
Mechanical Barrier as a Protection against Rejection of Allogeneic Cartilage Formed in Joint Surface Defects in Rats
Authors: Moskalewski Stanislaw , Osiecka-Iwan Anna , Hyc Anna , Jozwiak Jaroslaw
Published: June 2017

Cartilage formed in transplants of allogeneic chondrocytes into joint cartilage defects in rats was infiltrated by immune cells migrating from the bone marrow while the surface on the side of the joint cavity remained free of infiltrations. This suggested that immunization occurred via bone marrow and not via joint cavity. Because articular cartilage is nourished exclusively by the synovial fluid, we have attempted to prevent cartilage rejection by protecting transplants from the contact with bone marrow. Defects in articular surface were filled with bone cement and chondrocytes were transplanted into a cavity prepared within the bone cement plug. Cartilage formed within the cement shell remained free of infiltrations and did not evoke systemic immunological response. However, distribution of glycosaminoglycans in the matrix of protected transplants was irregular. Cultures of chondrocytes growing in vitro on cement contained less glycosaminoglycans than the controls. This suggests that some factor(s) released from the cement unfavorably influenced chondrocytes and matrix production in protected transplants.
Enhancement of Adult Muscle Regeneration by Primary Myoblast Transplantation
Authors: Derosimo John F. , Washabaugh Charles H. , Ontell Martin P. , Daood Monica J. , Watchko Jon F. , Watkins Simon C. , Ameredes Bill T. , Ontell Marcia
Published: June 2017

Extensor digitorum longus muscles (EDL) of SCID mice were induced to undergo degeneration–regeneration subsequent to orthotopic, whole-muscle transplantation. Two days after transplantation some of these muscles received injections of primary myoblasts derived from EDL muscles of transgenic mice, which express nuclear localizing β-galactosidase under the control of the myosin light-chain 3F promoter and enhancer. Nine weeks after transplantation, regenerated muscles that received exogenous myoblasts were compared to similarly transplanted muscles that received no further treatment and to unoperated EDL muscles in order to determine the effect of myoblast transfer on muscle regeneration. Many myofibers containing donor-derived myonuclei could be identified in the regenerated muscles that had received exogenous myoblasts. The mass of the muscles subjected to transplantation only was significantly less (31 % less) than that of unoperated muscles. The addition of exogenous myoblasts to the regenerating EDL resulted in a muscle mass similar to that of unoperated muscles. The absolute twitch and tetanic tensions and specific twitch and tetanic tensions of transplant-only muscles were 28%, 36%, 32%, and 41%, respectively, of those of unoperated muscles. Myoblast transfer increased the absolute twitch and tetanic tensions of the regenerated muscles by 65% and 74%, respectively, and their specific twitch and tetanic tensions were increased by 41% and 48%, respectively. These data suggest a possible role for the addition of exogenous, primary myoblasts in the treatment of traumatized and/or diseased muscles that are characterized by myofiber loss.
Effects of Short-Term Hypoxia on a Transformed Cell-Based Bioartificial Pancreatic Construct
Authors: Papas K. K. , Long R. C. , Constantinidis I. , Sambanis A.
Published: June 2017

Hypoxia is an adverse condition that can jeopardize the function of a bioartificial pancreatic construct. In this study we have investigated the effects of short-term hypoxic exposure (up to 24 h) on the bioenergetic status, metabolism, and insulin secretion of perfused pancreatic constructs composed of alginate/poly-L-lysine/alginate (APA) encapsulated mouse insulinoma βTC3 cells. The bioenergetic status of the encapsulated cells was monitored noninvasively with the aid of 31P NMR spectroscopy, while glucose, lactate, and insulin concentrations were measured with off-line assays from media samples removed from the perfusion loop. Our results demonstrate that in freshly prepared constructs insulin secretion was not affected by the hypoxic conditions, although intracellular ATP concentration decreased and glucose consumption increased. Alternatively, in constructs that were maintained in our perfusion system for at least 10 days, identical hypoxic conditions resulted in a decreased insulin secretion concomitant to a decreased intracellular ATP concentration and increased glucose consumption. These results suggest that the effects of hypoxia on a transformed cell-based pancreatic construct are not constant throughout the duration of an in vitro culture. The observed differences are attributed to the significant cell growth and rearrangement that occurs with time during an in vitro culture of the constructs.
Pretransplant Induction of HSP-70 in Isolated Adult Pig Islets Decreases Early Islet Xenograft Survival
Authors: Brandhorst Daniel , Hammes Hans-Peter , Brandhorst Heide , Zwolinski Anke , Nahidi Fariborz , Alt Alexandra , Bretzel Reinhard G.
Published: June 2017

