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Biomedical subjects

J Si

Publications and source records attributed to J Si.

At least 19 recordsLinked to original sources
PubMed1999

Regulation of neuregulin-mediated acetylcholine receptor synthesis by protein tyrosine phosphatase SHP2.

Synapse-specific expression of the nicotinic acetylcholine receptor (AChR) is believed to be mediated by neuregulin, an epidermal growth factor-like trophic factor released by somatic motoneurons at the neuromuscular junction (NMJ). Neuregulin stimulates ErbB2, ErbB3, and ErbB4, members of the ErbB family of receptor tyrosine kinases. SHP2 is a cytoplasmic protein tyrosine phosphatase containing two Src homology 2 domains near its N terminus, and has been shown to be a positive mediator of mitogenic responses to various growth factors. We found that SHP2 interacted with ErbB2 and ErbB3 after neuregulin stimulation of muscle cells. Expression of SHP2 in C2C12 mouse muscle cells attenuated the neuregulin-induced expression of an AChR epsilon-promoter reporter gene, whereas a catalytically inactive SHP2 mutant or a mutant lacking the N-terminal Src homology 2 (SH2) domain enhanced reporter expression, suggesting that SHP2 negatively regulates the neuregulin signaling pathway. In fibroblast cells that express a mutant SHP2 with a targeted deletion of the N-terminal SH2 domain, neuregulin-mediated activation of the Ras/Raf/extracellular signal-regulated kinase cascade was enhanced. Furthermore, we found that SHP2 immunoreactivity colocalized with the staining of alpha-bungarotoxin, a marker of the NMJ. These results demonstrate a negative role of SHP2 in the neuregulin signal that leads to AChR gene expression at the NMJ.

Animals↗
PubMed1999

Essential roles of c-JUN and c-JUN N-terminal kinase (JNK) in neuregulin-increased expression of the acetylcholine receptor epsilon-subunit.

Neuregulin is a neural factor implicated in upregulation of acetylcholine receptor (AChR) synthesis at the neuromuscular junction. Previous studies have demonstrated that the extracellular signal-regulated kinase (ERK) subgroup of MAP kinases is required for neuregulin-induced AChR gene expression. We report here that the neuregulin-mediated increase in AChR epsilon-subunit mRNA was a delayed response in C2C12 muscle cells. Neuregulin induced expression of immediate early genes c-jun and c-fos, which followed and depended on the ERK activation. Treatment of muscle cells with cycloheximide to inhibit c-JUN synthesis at the protein level and suppression of c-JUN function by a dominant-negative mutant blocked neuregulin-induced expression of the epsilon-subunit gene, indicating an essential role of c-JUN in neuregulin signaling. Furthermore, neuregulin activated c-JUN N-terminal kinase (JNK) in C2C12 muscle cells. Blockade of JNK activation by overexpressing dominant-negative MKK4 inhibited epsilon-promoter activation. Moreover, overexpression of the JNK dominant-negative mutant inhibited neuregulin-mediated expression of the epsilon-transgene and endogenous epsilon-mRNA. Taken together, our results demonstrate important roles of c-JUN and JNK in neuregulin-mediated expression of the AChR epsilon-subunit gene and suggest that neuregulin activates multiple signaling cascades that converge to regulate AChR epsilon-subunit gene expression.

Animals↗
PubMed1999

ERK MAP kinase activation is required for acetylcholine receptor inducing activity-induced increase in all five acetylcholine receptor subunit mRNAs as well as synapse-specific expression of acetylcholine receptor epsilon-transgene.

The AChR is a pentamer of four different subunits in a stoichiometry of alpha2betagammadelta in embryonic and alpha2betaepsilondelta in adult animals. Transcription of AChR subunit genes is most active in synaptic nuclei in adult skeletal muscle cells, and is regulated by neural factors such as ARIA. We report here that ARIA up-regulated specifically the expression of all five AChR subunits in C2C12 cells. The mRNA level of erbB2, erbB3, rapsyn, MuSK, SHP-2 and beta-actin remained unchanged in response to ARIA stimulation in C2C12 cells. The ARIA-induced increase in AChR subunit expression in C2C12 cells was inhibited by the erbB kinase inhibitor tyrphostin AG1478 and the MEK inhibitor PD98059, but not by the PI3 kinase inhibitor wortmannin, suggesting an important role of the erbB protein tyrosine kinases and MAP kinase in the regulation of the expression of the five different AChR subunits. To determine the signaling pathways in vivo, we studied the expression of reporter genes driven by the epsilon-promoter in injected muscles. The in vivo expression of the epsilon-transgene was inhibited by co-expression of dominant negative mutants of key components in the MAP kinase pathway including ras, raf and MEK, but not the dominant negative mutant of PI3 kinase. These results suggest that ERK MAP kinase activation is required for ARIA-induced increase in all five AChR subunit mRNAs as well as synapse-specific expression of AChR epsilon-transgene.

