PTTG is a Secretory Protein in Human Pituitary Adenomas and in Mouse Pituitary Tumor Cell Lines

The pituitary tumor-transforming gene (PTTG) is a homolog of yeast Securin, which arrests the activation of Separin to induce sister chromatid separation in the transition from metaphase to anaphase. Pituitary tumor-transforming gene is also known to induce angiogenesis during pituitary tumorigenesis. It has not been clarified whether PTTG functions as a cytoplasmic or a nuclear protein. Our immunohistochemical study indicated that PTTG is localized in the cytoplasm of pituitary tumor cells. In the present study, confocal laser scanning microscopy (CLSM) analysis of human pituitary adenomas and immunoelectron microscopy of the mouse pituitary cell line, AtT-20, demonstrated the localization of PTTG in the Golgi apparatus and vesicles. Secreted PTTG was detected by immunoblotting from culture medium of mouse pituitary tumor cell lines. Our results suggested that PTTG is a secretory protein produced by pituitary tumor cells. In addition, PTTG may exert autocrine and/or paracrine functions as a newly proposed important pathway for the action of PTTG.     

Development-dependent expression of cyclin D3 in precursor T-cell lymphoblastic leukemia/lymphoma

In contrast to the clear oncogenic role of cyclins D1 and D2, cyclin D3 is suggested to have a role in the initiation and/or maintenance of differentiation in a lineage-associated manner in addition to its basic role in proliferation. Recently, it has been reported that in cyclin D3-deficient mice, normal expansion of T lymphocytes is impaired because of maturation arrest at the double-negative thymocyte stage, suggesting a crucial role for cyclin D3 in early T-cell development. Therefore, cyclin D3 expression was examined in 36 human precursor T-lymphoblastic leukemia/lymphomas (T-LBLL), a neoplastic counterpart of T cells at the early developmental stages of differentiation. Using a standard panel of differentiation markers, all T-LBLL were categorized into four stages according to differentiation: progenitor, double-negative, double-positive, and single-positive stages. Cyclin D3 expression was initiated at the boundary between double-negative and double-positive stages, and was sustained in the single-positive stage. T-cell receptor was expressed simultaneously with cyclin D3, whereas CD79a expression was specific in the double-negative stage, and thus it was inversely correlated with that of cyclin D3. Taken together with the crucial and non-redundant role in T-cell development in mice, this molecule is suggested to play an important role in human T-cell development.     

The induced RNA‐binding protein,HuR, targets 3′‐UTR region of IL‐6 mRNA and enhances its stabilization in periodontitis

RNA‐binding proteins (RBPs) regulate mRNA stability by binding to the 3′‐untranslated region (UTR) region of mRNA. Human antigen‐R (HuR), one of the RBPs, is involved in the progression of diseases, such as rheumatoid arthritis, diabetes mellitus and some inflammatory diseases. Interleukin (IL)‐6 is a major inflammatory cytokine regulated by HuR binding to mRNA. Periodontal disease (PD) is also an inflammatory disease caused by elevations in IL‐6 following an infection by periodontopathogenic bacteria. The involvement of HuR in the progression of PD was assessed using in‐vitro and in‐vivo experiments. Immunohistochemistry of inflamed periodontal tissue showed strong staining of HuR in the epithelium and connective tissue. HuR mRNA and protein level was increased following stimulation with Porphyromonas gingivalis (Pg), one of the periodontopathogenic bacteria, lipopolysacchride (LPS)‐derived from Pg (PgLPS) and tumour necrosis factor (TNF)‐α in OBA‐9, an immortalized human gingival epithelial cell. The luciferase activity of 3′‐UTR of IL‐6 mRNA was increased by TNF‐α, Pg and PgLPS in OBA‐9. Luciferase activity was also increased in HuR‐over‐expressing OBA‐9 following a bacterial stimulation. Down‐regulation of HuR by siRNA resulted in a decrease in mRNA expression and production of IL‐6. In contrast, the over‐expression of HuR increased IL‐6 mRNA expression and production in OBA‐9. The HuR inhibitor, quercetin, suppressed Pg‐induced HuR mRNA expression and IL‐6 production in OBA‐9. An oral inoculation with quercetin also inhibited bone resorption in ligature‐induced periodontitis model mice as a result of down‐regulation of IL‐6. These results show that HuR modulates inflammatory responses by regulating IL‐6.     

VEGF-A induces tumor and sentinel lymph node lymphangiogenesis and promotes lymphatic metastasis

The mechanisms of tumor metastasis to the sentinel lymph nodes are poorly understood. Vascular endothelial growth factor (VEGF)-A plays a principle role in tumor progression and angiogenesis; however, its role in tumor-associated lymphangiogenesis and lymphatic metastasis has remained unclear. We created transgenic mice that overexpress VEGF-A and green fluorescent protein specifically in the skin, and subjected them to a standard chemically-induced skin carcinogenesis regimen. We found that VEGF-A not only strongly promotes multistep skin carcinogenesis, but also induces active proliferation of VEGF receptor-2-expressing tumor-associated lymphatic vessels as well as tumor metastasis to the sentinel and distant lymph nodes. The lymphangiogenic activity of VEGF-A-expressing tumor cells was maintained within metastasis-containing lymph nodes. The most surprising finding of our study was that even before metastasizing, VEGF-A-overexpressing primary tumors induced sentinel lymph node lymphangiogenesis. This suggests that primary tumors might begin preparing their future metastatic site by producing lymphangiogenic factors that mediate their efficient transport to sentinel lymph nodes. This newly identified mechanism of inducing lymph node lymphangiogenesis likely contributes to tumor metastasis, and therefore, represents a new therapeutic target for advanced cancer and/or for the prevention of metastasis.     

