The long belief that dental primary afferent (DPA) neurons are entirely composed of nociceptive neurons has been challenged by several anatomical and functional investigations. In order to characterize non-nociceptivepopulation among DPA neurons, retrograde transport fluorescent dye was placed in upper molars of rats and immunohistochemical detection of peripherin and neurofilament 200 in the labeled trigeminal ganglia was performed. As the results, majority ofDPA neurons were peripherin-expressing small-sized neurons, showing characteristic ofnociceptive C-fibers. However, 25.7% of DPA were stained with antibody against neurofilament 200, indicating significant portion of DPA neurons are related to large myelinated Abeta fibers. There were a small number of neurons thatexpressed both peripherin and neurofilament 200, suggestive of Adelta fibers. The possible transition of neurochemical properties by neuronal injury induced by retrograde labeling technique was ruled out by detection of minimal expression of neuronal injury marker, ATF-3. These results suggest that in addition to the large population of C-fiber-related nociceptive neurons, a subset of DPA neurons is myelinated large neurons, which is related to low-threshold mechanosensitive Abeta fibers. We suggest that these Abeta fiber-related neurons might play a role as mechanotransducers of fluid movement within dentinal tubules.
Animals ; Dentin ; Intermediate Filament Proteins ; Membrane Glycoproteins ; Molar ; Myelin Sheath ; Nerve Tissue Proteins ; Neurofilament Proteins ; Neurons ; Neurons, Afferent ; Nociceptors ; Rats ; Trigeminal Ganglion

BACKGROUND: Tumor angiogenesis plays an important role in tumor growth, maintenance and metastatic potential. Tumor tissue produces many types of angiogenic growth factors. Vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) have both been implicated to have roles in tumor angiogenesis. In this study, the expression of tissue VEGF and bFGF from non-small cell lung cancer (NSCLC) patients were analyzed. METHODS: We retrospectively investigated 35 patients with a histologically confirmed adenocarcinoma or squamous cell carcinoma of the lung, where the primary curative approach was surgery. An ELISA was employed to determine the expression of VEGF and bFGF in extracts prepared from 35 frozen tissue samples taken from the cancer patients. RESULTS: VEGF and bFGF concentrations were significantly increased in lung cancer tissue as compared with control (non-cancerous) tissue. The VEGF concentration was significantly increased in T2 and T3 cancers as compared with T1 cancer. Expression of VEGF was increased in node-positive lung cancer tissue as compared with node-negative lung cancer tissue (p=0.06). VEGF and bFGF expression were not directly related to the stage of lung cancer and patient survival. CONCLUSION: Expression of VEGF and bFGF were increased in lung cancer tissue, and the expression of VEGF concentration in lung cancer tissue was more likely related with tumor size and the presence of a lymph node metastasis than the expression of bFGF. However, in this study, expression of both VEGF and bFGF in tissue were not associated with patient prognosis.
Adenocarcinoma ; Carcinoma, Non-Small-Cell Lung ; Carcinoma, Squamous Cell ; Enzyme-Linked Immunosorbent Assay ; Fibroblast Growth Factor 2 ; Humans ; Intercellular Signaling Peptides and Proteins ; Lung ; Lung Neoplasms ; Lymph Nodes ; Neoplasm Metastasis ; Prognosis ; Retrospective Studies ; Vascular Endothelial Growth Factor A

