bluuebunny/crossref_metadata_2025 huggingface dataset:
The Anion Gap and Routine Serum Protein Measurements in Monoclonal Gammopathies
Authors: van Hoeven Karen H. , Joseph Rosy E. , Gaughan William J. , McBride Laura , Bilotti Elizabeth , McNeill Ann , Schmidt Linda , Schillen Danielle , Siegel David S.
Published: December 2011

Background and objectives

An abnormal anion gap and an increased total protein and globulin are clues to the diagnosis of monoclonal gammopathy. We explored the utility of these markers in IgG, IgA, IgM, and free light chain monoclonal gammopathies.

Design, Setting, Participants, & Measurements

The anion gap, Na+ – (Cl– + HCO3
–), corrected for hypoalbuminemia, was calculated in patients with monoclonal gammopathies. Exclusion criteria were serum calcium >10.5 mg/dl and/or creatinine >2 mg/dl.

Results

Among 287 patients, 242 remained after applying exclusion criteria (109 IgG, 64 IgA, 21 IgM, and 48 light chain); 36% of 242 patients required correction for hypoalbuminemia. The anion gap was decreased (<10) in 22% of IgG and increased (>15) in 31% of IgA monoclonal gammopathies. IgM did not affect the gap. In light chain gammopathies, the anion gap showed no consistent trend (15% increased, 17% decreased). Mean clonal IgG, IgA, and IgM concentrations were 10-fold higher than mean clonal free light chain concentrations in the respective monoclonal gammopathies (P < 0.001). These paraprotein level disparities were reflected in significantly increased mean serum total protein and globulin concentrations in IgG, IgA, and IgM versus free light chain monoclonal gammopathies, where mean total protein and globulin levels were within normal limits (P < 0.001).

Conclusions

The anion gap was significantly altered in IgG and IgA monoclonal gammopathies, but it was not a sensitive tool for suspecting the diagnosis. In light chain monoclonal gammopathies, the anion gap, total protein, and globulin did not provide reliable diagnostic clues.
Renal Monoclonal Immunoglobulin Deposition Disease
Authors: Nasr Samih H. , Valeri Anthony M. , Cornell Lynn D. , Fidler Mary E. , Sethi Sanjeev , D’Agati Vivette D. , Leung Nelson
Published: December 2011

Background and objectives

To better define the clinical-pathologic spectrum and prognosis of monoclonal immunoglobulin deposition disease (MIDD), this study reports the largest series.

Design, setting, participants, & measurements

Characteristics of 64 MIDD patients who were seen at Mayo Clinic are provided.

Results

Of 64 patients with MIDD, 51 had light chain deposition disease, 7 had heavy chain deposition disease, and 6 had light and heavy chain deposition disease. The mean age at diagnosis was 56 years, and 23 patients (36%) were ≤50 years of age. Clinical evidence of dysproteinemia was present in 62 patients (97%), including multiple myeloma in 38 (59%). M-spike was detected on serum protein electrophoresis in 47 (73%). Serum free light chain ratio was abnormal in all 51 patients tested. Presentation included renal insufficiency, proteinuria, hematuria, and hypertension. Nodular mesangial sclerosis was seen in 39 patients (61%). During a median of 25 months of follow-up (range, 1–140) in 56 patients, 32 (57%) had stable/improved renal function, 2 (4%) had worsening renal function, and 22 (39%) progressed to ESRD. The mean renal and patient survivals were 64 and 90 months, respectively. The disease recurred in three of four patients who received a kidney transplant.

Conclusions

Patients with MIDD generally present at a younger age than those with light chain amyloidosis or light chain cast nephropathy. Serum free light chain ratio is abnormal in all MIDD patients, whereas only three-quarters have abnormal serum protein electrophoresis. The prognosis for MIDD is improving compared with historical controls, likely reflecting earlier detection and improved therapies.
Maternal Deletion Allele of Angiotensin-Converting Enzyme Gene is Associated with Fetal Growth Restriction
Authors: Nakamura Eiki , Okamoto Toshio , Nagaya Ken , Hayashi Tokitsugi
Published: December 2011

To investigate the correlation between maternal and fetal angiotensin-converting enzyme gene insertion or deletion polymorphism and birth size, angiotensin-converting enzyme gene insertion or deletion polymorphisms of 470 Japanese pairs of mothers and infants were genotyped. The relationships between maternal and fetal angiotensin-converting enzyme gene insertion or deletion polymorphisms and birth sizes (weight, length, head circumference, and the incidence of small-for–gestational age status) were analyzed. No relationship between fetal angiotensin-converting enzyme genotype and birth size was observed. In contrast, the incidence of small-for–gestational age status in babies whose mothers had 1 or 2 deletion alleles was greater than that in babies whose mothers had 2 insertion alleles (7% vs 3%, P = 0.045). Maternal deletion allele of angiotensin-converting enzyme is associated with fetal growth restriction in the Japanese normal population.
Role of the APOB Gene Polymorphism (c.12669G>A, p. Gln4154Lys) in Coronary Artery Disease in the Indian Punjabi Population
Authors: Sharma R , Mahajan M , Singh B , Singh G , Singh P
Published: December 2011

Role of theAPOBGene Polymorphism (c.12669G>A, p. Gln4154Lys) in Coronary Artery Disease in the Indian Punjabi Population

