Biomedical subjects

Sang-Hun Lee

Publications and source records attributed to Sang-Hun Lee.

At least 19 recordsLinked to original sources

PubMed2006

Tophaceous gout of the lumbar spine mimicking pyogenic discitis.

BACKGROUND CONTEXT: Gout of the spine is very rare. Forty-one cases of gout affecting the spine have been reported. PURPOSE: To present a patient with tophaceous gout of the lumbar spine mimicking pyogenic discitis. STUDY DESIGN: Case report. METHODS: The medical record, including operative notes, progress notes, discharge summary, clinical notes, radiological study, and pathological studies was reviewed. RESULTS: The patient had acute low back pain and radiculopathy with high spiking fever. Therefore, pyogenic discitis was suspected. However, histological diagnosis revealed tophaceous gout. CONCLUSIONS: Tophaceous gout of the spine is very rare. However, when a patient presents with acute back pain and fever, spinal gout should be considered, particularly in a patient with a previous history of hyperuricemia or gout.

Allopurinol↗

PubMed2006

Acquisition of in vitro and in vivo functionality of Nurr1-induced dopamine neurons.

Neural precursor cells provide an expandable source of neurons and glia for basic and translational applications. However, little progress has been made in directing naive neural precursors toward specific neuronal fates such as midbrain dopamine (DA) neurons. We have recently demonstrated that transgenic expression of the nuclear orphan receptor Nurr1 is sufficient to drive dopaminergic differentiation of forebrain embryonic rat neural precursors in vitro. However, Nurr1-induced DA neurons exhibit immature neuronal morphologies and functional properties and are unable to induce behavioral recovery in rodent models of Parkinson's disease (PD). Here, we report on the identification of key genetic factors that drive morphological and functional differentiation of Nurr1-derived DA neurons. We show that coexpression of Nurr1, Bcl-XL, and Sonic hedgehog (SHH) or Nurr1 and the proneural bHLH factor Mash1 is sufficient to drive naive rat forebrain precursors into neurons exhibiting the biochemical, electrophysiological, and functional properties of DA neuron in vitro. On transplantation into the striatum of Parkinsonian rats, precursor cells engineered with Nurr1/SHH/Bcl-XL or Nurr1/Mash1 survived in vivo and differentiated into mature DA neurons that can reverse the behavioral deficits in the grafted animals.

Animals↗

PubMed2006

Differential actions of the proneural genes encoding Mash1 and neurogenins in Nurr1-induced dopamine neuron differentiation.

The steroid receptor-type transcription factor Nurr1 has a crucial role in the development of the mesencephalic dopamine (DA) neurons. Although ectopic expression of Nurr1 in cultured neural precursor cells is sufficient in establishing the DA phenotype, Nurr1-induced DA cells are morphologically and functionally immature, suggesting the necessity of additional factor(s) for full neuronal differentiation. In this study, we demonstrate that neurogenic basic helix-loop-helix (bHLH) factors Mash1, neurogenins (Ngns) and NeuroD play contrasting roles in Nurr1-induced DA neuronal differentiation. Mash1, but not Ngn2, spatially and temporally colocalized with aldehyde dehydrogenase 2 (AHD2), a specific midbrain DA neuronal progenitor marker, in the early embryonic ventral mesencephalon. Forced expression of Mash1 caused immature Nurr1-induced DA cells to differentiate into mature and functional DA neurons as judged by electrophysiological characteristics, release of DA, and expression of presynaptic DA neuronal markers. By contrast, atonal-related bHLHs, represented by Ngn1, Ngn2 and NeuroD, repressed Nurr1-induced expression of DA neuronal markers. Domain-swapping experiments with Mash1 and NeuroD indicated that the helix-loop-helix domain, responsible for mediating dimerization of bHLH transcription factors, imparts the distinct effect. Finally, transient co-transfection of the atonal-related bHLHs with Nurr1 resulted in an E-box-independent repression of Nurr1-induced transcriptional activation of a reporter containing Nurr1-binding element (NL3) as well as a reporter driven by the native tyrosine hydroxylase gene promoter. Taken together, these findings suggest that Mash1 contributes to the generation of DA neurons in cooperation with Nurr1 in the developing midbrain whereas atonal-related bHLH genes inhibit the process.

