bluuebunny/crossref_metadata_2025_split huggingface dataset:
Reference distributions for alpha2‐macroglobulin: A practical, simple and clinically relevant approach in a large cohort
Authors: Ritchie Robert F. , Palomaki Glenn E. , Neveux Louis M. , Navolotskaia Olga , Ledue Thomas B. , Craig Wendy Y. 
Published: March 2004

AbstractIn this 11th article in a series, reference values of serum levels alpha2‐macroglobulin α2M) are examined. The study is based on a cohort of 40,420 Caucasian individuals from northern New England that were tested in our laboratory between 1994 and 2000. Measurements were standardized against Certified Reference Material (CRM 470)/Reference Preparation for Proteins in Human Serum (RPPHS) and the results analyzed using a previously described statistical approach. Individuals with unequivocal laboratory evidence of inflammation (C‐reactive protein >10 mg/L) were excluded in one leg of the study and included in the other, confirming that α2M does not respond to acute phase drive in man. Nephrotic syndrome, diabetes mellitus, and chronic liver disease have significant effect on levels of α2M. Dramatic changes occur during life with males higher from birth to age 12, females thereafter have higher values until the ninth decade. When values were expressed as multiples of the age‐ and gender‐specific median levels, the resulting distributions fitted a log‐Gaussian distribution well over a broad range. When patient data are normalized in this manner, the distribution parameters can be used to assign a centile corresponding to an individual's measurement thus simplifying interpretation. J. Clin. Lab. Anal. 18:139–147, 2004. © 2004 Wiley‐Liss, Inc.
Lamprey contractile protein genes mark different populations of skeletal muscles during development
Authors: Kusakabe Rie , Takechi Masaki , Tochinai Shin , Kuratani Shigeru 
Published: March 2004

AbstractAgnathan lampreys retain ancestral characteristics of vertebrates in the morphology of skeletal muscles derived from two mesodermal regions: trunk myotomes and unsegmented head mesoderm. During lamprey development, some populations of myoblasts migrate via pathways that differ from those of gnathostomes. To investigate the evolution of skeletal muscle differentiation in vertebrates, we characterize multiple contractile protein genes expressed in the muscle cells of the Japanese lamprey, Lethenteron japonicum. Lamprey actin gene LjMA2, and myosin heavy chain (MyHC) genes LjMyHC1 and LjMyHC2 are all expressed in the developing skeletal muscle cells of early embryos. However, LjMyHC1 and LjMyHC2 are expressed only in cells originating from myotomes, while LjMA2 is expressed in both myotomal and head musculature. Thus, in lampreys, myotomes and head mesoderm differ in the use of genes encoding contractile protein isoforms. Phylogenetic tree analyses including lamprey MyHCs suggest that the variety of muscle MyHC isoforms in different skeletal muscles may correspond to the morphological complexity of skeletal muscles of different vertebrate species. Another lamprey actin gene LjMA1 is likely to be the first smooth muscle actin gene isolated from non‐tetrapods. We conclude that, in vertebrate evolution, the different regulatory systems for striated and smooth muscle‐specific genes may have been established before the agnathan/gnathostome divergence. J. Exp. Zool. (Mol. Dev. Evol.) 302B:121‐133, 2004. © 2004 Wiley‐Liss, Inc.
S100A1 gene transfer: a strategy to strengthen engineered cardiac grafts
Authors: Remppis Andrew , Pleger Sven T. , Most Patrick , Lindenkamp Jan , Ehlermann Philipp , Schweda Christopher , Löffler Eva , Weichenhan Dieter , Zimmermann Wolfram , Eschenhagen Thomas , Koch Walter J. , Katus Hugo A. 
Published: March 2004

