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The presence of protein inclusions, called Lewy bodies (LBs) and Lewy neurites (LNs), in the brain is the main feature of Parkinson’s disease (PD). Recent evidence that the prion-like propagation of α-synuclein (α-syn), as a major component of LBs and LNs, plays an important role in the progression of PD has gained much attention, although the molecular mechanism remains unclear. In this study, we evaluated whether neuronal ApoE regulates the cell-to-cell transmission of α-syn and explored its molecular mechanism using in vitro and in vivo model systems. We demonstrate that neuronal ApoE deficiency attenuates both α-syn uptake and release by downregulating LRP-1 and LDLR expression and enhancing chaperone-mediated autophagy activity, respectively, thereby contributing to α-syn propagation. In addition, we observed that α-syn propagation was attenuated in ApoE knockout mice injected with pre-formed mouse α-syn fibrils. This study will help our understanding of the molecular mechanisms underlying α-syn propagation.

This paper investigates the concrete breakout shear capacity of a single cast-in headed anchor embedded in steel fiber-reinforced concrete (SFRC). Experiments were carried out for anchors with a diameter of 30 mm, embedment depth of 240 mm, and edge distance of 150 mm. Fiber volume fractions of 0.33, 0.67, and 1.00% were examined. In addition to shear tests, four-point bending tests were carried out to evaluate the relationship between the material properties of SFRC and the shear capacity of anchors. The experimental results show that the current design formulas are valid for anchors in conventional plain concrete but not appropriate for the shear capacity of anchors in SFRC. Using the equivalent flexural strength ratio, a revised design equation is proposed to incorporate the effect of the steel fiber content on the concrete breakout shear capacity and provide reasonably similar safety for anchors in both un-reinforced and SFRC.

This study aimed to apply fluoride formulations to enamel with cold atmospheric plasma (CAP) and analyze the fluoride uptake, retention, and acid resistance quantitatively. Human enamel specimens were divided randomly into 2 groups: group APF1, 1.23% acidulated phosphate fluoride (APF) gel; group APF2, 1.23% APF gel with CAP. Fluoride and CAP were applied to the samples 4 times at 1-wk intervals. The specimens were also stored in artificial saliva for 4 wk to evaluate the retention of fluoride. The fluoride content on the fluoride-treated enamel was measured by an electron probe microanalyzer. To detect the resistance to demineralization, the calcium-to-phosphate ratio of the enamel samples was measured after the application of APF gel with or without CAP, followed by soaking in the demineralization solution. In groups APF1 and APF2, the amount of fluoride detected increased depending on the application frequency, and more fluoride was detected in group APF2 than in group APF1. In the experiment examining the maintenance effect, fluoride was not detected in group APF1, whereas fluoride was detected in group APF2 up to the fourth week. As for the resistance to demineralization, the calcium-to-phosphate ratio of the enamel treated with APF and CAP was higher than that treated with APF alone, and it increased with the frequency of treatment. This study suggests that the combination treatment of CAP and fluoride improves retention of fluoride on the enamel and resistance to demineralization when compared with treatment with fluoride alone.

Tests of the lateral and torsional stability are quite sensitive to the experimental conditions, such as support conditions and loading system. Controlling all of these conditions in a full-size test is a very challenging task. Therefore, in this paper, an experimental measurement method that can control the experimental conditions using a small-scale model was proposed to evaluate the lateral and torsional stability of beams. For this, a loading system was provided to maintain the vertical direction of the load applied to the beam, and a support frame was produced to satisfy the in-plane and out-of-plane support conditions. The experimental method using a small-scale model was applied successively to the lateral and torsional behavior and stability of I-shaped beams. The proposed experimental methods, which effectively accommodate the changes in the geometry and length of the beam, could contribute to further experimental studies regarding the lateral and torsional stability of flexural members.

Among autologous breast reconstruction techniques, breast reconstruction using the latissimus dorsi musculocutaneous flap is widely used, offering advantages including the relative simplicity of the procedure and the reliable and consistent vascularity of the flap. Accordingly, more than 500 cases have been performed in the past 8 years at Kyungpook National University Medical Center. This study reports on a rare case involving a radial nerve neuropathy complication which was experienced for the first time at the medical center. The current case demonstrates that in addition to common complications, such as seroma of the donor site and scarring, additional intraoperative complications in areas unrelated to the surgical site can occur, including radial nerve neuropathy in the opposite arm.