Aldehyde Dehydrogenase↗

PubMed2006

Clinical outcomes of 3 fusion methods through the posterior approach in the lumbar spine.

STUDY DESIGN: This prospective randomized study compared 3 fusion methods: posterolateral fusion (PLF), posterior lumbar interbody fusion (PLIF), and PLIF combined with PLF (PLF+PLIF). OBJECTIVES: To compare the outcomes of the 3 fusion methods and find a useful fusion method. SUMMARY OF BACKGROUND DATA: Many studies have shown clinical results, advantages, and postoperative complications of each fusion method, but few have compared the 3 fusion methods prospectively. METHODS: A total of 167 patients who underwent 1 or 2-level fusion surgery because of degenerative lumbar disease from January 1996 to September 2000 were studied. Minimum follow-up was 3 years. The patients were randomized into 1 of 3 treatment groups: group 1 (PLF; n = 62); group 2 (PLIF; n = 57); and group 3 (PLF+PLIF; n = 48). A visual analog scale, the Oswestry Disability Questionnaire, and Kirkaldy-Willis criteria were used to measure low back pain, leg pain, and disability. For radiologic evaluation, disc height, lumbar lordosis, segmental angle, and bone union were examined. Postoperative complications were also analyzed. RESULTS: At the last follow-up, good or excellent results were obtained in 50 cases of PLF (80.7%), 50 cases of PLIF (87.8%), and 41 cases of PLF+PLIF (85.5%). No statistical differences were found among the 3 groups (P = 0.704). All methods indicated significant improvement in the disc height (P < 0.05), with PLF having the highest loss in disc height. Lumbar lordosis and segmental angle increased significantly, and improvement of the segmental angle in the 3 fusion methods had statistically significant differences. The nonunion rates at the last follow-up in the 3 fusion groups were not statistically significant, with 8% in group 1, 5% in group 2, and 4% in group 3 (P > 0.05). Complications included deep infection in 3 cases, transient nerve palsy in 4, permanent nerve palsy in 1, and donor site pain in 6. CONCLUSIONS: No significant differences in clinical results and union rates were found among the 3 fusion methods. PLIF had better sagittal balance than PLF. PLIF without PLF had advantages of the elimination of donor site pain, shorter operating time, and less blood loss.

Adult↗

PubMed2006

Real-time detection of mesothelin in pancreatic cancer cell line supernatant using an acoustic wave immunosensor.

BACKGROUND: An acoustic wave immunosensor was developed to illustrate the viability of such devices in early detection of molecular cancer biomarkers. The methods described here involve a real-time, less invasive technique for detecting mesothelin, a protein that has been linked to pancreatic and ovarian cancer. METHODS: Antibodies were immobilized on the gold surface of the device via a self-assembled alkanethiol monolayer. Supernatant from two different pancreatic cancer cell-lines (PL1 and CAPAN2) containing an unknown concentration of mesothelin was tested for the protein by a flow-through analytical technique in three types of experiments. Binding of the mesothelin to the immobilized antibody layer caused a shift in the device's resonant frequency, which was correlated to the concentration of supernatant. A reference sensor was used to correct for frequency shifts caused by pressure or viscosity effects from the injection of the supernatant solution. RESULTS: Repeated experiments indicate that the sensors are capable of nanogram detection thresholds of mesothelin proteins at room temperature and in complex mixture. CONCLUSIONS: Acoustic wave device biosensors have the potential to become a valuable tool in screening for pancreatic as well as other types of cancers. The main features include real-time detection, high sensitivity, and ease of use.

Antibodies, Monoclonal↗

PubMed2006

Excitatory actions of vasoactive intestinal peptide on mouse thalamocortical neurons are mediated by VPAC2 receptors.