AbstractBackgroundCardiac tissue replacement therapy, although a promising novel approach for the potential treatment of heart failure, still suffers from insufficient contractile support to the failing myocardium. Here, we explore a strategy to improve contractile properties of engineered heart tissue (EHT) by S100A1 gene transfer.MethodsEHTs were generated from neonatal rat cardiomyocytes and transfected (MOI 10 PFU) with the S100A1 adenovirus (AdvS100A1, n = 25) while an adenovirus devoid of the S100A1 cDNA served as a control (AdvGFP, n = 30). Contractile properties of transfected EHTs were measured 7 days following gene transfer.ResultsWestern blot analysis confirmed a 8.7 ± 3.6‐fold S100A1 protein overexpression in AdvS100A1‐transfected EHTs (n = 4; P < 0.01) that increased maximal isometric force (mN; AdvGFP 0.175 ± 0.03 vs. AdvS100A1 0.47 ± 0.06; P < 0.05) at 0.4 mmol/L extracellular calcium concentration [Ca2+]e. In addition, S100A1 overexpression enhanced both maximal Ca2+‐stimulated force generation (+81%; P < 0.05) and Ca2+‐sensitivity of EHTs (EC50% [Ca2+]e mM; AdvGFP 0.33 ± 0.04 vs. AdvS100A1 0.21 ± 0.0022; P < 0.05). The S100A1‐mediated gain in basal graft contractility was preserved throughout a series of isoproterenol interventions (10−9 to 10−6 M). Physiological properties of EHTs resembling intact heart preparations were preserved.ConclusionsS100A1 gene transfer in EHT is feasible and augments contractile performance, while characteristic physiological features of EHT remain unchanged. Thus, specific genetic manipulation of tissue constructs prior to implantation should be part of an improved tissue replacement strategy in heart failure. Copyright © 2004 John Wiley & Sons, Ltd.
Transcriptional silencing is associated with extensive methylation of the CMV promoter following adenoviral gene delivery to muscle
Authors: Brooks Alan R. , Harkins Richard N. , Wang Peiyin , Qian Hu Sheng , Liu Pengxuan , Rubanyi Gabor M. 
Published: March 2004

AbstractBackgroundAlthough the transient nature of transgene expression using first‐generation adenovirus (Ad) vectors is well known, the exact mechanisms responsible for this phenomenon are uncertain.MethodsRats were given intramuscular (i.m.) injections of a first‐generation Ad containing the human fibroblast growth factor 4 (hFGF‐4) gene driven by the cytomegalovirus (CMV) promoter and enhancer (CMV‐PE). The copy number of hFGF‐4 mRNA and viral DNA was measured in the same muscles by quantitative RT‐PCR and quantitative PCR at times between 1 h and 84 days after virus injection. Quantitative Southern blot analysis for the intact hFGF‐4 transcription unit DNA was also performed, and the methylation status of the CMV‐PE DNA in the muscle was determined using bisulfite sequencing.ResultsThe copy number of hFGF‐4 mRNA peaked at 6 h then decreased 56‐fold by 24 h, and a further 240‐fold between days 3 and 28. Although the viral DNA copy number also decreased 23‐fold between 6 h and 28 days, the ratio of copies of hFGF‐4 mRNA per copy of viral DNA decreased 385‐fold over this period. Methylation of the CMV‐PE DNA in the muscle at both CpG and non‐CpG sites was observed 24 h after virus administration and had increased at day 7.ConclusionsDecreased transcription associated with extensive methylation of the CMV‐PE was the major mechanism responsible for the decrease in transgene mRNA levels. Strategies for preventing transcriptional silencing will be valuable for improving the duration of transgene expression from adenoviral vectors. Copyright © 2004 John Wiley & Sons, Ltd.
CTb targeted non‐viral cDNA delivery enhances transgene expression in neurons
Authors: Barrett L. B. , Berry M. , Ying W.‐B. , Hodgkin M. N. , Seymour L. W. , Gonzalez A.‐M. , Read M. L. , Baird A. , Logan A. 
Published: March 2004