The heat-induced HSP-70 expression protects rat islet single cells against lysis mediated by nitric oxide (NO), reactive oxygen, and streptozotocin. The present study was performed to investigate the potential antiinflammatory effect of pretransplant heat shock in adult pig islets for subsequent early islet xenograft survival. Maximum HSP-70 expression in freshly isolated pig islets was induced by hyperthermia at 43°C for 90 min prior to islet regeneration at 37°C for 4–6 h. Heat-stressed and sham-treated islets were incubated in 0.6 mM H2O2 or 1.5 mM Na-nitroprusside at 37°C for 20 h. Early graft survival was evaluated in normoglycemic Lewis rats after simultaneous, contralateral transplantation of heat-shocked islets and sham-treated islets into the renal subcapsular space of the same recipient. Prior hyperthermia significantly reduced specific lysis of islets exposed to NO or H2O2, although protection was only marginal. No differences were observed between viability of heat-shocked and sham-treated islets after NO exposure. In contrast, prior heat shock increased islet viability after H2O2 treatment. The finding that hyperthermia reduced recovery of initially grafted pig insulin 48 h after transplantation by 30% compared to controls contrasted significantly with an increased insulin recovery in heat-exposed islets at the end of simultaneous 37°C culture. The observation, that the heat-induced HSP-70 expression decreases early islet xenograft survival as reflected by recovery of grafted insulin, implies an enhancement of islet immunogenicity and the induction of apoptosis. Future experiments aiming at augmentation of intrinsic defense mechanims should consider detrimental effects associated with induction of heat shock proteins.
The Efficiency of Muscle-Derived Cell-Mediated Bone Formation
Authors: Bosch Patrick , Musgrave Doug , Ghivizzani Steven , Latterman Christian , Day Charles S. , Huard Johnny
Published: June 2017

The development of new clinically applicable methods for the delivery of bone morphogenic protein (BMP) is an area of intensive research. Cell-mediated gene therapy approaches are being explored as a potential delivery vehicle. Primary muscle-derived cells isolated from an adult mouse were transduced with an adenoviral–BMP-2 construct. These cells were injected into the triceps surae of severe combined immune deficient (SCID) mice where they induced heterotopic bone formation. BMP-2 expression by these muscle-derived cell constructs was measured in vitro to estimate in vivo BMP-2 delivery. In vitro expression of BMP-2 by 3 × 105muscle-derived cells was 87.89 ng/72 h. These results suggest that the efficiency of muscle cell-based gene delivery of BMP-2 exceeds the direct delivery of recombinant BMP-2 protein.
Treatment of Carbon Tetrachloride and Phenobarbital-Induced Chronic Liver Failure with Intrasplenic Hepatocyte Transplantation
Authors: Kobayashi Naoya , Ito Masahiro , Nakamura Junta , Cai Jin , Hammel James M. , Fox Ira J.
Published: June 2017

Hepatocyte transplantation (HTx) has been shown to improve the survival of laboratory animals with experimentally induced acute liver failure and to ameliorate the physiologic abnormalities associated with liver-based metabolic deficiencies. However, the role of HTx in the treatment of liver cirrhosis (LC) has not been adequately studied. In order to address this issue, HTx was performed in rats following induction of stable LC using phenobarbital (PhB) and carbon tetrachloride (CCl4). Intrasplenic transplantation of 50 × 106 primary hepatocytes could significantly improve liver functions and prolong the survival of rats with irreversible, decompensated LC.
Establishment of Fluorescein Diacetate and Ethidium Bromide (FDAEB) Assay for Quality Assessment of Isolated Islets
Authors: Miyamoto Masaaki , Morimoto Yoshihiko , Nozawa Yuka , Balamurugan A. N. , Xu Baoyou , Inoue Kazutomo
Published: June 2017