Androstadienes↗
PubMed1999

Neuregulin-increased expression of acetylcholine receptor epsilon-subunit gene requires ErbB interaction with Shc.

Selective transcription of acetylcholine receptor (AChR) subunit genes by neuregulin is one of the mechanisms involved in the synaptic localization of AChRs to the neuromuscular junction. Neuregulin stimulates ErbB receptor tyrosine kinases and subsequently activates the Ras/ERK pathway, which is required for neuregulin-mediated induction of AChR subunit genes in muscle cells and synapse-specific expression in vivo. Here we investigated the neuregulin transduction mechanism that leads to ERK activation after ErbB receptor tyrosine phosphorylation. Neuregulin increases the association of the adaptor proteins Grb2 and Shc with both ErbB2 and ErbB3 in C2C12 muscle cells. Dephosphorylation of the tyrosine-phosphorylated ErbB proteins abolished their association with both Grb2 and Shc, suggesting a tyrosine phosphorylation-dependent interaction. The interaction of Shc with the ErbB receptors is mediated by Shc's phosphotyrosine-binding domain. In addition, neuregulin increased tyrosine phosphorylation of Shc. Mutagenesis approaches demonstrated that tyrosine phosphorylation of Shc is required for neuregulin induction of AChR subunit gene expression. Taken together, these data indicate that the interaction of ErbB receptors with Grb2 alone is insufficient for neuregulin-activated transcription, but that ErbB receptor signaling via Shc is necessary and important.

Adaptor Proteins, Signal Transducing↗
PubMed1998

Weight-value convergence of the SOM algorithm for discrete input.

Some insights on the convergence of the weight values of the self-organizing map (SOM) to a stationary state in the case of discrete input are provided. The convergence result is obtained by applying the Robbins-Monro algorithm and is applicable to input-output maps of any dimension.

Algorithms↗
PubMed1998

A systematic and effective supervised learning mechanism based on Jacobian rank deficiency.

Most neural network applications rely on the fundamental approximation property of feedforward networks. Supervised learning is a means of implementing this approximate mapping. In a realistic problem setting, a mechanism is needed to devise this learning process based on available data, which encompasses choosing an appropriate set of parameters in order to avoid overfitting, using an efficient learning algorithm measured by computation and memory complexities, ensuring the accuracy of the training procedures as measured by the training error, and testing and cross-validation for generalization. We develop a comprehensive supervised learning algorithm to address these issues. The algorithm combines training and pruning into one procedure by utilizing a common observation of Jacobian rank deficiency in feedforward networks. The algorithm not only reduces the training time and overall complexity but also achieves training accuracy and generalization capabilities comparable to more standard approaches. Extensive simulation results are provided to demonstrate the effectiveness of the algorithm.

Algorithms↗
PubMed1998

Regulation by ARIA/neuregulin of acetylcholine receptor gene transcription at the neuromuscular junction.

Transcription of genes encoding nicotinic acetylcholine receptor (AChR) subunits (alpha, beta, gamma or epsilon, and delta) is highest in nuclei localized to the synaptic region of the muscle which contributes to maintain a high density of AChRs at the postjunctional membrane. ARIA (AChR inducing activity) is believed to be the trophic factor utilized by motor neurons to stimulate AChR synthesis in the subsynaptic area. ARIA stimulates tyrosine phosphorylation of the erbB proteins and activates the MAP kinase pathway which is required for the ARIA-mediated induction of AChR genes.

Animals↗
PubMed1997

Identification of an element required for acetylcholine receptor-inducing activity (ARIA)-induced expression of the acetylcholine receptor epsilon subunit gene.

Acetylcholine Receptor (AChR)-inducing activity (ARIA) is believed to be the trophic factor utilized by motoneurons to stimulate AChR synthesis in the subsynaptic area. Among the four AChR subunit genes, the epsilon subunit gene is strictly expressed in nuclei localized to the synaptic region of the muscle. To understand mechanisms of the regulation of synapse-specific transcription, we studied the promoter activity of the 5'-flanking region of the AChR epsilon subunit gene in response to ARIA. Transgenes containing the wild type or mutant 5'-flanking regions upstream of a luciferase gene were transfected in C2C12 muscle cells. The promoter activity of these transgenes was determined by assaying activity of expressed luciferase. Analyzing a combination of 5' deletion and site-directed mutants, we identified a 10-nucleotide element (position -55/-46), which was crucial for ARIA-induced expression from the epsilon subunit promoter. This element was named ARE for ARIA-responsive element. Mutation of ARE greatly diminished ARIA-induced transgene expression and deletion of ARE abolished completely the ARIA response. Electrophoretic mobility shift analyses revealed a DNA binding activity in muscle nuclear extract that interacted with ARE. Such interaction was enhanced by ARIA stimulation of muscle cells and appeared to be dependent on nuclear protein phosphorylation.