High concentration of apolipoprotein B (apoB) is a risk factor for coronary artery disease (CAD). The association of theAPOBgene polymorphism c.12669G>A, p. Gln4154Lys with the risk of CAD varies considerably in different populations. The present study represents the first investigation regarding the role of thisAPOBgene polymorphism with CAD in the Indian Punjabi population. We have studied theAPOBgene polymorphism c.12669G>A, p. Gln4154Lys and its relationship with lipid, apoB, low-density lipoprotein (LDL) heterogeneity and oxidation in subjects suffering from CAD. The study was conducted on 87 patients with CAD; 75 healthy subjects served as controls. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was used to determine the DNA polymorphism in theAPOBgene. Frequency of R-(mutant) allele was significantly high (p<0.05) in CAD patients when compared to controls. Variations in serum lipid levels in the R+R+ and R+R-APOBgenotypes were insignificant (p>0.05). However, serum apoB levels were significantly raised (p<0.05) in CAD patients with the R+R- genotype as compared to those with the R+R+APOBgenotype. Coronary artery disease patients had raised significantly raised (p<0.01) Log triglyceride/high density lipoprotein-cholesterol (HDL-C) ratio, apoB carbonyl content and increased malondialdehyde-low density lipoprotein (MDA-LDL levels, irrespective ofAPOBgenotype as compared to controls. Carriers of the R- allele are at higher risk of CAD, probably because of elevated serum apoB levels in the Indian Punjabi population. Overall, it may be concluded that the R- allele might be associated with increased susceptibility towards CAD development in the Indian Punjabi population, and one of the linking factor is the elevation in serum apoB levels. However, this association needs further evaluation in a larger population. Secondly, the robust mechanism behind the positive association of the R- allele with raised serum apoB levels needs to be explored, which might be helpful in the strengthening the observed results.
Global and distributed torque calculations using the CDSA approach
Authors: Li M. , Lowther D.
Published: December 2011

Global and distributed torque calculations using the CDSA approach

Accurate force and torque calculations are fundamental to being able to predict the operation of an electromechanical device or system. The Maxwell stress tensor and the virtual work principle are the two major theories for force and torque calculation. However, if local distributions of torque are needed to couple to structural and vibration analyses, the conventional Maxwell stress approach cannot provide this easily. A recently developed approach based on sensitivity analysis has the capability to deliver local stress and torque as well as accurate global results. In addition, this approach divides the total torque into different components which are essential to the design of electrical devices. This paper includes several numerical examples of torque calculations of different electrical machines. The results are verified by a commercial software package using the Maxwell stress based force calculation.
Two-level approach for solving the inverse problem of defects identification in Eddy Current Testing - type NDT
Authors: Putek Piotr , Crevecoeur Guillaume , Slodička Marian , Gawrylczyk Konstanty , van Keer Roger , Dupré Luc
Published: December 2011

Two-level approach for solving the inverse problem of defects identification in Eddy Current Testing - type NDT

This work deals with the inverse problem associated to 3D crack identification inside a conductive material using eddy current measurements. In order to accelerate the time-consuming direct optimization, the reconstruction is provided by the minimization of a last-square functional of the data-model misfit using space mapping (SM) methodology. This technique enables to shift the optimization burden from a time consuming and accurate model to the less precise but faster coarse surrogate model. In this work, the finite element method (FEM) is used as a fine model while the model based on the volume integral method (VIM) serves as a coarse model. The application of the proposed method to the shape reconstruction allows to shorten the evaluation time that is required to provide the proper parameter estimation of surface defects.
Wedge functions applied to 2D magnetostatic problems
Authors: Huijer Ernst , Karaki Sami
Published: December 2011

Wedge functions applied to 2D magnetostatic problems

In this paper the application of so called wedge functions is presented to solve two-dimensional simple geometries of magnetostatic and electrostatic problems, e.g. rectangles of varying aspect ratio and with different values of the magnetic permeability μ. Such problems require the use of surface charge density, or segment source, functions of the form ρs = σa-1, where the power parameters, a, have special fractional values. A methodology is presented to determine these special values of a and use them in segment sources on simple geometries, i.e. rectangles of varying aspect ratio, and with different values of the magnetic permeability μ. Wedge solutions are obtained by coupling the strength coefficients of source segments of the same power around an edge. These surface source functions have been used in the analysis of conducting and infinite permeability structures. Here we apply such functions in a boundary integral analysis method to problems having regions of finite permeability.
Synthesis of nano-crystalline Zn – CoO pigment by the chemical combustion method: Effect of fuel to oxidizer ratio on structure, microstructure and color properties
Authors: Rasouli Sousan , Arabi Amir Masoud
Published: December 2011

The aim of this study was to investigate the effect of fuel/oxidizer ratio on the combustion synthesis of nano-crystalline Zn – CoO green pigment. The fuel and oxidizer used in this study were glycine and nitrate respectively. Products were characterized by means of X-ray diffraction, Fourier transformation infra red spectroscopy, scanning electron microscopy and diffuse reflectance spectroscopy. X-ray diffraction patterns (XRD) showed that the formation of pure hexagonal ZnO phase was insensitive to fuel/oxidizer ratio variations. Well-crystalline single-phase ZnO was observed for low and stoichiometric fuel/oxidizer ratios. Average crystallite size determined from the Scherrer formula varied from 18 to 75 nm. The morphology of the obtained samples changed from hexagonal pyramidal to irregular porous sponge-like particles. The colorimetric results showed a broad reflectance band around 530 nm (green region) for all samples. The observation of green appearance of different samples synthesized using different fuel/oxidizer ratios was related to fade of Co3O4 X-ray diffraction patterns and entrance of Co into ZnO structure.
The viscosity of eutectic Pd – Si alloys – a case study
Authors: Egry Ivan , Schick Michael
Published: December 2011