Thalamic nuclei can generate intrathalamic rhythms similar to those observed at various arousal levels and pathophysiological conditions such as absence epilepsy. These rhythmic activities can be altered by a variety of neuromodulators that arise from brain stem regions as well as those that are intrinsic to the thalamic circuitry. Vasoactive intestinal peptide (VIP) is a neuropeptide localized within the thalamus and strongly attenuates intrathalamic rhythms via an unidentified receptor subtype. We have used transgenic mice lacking a specific VIP receptor, VPAC(2), to identify its role in VIP-mediated actions in the thalamus. VIP strongly attenuated both the slow, 2-4 Hz and spindle-like 5-8 Hz rhythmic activities in slices from wild-type mice (VPAC(2)(+/+)) but not in slices from VPAC(2) receptor knock-out mice (VPAC(2)(-/-)), which suggests a major role of VPAC(2) receptors in the antioscillatory actions of VIP. Intracellular recordings revealed that VIP depolarized all relay neurons tested from VPAC(2)(+/+) mice. In VPAC(2)(-/-) mice, however, VIP produced no membrane depolarization in 80% of neurons tested. In relay neurons from VPAC(2)+/+ mice, VIP enhanced the hyperpolarization-activated mixed cation current, I(h), via cyclic AMP activity, but VIP did not alter I(h) in VPAC(2)-/- mice. In VPAC(2)-/- mice, pituitary adenylate cyclase activating-polypeptide (PACAP) depolarized the majority of relay neurons via I(h) enhancement presumably via PAC(1) receptor activation. Our findings suggest that VIP-mediated actions are predominantly mediated by VPAC(2) receptors, but PAC(1) receptors may play a minor role. The excitatory actions of VIP and PACAP suggest these peptides may not only regulate intrathalamic rhythmic activities, but also may influence information transfer through thalamocortical circuits.

Animals↗

PubMed2006

The quantitative analysis of tissue injury markers after mini-open lumbar fusion.

STUDY DESIGN: A prospective and comparative analysis of tissue injury after mini-open lumbar fusion and conventional technique. OBJECTIVE: To evaluate the tissue injury quantitatively after mini-open lumbar fusion. SUMMARY OF BACKGROUND DATA: The advantages of minimally invasive techniques demonstrated by clinical reports have been a smaller scar, lesser pain, and rapid rehabilitation. It has not been established, however, that the minimally invasive technique has less tissue injury and less systemic response than the conventional spine surgery. METHODS: Twenty patients who had undergone posterior decompression and the fusion of the L4-L5 segment for spinal stenosis were studied prospectively. Ten patients that had the conventional posterior decompression and posterior lumbar interbody fusion (PLIF) at L4-L5 were enrolled in a control group. The remaining 10 patients who underwent MISS techniques (paramedian mini-open approach using a tubular retractor) were enrolled in a study group. The serum enzymes representing skeletal muscle injury (creatinine kinase and aldolase), pro-inflammatory cytokines (IL-6, IL-8), and anti-inflammatory cytokines (IL-10, IL-1 receptor antagonist) were analyzed with ELISA techniques. They were checked on the day before operation and 1, 3, 7, and 14 days after operation. RESULTS: Serum creatinine kinase and most of the inflammatory cytokines were significantly high in the control group on postoperative days 1 and 3 and returned to the normal level on postoperative day 7. Serum aldolase was significantly high on postoperative day 1, and IL-8 remained elevated until postoperative day 7 in the control group. CONCLUSION: Mini-open lumbar fusion may significantly contribute to the reduction of muscle injury and systemic inflammatory reactions during the acute postoperative period. This study suggests that mini-open lumbar fusion may also play an important role in preventing medical morbidity after spinal surgery.

Aged↗

PubMed2005

Effect of cell-density on in-vitro dopaminergic differentiation of mesencephalic precursor cells.

Neural precursor cells isolated from early embryonic mesencephalon are in-vitro expanded and differentiated toward dopamine neurons. However, conditions for controlled conversion of the precursors into dopamine neurons largely remained to be determined. We here examined the effects of plating cell density and duration of in-vitro cell expansion on the precursors-derived dopamine differentiation. The yield of dopamine neurons from cultured mesencephalic precursors was greater when the cells were initially plated at higher density. Soluble factors secreted from the precursors appeared to be responsible for the cell density effect. We further demonstrated that the dopamine differentiation potential of the precursors was lost after a long-term cell expansion. Therefore, in order to attain high percentage of dopamine neuron population in mesencephalic precursor cultures, cultures need to be seeded at high cell density and to be expanded for a short period of time.

