Overexpression of stathmin1 in the diffuse type of gastric cancer and its roles in proliferation and migration of gastric cancer cells.

BACKGROUND: Stathmin1 is a microtubule-regulating protein that has an important role in the assembly and disassembly of the mitotic spindle. The roles of stathmin1 in carcinogenesis of various cancers, including prostate and breast cancer, have been explored. However, its expression and roles in gastric cancer have not yet been described. METHODS: Stathmin1 expression in paraffin-embedded tissue sections from 226 patients was analysed by immunohistochemistry. Roles of stathmin1 were studied using a specific small interfering RNA (siRNA). RESULTS: The expression of stathmin1 was positively correlated with lymph node metastasis, TNM stages and vascular invasion, and negatively with recurrence-free survival, in the diffuse type of gastric cancer. The median recurrence-free survival in patients with a negative and positive expression of stathmin1 was 17.0 and 7.0 months, respectively (P=0.009). When the expression of stathmin1 was knocked down using siRNA, the proliferation, migration and invasion of poorly differentiated gastric cancer cells in vitro were significantly inhibited. Moreover, stathmin1 siRNA transfection significantly slowed the growth of xenografts in nude mice. CONCLUSION: These results suggest that stathmin1 can be a good prognostic factor for recurrence-free survival rate and is a therapeutic target in diffuse-type gastric cancer.

Distribution and fate of polybrominated diphenyl ethers in indoor environments of elementary schools.

UNLABELLED: Polybrominated diphenyl ethers (PBDEs) are considered harmful to human health because of their toxicities and persistence in environments. In the current study, the distribution and fate of PBDEs in classrooms and computer rooms in 17 elementary schools in South Korea have been described. Eight congeners (brominated diphenyl ether-28, -47, -99, -100, -153, -154, -183, and -209) in air, floor dust, and product surface dust were measured. While Sigma(8)PBDEs in the air in classrooms showed considerable variations (0.659-1600 pg/m(3), arithmetic mean +/- s.d.: 377 +/- 441 pg/m(3)), those in computer rooms were somewhat similar (134-220 pg/m(3), arithmetic mean +/- s.d.: 169 +/- 40 pg/m(3)). Sigma(8)PBDEs in floor dust varied over a wide range, from 453 to 45,700 ng/g, for all rooms. Based on congener patterns, two groups were created--CL-1 that is dominated by high-brominated congeners and CL-2 primarily comprising low-brominated congeners--for both air and floor dust of classrooms. Surface dust had low concentrations, ranged from ND to 181, from ND to 128, and from ND to 256 pg/cm(2) for desk/chair sets, lockers, and playing tools, respectively. Pearson correlation coefficients were calculated individually for air, floor dust, and surface dust. The results indicate that both surface dust and floor dust may act as a secondary source of PBDEs in indoor environments after emission from facilities. PRACTICAL IMPLICATIONS: Children have been estimated to have a higher potential exposure to PBDEs than adults. Since children spend most of their day time at school, PBDE distributions in school environments should be a matter of great concern.

Manganese does not alter the severe neurotoxicity of MPTP.

We utilized a mice model of Parkinsonism: (1) to evaluate 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced neurotoxicity; and (2) to evaluate whether manganese (Mn) exposure can affect MPTP-induced neurotoxicity. A 2 x 3 experimental design (MPTP x+/- Mn) was as follows: SS, MPTP(-) x Mn(-); SLMn, MPTP(-) x low Mn(+); SHMn, MPTP(-) x high Mn(+); MpS, MPTP(+) x Mn(-); MpLMn, MPTP(+) x low Mn(+); MpHMn, MPTP(+) x high Mn(+). We administered MPTP (30 mg/kg per day) to male C57BL/6 mice intraperitoneally, once a day for 5 days. Subsequently, mice were treated with either 2 or 8 mg/kg of MnCl(2).4H(2)O intraperitoneally, once a day for 3 weeks. Blood and striatal Mn levels were elevated in the Mnexposed groups. The number of tyrosine hydroxylase (TH)-immunoreactive (ir) neurons in the substantia nigra pars compacta were decreased significantly in the MPTP-exposed groups. The densities of TH-ir axon terminals in caudate-putamen (CPU) were significantly decreased in the MPTP-treated groups. However, Mn treatment did not affect MPTP neurotoxicity. The densities of glial fibrillary acidic protein (GFAP)-ir astrocytes in the CPU or globus pallidus were significantly increased in the MPTP-treated groups. Concentrations of dopamine in the striatum were decreased significantly in the MPTP-exposed groups only, but Mn had no effect.