One of the most important factors in clinical islet transplantation is isolation of a great number of islets with good viability. According to viability assessments of isolated islets, the static incubation test and the perifusion test of islets, which are used retrospectively, take much time and need various apparatus. But viability assessments of isolated islets for clinical islet transplantation require a simple, rapid, sensitive, and prospective method. We have developed a microfluorometric viability assay for isolated human, porcine, and dog islets of pancreata using fluorescein diacetate and ethidium bromide (FDAEB). Fluorescein diacetate (FDA) causes live cells to fluoresce green under blue light excitation (490 nm) and ethidium bromide (EB) causes dead cells to fluoresce red. In this study, we investigated the applicability of FDAEB staining to quality assessment of isolated islets for clinical use by correlation with the counting method with insulin secretion of islets. Discrimination of living from dead islets by insulin secretion correlated well with viability as determined by FDAEB staining. The proportion of living islets within isolated canine islets, as measured by microfluorometric counting, was found to correlate highly significantly on low-temperature (24°C) culture (R = 0.831, p < 0.001) and on 37°C culture (R = 0.553, p < 0.05) with the insulin contents of the same islets. Therefore, it is possible to differentiate degrees of viability, and a scoring system is described for this purpose. The FDAEB assay prospectively and easily provides a rapid, accurate, and objective measurement of the proportion of living cells and dead cells in isolated islets for clinical islet transplantation.
Grafting of Mitomycin C-Treated Islet Xenograft under the Kidney Capsule Produces a Clear Bleb
Authors: Sato Yoshihiro , Grochowiecki Tadeusz R. , Takeda Yutaka , Dono Keizo , Ise Kazuya , Kanazawa Yukio , Saito Takuro , Abe Tsuyoshi , Gotoh Mitsukazu
Published: June 2017

We have previously shown that mitomycin C (MMC) treatment of donor tissue resulted in significant prolongation of graft survival in allo- and xenotransplantation models. However, the mechanisms involved in this prolongation are not clearly understood. This study aims to shed light on the immune responses to MMC-treated islet xenografting under the kidney capsule. Collagenase-digested WS (RT1k) rat islets incubated for 30 min with MMC and subsequently cultured for 20 h were transplanted into the renal subcapsular space of streptozotocin-induced diabetic C57BL/6 (B6;H-2b) mice. The grafts were harvested on postgrafting day 7 and sections were prepared and stained by hematoxylin and eosin (H&E). Histological study of the grafts in a group not treated with MMC showed marked cellular infiltration and destruction of islet clusters, whereas that of MMC-treated grafts demonstrated a bleb formation under the kidney capsule, in which islet cell clusters were reorganized, creating a layer of cells fixed to the interior of the bleb. Minimal invasion by inflammatory cells was observed only at the edge of the bleb, and most islet cells were protected from these infiltrating cells. In conclusion, MMC treatment induces remodeling of islet structure and forms a bleb under the kidney capsule, where no inflammatory cell infiltration occurs, suggesting that this site is a kind of immunologically privileged environment for xenografted islets.
Long-Term Culture of Glutamine Synthetase-Transfected HepG2 Cells in Circulatory Flow Bioreactor for Development of a Bioartificial Liver
Authors: Enosawa Shin , Miyashita Tomoyuki , Suzuki Seiichi , Li Xiao-Kang , Tsunoda Miyuki , Amemiya Hiroshi , Yamanaka Mitsugu , Hiramatsu Shinya , Tanimura Naoko , Omasa Takeshi , Suga Kenichi , Matsumura Toshiharu
Published: June 2017

Glutamine synthetase (GS) is involved in an accessory pathway of ammonia removal in mammals. To develop a bioartificial liver with a human cell line, GS gene was transfected into HepG2 cells, which had no ammonia removal activity. After culturing in the presence of methionine sulfoximine (MSX), a GS inhibitor, we obtained a MSX-resistant HepG2 subline (GS-HepG2), which had amplified GS gene; ammonia removal activity was estimated to be 1/7 of that of rat primary culture hepatocytes. The cells were cultured in a circulatory flow bioreactor for 109 days, while they multiplied from 5 × 107 to 4 × 109 cells. Three days after inoculation, the ammonia level of the culture medium was lowered to a level maintained thereafter, suggesting that using recombinant cell lines for bioartificial livers enables long-term repeated treatment for hepatic failure patient. Judging from the rate of decrease in the amount of the added ammonia, the ammonia removal capability of 4 × 109 GS-HepG2 cells was almost equivalent to 5 × 108 porcine hepatocytes inoculated into the circulatory flow bioreactor. Apart from their ammonia removal activity, GS-HepG2 cells eliminated human tumor necrosis factor-α (TNF-α). Cytokine removal therefore promises to be another useful property of bioreactor cells.
The Effects of Taurine on hNT Neurons Transplanted in Adult Rat Striatum
Authors: Rivas-Arancibia Selva , Willing Alison E. , Zigova Tanja , Rodriguez Alba I. , Cahill David W. , Sanberg Paul R.
Published: June 2017