Pseudoangiomatous stromal hyperplasia (PASH) is a benign proliferative mesenchymal lesion of the breast. PASH presents as a palpable or mammographically detectable mass or is an incidental microscopic finding in up to 23% of breast biopsies. Its pathogenesis is unclear but may involve hormonal factors, as recently postulated. Its clinical, cytological, and radiological characteristics resemble those of other benign tumors, including low-grade angiosarcomas and phyllodes tumors. Because of its uncertain pathogenesis, PASH is usually treated via surgical excision, with post-excision recurrence rates ranging from 7% to 22% [1]. Although several cases of PASH have been recently reported, cases of PASH involving large, rapidly growing lesions refractory to surgical incision are rare [2]. In this study, we report our experience with a case of PASH that presented as rapidly growing bilateral breast enlargement refractory to surgical excision. We also review the pertinent literature, including the clinical and histopathological features, pathogenesis, imaging findings, treatment, and prognosis of PASH.

Background The emergence of breast-conserving surgery combined with radiotherapy as the treatment of choice for early stage breast cancer has resulted in greater focus on oncoplastic breast surgery. The use of perforator flaps has particularly gained in reputation for its effectiveness in the reconstruction of partial breast defects in Korean women. Herein, we present our experience with the use of thoracodorsal artery perforator (TDAP) and lateral intercostal artery perforator (LICAP) flaps. Methods This study included 33 patients who underwent breast reconstruction using TDAP or LICAP flaps at our hospital from January 2011 to December 2014. Data from patient medical records, and patient satisfaction surveys, which were conducted 12 months postoperatively, were retrospectively evaluated. Results TDAP and LICAP flap-based reconstructions were performed in 14 and 19 patients, respectively. Five patients developed complications that required additional intervention. Overall patient satisfaction was observed to be excellent in 15 (46%) patients, and good in 12 (36%). Conclusions Based on our experience, oncoplastic breast surgery using TDAP or LICAP flap is an effective remodeling technique for small-to-moderate breast defects in Korean women with smaller breasts.

Summary: Recent evidence of prion-like propagation of α-synuclein (α-syn) into neighboring neurons set up a paradigm to elucidate the mechanism of progression of Parkinson’s disease (PD) and to develop therapeutic strategies. Here, we show that FcγRIIB expressed in neurons functions as a receptor for α-syn fibrils and mediates cell-to-cell transmission of α-syn. SHP-1 and 2 are activated downstream by α-syn fibrils through FcγRIIB and play an important role in cell-to-cell transmission of α-syn. Also, taking advantage of a co-culture system, we show that cell-to-cell transmission of α-syn induces intracellular Lewy body-like inclusion body formation and that the FcγRIIB/SHP-1/2 signaling pathway is involved in it. Therefore, the FcγRIIB-SHP-1/-2 signaling pathway may be a therapeutic target for the progression of PD. The in vitro system is an efficient tool for further high-throughput screening that can be used for developing a therapeutic intervention in PD. : Prion-like propagation of α-synuclein (α-syn) may contribute to the progression of Parkinson’s disease. Choi et al. demonstrate that FcγRIIB functions as a receptor for α-syn fibrils and that the FcγRIIB-SHP-1/2 signaling pathway mediates cell-to-cell transmission of α-syn. Blocking this signaling pathway attenuates transmission, suppressing Lewy body-like inclusion body formation. Keywords: α-synuclein, cell-to-cell transmission, FcγRIIB, SHP-1, SHP-2

*Department of Biochemistry, Hallym University Medical School, Chuncheon 200-702, Korea.**Institute of Natural Medicine, Hallym University Medical School, Chuncheon 200-702, Korea.***Department of Food and Nutrition, College of Natural Sciences, Hallym University, Chuncheon 200-702, Korea.****Institute of Cell Differentiation and Aging, Hallym University Medical School, Chuncheon 200-702, Korea.ABSTRACT : Chelidonium majus (CM) contains several isoquinoline alkaloids that have been reported to have various bio-logical activities such as anti-inflammatory, antimicrobial, antioxidant, immune-modulatory, and antitumoral. It has beenreported that the extract of CM had an antioxidant potential, however the mechanism has not been verified. In this study,we found that CM extract activated FOXO3a. FOXO3a is a transcription factor that involved in various biological processessuch as cell cycle arrest, apoptosis, DNA repair, and ROS detoxification. Transcriptional activities of FOXO3a were regu-lated by post-translational modifications including phosphorylation, acetylation, and ubiquitination. Protein level ofFOXO3a was increased by CM extract. Promoter activities of FOXO-transcriptional target genes such as MnSOD, p27 andGADD45 were activated by CM extract in a dose dependent manner. In addition, protein level of MnSOD, major antioxi-dant enzyme, was increased by CM extract. Thereby ROS level was decreased by CM in old HEF cells. These results suggestthat CM extract has an antioxidant activity through FOXO activation. Key Words : Chelidonium majus, FOXO3a, Antioxidant