The viscosity of eutectic PdSi alloys has been measured using different techniques. Conventional oscillating cup viscosimetry was used in the ground-based laboratory, while the oscillating drop technique was applied in a microgravity experiment. Different working equations were utilised to derive the viscosity from the damping of the oscillations, and the results thus obtained were compared to each other and available literature data. An overall satisfactory agreement was found, while, at lower temperatures, an increasing difference between the viscosities obtained using the two models was observed.
The use of ground silica sand, ground granulated blast furnace slag and ground basaltic pumice in cementitious adhesives for tiles
Authors: Binici Hanifi
Published: December 2011

The aim of this study was to use ground silica sand, ground granulated blast furnace slag and ground basaltic pumice in the production of adhesives for tiles. The main purpose was to improve the durability of adhesives. Furthermore, an investigation was carried out to determine adhesive resistance to aggressive conditions. The study examines the effects of acids, shrinkage and freezing thawing cycles. The results showed that the adhesives were prepared and applied easily due to improved initial adhesion. The durability improvement can be explained by the low water absorption and high permeability. All specimens were observed to fulfill the shear and tensile adhesion strength requirements of both European and Turkish Standards.
A simple hydrothermal route to bimodal mesoporous nanorod γ-alumina with high thermal stability
Authors: Li Xiang , Han Dezhi , Xue Hongxia , Liu Xinmei , Yan Zifeng
Published: December 2011

In the presence of polyethylene glycol, bimodal mesoporous nanorod γ-alumina was successfully synthesized via the thermal decomposition of ammonium aluminium carbonate hydroxide precursor which was prepared via hydrothermal processing with inorganic aluminium salt. The alumina exhibits high surface area (494 m2g–1), large porosity (1.1 m3g–1) and a particular double-pore structure after calcination at 500 °C. The smaller pore diameter is concentrated on about 3 nm and the larger one is exhibited in the range of 10 – 38 nm. The scaffold-like aggregation of γ-alumina nanorods endows this novel material with excellent thermal stability. A possible formation mechanism of bimodal mesoporous structure is also proposed in this study.
Synchrotron measurement of the 3D shape of X-ray reflections from the γ/γ′-microstructure of nickel-base superalloys
Authors: Epishin Alexander , Link Thomas , Ulbricht Alexander , Zizak Ivo , Bansal Mamta
Published: December 2011

The 3D shape of X-ray reflections from the γ/γ′-microstructure of a nickel-base superalloy was investigated using synchrotron X-ray radiation and a position sensitive area detector. The measurements were performed on the 4th generation single-crystal nickel-base superalloy TMS138. The results show that X-ray reflections from non-cubic crystallographic planes have a complex 3D shape which changes during rafting. The 3D intensity distributions contain information about the spacing of the planes and their orientation as well. Whereas h00 reflections show the usual splitting into a γ′ and one γ-subreflection, the hh0 and hhh reflections show two and three γ-peaks respectively, resulting from the different types of {100} matrix channels. Therefore, these 3D diffraction measurements supply additional information about the spatial distribution of microstrains.
Time-dependent corrosion resistance of zinc phosphate coating on AZ31B magnesium alloy in sodium chloride solution
Authors: Huanfang Gao , Shengtao Zhang , Chenglong Liu
Published: December 2011

The composition, morphology and the long-term corrosion resistance in sodium chloride solution of zinc phosphate coating on AZ31B magnesium alloy were investigated using X-ray diffraction, scanning electron microscopy, energy dispersive X-ray spectroscopy, hydrogen evolution and potentiodynamic polarization curve experiments. The results indicate that the phosphate coating consists mainly of Zn3(PO4)2 · 4 H2O, Zn2Mg(PO4)2 and metallic Zn. The variations of the hydrogen evolution, i
corr and corrosion morphologies indicate that the corrosion process of zinc phosphate has three stages. At the first stage, the corrosion resistance of the phosphate sample is higher than that of the uncoated sample. At the second stage, the corrosion rate basically remains constant. At the third stage, a Zn – Mg galvanic cell and galvanic corrosion are formed between zinc and the AZ31B substrate, and the corrosion resistance of the phosphate sample is lower than that of the uncoated sample.
Nanobiotechnology approach to fabricate polycaprolactone nanofibers containing solid titanium nanoparticles as future implant materials
Authors: Sheikh Faheem A. , Kanjwal Muzafar A. , Cha Jaegwan , Kim Namsoo , Barakat Nasser A. M. , Kim Hak Yong
Published: December 2011

In this study, a good combination of electrospun poly(caprolactone) nanofibers incorporated with high purity titanium nanoparticles is introduced for hard tissue engineering applications. A simple approach to utilize the colloidal properties of poly(caprolactone) and titanium nanoparticles are exploited to form nanofibers by the simple electrospinning process. The prepared colloidal solutions were characterized using dynamic light scattering and electrophoratic light scattering which indicated unimodal size distribution and negative zeta potential. To investigate the bioactivity of the resultant nanofiber mats, they were incubated in simulated body fluid at 37 °C for 10 days. Field emission scanning electron microscopy in combination with energy-dispersive X-ray spectroscopy indicated that incorporation of titanium strongly activates precipitation of the apatite-like materials from the utilized simulated body fluid. Moreover, in-vivo experiments using experimental dogs revealed that nanofibers can yield good tissue regeneration on the surfaces of nanofibers.
A novel method of intracranial injection via the postglenoid foramen for brain tumor mouse models
Authors: Iwami Kenichiro , Momota Hiroyuki , Natsume Atsushi , Kinjo Sayano , Nagatani Tetsuya , Wakabayashi Toshihiko
Published: December 2011

Object

Mouse models have been widely used in developing therapies for human brain tumors. However, surgical techniques such as bone drilling and skin suturing to create brain tumors in adult mice are still complicated. The aim of this study was to establish a simple and accurate method for intracranial injection of cells or other materials into mice.