AbstractBackgroundEfficient neuronal gene therapy is a goal for the long‐term repair and regeneration of the injured central nervous system (CNS). We investigated whether targeting cDNA to neurons with cholera toxin b chain conjugated non‐viral polyplexes led to increased efficiency of non‐viral gene transfer in the CNS. Here, we illustrate the potential for this strategy by demonstrating enhanced transfection of a differentiated neuronal cell type, PC12.MethodsIn vitro transfection efficiency of a cholera toxin b chain–poly(D‐lysine) molecular conjugate (CTb‐K100) was compared by fluorescence‐activated cell sorting (FACS) analysis of green fluorescent protein (GFP) expression and luminometric measurement of β‐galactosidase (β‐gal) expression, to untargeted poly(D‐lysine) (K100) in undifferentiated and NGF‐differentiated PC12 cells.ResultsTransfection of undifferentiated PC12 cells with CTb‐K100 polyplexes resulted in a 36‐fold increase in levels of pCMV‐DNALacZ expression and a 20‐fold increase in the frequency of transduction with pCMV‐DNAGFP, compared with untargeted K100 polyplexes. Treatment of PC12 cells with 50 ng/ml/day of NGF for 14 days led to differentiation to a neuronal phenotype. Transfection of NGF‐differentiated cells with CTb‐K100 polyplexes resulted in a 133‐fold increase in levels of pCMV‐DNALacZ expression and a 11‐fold increase in the percentage of cells transduced with pCMV‐DNAGFP, compared with untargeted K100 polyplexes. Transfection was dependent on CTb, with CTb‐K100‐mediated transfections competitively inhibited with free CTb in both PC12 phenotypes.ConclusionsNon‐viral systems for gene transfer in damaged CNS show superior toxicological profiles to most viruses but are limited by inefficient and non‐selective gene expression in target tissue. Cholera toxin is known to interact preferentially with neuronal cells of the central and peripheral nervous systems, mediating binding through the b subunit, CTb, and the pentasaccharide moiety of the gangliosaccharide, GM1, which is present at high levels on the neuronal cell surface. Here, we show that a molecular conjugate of the CTb subunit, covalently linked to poly(D‐lysine), is able to successfully target and significantly enhance transfection of a neuronal cell type, NGF‐differentiated rat PC12 pheochromocytoma cells. This observation encourages the further development of non‐viral strategies for the delivery of therapeutic genes to neurons. Copyright © 2004 John Wiley & Sons, Ltd.
Efficient transduction and selection of human T‐lymphocytes with bicistronic Thy1/HSV1‐TK retroviral vector produced by a human packaging cell line
Authors: Lemoine François M. , Mesel‐Lemoine Mariana , Cherai Mustapha , Gallot Géraldine , Vié Henri , Leclercq Virginie , Trébèden‐Negre Hélène , Mammès Olivier , Boyer Olivier , Noguiez‐Hellin Patricia , Klatzmann David 
Published: March 2004

AbstractBackgroundT‐cells expressing the HSV1‐TK suicide gene can be used for the control of graft‐versus‐host disease following allogeneic stem cell transplantation. To develop clinical trials based on such a strategy, we have generated under good manufacturing procedures a novel ‘split genome’ human packaging cell line (1704 cells).MethodsTo minimize the risk of generating replication‐competent retroviruses, pol was truncated to remove sequences overlapping with env. To improve retroviral infection and selection of transduced T‐cells, high titers of GALV‐pseudotyped retroviral particles harboring a bicistronic Thy1‐IRES‐TK vector coding for the CD90 GPI‐anchored membrane molecule were produced by 1704 cells.ResultsUsing 1704 cell supernatant and an optimized transduction protocol, approximately 50% of primary T‐cells were transduced and could then be purified (∼95%) using clinical‐grade immunomagnetic beads directed against CD90. Over 96% of these OKT3/IL‐2‐activated CD90+‐selected T‐cells were killed by ganciclovir. Cell proliferation and cytokine production of transduced T‐cells and HLA‐restricted cytotoxicity of transduced T‐cell clones were identical to those of their non‐transduced counterparts cultured under the same conditions.ConclusionsGALV‐pseudotyped retroviral particles harboring a bicistronic Thy1‐IRES‐TK vector allow efficient transduction and rapid selection of human T‐cells under conditions applicable for clinical trials using the new human 1704 packaging cell line. Copyright © 2004 John Wiley & Sons, Ltd.
1H‐MRSI of radiation effects in normal‐appearing white matter: Dose‐dependence and impact on automated spectral classification
Authors: Lee Michael C. , Pirzkall Andrea , McKnight Tracy R. , Nelson Sarah J. 
Published: March 2004