The purpose of this study is to review and suggest coastal shipping policies for introduction of WIG craft into domenstic passenger shipping market. Two Korean companies are leading in the development of WIG craft as an innovative sea transportation vehicle, and it is awaiting for commercialization. WIG craft is expected to be commercialized from coastal passenger market and we used AHP method to investigate the major factors and its` priority for smooth market entry. The results shows that priorities are on the WIG craft legislation, dock installation and operation, port state control, and pilot hiring and training, craft maintenance, and ship`s certification in order.

Logistics in a supply chain network has become an important operational strategy in a competitive market. A number of internal and external risks involved in the logistics operations in a company tend to create problems in fulfilling customer orders. This research presents how ERM (enterprise risk management) can be used to identify, assess, and control logistics risks. An electronic company's logistics activities were used as an industry case to demonstrate a way to identify and assess risks surrounding global logistics function. This paper has further presented action plans to mitigate the impact of the risks that occur.

In the central Busan area from Haeundae to Saha, fishing villages are vanished actually in the progress of urbanization, and 17 fishing ports surrounded by urbanized area are left alone and isolated. But, we think these urbanized fishing ports are the very resource which grant the identity as the marine city and they have considerable potentials to enhance the value of neighboring community depending on the proper development of it. In this study, we defined the fishing ports in central Busan area as the representatives of `urban fishing port`, and tried to find the way of coexistence and harmony between `city` and `fishing port`. For this purpose, we investigated the fishery potentials, physical environments and development demands of 17 fishing ports in central Busan area, and composed opinions from fishing communities and neighboring inhabitants to set up the long term directions of urban fishing ports development for harmonious coexistence of city and fishing port.

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.

Hedgehog (Hh) signaling is an essential pathway in embryonic development of prostate. Hh also plays roles in the proliferation of progenitor cells and cancer cells of adult prostate. However, how Hh signaling contributes to carcinogenesis of prostate is poorly understood. Stathmin1 is a microtubule-regulating protein that plays an important role in the assembly and disassembly of the mitotic spindle. Stathmin1 is expressed in normal developing mouse prostate and in prostate cancer. The expression pattern of stathmin1 is similar to that of Shh in prostate development and cancer, suggesting a connection between these two proteins. In this study, we examined the relationship between stathmin1 and Hh signaling. Here, we show that stathmin1 expression is regulated by Hh signaling in prostate cancer cells. Cyclopamine, a specific inhibitor of Hh signaling, reduced the expression of stathmin1 in prostate cancer cells. However, the Shh peptide induced stathmin1 expression. Overexpression of Gli1 further confirmed the relationship. Co-expression of stathmin1 and Patched 1, a receptor for Hh signaling was observed in prostate cancer tissues. Cyclopamine and stathmin1 siRNA both decreased proliferation of prostate cancer cells but did not produce an additive effect, suggesting a common pathway. These results suggest that Hh signaling regulates proliferation of prostate cancer cells by controlling stathmin1 expression.

Thymus can regenerate to its normal mass within 14 days after acute involution induced by cyclophosphamide (CY) in adult rat. Despite the established role of Wnt pathways in the process of thymus development, they have not yet been associated with the regeneration of adult thymus. The purpose of this study was to investigate whether Wnt7b, which is expressed in developing thymic epithelial cells rather than in thymocytes, is modulated during thymic regeneration in adult rat. Here, we show that Wnt7b expression was up-regulated in the regenerating thymus. Cells immunolabeled for the Wnt7b were identified as macrophages. Furthermore, Wnt7b gene expression was decreased by the treatment of receptor activator of NF-kappaB ligand (RANKL). Taken together, our results demonstrate that Wnt7b gene expression was increased in macrophages during thymic regeneration and negatively regulated by RANKL.