Animals↗

PubMed2005

In vitro and in vivo analyses of human embryonic stem cell-derived dopamine neurons.

Human embryonic stem (hES) cells, due to their capacity of multipotency and self-renewal, may serve as a valuable experimental tool for human developmental biology and may provide an unlimited cell source for cell replacement therapy. The purpose of this study was to assess the developmental potential of hES cells to replace the selectively lost midbrain dopamine (DA) neurons in Parkinson's disease. Here, we report the development of an in vitro differentiation protocol to derive an enriched population of midbrain DA neurons from hES cells. Neural induction of hES cells co-cultured with stromal cells, followed by expansion of the resulting neural precursor cells, efficiently generated DA neurons with concomitant expression of transcriptional factors related to midbrain DA development, such as Pax2, En1 (Engrailed-1), Nurr1, and Lmx1b. Using our procedure, the majority of differentiated hES cells (> 95%) contained neuronal or neural precursor markers and a high percentage (> 40%) of TuJ1+ neurons was tyrosine hydroxylase (TH)+, while none of them expressed the undifferentiated ES cell marker, Oct 3/4. Furthermore, hES cell-derived DA neurons demonstrated functionality in vitro, releasing DA in response to KCl-induced depolarization and reuptake of DA. Finally, transplantation of hES-derived DA neurons into the striatum of hemi-parkinsonian rats failed to result in improvement of their behavioral deficits as determined by amphetamine-induced rotation and step-adjustment. Immunohistochemical analyses of grafted brains revealed that abundant hES-derived cells (human nuclei+ cells) survived in the grafts, but none of them were TH+. Therefore, unlike those from mouse ES cells, hES cell-derived DA neurons either do not survive or their DA phenotype is unstable when grafted into rodent brains.

Animals↗

PubMed2005

Retrovirally transduced NCAM140 facilitates neuronal fate choice of hippocampal progenitor cells.

Neural cell adhesion molecule (NCAM) influences proliferation and differentiation of neuronal cells. However, only a little is known about the downstream effects of NCAM signalling, such as alterations in gene transcription, which are associated with cell fate choice. To examine whether NCAM plays a role in cell fate choice during hippocampal neurogenesis, we performed a gain-of-function study, using a retroviral vector which contained full-length NCAM140 cDNA and the marker gene EGFP, and found that NCAM140 promoted neurogenesis by activating proneural transcription activators with concurrent inhibition of gliogenesis. The enhanced transcript levels of proneural transcription factors in NCAM140-transduced cells were down-regulated by treatment of the cells with mitogen-activated protein kinase kinase (MEK) inhibitor PD098059. Overall, these findings suggest that NCAM140 may facilitate hippocampal neurogenesis via regulation of proneurogenic transcription factors in an extracellular signal-regulated kinase (ERK)-dependent manner.

Animals↗

PubMed2005

The role of temporal structure in human vision.

Gestalt psychologists identified several stimulus properties thought to underlie visual grouping and figure/ground segmentation, and among those properties was common fate: the tendency to group together individual objects that move together in the same direction at the same speed. Recent years have witnessed an upsurge of interest in visual grouping based on other time-dependent sources of visual information, including synchronized changes in luminance, in motion direction, and in figure/ ground relations. These various sources of temporal grouping information can be subsumed under the rubric temporal structure. In this article, the authors review evidence bearing on the effectiveness of temporal structure in visual grouping. They start with an overview of evidence bearing on temporal acuity of human vision, covering studies dealing with temporal integration and temporal differentiation. They then summarize psychophysical studies dealing with figure/ground segregation based on temporal phase differences in deterministic and stochastic events. The authors conclude with a brief discussion of neurophysiological implications of these results.

Humans↗

PubMed2004

Traveling waves of activity in primary visual cortex during binocular rivalry.

When the two eyes view large, dissimilar patterns that induce binocular rivalry, alternating waves of visibility are experienced as one pattern sweeps the other out of conscious awareness. Here we combine psychophysics with functional magnetic resonance imaging to show tight linkage between dynamics of perceptual waves during rivalry and neural events in human primary visual cortex (V1).

Functional Laterality↗

PubMed2004

Temporal expression changes during differentiation of neural stem cells derived from mouse embryonic stem cell.