AbstractPurposeTo identify radiation‐induced changes in healthy white‐matter spectra in the first six months following radiotherapy, and assess the impact of these changes on an automated algorithm for detecting spectral abnormalities.Materials and Methods1H‐MRSI was performed on 10 patients with grade IV gliomas who were to undergo radiation therapy. Choline (Cho), creatine (Cr), and N‐acetylaspartate (NAA) ratios were studied as a function of dose and time. The impact of these spectral changes on a spectral analysis algorithm was evaluated.ResultsThe Cho/NAA ratios rose to values of 0.66 ± 0.15, 0.75 ± 0.21, and 0.73 ± 0.15 two months after therapy, compared to immediate post‐therapy values of 0.56 ± 0.15, 0.60 ± 0.16, and 0.61 ± 0.15 for the < 25, 25–50, and > 50 Gy dose groups, respectively. These maxima were followed by a dose‐dependent recovery. A similar trend was found in the Cho/Cr ratio. The automated spectral analysis system incorporated the changing Cho/NAA ratio into a global redefinition of healthy tissue, but did not account for dose‐dependent spatial variations in Cho/NAA ratios.ConclusionRadiation significantly alters the spectra of healthy tissues in the first six months after radiotherapy. This suggests that the radiation dose distribution should be considered during analysis of post‐therapy spectra. J. Magn. Reson. Imaging 2004;19:379–388. © 2004 Wiley‐Liss, Inc.
Comparison of TCA and ICA techniques in fMRI data processing
Authors: Zhao Xia , Glahn David , Tan Li Hai , Li Ning , Xiong Jinhu , Gao Jia‐Hong 
Published: March 2004

AbstractPurposeTo make a quantitative comparison of temporal cluster analysis (TCA) and independent component analysis (ICA) techniques in detecting brain activation by using simulated data and in vivo event‐related functional MRI (fMRI) experiments.Materials and MethodsA single‐slice MRI image was replicated 150 times to simulate an fMRI time series. An event‐related brain activation pattern with five different levels of intensity and Gaussian noise was superimposed on these images. Maximum contrast‐to‐noise ratio (CNR) of the signal change ranged from 1.0 to 2.0 by 0.25 increments. In vivo visual stimulation fMRI experiments were performed on a 1.9 T magnet. Six human volunteers participated in this study. All imaging data were analyzed using both TCA and ICA methods.ResultsBoth simulated and in vivo data have shown that no statistically significant difference exists in the activation areas detected by both ICA and TCA techniques when CNR of fMRI signal is larger than 1.75.ConclusionTCA and ICA techniques are comparable in generating functional brain maps in event‐related fMRI experiments. Although ICA has richer features in exploring the spatial and temporal information of the functional images, the TCA method has advantages in its computational efficiency, repeatability, and readiness to average data from group subjects. J. Magn. Reson. Imaging 2004;19:397–402. © 2004 Wiley‐Liss, Inc.
Tumor R2* is a prognostic indicator of acute radiotherapeutic response in rodent tumors
Authors: Rodrigues Loreta M. , Howe Franklyn A. , Griffiths John R. , Robinson Simon P. 
Published: March 2004