Methods

The authors performed micro CT scans and skull dissection to assess the anatomical characteristics of the mouse postglenoid foramen. They then used xenograft and genetically engineered mouse models to evaluate a novel technique of percutaneous intracranial injection via the postglenoid foramen. They injected green fluorescent protein–labeled U87MG cells or virus-producing cells into adult mouse brains via the postglenoid foramen and identified the location of the created tumors by using bioluminescence imaging and histological analysis.

Results

The postglenoid foramen was found to be a well-conserved anatomical structure that allows percutaneous injection into the cerebrum, cerebellum, brainstem, and basal cistern in mice. The mean (± SD) time for the postglenoid foramen injection technique was 88 ± 15 seconds. The incidence of in-target tumor formation in the xenograft model ranged from 80% to 100%, depending on the target site. High-grade gliomas were successfully developed by postglenoid foramen injection in the adult genetically engineered mouse using virus-mediated platelet-derived growth factor B gene transfer. There were no procedure-related complications.

Conclusions

The postglenoid foramen can be used as a needle entry site into the brain of the adult mouse. Postglenoid foramen injection is a less invasive, safe, precise, and rapid method of implanting cells into the adult mouse brain. This method can be applied to both orthotopic xenograft and genetically engineered mouse models and may have further applications in mice for the development of therapies for human brain tumors.
Multi-Location Study of Soil Enzyme Activities as Affected by Types and Rates of Manure Application and Tillage Practices
Authors: Acosta-Martinez Veronica , Mikha Maysoon M. , Sistani Karamat R. , Stahlman Phillip W. , Benjamin Joseph G. , Vigil Merle F. , Erickson Richie
Published: December 2011

Significant amounts of manure are produced in the USA; however, information on the changes in ecosystem services related to soil biogeochemical cycling for agroecosystems supported with organic amendments such as manure is limited. A multi-location field study was initiated in Colorado (CO), Kansas (KS) and Kentucky (KY), USA in loam soils to evaluate the effects of manure and tillage practices on enzyme activities that are key to biogeochemical cycling such as β-glucosidase (C cycling), α-galactosidase (C cycling), β-glucosaminidase (C and N cycling) and phosphomonoesterases (P cycling). The treatments were as follows: (i) two years of beef manure applications to a fine sandy loam at different rates (control: 0, low: 34 kg N ha−1 and high: 96 kg N ha−1) and tillage practices in CO; (ii) three years of beef manure applications to a silt loam at different rates (0, low: 67 kg N ha−1 and high: 134 kg N ha−1) and tillage practices in KS and; (iii) three years of poultry and dairy manure applications to a silt loam with different tillage practices at the same rate (403 kg N ha−1) in KY. Tillage practices (none vs. conventional) had no effect on the enzyme activities. Principal Component Analyses (PCA) grouped all enzyme activities with the high beef manure application rate after the first year in CO at 0–5 cm. By the second year, the low and high beef manure rates differed in enzyme activities for the KS soil with no difference between the low rate and control in CO. Since the first year of the KY study, acid phosphatase activity was greater in the poultry treated soil compared to dairy or the control; whereas, C cycling enzyme activities were similar in soil treated with dairy or poultry manure. For all studies, PCAs for soil samples from 5–10 cm depth did not reveal treatment separation until the second year, i.e., only high application rate differed from the other treatments. Results of the study indicated significant responses in C and P cycling enzyme activities to manure applications within two years, suggesting potential benefits to soil biogeochemical cycling essential for the productivity of agroecosystems supported with organic fertilizers.
Road Expansion and Its Influence on Trail Sustainability in Bhutan
Authors: Ito Taiichi
Published: December 2011

Bhutan was an inhabited wilderness until 1961, when road construction started after the closure of the Tibetan border. Since then, the road network has expanded from the Indian boarder, often tracing traditional trails. This has accelerated commerce as well as movement of people from India, benefitting both the Bhutanese and foreign tourists. At the same time, dependence on imported automobiles and fossil fuel has risen, and roadless areas have begun to shrink. This brought an inevitable loss of traditional environmental knowledge, such as the care of mules for packing, and reduction in physical and mental health among the Bhutanese. People who lost jobs as horsemen moved into towns to find jobs. Road extension is also a double-edged sword for visitors. It has resulted in shrinking trekking areas and loss of traditional culture, both of which have been sacrificed for easy access. Protected areas often function as fortifications against mechanical civilization. However, protected-area status or its zoning does not guarantee that an area will remain roadless where there is considerable resident population. An analysis in Jigme Dorji National Park showed the gradual retreat of trailheads and increasing dependence on automobiles among residents and trekkers. B. MacKaye, a regional planner in the Eastern United States, proposed using trails as a tool to control such mechanical civilization. His philosophy of regional planning suggests two measures; one is consolidated trailheads as dams, and the other is confinement of roads by levees, consisting of new trails and wilderness belts. According to case studies, the author proposed six options for coexistence of trails with roads.
Multitract Microtransplantation Increases the Yield of DARPP-32-Positive Embryonic Striatal Cells in a Rodent Model of Huntington's Disease
Authors: Jiang Wei , Büchele Fabian , Papazoglou Anna , Döbrössy Máté , Nikkhah Guido
Published: December 2011