Background Lymphatic malformation (LM) is a form of congenital vascular malformation with a low incidence. Although LM has been studied, no consensus has emerged regarding its cause or treatment. Methods In this study, we retrospectively evaluated 40 patients who visited our vascular anomalies center for the treatment of cervicofacial LM, which is a common manifestation of LM. The medical records of patients over a period of 12 years were reviewed and analyzed for commonalities regarding the diagnosis and the results of treatment. Results Suspected cervicofacial LM was confirmed through imaging studies. No difference in incidence was observed according to sex, and 73% of patients first presented with symptoms before the age of two years. The left side and the V2–V3 area were most commonly affected. No significant differences in incidence were observed among the macrocystic, microcystic, and combined types of LM. A total of 28 out of 36 patients received sclerotherapy as the first choice of treatment, regardless of the type of lesion. Complete resolution was achieved in only 25% of patients. Conclusions LM is important to confirm the diagnosis early and to choose an appropriate treatment strategy according to the stage of the disease and each individual patient's symptoms. When treatment is delayed or an incorrect treatment is administered, patient discomfort increases as the lesion gradually spreads. Therefore, more so than is the case for most other diseases, a team approach on a case-by-case basis is important for the accurate and appropriate treatment of LM.

Gastric cancer is the second most common cause of cancer deaths worldwide. The underlying molecular mechanisms of its carcinogenesis are relatively poorly characterized. Hedgehog (Hh) signaling, which is critical for development of various organs including the gastrointestinal tract, has been associated with gastric cancer. The present study was undertaken to reveal the underlying mechanism by which Hh signaling controls gastric cancer cell proliferation. Treatment of gastric cancer cells with cyclopamine, a specific inhibitor of Hh signaling pathway, reduced proliferation and induced apoptosis of gastric cancer cells. Cyclopamine treatment induced cytochrome c release from mitochondria and cleavage of caspase 9. Moreover, Bcl-2 expression was significantly reduced by cyclopamine treatment. These results suggest that Hh signaling regulates the survival of gastric cancer cells by regulating the expression of Bcl-2.

Background Caffeine is the most commonly consumed psycho-stimulant in the world. The effects of caffeine on the body have been extensively studied; however, its effect on the structure of the brain has not been investigated to date. Results In the present study we found that the long-term consumption of caffeine can induce ventriculomegaly; this was observed in 40% of the study rats. In the caffeine-treated rats with ventriculomegaly, there was increased production of CSF, associated with the increased expression of Na+, K+-ATPase and increased cerebral blood flow (CBF). In contrast to the chronic effects, acute treatment with caffeine decreased the production of CSF, suggesting 'effect inversion' associated with caffeine, which was mediated by increased expression of the A1 adenosine receptor, in the choroid plexus of rats chronically treated with caffeine. The involvement of the A1 adenosine receptor in the effect inversion of caffeine was further supported by the induction of ventriculomegaly and Na+, K+-ATPase, in A1 agonist-treated rats. Conclusion The results of this study show that long-term consumption of caffeine can induce ventriculomegaly, which is mediated in part by increased production of CSF. Moreover, we also showed that adenosine receptor signaling can regulate the production of CSF by controlling the expression of Na+, K+-ATPase and CBF.

In many clinical situations which cause thymic involution and thereby result in immune deficiency, T cells are the most often affected, leading to a prolonged deficiency of T cells. Since only the thymic-dependent T cell production pathway secures stable regeneration of fully mature T cells, seeking strategies to enhance thymic regeneration should be a key step in developing therapeutic methods for the treatment of these significant clinical problems. This study clearly shows that receptor activator of NF-kappaB ligand (RANKL) stimulates mouse thymic epithelial cell activities including cell proliferation, thymocyte adhesion to thymic epithelial cells, and the expression of cell death regulatory genes favoring cell survival, cell adhesion molecules such as ICAM-1 and VCAM-1, and thymopoietic factors including IL-7. Importantly, RANKL exhibited a significant capability to facilitate thymic regeneration in mice. In addition, this study demonstrates that RANKL acts directly on the thymus to activate thymus regeneration regardless of its potential influences on thymic regeneration through an indirect or systemic effect. In light of this, the present study provides a greater insight into the development of novel therapeutic strategies for effective thymus repopulation using RANKL in the design of therapies for many clinical conditions in which immune reconstitution is required.