Taurine acts as an antioxidant able to protect neurons from free radical-mediated cellular damage. Moreover, it modulates the immune response of astrocytes that participate in neurodegenerative processes. The objective of this study was to examine whether taurine can prevent or attenuate the host inflammatory response induced by the xenotransplantation of neurons derived from the human teratocarcinoma cell line (hNT neurons). Male Sprague-Dawley rats were treated IP with either saline or taurine. Animals from both groups were perfused on the 4th or 11th day and the saline or taurine was administered from the start of the study until the day prior to sacrifice. The brains were processed immunohistochemically using antibodies against glial fibrillary acidic protein (GFAP), microglia (OX42), and human nuclear matrix antigen (NuMA). In the saline group, NuMA labeling revealed small grafts on the 4th day and no surviving cells on the 11th day. However, in the group that received taurine there were surviving grafts at both time points. Strong immunoreactivity for GFAP and OX42 was detected in the saline group surrounding the transplant. These effects were reduced in animals receiving taurine. Taken together, these results demonstrated that taurine was able to facilitate graft survival and attenuate the immune response generated by the xenograft.
A New Type of Biocompatible Bridging Structure Supports Axon Regrowth after Implantation into the Lesioned Rat Optic Tract
Authors: Plant Giles W. , Harvey Alan R.
Published: June 2017

We have developed a new type of polymer/cell/matrix implant and tested whether it can promote the regrowth of retinal ganglion cell (RGC) and other axons across surgically induced tissue defects in the CNS. The constructs, which consisted of 2–2.5-mm-long polycarbonate tubes filled with lens capsule-derived extracellular matrix coated with cultured neonatal Schwann cells, were implanted into lesion cavities made in the left optic tract (OT) of 18–21-day-old rats. In one group, to promote Schwann cell proliferation and perhaps also to stimulate axon regrowth, basic fibroblast growth factor (bFGF) was added to the lens capsule matrix prior to implantation. In another group, to determine whether application of growth factors to the somata of cells enhances the regrowth of distally injured axons, the neurotrophin NT-4/5 was injected into the eye contralateral to the OT lesion. NT-4/5 and bFGF treatments were combined in some rats. After medium-term (4–10 weeks) or long-term (15–20 weeks) survivals, axon growth into implants was assessed immunohistochemically using a neurofilament (RT97) antibody. RGC axons were visualized after injection of WGA/HRP into the right eye. Viable Schwann cells were present in implants at all times after transplantation. Large numbers of RT97+ axons were consistently found within the bridging implants, often associated with the peripheral glia. Axons were traced up to 1.7 mm from the nearest CNS neuropil and there was immunohistochemical evidence of myelination by Schwann cells and by host oligodendrocytes. There were fewer RGC axons in the implants, fibers growing up to 1.6 mm from the thalamus. Neither NT-4/5 nor bFGF, alone or in combination, significantly increased the extent of RGC axon growth within the implants. A group of OT-lesioned rats was implanted with polymer tubes filled with 2–2.5-mm-long pieces of predegenerate peripheral nerve. Surprisingly, polymer/cell/matrix constructs contained comparatively greater numbers of RGC and other axons and supported more extensive axon elongation. Thus, implants of this type may potentially be useful in bridging large tissue defects in the CNS.
Alginate-Encapsulated Producer Cells: A Potential New Approach for the Treatment of Malignant Brain Tumors
Authors: Thorsen Frits , Read Tracy-Ann , Lund-Johansen Morten , Tysnes Berit Bølge , Bjerkvig Rolf
Published: June 2017