Animals↗
PubMed1997

Self-organization of firing activities in monkey's motor cortex: trajectory computation from spike signals.

The population vector method has been developed to combine the simultaneous direction-related activities of a population of motor cortical neurons to predict the trajectory of the arm movement. In this article, we consider a self-organizing model of a neural representation of the arm trajectory based on neuronal discharge rates. As self-organizing feature map (SOFM) is used to select the optimal set of weights in the model to determine the contribution of an individual neuron to an overall movement representation. The correspondence between movement directions and discharge patterns of the motor cortical neurons is established in the output map. The topology-preserving property of the SOFM is used to analyze the recorded data of a behaving monkey. The data used in this analysis were taken while the monkey was tracing spirals and doing center-->out movements. The arm trajectory could be well predicted using such a statistical model based on the motor cortex neuronal firing information. The SOFM method is compared with the population vector method, which extracts information related to trajectory by assuming that each cell has a fixed preferred direction during the task. This implies that these cells are acting along lines labeled only for direction. However, extradirectional information is carried in these cell responses. The SOFM has the capability of extracting not only direction-related information but also other parameters that are consistently represented in the activity of the recorded population of cells.

Algorithms↗
PubMed1996

Induction of acetylcholine receptor gene expression by ARIA requires activation of mitogen-activated protein kinase.

Transcription of genes encoding nicotinic acetylcholine receptor (AChR) subunits (alpha, beta, gamma or epsilon, and delta) is highest in nuclei localized to the synaptic region of the muscle, which contributes to maintain a high density of AChRs at the postjunctional membrane. ARIA (AChR inducing activity) is believed to be the trophic factor utilized by motor neurons to stimulate AChR synthesis in the subsynaptic area. To elucidate the signaling mechanism initiated by ARIA, we established stable C2C12 cell lines carrying the nuclear lacZ gene under the control of the mouse epsilon subunit promoter or chicken alpha subunit promoter. ARIA stimulated tyrosine phosphorylation of erbB proteins in these C2C12 cells within 15 s with a peak at 5 min. Immediately following tyrosine phosphorylation of erbB proteins, mitogen-activated protein (MAP) kinase was activated which occurred within 30 s and peaked at 8 min after ARIA stimulation. Concomitantly, expression of AChR genes was induced by ARIA. ARIA-induced AChR subunit transgene expression was observed only in differentiated myotubes and not in myoblasts, suggesting that downstream signaling component(s) are regulated in a manner dependent on the myogenic program. Inhibition of the MAP kinase activity by using a specific MAP kinase kinase inhibitor or by overexpressing dominant negative mutants of Raf or MAP kinase kinase attenuated or abolished the ARIA-induced activation of AChR alpha and epsilon subunit gene expression. These results indicate that regulation of AChR gene expression by ARIA in C2C12 cells requires activation of the MAP kinase signaling pathway.

Androstadienes↗
PubMed1996

Differentiation of pseudocondyloma of vulva and condyloma acuminata by dot blot hybridization and polymerase chain reaction.

This study differentiated pseudocondyloma of vulva from condyloma acuminata using dot blot hybridization and polymerase chain reaction (PCR). A total of 27 cases of pseudocondyloma of vulva and 65 cases of condyloma acuminata were selected for the study. The genital lesions were examined clinically and were biopsied. Each biopsy was subjected to histological examination and HPV DNA analysis by dot blot hybridization and PCR. Dot blot analysis detected HPV DNA in 19(82.6%) out of 23 cases of condyloma acuminata and 2(25%) out of 8 cases pseudocondyloma of vulvae (P < 0.05). PCR detected HPV DNA in 51(92.7%) out of 55 cases of condyloma acuminata, compared with none in 23 cases of pseudocondyloma (P < 0.001). HPV DNA was present in the majority of condyloma acuminata specimens, HPV 6 and 11 were the predominant types. Peudocondyloma is probably not associated with HPV. PCR was the most sensitive and useful technique for HPV DNA detection.

Adolescent↗
PubMed1995

A computationally efficient method for solving the redundant problem in biomechanics.