PURPOSE: This study was undertaken to investigate in detail the xenograft mouse orthotopic lung cancer model induced by PC14PE6 adenocarcinoma cells. MATERIALS AND METHODS: Three cell doses (0.5x10(6); 1x10(6); 2x10(6)) of PC14PE6 cells were injected into the lungs of male BALB/c nude mice by the intrathoracic injection method. The lung and other organs, including brain, liver, spleen, kidney, muscle, adrenal gland, and lymph node on knee, were removed and stained with H/E to detect the presence of tumor cells. RESULTS: The reliable tumorigenicity time in the PC14PE6 adenocarcinoma cell-inoculated BALB/c nude mouse was 10 days after intrathoracic injection. The average life span of the three groups after inoculation was 14 days in the 2x10(6) cells inoculum group; 25 days in the 1x10(6) cells inoculum group; and 32 days in the 0.5x10(6) cells inoculum group. The PC14PE6 adenocarcinoma cells induced orthotopic lung cancer limited within the thorax. CONCLUSIONS: This orthotopic lung cancer model is an efficient cancer model with easy inoculation methods, rapid and high tumorigenicity, and simple monitoring methods for metastasis.
Adenocarcinoma ; Adrenal Glands ; Animals ; Brain ; Heterografts ; Humans ; Kidney ; Knee ; Liver ; Lung Neoplasms* ; Lung* ; Lymph Nodes ; Male ; Mice* ; Mice, Nude ; Models, Animal ; Neoplasm Metastasis ; Spleen ; Thorax

The Kinesin superfamily proteins (KIFs) make up a large superfamily of molecular motors that transport cargo such as vesicles, protein complexes, and organelles. KIF1Balpha is a monomeric motor that conveys mitochondria and plays an important role in cellular function. Here, we used the yeast two-hybrid system to identify the proteins that interacts with KIF1Balpha and found a specific interaction with the mammalian LIN-7 (MALS)-3/vertebrate homology of LIN-7 (Veri) and synaptic scaffolding molecule (S-SCAM). MALS-3 protein bound to the tail region of KIF1Balpha but not to other kinesin family members in the yeast two-hybrid assay. The "T-X-V" motif at the C-terminal end of KIF1Balpha is essential for interaction with MALS-3. In addition, this protein showed specific interactions in the Glutathione S-transferase (GST) pull-down assay. An antibody to MALS-3 specifically coimmunoprecipitated KIF1Balpha associated with MALS-3 from mouse brain extracts. These results suggest that MALS-3, as KIF1Balpha receptor, is involved in the KIF1Balpha-mediated transport.
Animals ; Brain ; Glutathione Transferase ; Humans ; Kinesin* ; Mice ; Microtubules ; Mitochondria ; Organelles ; PDZ Domains* ; Two-Hybrid System Techniques

OBJECTIVE: The chemotherapeutic agent cisplatin (cis-diamminedichloroplatinum (II)) is particularly effective against cervical cancer. The purpose of this study is to elucidate combination effect of cisplatin and green tea extracts on the growth inhibition of TC-1 cell. METHODS: To observe the anti-proliferative effects, we treated different doses of cisplatin (0.1, 0.5, 2.5 uM), GTP (1, 5, 25 ug/ml) and EGCG (25, 50, 100 uM). to TC-1 cells. Also, we treated 0.5 uM of cisplatin and different doses of GTP (1 and 5 ug/ml) or EGCG (25 and 50 uM). Cell viability was scored by use of MTT assay. In addition, E6 gene expression patterns in TC-1 cell were investigated by using RT-PCR. RESULTS: Cell growth inhibition in a dose dependent was observed at the different concentration of ciaplatin, GTP and EGCG. Also, in the groups treated by 0.5 uM of cisplatin and GTP (1 and 5 ug/ml) or EGCG (25 and 50 uM), the inhibition of cell growth showed with 12.2%, 6.9% and 63.4%, 72.2% as compared to the group treated by cisplatin only. In RT-PCR, down regulation of E6 was shown. CONCLUSION: Additive effect of the combination of cisplatin with GTP or EGCG on the inhibition of cell growth was observed. This effect suggests the possibility lowering the concentration of chemotherapeutic drugs, which alleviate the side effect of drugs.
Cell Survival ; Cisplatin* ; Down-Regulation ; Gene Expression ; Guanosine Triphosphate ; Tea* ; Uterine Cervical Neoplasms