AbstractPurposeTo test the prognostic potential of tumor R2* with respect to radiotherapeutic outcome. Blood oxygenation level dependent (BOLD) MRI images are sensitive to changes in deoxyhemoglobin concentration through the transverse MRI relaxation rate R2* of tissue water, hence the quantitative measurement of tumor R2* may be related to tissue oxygenation.Methods and MaterialsTumor growth inhibition in response to radiation was established for both GH3 prolactinomas and RIF‐1 fibrosarcomas with animals breathing either air or carbogen during radiation. In a separate cohort, the baseline R2* and carbogen (95% O2, 5% CO2)‐induced ΔR2* of rat GH3 prolactinomas and murine RIF‐1 fibrosarcomas were quantified using multigradient echo (MGRE) MRI prior to radiotherapy, and correlated with subsequent tumor growth inhibition in response to ionizing radiation, while the animals breathed air.ResultsA radiation dose of 15 Gy caused pronounced growth delay in both tumor models and transient regression of the GH3 prolactinomas. When the animals breathed carbogen during radiation, the growth delay/regression was enhanced only in the GH3 prolactinomas. The GH3 prolactinomas, which exhibit a relatively fast baseline R2* and large ΔR2* in response to carbogen breathing prior to radiotherapy, showed a substantial reduction in normalized tumor volume to 66 ± 3% with air breathing and 36 ± 5% with carbogen seven days after 15 Gy irradiation. In contrast, the effect of 15 Gy on the RIF‐1 fibrosarcomas, which give a relatively slow baseline R2* and negligible ΔR2* response to carbogen prior to treatment, showed a much smaller growth inhibition (143 ± 3% with air, 133 ± 12% with carbogen).ConclusionQuantitation of tumor R2* and carbogen‐induced ΔR2* by MGRE MRI provides completely noninvasive prognostic indicators of a potential acute radiotherapeutic response. J. Magn. Reson. Imaging 2004;19:482–488. © 2004 Wiley‐Liss, Inc.
Time‐of‐flight quantitative measurements of blood flow in mouse hindlimbs
Authors: Wagner Shawn , Helisch Armin , Bachmann Georg , Schaper Wolfgang 
Published: March 2004

AbstractPurposeTo evaluate the feasibility of using time‐of‐flight (TOF) imaging to directly measure hindlimb blood flow in a mouse model of peripheral vascular disease.Materials and MethodsFour tubes were imaged simultaneously (diameters = 0.39 mm, 0.59 mm, and two at 1.46 mm) with a 1.0 mM copper sulfate solution for 19 flow velocities. In vivo measurements were performed in the hindlimbs of three mouse strains—C57BL/6 (N = 5), BALB/c (N = 5), and 129S2/Sv (N = 5)—three weeks after femoral artery ligation with a calibration standard.ResultsThe flow phantom showed that the intensity was linear (r2 = 0.92) over the pertinent blood flow velocities in the mouse hindlimbs. Measurements of the blood flow in the distal hindlimbs in different strains of mice (combination of both the venous and arterial flows) were obtained 21 days after right‐sided femoral artery occlusion. The results showed that under similar conditions of anesthesia and temperature, SV129 mice on the nonligated side had the highest flows (0.50 ± 0.07 mL/minute), followed by C57BL/6 (0.28 ± 0.04 mL/minute) and BALB/c (0.23 ± 0.05 mL/minute), P < 0.02. The ligated side measurements (SV129, 0.31 ± 0.05 mL/minute (P = 0.02); C57BL/6, 0.21 ± 0.02 mL/minute (P = 0.13); and BALB/c, 0.12 ± 0.02 mL/minute (P= 0.06)) showed a trend in BALB/c and C57BL/6 and significant differences in SV129 for incomplete recovery three weeks after surgery, compared to the nonligated side.ConclusionTwo‐dimensional TOF imaging permits quantitative in vivo measurements of hindlimb blood flow in a mouse model of peripheral vascular disease without the need of contrast injection, offering advantages of serial imaging not limited by tissue penetration. J. Magn. Reson. Imaging 2004;19:468–474. © 2004 Wiley‐Liss, Inc.
Screening of colonic tumors by air‐inflated magnetic resonance (MR) colonography
Authors: Lam Wynnie W.M. , Leung Wai K. , Wu Justin K.L. , So Nina M.C. , Sung Joseph J.Y. 
Published: March 2004

