Association Between Nonrestorative Sleep and Risk of Diabetes: A Cross-Sectional Study.

Although insufficient sleep or poor sleep quality has been reported to be associated with the development of type 2 diabetes, the relation of type 2 diabetes with nonrestorative sleep (NRS), a subjective feeling, has been overlooked. We used a large-scale medical checkup database to investigate whether there is a cross-sectional association between NRS and type 2 diabetes risk in Japanese individuals. We extracted data for 14,476 individuals who were not receiving therapeutic drugs for diabetes. About 36.8% of individuals were identified as having NRS. In a multiple logistic regression analysis, NRS was significantly associated with the risk of developing diabetes. Thus, this line of research may have implications for diabetes prevention.

Association between novel arterial stiffness indices and risk factors of cardiovascular disease.

BACKGROUND: Prevention and early detection of arterial stiffness are required to avoid severe cardiovascular events. Recently, new noninvasive arterial stiffness indices, the arterial pressure volume index (API) and the arterial velocity pulse index (AVI), have been developed. The purpose of this study was to examine the clinical validity of these new indices by investigating the association between known risk factors of cardiovascular disease (CVD) and API or AVI in a large population. METHODS: This cross-sectional survey included 7248 adults who underwent an annual medical checkup at a single medical institution. API and AVI were measured using cuff oscillometry by trained nurses. We used correlation coefficients, t-tests, and multiple regression analyses to evaluate associations, and calculated intraclass correlation coefficients (ICC) to examine test-retest reliabilities of these indices. RESULTS: Mean age was 45.5 years (SD = 5.8), and 4083 (56.3 %) participants were men, while 3165 were women. Mean values of API and AVI were 25.1 (SD = 7.0) and 16.6 (SD = 5.4), respectively. API was strongly correlated with body mass index (BMI), systolic blood pressure (sBP), and diastolic blood pressure (dBP) (r > 0.3, p < 0.001). AVI was strongly correlated with age, sBP, and API (r > 0.3, p < 0.001). Multiple regression analyses showed that sex, age, BMI, and sBP were independently associated with API. Sex, age, BMI, sBP, fasting plasma glucose (FPG), and smoking condition were also independently associated with AVI. As reliabilities of measurements, the ICC of API was 0.74, and the ICC of AVI was 0.80. CONCLUSIONS: These new noninvasive arterial stiffness indices, which had high test-retest reliabilities, were associated with known risk factors of CVD. Further study is warranted to determine the clinical validity of these indices.

Erythropoietin Receptor Antagonist Suppressed Ectopic Hemoglobin Synthesis in Xenografts of HeLa Cells to Promote Their Destruction.

The aim of this study is to explore a cause-oriented therapy for patients with uterine cervical cancer that expresses erythropoietin (Epo) and its receptor (EpoR). Epo, by binding to EpoR, stimulates the proliferation and differentiation of erythroid progenitor cells into hemoglobin-containing red blood cells. In this study, we report that the HeLa cells in the xenografts expressed ε, γ, and α globins as well as myoglobin (Mb) to produce tetrameric α2ε2 and α2γ2 and monomeric Mb, most of which were significantly suppressed with an EpoR antagonist EMP9. Western blotting revealed that the EMP9 treatment inhibited the AKT-pAKT, MAPKs-pMAPKs, and STAT5-pSTAT5 signaling pathways. Moreover, the treatment induced apoptosis and suppression of the growth and inhibited the survival through disruption of the harmonized hemoprotein syntheses in the tumor cells concomitant with destruction of vascular nets in the xenografts. Furthermore, macrophages and natural killer (NK) cells with intense HIF-1α expression recruited significantly more in the degenerating foci of the xenografts. These findings were associated with the enhanced expressions of nNOS in the tumor cells and iNOS in macrophages and NK cells in the tumor sites. The treated tumor cells exhibited a substantial number of perforations on the cell surface, which indicates that the tumors were damaged by both the nNOS-induced nitric oxide (NO) production in the tumor cells as well as the iNOS-induced NO production in the innate immune cells. Taken together, these data suggest that HeLa cells constitutively acquire ε, γ and Mb synthetic capacity for their survival. Therefore, EMP9 treatment might be a cause-oriented and effective therapy for patients with squamous cell carcinoma of the uterine cervix.

Hypoxia correlates with angiogenesis in cervical cancers.