Embryonic striatal graft-mediated functional recovery in the rodent lesion model of Huntington's disease (HD) has been shown to correlate with the proportion of dopamine- and adenosine 3′,5′-monophosphate-regulated phosphoprotein with a molecular weight of 32 kDa (DARPP-32)-positive neurons in the graft. The current study investigated the impact of graft distribution on the yield of DARPP-32-positive cells in the grafts following either single-tract or multitract cell delivery protocols using the microtransplantation approach. Cells derived from the whole ganglionic eminence of E15 rat embryos, ubiquitously expressing green fluorescent protein (GFP), were implanted into unilaterally QA-lesioned rat striatum either as 2 × 1.8 μl macrodeposits in a single tract, or as 18 × 0.2 μl microdeposits disseminated over six needle, multitract, penetrations. For both groups, an ultrathin glass capillary with an outer diameter of 50 μm was used. Histological assessment at 4 months after transplantation showed nearly twofold increase of DARRP-32-positive striatal-like neurons in the multitract compared to the single-tract group. However, the cellular make-up of the grafts did not translate into functional differences as tested in a basic spontaneous behavior test. Furthermore, the volumetric values for overall volume, DARPP-32-positive patches, and dopaminergic projection zones were similar between both groups. The results show that distribution of fetal striatal tissue in multiple submicroliter deposits provides for an increased yield of striatal-like neurons, potentially due to the enlargement of the graft–host border area intensifying the graft's exposure to host-derived factors. Furthermore, the use of embryonic tissue from GFP donors was validated in cell-based therapy studies in the HD model.
The Effect of Two- and Three-Dimensional Cell Culture on the Chondrogenic Potential of Human Adipose-Derived Mesenchymal Stem Cells after Subcutaneous Transplantation with an Injectable Hydrogel
Authors: Merceron Christophe , Portron Sophie , Masson Martial , Lesoeur Julie , Fellah Borhane Hakim , Gauthier Olivier , Geffroy Olivier , Weiss Pierre , Guicheux Jérôme , Vinatier Claire
Published: December 2011

Articular cartilage is an avascular tissue composed of chondrocytes, a unique cell type responsible for abundant matrix synthesis and maintenance. When damaged, it never heals spontaneously under physiological circumstances. Therefore, the delivery of mesenchymal stem cells using hydrogel has been considered for cartilage repair. This study aims at investigating the influence of in vitro chondrogenic differentiation of human adipose tissue-derived stem cells (hATSCs) on in vivo cartilage formation when associated with a cellulose-based self-setting hydrogel (Si-HPMC). hATSCs were characterized for their proliferation, surface marker expression, and multipotency. The in vitro chondrogenic potential of hATSCs cultured within Si-HPMC in control or chondrogenic medium was evaluated by measuring COL2A1, ACAN, SOX9, and COMP expression by real-time PCR. Alcian blue and type II collagen staining were also performed. To determine whether in vitro chondrogenically differentiated hATSCs may give rise to cartilage in vivo, cells differentiated as a monolayer or in pellets were finally associated with Si-HPMC and implanted subcutaneously into nude mice. Cartilage formation was assessed histologically by alcian blue and type II collagen staining. Our data demonstrate that hATSCs exhibited proliferation and self-renewal. hATSCs also expressed typical stem cell surface markers and were able to differentiate towards the adipogenic, osteogenic, and chondrogenic lineages. Real-time PCR and histological analysis indicated that Si-HPMC enabled chondrogenic differentiation of hATSCs in inductive medium, as demonstrated by increased expression of chondrogenic markers. In addition, histological analysis of implants showed that chondrogenically differentiated hATSCs (monolayers or pellets) have the ability to form cartilaginous tissue, as indicated by the presence of sulphated glycosaminoglycans and type II collagen. This study therefore suggests that an in vitro induction of hATSCs in 2D was sufficient to obtain cartilaginous tissue formation in vivo. Si-HPMC associated with autologous hATSCs could thus be a significant tool for regenerative medicine in the context of cartilage damage.
Alterations of the Female Reproductive System in Islet Recipient Receiving Immunosuppression
Authors: Del Olmo Garcia Maria Isabel , Lauriola Vincenzo , Aracena Altagracia Gomez , Messinger Shari , Corrales Andrea , Ricordi Camillo , Alejandro Rodolfo
Published: December 2011

Pancreatic islet allotransplantation is an option for patients with unstable type 1 diabetes mellitus (T1DM). Major improvements in islet isolation techniques and the implementation of steroid-free immunosuppressive regimens can maintain insulin independence in the majority of T1DM for at least 1 year after transplantation. Recent studies have emphasized the impact of sirolimus on female reproductive tract. In this communication we report on the alterations of the female reproductive tract in 18 chronically immunosuppressed patients with T1DM following allogenic islet transplantation. Previous research has shown development of ovarian cysts in islet transplant patients receiving sirolimus. We extensively reevaluated this and other possible side effects on the female reproductive system. These side effects have been underestimated, although they are significant, requiring surgical or intensive medical treatment. Pre- and posttransplant gynecological evaluation should be performed to address the development of complications secondary to sirolimus in order to intervene sooner with alternative therapies.
Neural Differentiation of Brain-Derived Neurotrophic Factor-Expressing Human Umbilical Cord Blood-Derived Mesenchymal Stem Cells in Culture via TrkB-Mediated ERK and β-Catenin Phosphorylation and following Transplantation into the Developing Brain
Authors: Lim Jung Yeon , Park Sang In , Kim Seong Muk , Jun Jin Ae , Oh Ji Hyeon , Ryu Chung Hun , Jeong Chang Hyun , Park Sun Hwa , Park Soon A. , Oh Wonil , Chang Jong Wook , Jeun Sin-Soo
Published: December 2011