The structural basis for DNA binding by an anti-DNA autoantibody.

We have used single and multiple site-directed mutagenesis, and molecular modeling, to identify critical residues in the DNA binding site of MAb 2C10, an IgG anti-dsDNA autoantibody from an MRL/lpr lupus mouse. Simultaneous replacement of four Arg residues in the CDR3H abolished binding activity. With one exception, replacement of any one of these Arg residues reduced the activity to 20-50% of the unmutated scFv activity. Arg to Asp replacements had a slightly greater effect than Arg to Ala replacements. In the one exceptional case, replacement of Arg99 with Ala actually increased DNA binding five-fold and replacement by Asp had little effect. Mutation of Phe32 and Asn35 to A1a in CDRIH decreased DNA binding, whereas replacement of Arg31 with A1a had negligible effect. Ala substitution of any one of a cluster of Asp residues in CDR1L increased DNA binding three to six-fold, confirming previous findings that the L-chain of MAb 2C10 is not favorable for DNA binding. The L-chain does participate in shaping the selectivity of antigen binding, and mutation of CDR3L residue Asp92 or Asn93 caused a decrease in DNA binding activity. Directed mutagenesis, consistent with a molecular model, indicates that: several CDR amino acids contribute to DNA binding, without one residue dominating; both VH and VL CDR3 domains contribute to specificity of binding whereas the CDR1L hinders DNA binding. The results suggest a significant role for electrostatics in the interaction of DNA with MAb 2C10.

Antitumor effect of intratumoral administration of dendritic cell combination with vincristine chemotherapy in a murine fibrosarcoma model.

A new antitumor therapeutic strategy utilizing the combined effect of chemotherapy and DC (dendritic cell)-based immunotherapy was designed, and the effect of intratumoral injections of unpulsed, immature DCs was evaluated after in vivo pretreatment of vincristine on tumor growth in a murine fibrosarcoma tumor model. Vincristine exerted a much more potent apoptosis/necrosis-inducing effect on MCA-102 tumor cells than on DCs both in vitro and in vivo. Moreover, CD11c, CD40, CD80 and CD86 molecules on DCs were not downregulated after treatment with vincristine either in vitro or in vivo. The growth of tumor significantly regressed in the group which received the combined vincristine chemotherapy with intratumoral administration of DCs in contrast to the untreated group, the group treated with DCs alone, and the group treated with vincristine alone. In particular, an upregulated expression of CD40, CD80 and CD86 molecules on DCs was found in the combination treatment group. Furthermore, the number of CD4+ and CD8+ T cells and the staining intensity of their CD4 and CD8 surface molecules also increased after the combination treatment. Therefore, our results indicate the feasibility of this combination therapy with vincristine chemotherapy and DC-based immunotherapy as an efficient antitumor strategy for the treatment of fibrosarcoma.

Analysis of the in vivo dendritic cell response to the bacterial superantigen staphylococcal enterotoxin B in the mouse spleen.