The kinesin proteins (KIFs) make up a large superfamily of molecular motors that transport cargo such as vesicles, protein complexes, and organelles. KIF1A is a monomeric motor that conveys synaptic vesicle precursors and plays an important role in neuronal function. Here, we used the yeast two-hybrid system to identify the neuronal protein (s) that interacts with the tail region of KIF1A and found a specific interaction with synaptotagmin XI. The amino acid residues between 830 and 1300 of KIF1A are required for the interaction with synaptotagmin XI. KIF1A also bound to the tail region of synaptotagmin IV but not to other synaptotagmin in the yeast two-hybrid assay. KIF1A interacted with GST-synaptotagim XI fusion proteins, but not with GST alone. An antibody to synaptotagmin XI specifically co-mmunoprecipitated KIF1A associated with synaptotagimin from mouse brain extracts. These results suggest that KIF1A motor protein transports of synaptotagmin XI-containing synaptic vesicle precursors along microtubule.
Animals ; Brain ; Kinesin* ; Mice ; Microtubules ; Neurons ; Organelles ; Protein Transport ; Synaptic Vesicles ; Synaptotagmins* ; Two-Hybrid System Techniques

OBJECTIVE: To know the effect of adenosine 5'-triphosphate (ATP) on intracellular calcium level and cell proliferation in cervical cancer cells. METHODS: Study design: Four different human cervical cancer cell lines (Caski, C33A, HeLaS3 and SiHa) were used in this study. The change of intracellular calcium level, cell proliferation and the activity of proliferation- and calcium-related transcription factors by extracellular ATP were examined in these cell lines. RESULTS: Extracellular ATP induced calcium mobilization, cell proliferation and the activation of NF-kappa B in all cell lines used. CONCLUSION: These results suggest that calcium mobilization and NF-kappa B dependent signaling pathway play an important role in the cell proliferation by ATP in cervical cancer.
Adenosine Triphosphate ; Adenosine* ; Calcium* ; Cell Line ; Cell Proliferation ; Humans ; NF-kappa B* ; Transcription Factors ; Uterine Cervical Neoplasms*

OBJECTIVES: This study was performed to evaluate the pregnancy rate following the transfers of frozen- thawed embryos which was derived from intracytoplasmic sperm injection (ICSI) using sperm obtained by ejaculated, testicular sperm extraction (TESE), and frozen-thawed testicular sperm extraction (t-TESE). METHODS: Frozen-thawed embryos were successfully transferred to the patients in 664 cycles among 695 cycles from January 1998 to December 2002, where ICSI was done with various origins of sperm. Subjects were divided into three groups according to the origin of sperm; ejaculated sperm group as a control (n=535), TESE group (n=98) and t-TESE group (n=62). After conventional ICSI, the supernumerary PN stage or developing embryos were cryopreserved by slow freezing protocol with 1, 2-propanediol as cryoprotectant. RESULTS: The survival rate of frozen-thawed embryos was 77.7% (2515/3236) in ejaculated sperm group, 76.6% (441/576) in TESE group and 83.9% (292/348) in frozen-thawed TESE group, respectively. The difference of survival rate of between t-TESE group and other two groups was statistically significant (p<0.01). The good embryo formation rate and positive beta-hCG rate was 46.3% (1164/2515), 28.8% (148/513) in ejaculated sperm group, 49.2% (217/441), 36.6% (34/93) in TESE group and 46.2% (135/292), 34.9% (22/63) in frozen-thawed TESE group, respectively. CONCLUSION: This study demonstrates that comparable pregnancy rate and implantation rate could be achieved after the transfer of frozen-thawed embryos following ICSI using various sources of sperm. As there was no statistically significant difference in pregnacy rate between ICSI with fresh testicular sperm and with frozen-thawed testicular sperm, the sequential cryopreservation of supernumerary testicular sperm and embryos may be a useful method for increasing pregnancy outcome in infertile couples with male factor.
Cryopreservation ; Embryonic Structures* ; Family Characteristics ; Female ; Freezing ; Humans ; Male ; Pregnancy ; Pregnancy Outcome* ; Pregnancy Rate ; Pregnancy* ; Sperm Injections, Intracytoplasmic* ; Spermatozoa* ; Survival Rate