The ability of mesenchymal stem cells (MSCs) to differentiate into neural cells makes them potential replacement therapeutic candidates in neurological diseases. Presently, overexpression of brain-derived neurotrophic factor (BDNF), which is crucial in the regulation of neural progenitor cell differentiation and maturation during development, was sufficient to convert the mesodermal cell fate of human umbilical cord blood-derived MSCs (hUCB-MSCs) into a neuronal fate in culture, in the absence of specialized induction chemicals. BDNF overexpressing hUCB-MSCs (MSCs-BDNF) yielded an increased number of neuron-like cells and, surprisingly, increased the expression of neuronal phenotype markers in a time-dependent manner compared with control hUCB-MSCs. In addition, MSCs-BDNF exhibited a decreased labeling for MSCs-related antigens such as CD44, CD73, and CD90, and decreased potential to differentiate into mesodermal lineages. Phosphorylation of the receptor tyrosine kinase B (TrkB), which is a receptor of BDNF, was increased significantly in MSC-BDNF. BDNF overexpression also increased the phosphorylation of β-catenin and extracellular signal-regulated kinases (ERKs). Inhibition of TrkB availability by treatment with the TrkB-specific inhibitor K252a blocked the BDNF-stimulated phosphorylation of β-catenin and ERKs, indicating the involvement of both the β-catenin and ERKs signals in the BDNF-stimulated and TrkB-mediated neural differentiation of hUCB-MSCs. Reduction of β-catenin availability using small interfering RNA-mediated gene silencing inhibited ERKs phosphorylation. However, β-catenin activation was maintained. In addition, inhibition of β-catenin and ERKs expression levels abrogated the BDNF-stimulated upregulation of neuronal phenotype markers. Furthermore, MSC-BDNF survived and migrated more extensively when grafted into the lateral ventricles of neonatal mouse brain, and differentiated significantly into neurons in the olfactory bulb and periventricular astrocytes. These results indicate that BDNF induces the neural differentiation of hUCB-MSCs in culture via the TrkB-mediated phosphorylation of ERKs and β-catenin and following transplantation into the developing brain.
Mesenchymal Stem Cells Restore Lung Function by Recruiting Resident and Nonresident Proteins
Authors: Jungebluth Philipp , Luedde Mark , Ferrer Elisabet , Luedde Tom , Vucur Mihael , Peinado Victor I. , Go Tetsuhiko , Schreiber Catharina , Von Richthofen Maximilian , Bader Augustinus , Haag Johannes , Darsow Kai H. , Bartel Sebastian J. , Lange Harald A. , Furlani Dario , Steinhoff Gustav , Macchiarini Paolo
Published: December 2011

Because human lungs are unlikely to repair or regenerate beyond the cellular level, cell therapy has not previously been considered for chronic irreversible obstructive lung diseases. To explore whether cell therapy can restore lung function, we administered allogenic intratracheal mesenchymal stem cells (MSCs) in the trachea of rats with chronic thromboembolic pulmonary hypertension (CTEPH), a disease characterized by single or recurrent pulmonary thromboembolic obliteration and progressive pulmonary vascular remodeling. MSCs were retrieved only in high pressure-exposed lungs recruited via a homing stromal derived factor-1α/ CXCR4 pathway. After MSC administration, a marked and long-lasting improvement of all clinical parameters and a significant change of the proteome level were detected. Beside a variation of liver proteome, such as caspase-3, NF-κB, collagen1A1, and α-SMA, we also identified more than 300 resident and nonresident lung proteins [e.g., myosin light chain 3 (P16409) or mitochondrial ATP synthase subunit alpha (P15999)]. These results suggest that cell therapy restores lung function and the therapeutic effects of MSCs may be related to protein-based tissue reconstituting effects.
Human Mesenchymal Stem Cells Are Susceptible to Lysis by CD8+ T Cells and NK Cells
Authors: Crop Meindert J. , Korevaar Sander S. , De Kuiper Ronella , Ijzermans Jan N. M. , Van Besouw Nicole M. , Baan Carla C. , Weimar Willem , Hoogduijn Martin J.
Published: December 2011