To investigate the in vivo effects of Staphylococcal enterotoxin B (SEB) on dendritic cells (DCs) in the spleen, a single dose of SEB (50 microg/kg) was administered to BALB/c mice by intraperitoneal injection. Afterwards, the mice were sacrificed at 2, 6 and 24 hr, 2, 4, 7 and 15 days, and the spleens were removed. The immunocytochemical characterization of the cells was carried out using various monoclonal antibodies in cryostat-cut sections. The distribution patterns of DCs and their major costimulatory molecules, CD80, CD86 and CD40 in the spleen were identified, and the evidence for maturation of DCs in vivo in response to SEB was obtained. It was found that systemic administration of SEB induced the migration of most of the immature, splenic DCs from the marginal zone to the periarterial lymphatic sheath within 6 hr. This movement paralleled a maturation process, as assessed by upregulation of CD40, CD80 and CD86 expression in the interdigitating dendritic cells (IDCs). The upregulation of costimulatory molecule expression was conspicuous only in DCs in contrast to other antigen-presenting cells (APCs) such as macrophages and B cells which did not show any significant alterations in their costimulatory molecule expression. We also demonstrated the temporal expression pattern of these costimulatory molecules on the activated DCs. The upregulation of costimulatory molecules on DCs reached a peak level 6 hr after SEB injection, while the increase in number of T cells expressing T cell receptor V138 reached a peak level on day 2 after SEB treatment. In conclusion, we demonstrated the in vivo DC response to SEB in the mouse spleen, especially a potent stimulative effect of SEB on DCs in vivo, a temporal distribution pattern of DCs as well as T cells including TCR Vbeta8+ T cells, and a differential expression pattern of costimulatory molecules on the activated DCs. The results of the present study indicate that DCs are the principal type of APCs which mediate T cell activation by SAg in vivo, and that each costimulatory molecule may have different role in the activation of DCs by SAg. Thus, it is plausible to speculate that DCs play a critical role in the T cell clonal expansion by SAgs and other SAg-induced immune responses in vivo.

Distribution and origin of vasoactive intestinal polypeptide-like immunoreactive fibers in the central amygdaloid nucleus of the rat: an immunocytochemical analysis.

We studied the distribution of vasoactive intestinal polypeptide-like immunoreactive (VIPLI) fibers in the central amygdaloid (AC) nucleus of the rat, using indirect immunofluorescence and the origins of such fibers using a combination of retrograde tracing with immunocytochemistry. VIPLI fibers formed a dense fiber plexus in the lateral subdivision of the AC nucleus, but other subdivisions showed little immunoreactivity. Destruction of the supramammillary (SuM) region and the adjacent lateral hypothalamus, both of which contained a group of VIPLI neurons, resulted in the marked reduction of VIPLI fibers in the ipsilateral AC nucleus, indicating that many of the fibers in the AC nucleus originate from these two areas. This assumption was supported by the finding that injection of fast blue dye into the AC nucleus labeled the VIPLI neurons in the SuM region and lateral hypothalamus.

Calcitonin gene-related peptide-like immunoreactive (CGRPI) elements in the circadian system of the mouse: an immunohistochemistry combined with retrograde transport study.

This study was based on immunohistochemical detection of calcitonin gene-related peptide-like immunoreactive (CGRPI) neurons and fibers in the suprachiasmatic nucleus (SCN) and intergeniculate leaflet (IGL) of the mouse. CGRPI neurons and fibers were found within the ventrolateral part of the SCN, in the whole extent of IGL and sparsely distributed in ventral lateral geniculate body. The presence of CGRPI structures in the SCN and IGL of the mouse further supports the hypothesis of differences in the content of neuroactive substances in the circadian clock between mammalian species. Fluorogold retrograde transport combined with CGRP immunofluorescence demonstrates that CGRPI neurons in the IGL constitute a part of IGL reciprocal connections.

Profile of Fos-like immunoreactivity induction by light stimuli in the intergeniculate leaflet is different from that of the suprachiasmatic nucleus.