Angiogenesis is important for adult tissue regeneration as well as normal development. Vascular endothelial growth factor (VEGF) is a unique potent angiogenic factor, and plays an essential role in regulating angiogenesis during embryonic development, normal tissue growth, and tissue regeneration. Recent evidence shows that nerve growth factor (NGF) also plays a role as an angiogenic regulator as well as a well-known neurotrophic factor. The aim of this study was to investigate whether thymus regeneration accompanies reparative angiogenesis and also to evaluate whether the thymic expression of VEGF is regulated by NGF in vivo and in vitro. Here, we show that high VEGF mRNA and protein levels are concomitant with reparative angiogenesis that occurs dramatically during regeneration following acute involution induced by cyclophosphamide (CY) in the rat thymus. Fluorescent thymus angiography using FITC-dextran showed that thymic regeneration is associated with a much denser capillary network compared with normal control thymus. Furthermore, the expressions of NGF and TrkA were highly increased during thymic regeneration. We also show that NGF mediates thymic epithelial induction of VEGF expression in vitro and in vivo. Taken together, our results suggest that NGF-mediated VEGF up-regulation in thymic epithelial cells may contribute to reparative angiogenesis during thymic regeneration in adult.

The marked induction of glial fibrillary acidic protein (GFAP) has been observed in astrocytes during neuropathological processes accompanying reactive gliosis; however, the precise molecular mechanism(s) underlying this GFAP induction remains poorly resolved. Therefore, in this study, we examined whether the change of nuclear factor-kappa B (NF-kappaB) activity can influence GFAP expression levels. Aspirin, widely used to prevent NF-kappaB activity, reduced the levels of GFAP mRNA and protein in human astroglial cells including human glioblastoma A172 cells and primary human brain astrocyte cells (HBAs). Furthermore, aspirin inhibited the effects of hypoxic injury on the up-regulation of GFAP expression in HBAs. We confirmed the repressive effect of aspirin on GFAP transcription by GFAP promoter-driven reporter assay and found that one NF-kappaB binding site conserved in the mouse and human GFAP gene promoters is critical for this effect. To further delineate whether NF-kappaB is directly involved in the regulation of GFAP gene expression, we transfected A172 cells with an expression vector encoding a super-repressor IkappaBalpha protein (IkappaBalpha-SR) to specifically inhibit NF-kappaB activity and found the marked reduction of GFAP protein levels in IkappaBalpha-SR-transfectant cells. Taken together, our results suggest that NF-kappaB may play pivotal roles in GFAP gene expression.

This study is performed to detect changes of gene expression in substantia nigra (SN) and striatum in manganese (Mn)-exposed mice brain. The cDNA array is a recently developed molecular biological method that can detect the differential expression of several hundreds of genes simultaneously and is therefore advantageous in the study of trace metal intoxication effect at the genetic level. Using this technology, we discovered 5 genes in the mouse striatum and 9 genes in SN changed by more than 50% following Mn exposure. Depression were observed in two genes (neural cell adhesion protein BIG2, heavy neurofilament subunit genes) in striatum and three genes (light neurofilament subunit, brain acyl-CoA synthetase II, heavy neurofilament subunit genes) in the SN. However three genes (N-acetylglucosaminyltransferase I, S100beta, and synaptonemal complex protein I genes) in striatum and six genes (noggin, striatin, Ost oncogene, S100beta, calcium/calmodulin-dependent protein kinase kinase beta, and N-acetylglucosaminyltransferase I genes) in SN were elevated following Mn exposure. Immunohistochemical study revealed that protein levels of S100beta also increased following Mn treatment. Activated astrocytes overexpressing S100beta are invariably and intimately associated with decreased expression of heavy and light neurofilament subunits which is a distinguishing feature of neurodegeneration by Mn exposure. All our findings suggested that neuronal degenerations occur in SN as well as striatum of mice exposed to Mn.

The acidic glycoconjugates of mouse ileum Paneth cells were examined with the aid of light and electron microscopy, using cationic colloidal gold (CCG) as a probe. Specimens of mouse ilea were fixed in half-strength Karnovsky's fixative and embedded in Lowicryl K4M resin. Semithin and ultrathin sections were cut of examination with light and electron microscopy, respectively. Examination of the sections using light microscopy revealed the positive staining of CCG at pH 1.0 and pH 2.5, which was detected at the rim of secretory granules and at the supranuclear regions of the Paneth cells. At pH 4.0, in addition to staining of the secretory granule rim, weak staining was observed in the granule core. At pH 7.2, the cytoplasm other than secretory granules exhibited positive CCG staining. Examination of the sections using electron microscopy, at pH 1.0, the trans lamellae of the Golgi apparatus, the rim of the secretory granules, and lysosomes were labeled selectively by CCG. At pH 2.5, labeling was also discernible over the same structures in the cells. However, at this pH, the labeling intensity was stronger than that at pH 1.0, due to the dual labeling of sulfated and sialylated glycoconjugates in these structures. At pH 4.0, the Golgi apparatus, rims and cores of secretory granules and ribosomes were labeled. Lysosomes and nuclei were also positively stained. At pH 7.2, the rims of secretory granules were not stained. The present results indicate that the CCG method gives good resolution and contrast when applied to staining, and therefore is useful for the specific staining of glycoconjugates such as sulfated, sialylated and phosphated glycoconjugates for light and electron microscopy.