There is growing interest in the use of mesenchymal stem cells (MSCs) to improve the outcome of organ transplantation. The immunogenicity of MSCs is, however, unclear and is important for the efficacy of MSC therapy and for potential sensitization against donor antigens. We investigated the susceptibility of autologous and allogeneic MSCs for lysis by CD8+ T-lymphocytes and NK cells in a kidney transplant setting. MSCs were derived from adipose tissue of human kidney donors and were CD90+, CD105+, CD166+, and HLA class I+. They showed differentiation ability and immunosuppressive capacity. Lysis of MSCs by peripheral blood mononuclear cells (PBMCs), FACS-sorted CD8+ T cells, and NK cells was measured by europium release assay. Allogeneic MSCs were susceptible for lysis by cytotoxic CD8+ T cells and NK cells, while autologous MSCs were lysed by NK cells only. NK cell-mediated lysis was inversely correlated with the expression of HLA class I on MSCs. Lysis of autologous MSCs was not dependent on culturing of MSCs in FBS, and MSCs in suspension as well as adherent to plastic were lysed by NK cells. Pretransplant recipient PBMCs did not lyse donor MSCs, but PBMCs isolated 3, 6, and 12 months after transplantation showed increasing lysing ability. After 12 months, CD8+ T-cell-mediated lysis of donor MSCs persisted, indicating there was no evidence for desensitization against donor MSCs. Lysis of MSCs is important to take into account when MSCs are considered for clinical application. Our results suggest that the HLA background of MSCs and timing of MSC administration are important for the efficacy of MSC therapy.
Characterization of Human Umbilical Cord Lining-Derived Epithelial Cells and Transplantation Potential
Authors: Zhou Yue , Gan Shu Uin , Lin Gen , Lim Yan Ting , Masilamani Jeyakumar , Mustafa Fatimah Bte , Phua Meow Ling , Rivino Laura , Phan Toan Thang , Lee Kok Onn , Calne Roy , Macary Paul A.
Published: December 2011

In this study we describe the derivation and immunological characterization of a primary epithelial cell type from the human umbilical cord membrane. These cord lining epithelial cells (CLECs) expressed and/or secreted isoforms of the nonclassical human leukocyte antigen class I (HLA-1b) glycoproteins, HLA-G and E. Conditioned media from CLECs inhibited mitogen-stimulated T-lymphocyte responses, and in a mixed leukocyte reaction (MLR) assay, cocultured CLECs inhibited allogeneic responses with a concomitant reduction in proinflammatory cytokines. Using a transwell coculture system, it was demonstrated that these immunoregulatory effects were mediated by soluble factors secreted by CLECs, in a dose-dependent manner. Functional studies using HLA-G blocking antibody showed that the effects of CLEC-secreted products could be inhibited, thus demonstrating a significant and important role for soluble HLA-G. In vivo, we show that transplanted CLECs could be maintained for extended periods in immunocompetent mice where xenorejection rapidly destroyed primary keratinocytes, a control human epithelial cell type. Additionally, CLECs delayed the rejection of keratinocytes and extended their survival when cotransplanted, indicating an ability to protect adjacent human cell types that would otherwise be rejected if transplanted alone. We also show that CLECs transduced with a modified human proinsulin gene were transplanted intraperitoneally into streptozotocin (STZ)-induced diabetic mice, resulting in significantly lower levels of serum glucose compared to control mice. This study has characterized the immunological properties of CLECs and tested a potential therapeutic application in the treatment of a type 1 diabetes mouse model.
Highly Purified versus Filtered Crude Collagenase: Comparable Human Islet Isolation Outcomes
Authors: Wang Yong , Paushter Daniel , Wang Shusen , Barbaro Barbara , Harvat Tricia , Danielson Kirstie K. , Kinzer Katie , Zhang Liza , Qi Meirigeng , Oberholzer Jose
Published: December 2011

This study was designed to retrospectively compare the impact of crude Sigma V collagenase (Sigma V, n = 52) with high-purified Serva NB1 collagenase (Serva NB1, n = 42) on human islet isolation outcomes. A three-step filtration was applied to the crude Sigma V to remove endotoxin contamination and impurities; in addition, this process was used as a lot prescreening tool. Isolation outcomes were determined by digestion efficacy, islet yields, purity, viability, glucose-stimulated insulin release, and endotoxin content. The digestion efficacy between Sigma V and Serva NB1 was statistically significant (Sigma V: 64.71% vs. Serva NB1: 69.71%, p = 0.0014). However, the islet yields were similar (Sigma V: 23422.58 vs. Serva NB1: 271097 IEq, p = 0.23) between groups. There was no significant purity difference observed in fractions with purities greater than 75%. Viability (Sigma V: 93.3% vs. Serva NB1: 94.8%, p = 0.061) and stimulation indexes (Sigma V: 3.41 vs. Serva NB1: 2.74, p = 0.187) were also similar between the two groups. The impact of cold ischemia and age on the isolation outcome in the Sigma V group was comparable to the Serva NB1 group. The endotoxin content of the final products in the filtered Sigma V group was significantly less than that in the high-purified Serva NB1 group (0.022 vs. 0.052 EU/ml, p = 0.003). Additionally, in the Sigma V group there was minimal lot to lot variation and no significant loss of enzymatic activity after filtration. These findings indicate that the use of Sigma V or other crude enzyme blends for research pancreata is warranted to reduce isolation costs and increase the amount of islets available for critical islet research. These findings also validate the need for a systematic enzyme analysis to resolve these inconsistencies in overall enzyme quality once and for all.
The Time is Crucial for Ex Vivo Expansion of T Regulatory Cells for Therapy
Authors: Marek Natalia , Bieniaszewska Maria , Krzystyniak Adam , Juścińska Jolanta , Myśliwska Jolanta , Witkowski Piotr , Hellmann Andrzej , Trzonkowski Piotr
Published: December 2011