Temporal analysis in gene expression during differentiation of neural stem cells (NSCs) was performed by using in-house microarrays composed of 10,368 genes. The changes in mRNA level were measured during differentiation day 1, 2, 3, 6, 12, and 15. Out of 10,368 genes analyzed, 259 genes were up-regulated or down-regulated by 2-fold or more at least at one time-point during differentiation, and were classified into six clusters based on their expression patterns by K-means clustering. Clusters characterized by gradual increase have large numbers of genes involved in transport and cell adhesion; those which showed gradual decrease have much of genes in nucleic acid metabolism, cell cycle, transcription factor, and RNA processing. In situ hybridization (ISH) validated microarray data and it also showed that Fox M1, cyclin D2, and CDK4 were highly expressed in CNS germinal zones and ectonucleotide pyrophosphatase/phosphodiesterase 2 (Enpp2) was highly expressed in choroid plexus where stem/progenitor cells are possibly located. Together, this clustering analysis of expression patterns of functionally classified genes may give insight into understanding of CNS development and mechanisms of NSCs proliferation and differentiation.

Animals↗

PubMed2004

Properties of cortical precursor cells cultured long term are similar to those of precursors at later developmental stages.

In vitro cultures of neural precursor cells are useful experimental tools for studies on the mechanisms of brain development, as well as for generating renewable sources in cell therapy for neurodegenerative disorders. The systematic characterization of cultured neural precursors is a prerequisite for obtaining basic information on brain development. Here, we examine the cell survival, proliferation, and differentiation potential of cultured neural precursors from different embryonic ages and those of the precursors expanded in vitro for different periods of time. Precursor cells were isolated at rat embryonic days 14 (E14) and 19 (E19) and cultured in the presence of a mitogen basic fibroblast growth factor (bFGF). The numbers of TUNEL+ and BrdU+ cells in E19 cortical precursor cultures were significantly lower than those in E14 cultures, indicating that the programmed cell death and proliferation potential of neural precursors are reduced during the progression of brain development. E14 cells tended to differentiate into neurons, and E19 cells into astrocytes. To determine whether the intrinsic properties of neural precursors are similarly altered during in vitro culture, E14 precursor cells were expanded for different periods. Precursor cells expanded for longer periods displayed lower apoptotic and proliferation indices, as well as astrogenic developmental potential. Clonal analysis data confirmed the transition of precursor differentiation potential from neurogenic to astrogenic over the culture period. Our findings collectively suggest that neural precursor cells undergo time-dependent changes in properties via an intrinsic program, both in vivo and in vitro.

Animals↗

PubMed2004

Changes of gene expression profiles during neuronal differentiation of central nervous system precursors treated with ascorbic acid.

Ascorbic acid (AA) has been shown to increase the yield of dopaminergic (DA) neurons derived from basic fibroblast growth factor (bFGF)-expanded mesencephalic precursors. To understand the molecular mechanisms underlying this phenomenon, we used cDNA microarray analysis to examine differential expression of neuronal genes following AA treatment. The putative precursor cells were isolated from E13 rat ventral mesencephalons and expanded in the presence of bFGF. Cells were incubated in mitogen-free media supplemented with 200 microM AA or were left untreated as a control, and total RNA was isolated at different time points (expansion stage and 1, 3, and 6 days after induction of differentiation) and subjected to cDNA microarray analysis. Differentiation was evaluated by Western blot analysis and immunocytochemistry of neuron-specific markers. AA treatment of the mesencephalic precursors increased the expression of neuronal (MAP2) and astrocytic (glial fibrillary acidic protein) markers and the percentage of tyrosine hydroxylase (TH)-positive cells. The microarray analysis revealed that 12 known genes were up-regulated and 20 known genes were down-regulated in expansion-stage AA-treated cells. Six days after the induction of differentiation, AA-treated cells showed up-regulation of 48 known genes and down-regulation of 5 known genes. Our results identified several proteins, such as transferrin, S-100, and somatostatin, as being differentially regulated in AA-treated mesencephalic precursors. This novel result may lead to a better understanding of the molecular mechanisms underlying the AA-induced differentiation of mesencephalic precursors into DA neurons and may form the basis for improved DA neuronal production for treatment of Parkinson's disease patients.