Rat pheochromocytoma (PC12) cells have been used to investigate neurite outgrowth. Nerve growth factor (NGF) has been well known to induce neurite outgrowth from PC12 cells. RhoA belongs to Ras-related small GTP-binding proteins, which regulate a variety of cellular processes, including cell morphology alteration, actin dynamics, and cell migration. NGF suppressed GTP-RhoA levels after 12 h in PC12 cells and was consistently required for a long time to induce neurite outgrowth. Constitutively active (CA)-RhoA suppressed neurite outgrowth from PC12 cells in response to NGF, whereas dominant-negative (DN)-RhoA stimulated it, suggesting that RhoA inactivation is essential for neurite outgrowth. Here, we investigated the mechanism of RhoA inactivation. DN-p190RhoGAP abrogated neurite outgrowth, whereas wild-type (WT)-p190RhoGAP and WT-Src synergistically stimulated it along with accelerating RhoA inactivation, suggesting that p190RhoGAP, which can be activated by Src, is a major component in inhibiting RhoA in response to NGF in PC12 cells. Contrary to RhoA, Rap1 was activated by NGF, and DN-Rap1 suppressed neurite outgrowth, suggesting that Rap1 is also essential for neurite outgrowth. RhoA was co-immunoprecipitated with Rap1, suggesting that Rap1 interacts with RhoA. Furthermore, a DN-Rap-dependent RhoGAP (ARAP3) prevented RhoA inactivation, abolishing neurite formation from PC12 cells in response to NGF. These results suggest that NGF activates Rap1, which, in turn, up-regulates ARAP3 leading to RhoA inactivation and neurite outgrowth from PC12 cells. Taken together, p190RhoGAP and ARAP3 seem to be two main factors inhibiting RhoA activity during neurite outgrowth in PC12 cells in response to NGF.

4-1BB, a member of the tumor necrosis factor receptor (TNFR) superfamily, is a major costimulatory receptor that is rapidly expressed on the surface of CD4(+) and CD8(+) T cells after antigen- or mitogen-induced activation. The interaction of 4-1BB with 4-1BBL regulates immunity and promotes the survival and expansion of activated T cells. In this study, the expression of 4-1BB and 4-1BBL was examined during regeneration of the murine thymus following acute cyclophosphamide- induced involution. Four-color flow cytometry showed that 4-1BB and 4-1BBL were present in the normal thymus and were preferentially expressed in the regenerating thymus, mainly in CD4(+)CD8(+) double-positive (DP) thymocytes. Furthermore, the CD4(lo)CD8(lo), CD4(+)CD8(lo) and CD4(lo)CD8(+) thymocyte subsets, representing stages of thymocyte differentiation intermediate between DP and single-positive (SP) thymocytes, also expressed 4-1BB and 4-1BBL during thymus regeneration but to a lesser degree. Interestingly, the 4-1BB and 4-1BBL positive cells among the CD4(+)CD8(+) DP thymocytes present during thymus regeneration were TCR(hi) and CD69(+) unlike the corresponding controls. Moreover, the 4-1BB and 4-1BBL positive cells among the intermediate subsets present during thymus regeneration also exhibited TCR(hi/int+) and CD69(+/int) phenotypes, indicating that 4-1BB and 4-1BBL are predominantly expressed by the positively selected population of the CD4(+)CD8(+) DP and the intermediate thymocytes during thymus regeneration. RT-PCR and Western blot analyses confirmed the presence and elevated levels of 4-1BB and 4-1BBL mRNA and protein in thymocytes during thymus regeneration. We also found that the interaction of 4-1BB with 4-1BBL promoted thymocyte adhesion to thymic epithelial cells. Our results suggest that 4-1BB and 4-1BBL participate in T lymphopoiesis associated with positive selection during recovery from acute thymic involution.