In recent years gene therapy has evolved as a new treatment for brain tumors, where genetically engineered cells can be used to deliver specific substances to target cells. However, clinical success has been limited due to insufficient gene transfer, lack of prolonged gene expression, and immunorejection of producer cells. These obstacles may be overcome by encapsulating producer cells into immunoisolating substances such as alginate. This may provide a stable in situ delivery system of specific proteins, which can interfere with tumor growth and differentiation. This article represents a fundamental study describing the in vitro and the in vivo behavior of alginate-encapsulated producer cells. The viability and cell cycle distribution of encapsulated NIH 3T3 cells was studied by confocal laser scanning microscopy (CLSM) and by flow cytometry. The CLSM study showed a high viability of the encapsulated NIH 3T3 cells during 9 weeks in culture. The flow cytometric analysis revealed a change in cellular ploidy after 1 week in culture, with normalization in ploidy after 3 and 9 weeks. The production of the bacterial E. coli β-galactosidase in alginate-encapsulated BT4CnVlacZ cells was studied by x-gal staining, and the cells expressed prolonged β-galactosidase activity. H528 hybridoma cells producing monoclonal antibodies (mAbs) against the human epidermal growth factor receptor (EGFR) were encapsulated in alginate, and the mAb release was determined. The release of mAbs stabilized around 400 ng/ml/h after 12 days in vitro. To actually demonstrate that alginate-encapsulated H528 cells potentially inhibit a heterogeneous glioma cell population, cell migration from human GaMg glioma spheroids was studied during stimulation with EGF in the presence of encapsulated H528 cells. The migration in vitro was totally inhibited in the presence of H528 encapsulated cells. Alginate beads with H528 cells were also implanted into rat brains, and after 9 weeks the distribution of mAbs within the brain was studied by immunohistochemistry. It is shown that the alginate entrapped H528 cells produce mAbs inside the brain for prolonged periods and that the mAbs are distributed within all CSF compartments. Encapsulated producer cells represent a potential delivery system for specific proteins to brain tumors. Different producer cells may be encapsulated in alginate to target phenotypic features and microenvironmental factors, which may influence the progressive growth of brain tumors.
A Large-Animal Model to Evaluate the Clinical Potential of Fetal Pig Pancreas Fragment Transplantation
Authors: Hawthorne Wayne J. , Cachia Adrian R. , Walters Stacey N. , Patel Anita T. , Clarke Janelle E. , O'connell Philip J. , Chapman Jeremy R. , Allen Richard D. M.
Published: June 2017

The long-term goal of this study is to assess the feasibility of using fetal pig pancreas fragment (FPPF) transplantation to treat patients with type I diabetes. Using the highly inbred Westran Pigs, our initial aim was to establish a rejection-free transplant model of FPPF grafted into sibling recipient pigs without immunosuppression. FPPFs were isolated from 80–100-day-old fetuses of either Westran Pigs or outbred pigs and transplanted into the thymus, spleen, liver, or kidney of the recipient Westran pig. Biopsies were taken from each transplant site at set time points and assessed histologically for islet viability, rejection, and endocrine function. Fifty-eight fetal donors were used to transplant 16 recipient pigs. A nonspecific inflammation was seen for both outbred and inbred FPPF donor tissue at day 3 and was considered a response to ischemic necrosis. However, all the transplanted outbred FPPF donor tissue was acutely rejected and lost by day 10–14. In contrast, inbred FPPF tissue showed little evidence of graft necrosis after 3 days, and growth and formation of epithelial islet cell nest-like structures were seen to 28 days after transplantation. With time after transplantation, increasing amounts of insulin immunoperoxidase staining was seen together with chromogranin and somatostatin staining. In summary, this study confirms the potential of the Westran pig to answer the unproven ability of fetal pancreatic tissue to reverse type I diabetes in a large animal model.
Long-Term Preservation of Rat Skin Tissue by Epigallocatechin-3-O-Gallate
Authors: Kim Hakhee , Kawazoe Takeshi , Matsumura Kazuaki , Suzuki Shigehiko , Hyon Suong-Hyu
Published: June 2017

Skin grafts can be preserved by cryopreservation and refrigerated storage at 4°C. Epigallocatechin-3- O-gallate (EGCG) enhances the viability of stored skin grafts and also extends the storage time up to 7 weeks at 4°C. EGCG, the major polyphenolic constituent present in green tea, has potent antioxidant, antimicrobial, antiproliferative, and free radical scavenging effects. This study examined the effects of EGCG on skin cryopreservation. Skin sample biopsy specimens from GFP rats were previously treated with/without EGCG then moved to −196°C. Skin samples were transplanted to nude mice after 2, 8, and 24 weeks of preservation. Glucose consumption was measured after thawing to assess the metabolic activity. Two weeks later the transplanted skin grafts were excised and histologically analyzed. Histological examinations revealed the degeneration of the epidermal and dermal layers in all groups. In the EGCG groups, the grafts showed higher integrity in the epidermal layer and dermal matrix. The present findings suggest the future clinical usefulness of EGCG for skin preservation; however, the mechanism by which EGCG promotes skin preservation still remains unclear.
Preservation of Platelets by Adding Epigallocatechin-3-O-Gallate to Platelet Concentrates
Authors: Matsumura Kazuaki , Takayama Hiroshi , Bae Jung Yoon , Kurihara Mitsuru , Tsutsumi Sadami , Hyon Suong-Hyu
Published: June 2017