Ex vivo expanded CD4+CD25highCD127- T regulatory cells (Tregs) are recognized as a promising candidate for immunosuppressive therapy in humans. However, due to the plasticity of Tregs lineage and artificial environment present during ex vivo expansion, Tregs easily lose suppressive activity. Here, we followed expanding CD4+CD25highCD127- Tregs and their naive (CD45RA+) and memory-like (CD45RA-) subsets in order to establish the best conditions of the expansion. We found that, regardless of the phenotype sorted, expanding Tregs were undergoing changes resembling homeostatic proliferation and transformed into effector memory-like cells which produced not only suppressive interleukin-10 (IL-10) but also IL-6, IL-17, and interferon-γ (IFN-γ). With the time ex vivo, Tregs were losing the expression of FoxP3 and suppressive activity both when stimulated and when at rest. The only variable that helped preserve suppressive abilities of Tregs was the limitation of the time of ex vivo cultures to 2 weeks only. According to our study, the highest number of highly suppressive Tregs could be yielded with CD4+CD25highCD127- Tregs cultured no longer than 2 weeks. Thorough quality check, preferentially with the assessment of FoxP3 expression and IFN-γ suppression assay, should be applied to assess suppressive activity of the cells.
Human CD47 Expression Permits Survival of Porcine Cells in Immunodeficient Mice that Express SIRPα Capable of Binding to Human CD47
Authors: Wang Chunfeng , Wang Hui , Ide Kentaro , Wang Yuantao , Van Rooijen Nico , Ohdan Hideki , Yang Yong-Guang
Published: December 2011

Signal regulatory protein α (SIRPα) is a critical immune inhibitory receptor on macrophages, and its interaction with CD47 prevents autologous phagocytosis. We have previously shown that pig CD47 does not interact with human SIRPα, and that human CD47 expression inhibits phagocytosis of porcine cells by human macrophages in vitro. In this study, we have investigated the potential of human CD47 expression to promote porcine cell survival in vivo. Human CD47-expressing and control porcine B-lymphoma cells were transplanted into T- and B-cell-deficient nonobese diabetic/severe combined immunodeficient (NOD/SCID) mice that express SIRPα capable of interacting with human CD47. Only the human CD47-expressing porcine lymphoma cells survived and were able to form tumors in NOD/SCID mice; however, both the control and human CD47-expressing porcine cells survived in macrophage-depleted NOD/SCID mice. These results indicate that transgenic expression of human CD47 may provide an effective approach to inhibiting macrophage-mediated xenograft rejection in clinical xenotransplantation.
Anti-Inflammatory Properties of Exenatide in Human Pancreatic Islets
Authors: Cechin S. R. , Pérez-Álvarez I. , Fenjves E. , Molano R. D. , Pileggi A. , Berggren P-O. , Ricordi C. , Pastori R. L.
Published: December 2011

Exenatide is an analog of the incretin hormone glucagon-like peptide (GLP-1) that is used for the treatment of T2D for their metabolic effects. In addition to its insulinotropic effects, exenatide increases functional islet mass and improves their survival. Improved outcomes have been reported in recent clinical islet transplantation trials for the treatment of type 1 diabetes. The purpose of this study was to investigate whether exenatide has anti-inflammatory properties in human islets. Exenatide treatment improved islet function, significantly reduced content of inflammation-related molecules (tissue factor, IFN-γ, IL-17, IL-1β, and IL-2) and caspase 3 activation, whereas increased phosphorylation of ERK1/2, STAT3, and Akt in vitro. Immunostaining showed expression of GLP-1R in β-cells but not in α-cells. IL-1β colocalized with GLP-1R in β-cells. Induction of serine proteinase inhibitor 9 (PI-9) was detected after exposure of human islets to exenatide in vitro and after transplantation into immunodeficient mice. GLP-1 induced PI-9 expression in vitro but to a lower extent than exenatide. This effect was partially blocked by the antagonist exendin-9 in vitro. As assessed by immunostaining PI-9 is mostly expressed in β-cells but not in α-cells. In conclusion, we describe anti-inflammatory and cytoprotective properties of exenatide in human islets. Exenatide-mediated PI-9 expression, the only known granzyme B inhibitor, unveils potential immunoregulatory properties.
Improvement of the Cold Storage of Isolated Human Hepatocytes
Authors: Pless Gesine , Sauer Igor M. , Rauen Ursula
Published: December 2011

Increasing amounts of human hepatocytes are needed for clinical applications and different fields of research, such as cell transplantation, bioartificial liver support, and pharmacological testing. This demand calls for adequate storage options for isolated human liver cells. As cryopreservation results in severe cryoinjury, short-term storage is currently performed at 2–8°C in preservation solutions developed for the storage of solid organs. However, besides slowing down cell metabolism, cold also induces cell injury, which is, in many cell types, iron dependent and not counteracted by current storage solutions. In this study, we aimed to characterize storage injury to human hepatocytes and develop a customized solution for cold storage of these cells. Human hepatocytes were isolated from material obtained from partial liver resections, seeded in monolayer cultures, and, after a preculture period, stored in the cold in classical and new solutions followed by rewarming in cell culture medium. Human hepatocytes displayed cold-induced injury, resulting in >80% cell death (LDH release) after 1 week of cold storage in University of Wisconsin solution or cell culture medium and 3 h of rewarming. Cold-induced injury could be significantly reduced by the addition of the iron chelators deferoxamine and LK 614. Experiments with modified solutions based on the new organ preservation solution Custodiol-N showed that ion-rich variants were better than ion-poor variants, chloride-rich solutions better than chloride-poor solutions, potassium as main cation superior to sodium, and pH 7.0 superior to pH 7.4. LDH release after 2 weeks of cold storage in the thus optimized solution was below 20%, greatly improving cold storage of human hepatocytes. The results were confirmed by the assessment of hepatocellular mitochondrial membrane potential and functional parameters (resazurin reduction, glucagon-stimulated glucose liberation) and thus suggest the use of a customized hepatocyte storage solution for the cold storage of these cells.