Simple self‐reported behavioral or psychological characteristics as risk factors for future type 2 diabetes in Japanese individuals: Toranomon Hospital Health Management Center Study 14

Aims/Introduction

Depression, anger, sleep disorders and cognitive impairment are regarded as presenting a high risk for diabetes. We investigated whether responses to single statements on a self-report questionnaire on the presence of each of these four factors were associated with the development of type 2 diabetes.

Materials and Methods

We investigated 3,211 Japanese individuals without diabetes. Cumulative incidence rate and hazard ratios (HRs) for future type 2 diabetes over 7–13 years were evaluated according to the presence of lack of perseverance, anger, memory loss or sleep disorders.

Results

Results of Cox regression analysis showed that lack of perseverance (age- and sex-adjusted HR 1.41, 95% confidence interval 1.07–1.84), anger, (HR 1.51, 95% confidence interval 1.07–2.12) or memory loss (HR 1.47, 95% confidence interval 1.14–1.90) was predictive of the development of diabetes. Even after adjustment for metabolic factors including glycemic measurements, anger was significantly associated with an increased risk of future diabetes. Individuals with both anger and memory loss had a 1.94-fold (95% confidence interval 1.19–3.15) increased risk of type 2 diabetes than those without those two symptoms.

Conclusions

Responses to a simple self-report questionnaire as to whether individuals were aware of anger or memory loss were associated with the development of type 2 diabetes independent of traditional risk factors for diabetes in this cohort of Japanese individuals.     

Micro-computed tomography for evaluating alveolar bone resorption induced by hyperocclusion

Purpose

Occlusal trauma, resulting in the destruction of alveolar bone, is a form of periodontal disease caused by excessive mechanical stress (MS) during hyperocclusion. Previously, we showed that CC chemokine ligand (CCL) 2/CCR2 receptor axis plays a crucial role in MS-dependent osteoclastogenesis. However, in the previous work, we were unable to precisely measure changes in alveolar bone profiles. In the present study, we sought to establish a precise method for evaluating alveolar bone resorption induced by hyperocclusion using micro-computed tomography.

FDG PET for grading malignancy in thymic epithelial tumors: Significant differences in FDG uptake and expression of glucose transporter-1 and hexokinase II between low and high-risk tumors: Preliminary study

Purpose

To evaluate ¹⁸F-fluorodeoxyglucose (FDG) uptake to predict the malignant nature and analyze the correlation between FDG uptake and expression of glucose transporter 1 (Glut-1) and hexokinase II (HK-II) in thymic epithelial tumors.

Materials and methods

Eleven patients with a thymic epithelial tumor who underwent FDG PET/CT before therapy were reviewed. The thymic tumors were classified by the WHO histological classification and Masaoka clinical staging. Comparison of maximum standardized uptake value (SUV_max) of the lesion was made between the low-risk (Type A, AB and B1) and high-risk {Type B2, B3 and C (thymic cancer)} groups and among clinical stages. Expression of Glut-1 and HK-II was analyzed immunohistochemically.

Results

All 11 tumors showed FDG uptake visually. SUV_max was significantly higher in the high-risk group (n = 5, 5.24 ± 2.44) than the low-risk group (n = 6, 3.05 ± 0.55) (P = 0.008). Staining scores of both Glut-1 and HK-II were significantly higher in the high-risk group than in the low-risk group (Glut1: P = 0.034 and HK-II: P = 0.036). There were no significant differences in SUV_max (P = 0.11), Glut-1 (P = 0.35) and HK-II scores (P = 0.29) among clinical stages. SUV_max was significantly correlated to each of the staining scores of Glut-1 (ρ = 0.68, P = 0.031) and HK-II (ρ = 0.72, P = 0.024).

Conclusion

These preliminary results support the previously published view that SUV_max may be useful to predict the malignant nature of thymic epitherial tumors and suggest that the degree of FDG uptake in the thymic epitherial tumors is closely related to the amount of Glut-1 and HK-II in the tumor.     

Rhesus diploid rabies vaccine (adsorbed): a new rabies vaccine using FRhL-2 cells

The FRhL-2 cell line, a diploid line derived from the lung of a fetal rhesus monkey, was used to prepare a potent rabies vaccine by adapting the Kissling strain of rabies virus to FRhL-2 cells, growing the virus in quantity, inactivating the virus with beta-propiolactone, and concentrating the virus by adsorption to aluminum phosphate. High levels of antibody to rabies virus, induced by the vaccine in both guinea pigs and humans at 14 days after immunization, were determined to be IgG. Data from postexposure protocols with guinea pigs and simulated postexposure protocols in humans showed protection and antibody response even when rabies immune globulin was administered at the time of vaccination.