The effect of epigallocatechin-3- O-gallate (EGCG), a major component of green tea, on platelet preservation was evaluated. Single donor platelets ( N = 10) were collected and preserved by the standard method. EGCG was added to the platelet concentrates before preservation and then the functional and biochemical parameters were monitored throughout the storage period. After 6 days of preservation, the aggregability of the platelets was significantly maintained by addition of 50 and 100 μg/ml of EGCG. Platelet prothrombinase activity was also significantly retained by the addition of EGCG. The accumulation of P-selectin and RANTES in the plasma preserved with EGCG was less than those preserved without EGCG, which indicated that EGCG might inhibit platelet activation. Furthermore, EGCG reduced the increase of LDH in plasma during preservation and inhibited the activation of caspase-3 and cleavage of gelsolin, thereby showing that EGCG could inhibit the apoptosis of platelets. These results suggest that EGCG may play an effective role in preserving platelets by inhibiting the activation and apoptosis of platelets.
A Study on the Perfusion Preservation, Resuscitation, and Transplantation of a Rat Heart Isolated for 96 Hours
Authors: Hatayama Naoyuki , Yoshida Yu , Seki Kunihiro
Published: June 2017

Krebs-Henseleit (KH) solution was used to fill the heart chamber of an isolated rat heart before it was immersed in perfluorocarbon (PFC), which is an inert fluid. A gas mixture (PCO2 = 150 hPa and PO2 = 850 hPa) was then aerated at a constant rate into the PFC solution, and the isolated heart was thereafter preserved for 96 h with KH solution perfused continuously at a rate of 0.1 ml/h from the aorta of the isolated heart through a cannula. After preservation, the preserved heart was heterotopically transplanted into the neck of a recipient rat and then it was resuscitated. Using this method for preserving mammalian organs, we attained reproducibility after perfusion preservation for 96 h.
Estimation of Donor Usability for Islet Transplantation in the United States with the Kyoto Islet Isolation Method
Authors: Matsumoto Shinichi , Noguchi Hirofumi , Hatanaka Nobuyo , Shimoda Masayuki , Kobayashi Naoya , Jackson Andrew , Onaca Nicholas , Naziruddin Bashoo , Levy Marlon F.
Published: June 2017

The quality of donor pancreata is important for successful islet isolation. However, in some countries like Japan, the number of donor pancreata is very low; therefore, marginal donors have been used with less restrictive donor criteria. In order to use marginal donor pancreata, we established the Kyoto islet isolation method (KIIM). According to United Network for Organ Sharing (UNOS) in 2005, more than 6,000 pancreata were not clinically used in the US. In this study, we applied the KIIM for brain-dead donors and reevaluated donor usability based on the Japanese islet donor criteria. Islets were isolated with the Ricordi method using pancreata stored in University of Wisconsin (UW) solution (UW group) or by the two-layer method (TLM group) or the TLM combined with ductal injection (DI group). We implemented the KIIM (KIIM group) to confirm the effect of the KIIM on brain-dead donors. Donor charts in Texas from 2005 to 2006 were reviewed. If pancreata were not used clinically, the reason was reviewed and donors were reevaluated based on Japanese criteria. There were no significant differences of islet yield, viability, and purity between the UW and TLM groups. The DI group significantly improved islet yields and isolations were further improved in the KIIM group [UW: 251,663 ± 60,217 islet equivalent (IE); TLM: 243,738 ± 54,170 IE; DI: 498,639 ± 28,853 IE; KIIM: 678,286 ± 55,853]. The KIIM provided high-quality islets in high numbers from islet isolations from brain-dead donors. A total of 236 donor charts were reviewed and 194 pancreata (82%) were not used. Of these, 185 cases identified the reasons that the pancreata were not used. When we applied the Japanese criteria, an additional 82 cases out of 185 (44%) seem to be suitable for islet isolations. With the KIIM, more than 2,500 additional donor pancreata can be used for islet isolation in the US every year when the Japanese criteria are applied.