BACKGROUND: Tissue hypoxia stimulates the induction of the angiogenic substances vascular endothelial growth factor and erythropoietin in the locus of the tissue. We have previously demonstrated that erythropoietin promotes angiogenesis by binding to its receptor in the endothelial cells of uterine and ovarian malignancies. In the present study, we examined whether malignant uterine cervix tissue showed hypoxia and whether hypoxia correlated with high vascular density through vascular endothelial growth factor. METHODS: To detect tissue hypoxia, we estimated the content of ATP in squamous cell carcinoma of the uterine cervix and in the normal cervix, using liquid chromatography columns. Surgically resected samples were fixed in Zamboni solution and processed for immunohistochemical microscopy to identify the endothelial cells and the location of vascular endothelial growth factor, with the use of anti-factor VIII and anti-vascular endothelial growth factor165 antibody, respectively. The microvessels in a definite area were counted in sections of each specimen. RESULTS: Significantly lower ATP levels and significantly higher vascular density were seen in squamous cell carcinoma than in the controls (P<0.05). The microvessel number in relation to ATP content was significantly higher in squamous cell carcinoma than in the controls (P<0.001). Moreover vascular endothelial growth factor, the hyperplastic epithelium of the squamous cell carcinoma contained the immunoreactivity, with characteristic histopathological features suggesting retention of tissue fluid. CONCLUSION: Squamous cell carcinoma of the uterine cervix showed hypoxia which correlated with abundant vascularity. Vascular endothelial growth factor expressed in the hyperplastic epithelium appears to promote angiogenesis in squamous cell carcinoma of the uterine cervix.

Erythropoietin regulates tumour growth of human malignancies.

In addition to the chief function of erythropoietin (Epo) in promoting erythropoiesis, some other roles have been found in the brain and uterus. We have reported that signalling pathways of Epo and Epo receptor (EpoR) are involved in the tumourigenesis of ovarian and uterine cancers. To determine whether Epo plays a similar role in other malignancies, we studied the expression of Epo in several malignant human cell lines. We found that 24 malignant human cell lines examined express Epo and EpoR regardless of their origins, types, genetic characteristics and biological properties and secrete a very small amount of Epo individually and that most of them respond to hypoxic stimuli by enhanced secretion of Epo. To determine whether the Epo-EpoR pathway operates in tumours of these cell lines, we transplanted several cell lines into nude mice and confirmed the presence of Epo-responsive sites in xenografts in which the phosphorylation of the STAT5 (signal transducer and activator of transcription) is detectable. Furthermore, in nude mice we blocked the Epo signalling in xenografts of two representative cell lines, stomach choriocarcinoma and melanoma, by i.p. injections of EpoR antagonist and found inhibition of angiogenesis and survival of tumour cells leading to destruction of tumour masses and disturbances of phosphorylation of STAT5. In contrast, Epo mimetic peptide promotes angiogenesis and tumour cell survival. These findings suggest that Epo is indispensable for the growth and viability of malignant tumour and also that the deprivation of Epo signalling may be a promising therapy for human malignancy.

Erythropoietin is involved in growth and angiogenesis in malignant tumours of female reproductive organs.

The accumulating evidence that erythropoietin and erythropoietin receptor are expressed in various non-haematopoietic organs suggests that erythropoietin signalling might be involved in the growth of tumours, but this possibility has never been examined. We found that mRNAs for erythropoietin and erythropoietin receptor are expressed in malignant tumours of female reproductive organs, where erythropoietin levels are higher than in normal tissues. Furthermore, tumour cells and capillary endothelium showed erythropoietin receptor immunoreactivity. To investigate the role of the erythropoietin/erythropoietin receptor pathway in these tumours, we injected mouse monoclonal antibody against erythropoietin or the soluble form of erythropoietin receptor into blocks of tumour specimens and cultured the blocks. After 12 h of injections, these blocks were examined and compared with control blocks injected with mouse monoclonal antibody, heat denatured soluble form of erythropoietin receptor, mouse serum or saline. Tumour cells and capillaries were markedly decreased in a dose-dependent manner after either injection. A marked increase of the cells containing fragmented DNA and the histopathological characteristics of these cells suggest that the decrease in tumour cells and capillary endothelial cells was due to apoptotic cell death. The co-existence of JAK2 and phosphorylated-JAK2, and STAT5 and phosphorylated STAT5, all of which are involved in the mitogenic signalling of erythropoietin, was found frequently in tumour cells and capillary endothelial cells in the untreated blocks. In contrast, most of the phosphorylated-JAK2- or phosphorylated-STAT5-positive cells had disappeared in the experimental blocks. Moreover, reduced tyrosine phosphorylation of STAT5 in the experimental blocks was confirmed by western blotting analysis. The results strongly indicate that erythropoietin signalling contributes to the growth and/or survival of both transformed cells and capillary endothelial cells in these tumours. Thus, deprivation of erythropoietin signalling may be a useful therapy for erythropoietin-producing malignant tumours.