Light stimuli induce Fos-like immunoreactivity (FLI) in suprachiasmatic nucleus (SCN) and intergeniculate leaflet (IGL). Short pulses of light stimuli that synchronize the circadian rhythms induce FLI in SCN. The characteristics of light induction of FLI in the IGL were studied using immunohistochemistry. In the IGL, at least 2 h of sustained light stimuli were necessary to show an increase of FLI. This FLI persisted while the light was turned on. FLI induction in the IGL by light stimuli was not circadian time specific response. These findings imply that the functional significance of Fos activation on circadian rhythms and mechanism of FLI induction in IGL would be different from that in SCN.

Developmental expression of 'RZR beta, a putative nuclear-melatonin receptor' mRNA in the suprachiasmatic nucleus of the rat.

RZR beta is a member of the retinoid Z receptor (RZR) family of orphan receptors, and its expression is brain-specific. Recently, it was reported that the distribution of RZR beta mRNA is partially coincident with melatonin binding sites and that RZR beta is a putative nuclear receptor of melatonin. Using in situ hybridization, we investigated the developmental expression of RZR beta mRNA in the suprachiasmatic nucleus (SCN) of the rat, which is considered as circadian pacemaker. The RZR beta mRNA was first found in the dorsomedial portion of the SCN at embryonic day 20, and RZR beta mRNA expression in the dorsomedial SCN continued until postnatal day 60. This result suggests that RZR beta plays specific roles as a transcription factor in the dorsomedial SCN but not in the ventrolateral SCN during development and throughout postnatal life.

Pancreatic serous cystadenoma associated with islet cell tumour.

We report the case of a 29-year-old female patient with a diffuse type of serous cystadenoma involving the entire pancreas except for part of the head, which was replaced by islet cell tumour. Ultrasound and CT showed multiple cysts in the entire pancreas and a solid mass with calcification in the head. MRI characterized the fluid content of the cysts and the extent of disease.

Incidence and characterization of Listeria monocytogenes from domestic and imported foods in Korea.

A total of 1,537 domestic and imported food products were examined for the incidence of Listeria monocytogenes between 1993 and 1997 in Korea. L. monocytogenes was detected using the U.S. Department of Agriculture isolation method. Isolated L. monocytogenes was confirmed by polymerase chain reaction with hly1 and hly2 primers designed from the listeriolysin O. Overall, 122 samples (7.9%) contained L. monocytogenes. The rate of isolation was 4.3% for beef, 19.1% for pork, 30.2% for chicken, 1.2% for shellfish, 4.4% for raw milk, 4.4% for frozen smoked mussels, and 6.1% for ice cream. No L. monocytogenes was found in pasteurized milk, pasteurized processed cheese, saltwater fish, dried seafoods, or ham. The overall incidence was lower than that reported in previous studies from other countries. Most isolates were serotype 1/2b except for chicken, in which serotype 1/2a was predominant. The serotyping results might imply the presence of food or geography-specific L. monocytogenes strains.

Serum immunoreactivity to S-100 in children with cerebral palsy and delayed development and in their healthy parents.

The passive immunization of pregnant female rats to S-100 protein often leads to ultra-structural abnormalities in the brain glial structures of the offspring of these rats and induces signs of delayed development in the fetal brain. Additionally passive immunization of pregnant animals with certain antigens induces permanent Ag-specific changes in the immune response of their offspring. The purpose of this study was to investigate serum immunoreactiviy (SIR) to S-100 in cerebral-palsied and developmentally-delayed children as well as in their healthy parents and to evaluate its significance related to radiologic findings of brain MRI and single photon emission computed tomography (SPECT). The subjects were children with cerebral palsy and delayed development that had abnormal findings on brain MRI or Brain SPECT. SIR to S-100 protein was measured by ELISA method in the patients, their healthy parents, 20 normal adult controls and 22 normally developed children. The SIR to S-100 protein was significantly higher in the cerebral-palsied and developmentally-delayed children when compared to that of the normal control group children. Increased SIRs were detected in healthy mothers but not in their fathers. There was no difference of SIR between the cerebral-palsied and developmentally-delayed children or any significant difference of SIRs according to the findings of the brain MRI or to developmental quotients. But, the SIRs to S-100 protein were higher in the group of more abnormal findings on brain SPECT.