The kinesin proteins (KIFs) make up a large superfamily of molecular motors that transport cargo such as vesicles, protein complexes, and organelles. KIF5 is a heterotetrameric motor that conveys vesicles and plays an important role in neuronal function. Here, we used the yeast two-hybrid system to identify the neuronal protein (s) that interacts with the tail region of KIF5 and found a specific interaction with betaIII spectrin. The amino acid residues between 1394 and 1774 of betaIII spectrin were required for the interaction with KIF5C. betaIII spectrin also bound to the tail region of neuronal KIF5A and ubiquitous KIF5B but not to other kinesin family members in the yeast two-hybrid assay. In addition, these proteins showed specific interactions, confirmed by GST pull-down assay and co-immunoprecipitation. betaIII spectrin interacted with GST-KIF5 fusion proteins, but not with GST alone. An antibody to betaIII spectrin specifically co-immunoprecipitated KIF5s associated with betaIII spectrin from mouse brain extracts. These results suggest that KIF5 motor proteins transport vesicles or organelles that are coated with betaIII spectrin.
Animals ; Brain ; Humans ; Immunoprecipitation ; Kinesin* ; Mice ; Microtubules ; Neurons ; Organelles ; Spectrin* ; Transport Vesicles ; Two-Hybrid System Techniques

OBJECTIVES: To evaluate the safety and efficacy of selective fetal reduction (SFR) and compare the outcome of twin pregnancy after SFR in multiple pregnancy induced by assisted reproductive technology (ART) with that of natural twin pregnancy. METHODS: From September 1995 to March 2002 in Ajou University Hospital, SFR was performed in 79 patients whose gestational sacs were more than 3. Of these 79 patients, 47 patents resulted in twin pregnancy after SFR. SFR was performed using transvaginal intracardiac KCl injection at gestational age of 6~9 weeks. Control group was composed of 264 patients with natural twin pregnancy, who delivered after intrauterine pregnancy at 24 weeks, from June 1994 through December 2002. We compared Obstetric and perinatal outcomes between SFR group and natural twin group. RESULTS: Among 47 patients with twin pregnancy after SFR, 2 spontaneous abortion were occurred at intrauterine pregnancy at 8 and 19 weeks. Obstetrical and perinatal outcomes were available in 43 patients. Single intrauterine fetal death was occurred in 1 of 43 (2.3%) patients in SFR group. incidence of preterm labor, premature rupture of membrane, preeclampsia and placenta previa were similar, but gestational diabetes mellitus (GDM) was occurred more frequently in SFR group (3 (7.0%) vs 4 (1.5%), p=0.02). Mean gestational age, mean birth weight, incidence of discordancy, use of intubation and ventilation, incidence of fetal anomaly, low (<7) Apgar score and intrauterine growth restriction were similar in both groups. CONCLUSION: Twin pregnancy after SFR has the increased incidence for GDM but other obstetric and perinatal outcome was similar compared with natural twin pregnancy. So SFR is a safe and effective procedure, so we suggest SFR is needed in multifetal pregnancy more than triplet.
Abortion, Spontaneous ; Apgar Score ; Birth Weight ; Diabetes, Gestational ; Female ; Fetal Death ; Gestational Age ; Gestational Sac ; Humans ; Incidence ; Intubation ; Membranes ; Obstetric Labor, Premature ; Placenta Previa ; Pre-Eclampsia ; Pregnancy ; Pregnancy Reduction, Multifetal* ; Pregnancy, Multiple ; Pregnancy, Twin* ; Reproductive Techniques, Assisted ; Rupture ; Triplets ; Ventilation