AbstractPurposeTo assess the accuracy of air‐inflated magnetic resonance (MR) colonography for the detection of colonic lesions.Materials and MethodsA total of 36 patients underwent both colonoscopy and air‐inflated MR colonography. Breath‐hold sequences (volumetric interpolated breath‐hold examination (VIBE) coronal, and half‐Fourier acquisition single‐shot turbo spin‐echo (HASTE) axial and coronal, both supine and prone) were performed with a 1.5T scanner. The detection of colonic lesions by MR colonography was then correlated with the findings from the colonoscopy performed on the same day.ResultsTwo patients were unable to complete the MR colonography examination. Analysis was based on the results from 34 patients (17 males and 17 females, 38–70 years old, mean age = 54.9 years) who completed both examinations. MR colonography depicted two of two colonic tumors, one of one P4 (>2 cm) polyp, one of two P2 (0.5–1 cm) polyps, and two of 11 P1 (<0.5 cm) polyps. False‐positive MR colonography interpretations were noted for one P1 polyp and two P2 polyps. The overall sensitivity, positive predictive value, and accuracy of MR colonography were 38%, 67%, and 46.2%, respectively. For the detection of endoluminal lesions >5 mm, air‐inflated MR colonography yielded a sensitivity of 75%, specificity of 93.3%, accuracy of 91.2%, positive predictive value of 60%, and negative predictive value of 96.6%.ConclusionAir‐inflated MR colonography is a new technique that deserves further investigation. J. Magn. Reson. Imaging 2004;19:447–452. © 2004 Wiley‐Liss, Inc.
Real‐time high‐resolution MRI for the assessment of gastric motility: Pre‐ and postpharmacological stimuli
Authors: Ajaj Waleed , Lauenstein Thomas , Papanikolaou Nickolas , Holtmann Gerald , Goehde Susanne C. , Ruehm Stefan G. , Debatin Joerg F. 
Published: March 2004

AbstractPurposeTo determine the practicality of MRI using a new real‐time sequence for the assessment of gastric motion, and quantify the effects of motility‐modifying substances.Materials and MethodsSix healthy volunteers ingested 400 mL of a high‐calorie liquid nutrient. Two‐dimensional real‐time TrueFISP sequences were acquired for up to 30 minutes following the ingestion. The acquisition plane was chosen parallel to the axis of the gastric antrum. The examination was performed on three separate days with and without i.v. administration of 10 mg metoclopramide or 20 mg scopolamine. A motility index was calculated for each real‐time data set.ResultsDelineation of the gastric lumen proved easy and robust. The intravenous application of motility‐modifying agents resulted in significant changes in the motility index. The administration of metoclopramide resulted in an average increase of the index by a factor of 1.5, whereas the application of scopolamine led to a decrease of the index by a factor of 3.0.ConclusionTrueFISP MRI performed well in depicting the gastric lumen and assessing gastric motility. Furthermore, we were able to evaluate and quantify the effect of motility‐modifying agents. The noninvasive nature of MRI makes this imaging modality an attractive alternative to conventional invasive diagnostic tools for gastric motility disorders and monitoring of therapy. J. Magn. Reson. Imaging 2004;19:453–458. © 2004 Wiley‐Liss, Inc.
Tolerance of MRI vs. SPECT myocardial perfusion studies—A patient survey
Authors: Sparrow Patrick , Plein Sven , Jones Tim R. , Thorley Penelope J. , Hale Claire , Sivananthan Mohan U. 
Published: March 2004

AbstractPurposeTo compare patients' perceived satisfaction and tolerance of magnetic resonance imaging (MRI) and single photon emission computed tomography (SPECT) myocardial perfusion imaging.Materials and MethodsA questionnaire was sent retrospectively to 41 patients who had undergone both SPECT and MRI myocardial perfusion scans at our institution. The questionnaire assessed SPECT and MRI separately, and in a separate section compared the tests directly. The answers were scored and analyzed for statistical significance by the use of Wilcoxon signed‐ranks and χ2 tests.ResultsThirty‐five completed questionnaires were returned. In a direct comparison, 12 patients (34%) preferred MRI overall, nine (26%) preferred SPECT, and 14 (40%) expressed no preference. The ratings for the overall comfort of the scans were similar, with a score of 5.8 for SPECT and 5.7 for MRI (on a scale of 1–10). More patients stated a preference for MRI on scan comfort, duration, and safety (no statistical significance), but it was less well rated than SPECT for space on the scanner (P = 0.008). Three patients (9%) stated that they would not have an MRI scan again, while two patients (6%) said they would not repeat a SPECT scan.ConclusionMRI myocardial perfusion imaging represents an acceptable alternative to SPECT with respect to patient tolerance and satisfaction. J. Magn. Reson. Imaging 2004;19:410–416. © 2004 Wiley‐Liss, Inc.