Usefulness of T1-weighted image with fast inversion recovery technique in intracranial lesions: comparison with T1-weighted spin echo image.

To evaluate the usefulness of T1-weighted images using the fast inversion recovery (T1FIR) technique as compared with routine T1-weighted spin echo (T1SE) images in various intracranial lesions. Routine spin echo and T1FIR images were performed in 15 consecutive patients with 18 lesions, cerebral infarction in five, astrocytoma in four, vascular lesion in three, encephalomalacia and hemorrhage in each two, arachnoid cyst and meningioma in each one. T1FIR images were performed with 1.5-T Signa [repetition time (TR)/echo time (TE)/inversion time (TI) was 2000/34/800 in 14, 4000/34/1200 in four lesions] and qualitatively compared with the T1SE images in signal intensity, lesion detectability, determination of lesion extent and conspicuity, contrast between lesion and background. Additionally, gray-to-white matter and cerebrospinal fluid (CSF)-to-white matter contrast were evaluated. The signal intensity of the lesions was similar on both T1FIR and T1SE images in all cases. The lesion detectability was similar on both sequences in 15 lesions, and the determination of the lesion extent was definitely higher in 16 lesions on the T1FIR images. Lesion conspicuity was superior in 11, similar in 5, and inferior in 2 patients on the T1FIR images. And also, contrast of lesion-to-background, gray-to-white matter, and CSF-to-white matter was superior on the T1FIR images. The T1FIR technique improved the determination of lesion extent and lesion conspicuity and was qualitatively superior for image contrast as compared with T1SE, but it takes more time than T1SE. The clinical application of T1FIR images depends on whether the superior aspect of the T1FIR images outweighs the disadvantage of the longer time required for this technique.

Significance of resistive index in color Doppler ultrasonogram: differentiation between benign and malignant breast masses.

The objective of this article is to evaluate the significance of resistive index in differentiation between benign and malignant breast lesions on duplex ultrasonographic examination. Resistive indices obtained in 106 breast lesions of 104 patients were included. Sixty-four were benign (mean age: 32.4 +/- 11.1 years), and 42 were malignant lesions (mean age: 47.8 +/- 11.4 years). The resistive index was classified as follows: below 0.49, from 0.5 to 0.59, 0.6 to 0.69, 0.7 to 0.79, and above 0.8. We analyzed and defined the optimal threshold value of RI between benign and malignant lesions. The mean values of the RI of benign and malignant lesions were 0.62 +/- 0.095 (range 0.44-0.86) and 0.74 +/- 0.097 (range, 0.50-0.92), respectively. The resistive index exceeded 0.7 in 80% of malignant lesions. The difference of the RI between malignant and benign lesions was statistically significant when the threshold value was 0.7 (P < 0.001). A resistive index over 0.7 may suggest malignant lesions. Due to the considerable overlap of the range of the RI, it may not be diagnostic in any single patient; however, it may be helpful in conjunct with gray-scale image.

US and CT findings of tuberculosis of the thyroid: three case reports.

With two ultrasonographic and two CT films of three cases of thyroid tuberculosis, we evaluate the ultrasonographic and CT findings and correlate them with the pathologic findings. They are demonstrated as heterogeneous hypoechoic mass on ultrasonogram and peripheral-enhancing low-density abscess on CT scan with regional lymphadenopathy. Ultrasonography (US) and CT can help the diagnosis of thyroid tuberculosis.

Hepatic arterial color Doppler signals in Caroli's disease.