Animals↗

PubMed2004

Differential effects of corticosterone and dexamethasone on hippocampal neurogenesis in vitro.

Prenatal stress during fetal development results in the blockade of neurogenesis in the dentate gyrus in adulthood. Present study was undertaken to investigate the dominant role of the glucocorticoid receptors in corticosterone actions on the neurogenesis of fetal hippocampal progenitor cells. For that purpose, expressions of key molecules affected by corticosterone and dexamethasone were compared during proliferation and differentiation of the hippocampal progenitor cells. Corticosterone (2 microM) significantly decreased the number of bromodeoxyuridine-labeled cells (about 50%) and caused the dendritic atrophy in microtubule-associated protein 2-labeled cells. The expressions of NeuroD, BDNF, and NR1 mRNA levels and protein levels of p-ERK and p-CREB were remarkably decreased by corticosterone in a dose-dependent manner. In contrast, dexamethasone, a glucocorticoid receptor (GR) specific agonist, had an inhibitory effect on proliferation, but not differentiation. It is concluded that corticosterone elicits its effects on neurogenesis including proliferation and differentiation whereas stimulation of the glucocorticoid receptor is sufficient to decrease only proliferation.

Animals↗

PubMed2004

Enhanced in vitro midbrain dopamine neuron differentiation, dopaminergic function, neurite outgrowth, and 1-methyl-4-phenylpyridium resistance in mouse embryonic stem cells overexpressing Bcl-XL.

Embryonic stem (ES) cells provide a potentially unlimited source of specialized cells for regenerative medicine. The ease of inducing stable genetic modifications in ES cells allows for in vitro manipulations to enhance differentiation into specific cell types and to optimize in vivo function of differentiated progeny in animal models of disease. We have generated mouse ES cells that constitutively express Bcl-XL, an antiapoptotic protein of Bcl-2 family. In vitro differentiation of Bcl-XL overexpressing ES (Bcl-ES) cells resulted in higher expression of genes related to midbrain dopamine (DA) neuron development and increased the number of ES-derived neurons expressing midbrain DA markers compared with differentiation of wild-type ES cells. Moreover, DA neurons derived from Bcl-ES cells were less susceptible to 1-methyl-4-phenylpyridium, a neurotoxin for DA neurons. On transplantation into parkinsonian rats, the Bcl-ES-derived DA neurons exhibited more extensive fiber outgrowth and led to a more pronounced reversal of behavioral symptoms than wild-type ES-derived DA neurons. These data suggest a role for Bcl-XL during in vitro midbrain DA neuron differentiation and provide an improved system for cell transplantation in a preclinical animal model of Parkinson's disease.

1-Methyl-4-phenylpyridinium↗

PubMed2004

Comprehensive transcriptome analysis of differentiation of embryonic stem cells into midbrain and hindbrain neurons.

Neurogenesis is one of the most complex events in embryonic development. However, little information is available regarding the molecular events that occur during neurogenesis. To identify regulatory genes and underlying mechanisms involved in the differentiation of embryonic stem (ES) cells to neurons, gene expression profiling was performed using cDNA microarrays. In mouse ES cells, we compared the gene expression of each differentiated cell stage using a five-stage lineage selection method. Of 10,368 genes, 1633 (16%) known regulatory genes were differentially expressed at least 2-fold or greater at one or more stages. At stage 3, during which ES cells differentiate into neural stem cells, modulation of nearly 1000 genes was observed. Most of transcription factors (Otx2, Ebf-3, Ptx3, Sox4, 13, 18, engrailed, Irx2, Pax8, and Lim3), signaling molecules (Wnt, TGF, and Shh family members), and extracellular matrix/adhesion molecules (collagens, MAPs, and NCAM) were up-regulated. However, some genes which may play important roles in maintaining the pluripotency of ES cells (Kruppel-like factor 2, 4, 5, 9, myeloblast oncogene like2, ZFP 57, and Esg-1) were down-regulated. The many genes identified with this approach that are modulated during neurogenesis will facilitate studies of the mechanisms underlying ES cell differentiation, neural induction, and neurogenesis.

Animals↗