Determining the optimal set of musculotendon forces with which to produce a forward dynamic simulation of movement typically involves a huge investment of time and computational resources. A new, computationally efficient method is proposed that simultaneously achieves the desired trajectory and the dynamically optimized set of muscle stresses, and hence forces, according to the maximal endurance criterion function of Crowninshield and Brand (1981). Muscle-induced accelerations of the system resulting from unit stress contractions of individual muscles are superposed via the new pseudoinverse method to yield the desired motion trajectory. The method is tested on a control problem involving a five degree-of-freedom (DOF), 30 muscle, upper extremity model, which incorporates a dual rigid-body forearm to represent pronation and supination more adequately. The pseudoinverse method delivered the desired motion to within 0.25 degrees for each DOF during a three-second simulation. It is anticipated that the methodology can be easily and accurately applied to other highly redundant optimal control problems in biomechanics.

Acceleration↗
PubMed1995

Tyrosine phosphorylation and synapse formation at the neuromuscular junction.

Protein tyrosine phosphorylation is prevalent throughout the nervous system. It has been implicated to play an important role in the development and maintenance of neuronal functions. In the past few years significant advances have been made in our understanding of the molecular mechanisms of synapse formation and synaptic plasticity. Protein tyrosine phosphorylation appears to be important in the neuron-induced synthesis of the nicotinic acetylcholine receptor and aggregation of synaptic proteins at the neuromuscular junction during development. In addition, protein tyrosine phosphorylation may regulate the ion channel activity of the nicotinic acetylcholine receptor.

Animals↗
PubMed1995

[Detection of p53 mutation using PCR-SSCP silver staining method].

A simple and sensitive technique to detect point mutation of the p53 gene using silver staining method for single-strand conformation polymorphism analysis of polymerase chain reaction (PCR-SSCP) is reported. In this study, the aberrations of the p53 gene (exon 6-8) in clinical specimens of primary cervical carcinoma were examined. Of 27 tumor tissues 2 samples showed mutations of p53 exon 7. Absence of abnormal bands in the p53 exon 6-8 in all ten normal cervical tissues. It appears that inactivation of p53 gene by point mutation is infrequent in clinical samples of human cervical carcinomas.

Base Sequence↗
PubMed1995

[The reversing effect of HPV-16 E6E7 gene antisense plasmid on the HPV-16 positive human cervical carcinoma cells].

Antisense reconstruct, p16as E6E7Neo, harboring HPV-16E6E7 gene of which the expression is regulatable by dexamethasone was constructed. Calcium phosphate mediated transfection was employed to transduce this antisense reconstruct into HPV-16 positive CasKi cell line and HPV negative C-33A cell line (both derived from human cervical carcinoma) respectively. After dexamethasone was added into the culture medium to induce the expression of E6E7 antisense gene, the malignity of CasKi cells were reversed, while the growth characteristics and malignity of C-33A cells were unchanged. The results show that the reversing effect on CasKi cell is specific and mediated by inhibition of expression of E6E7 gene. It also demonstrated a more profound pathogenic role of HPV-16 E6E7 gene in the tumorogenesis of cervical carcinoma and provides a possibility for gene therapy of this tumor.

Animals↗
PubMed1995

[Human papillomavirus, human cytomegalovirus and p53 gene in cervical carcinoma].

OBJECTIVE: To investigate the role of human papillomavirus (HPV) human cytomegalovirus (HCMV) infections and p53 gene mutations in the oncogenesis of cervical cancer and to clarify the association between p53 inactivation and the presence of HPV DNA. METHODS: We examined 38 primary cervical carcinomas and 21 normal cervical specimens for the presence of HPV and HCMV DNA sequences by multiple primers PCR and nest primers PCR. The structure of p53 gene (exons 6-8) was also analyzed by PCR-SSCP silver staining method. RESULTS: Mutations of p53 gene (exon 7) were detected in 2 of 38 tumors. One of the cases with p53 mutation was positive for HPV 16 and two positive for HCMV. HPV 16 and 18 infections were noted in 63.2% (24/38) of the tumors, the positive rate of HCMV was 84.2% (32/38). However, HPV 16,18 and HCMV infection occurred in 4.8% and 38.1% respectively in 21 normal cervical specimens. 21 of the 24 HPV 16,18 positive tumors were also HCMV positive, but none of the normal cervical tissues was infected with both HPV and HCMV. CONCLUSION: Detection of p53 mutations in cervical carcinoma is infrequent and apparently independent of HPV infection. Cervical carcinoma is strongly associated with HPV 16 and 18 infection. A synergistic interaction may occur between HPV and HCMV infections in the oncogenesis of cervical cancer.

Carcinoma, Squamous Cell↗