Three siblings with congenital dilatation of the intrahepatic bile ducts (Caroli's disease) are presented. Bile duct pathology was associated with congenital hepatic fibrosis and polycystic renal disease in all three patients. On color Doppler imaging (CD imaging), multiple small color Doppler signals were observed in the vascular radicles within the dilated bile ducts or in the center of the lumen apart from the vascular radicles, as well as in other well-known sonographic findings such as bile duct dilatations and bilary calculi. Doppler frequency spectral analysis confirmed all these color signals as arterial in origin in all patients, revealing pulsatile wave patterns. In spite of the fact that portal venous radicles have been well described on conventional sonograms or computed tomography (CT), continuous wave patterns of venous flow on spectral analysis were not detected in all patients. Identification of such less emphasized arterial flow may add another clue in the diagnosis and pathogenesis of this rare disease entity. In conclusion, color Doppler signals of arterial wave pattern within the dilated bile ducts are another helpful diagnostic criteria in previously reported sonographic findings, and these color signals are easily depicted on sonograms with color mapping.

Character comparison of abdomen-derived and eyelid-derived mesenchymal stem cells.

OBJECTIVES: While most human adipose tissues, such as those located in the abdomen, hip and thigh, are of mesodermal origin, adipose tissues located in the face are of ectodermal origin. The present study has compared stem cell-related features of abdomen-derived adult stem cells (A-ASCs) with those of eyelid-derived adult stem cells (E-ASCs). MATERIALS AND METHODS: Adipose tissue-derived cells were maintained in DMEM supplemented with 10% FBS. Before passage 6, cells were analysed using FACS, immunocytochemistry and quantitative real time PCR (qRT-PCR). To examine multi-differentiational potential, early passage ASCs were cultivated in each of a commercial Stempro(®) Differentiation kit. RESULTS: Unlike fibroblast-like morphology of A-ASCs, E-ASCs had bipolar morphology. Both types of cell exhibited similar surface antigens, and neuronal cell-related genes and proteins. However, there were differences in mRNA expression levels of CD90 and CD146; neuron-specific enolase (NSE) and nuclear receptor-related protein 1 (Nurr1) were different between the two cell types. There was no difference in multi-differentiational potential between 3 E-ASCs lines, however, E-ASCs had higher expression levels of chondrocyte-related genes compared to A-ASCs. These cells underwent senescence and maintained normal karyotypes. CONCLUSIONS: Although isolated from similar adipose tissues, both types of cells displayed many contrasting characteristics. Understanding defining phenotypes of such cells is useful for making suitable choices in differing clinical indications.

Human insulin secreted from insulinogenic xenograft restores normoglycemia in type 1 diabetic mice without immunosuppression.

In the present study, we examined the therapeutic potential of human amnion-derived insulin-secreting cells for type 1 diabetes. Human amniotic mesenchymal stem cells (hAMs) were isolated from amnion and cultivated to differentiate into insulin-secreting cells in vitro. After culture in vitro, the differentiated cells (hAM-ISCs) were intensively stained with dithizone and secreted insulin and c-peptide in a high-glucose-dependent manner. They expressed mRNAs of pancreatic cell-related genes, including INS, PDX1, Nkx6-1, NEUROG3, ISL1, NEUROD1, GLUT1, GLUT2, PC1/3, PC2, GCK, PPY, SST, and GC, and were positive for human insulin and c-peptide. Transplantation of hAM-ISCs into the kidneys of mice with streptozotocin-induced diabetes restored body weight and normalized the blood glucose levels, which lasted for 210 days. Only human insulin and c-peptide were detected in the blood of normalized mice after 2 months of transplantation, but little mouse insulin and c-peptide. Removal of graft-bearing kidneys from these mice resulted in causing hyperglycemia again. Human cell-specific gene, hAlu, and human pancreatic cell-specific genes, insulin, PDX1, GLUT1, GLP1R, Nkx6-1, NEUROD1, and NEUROG3, were detected in the graft-bearing kidneys. Colocalization of human insulin and human nuclei antigen was also observed. These results demonstrate that hAMs could differentiate into functional insulin-secreting cells in vitro, and human insulin secreted from hAM-ISCs following transplantation into type 1 diabetic mice could normalize hyperglycemia, overcoming immune rejection for a long period.