To study the existence of the erythropoietin receptor (Epo-R) mRNA in brain capillary endothelial cells, the reverse transcription (RT) PCR was performed using total RNAs from rat brain capillary endothelial cells (RBECs) and MBEC4, which is one of the established mouse brain capillary endothelial cell lines. Southern analysis of the RT-PCR products indicated that both RBECs and MBEC4 expressed an authentic form of Epo-R mRNA as a minor form and an intron-5-inserted form of Epo-R mRNA, thus a soluble form of Epo-R mRNA, as a major form. Furthermore, the effect of recombinant human erythropoietin (rHuEpo) on the DNA synthesis in RBECs was analyzed. rHuEpo showed a dose-dependent mitogenic action on RBECs as a competence factor. Radioiodinated rHuEpo was bound specifically to RBECs with time, cell number and dose dependencies. Binding studies with "'I-rHuEpo showed that RBECs had a single class of receptors with low-affinity (K,, = 860 pM) and that the number of siteskell (10300) was abundant. These results suggest that brain capillary endothelial cells express not only an authentic form of Epo-R but also a soluble form of Epo-R and that erythropoietin acts directly on brain capillary endotheha1 cells as a competence factor.Keywords: erythropoietin ; recombinant human erythropoietin ; erythropoietin receptor; brain capillary endothelial cells.Hypoxemia resulting from lung and heart diseases, anemia and high-altitude residence induces a series of modifications in the mammals. As an adaptation mechanism to hypoxia, mammals increase the number of the capillaries/tissue mass [I] and the erythrocytes in the blood (21 to maintain an adequate 0, delivery.Erythropoietin (Epo) is a serum glycoprotein hormone required for survival, proliferation, and differentiation of committed erythroid progenitor cells and its production is accelerated in the kidney by hypoxia [3-51. Recently, i t has been reported that Epo may act on such non-erythroid cells as endothelial cells [6-91, smooth Chemistry, Osaka Prefecture University, Sakai, Osaka, Japan 593 [9]. Although these results suggest that Epo may function as an angiogenic factor for adapting to hypoxia, after embryogenesis, angiogenesis proceeds by the growth of new capillary vessels from an established microvasculature following stimulation by various physiological or pathological processes [18, 191. Furthermore, the mean microvessel density increases in brain [20] and no new capillaries develop in muscle [21] under hypoxic conditions such as exposure to high altitudes, implying that capillary endothelial cells from various tissues each has specific characteristics.HUVECs express Epo-R mRNA [7]. In the present study, we use rat brain capillary endothelial cells (RBECs) and MBEC4, which is one of the established mouse brain capillary endothelial cell lines [22], to analyze the existence of Epo-R mRNA in brain capillary endothelial cells, which have been thought to be associated with angiogenesis under hypoxic conditions, and provide evidence that both RBECs and MBEC4 express ...

Phthalate esters have been used extensively as plasticizers of synthetic polymers. Recent studies have revealed that these esters induce atrophy of the testis, although its pathogenesis remains unknown. The present study describes the possible involvement of oxidative stress in the pathogenesis of atrophy of the rat testis induced by di(2-ethylhexyl)phthalate (DEHP). Biochemical and immunohistochemical analysis revealed that oral administration of DEHP increased the generation of reactive oxygen species, with concomitant decrease in the concentration of glutathione and ascorbic acid in the testis, and selectively induced apoptosis of spermatocytes, thereby causing atrophy of this organ. Oxidative stress was selectively induced in germ cells, but not in Sertoli cells, treated with mono(2-ethylhexyl)phthalate (MEHP), a hydrolysed metabolite of DEHP. Furthermore, MEHP selectively induced the release of cytochrome c from mitochondria of the testis. These results indicate that oxidative stress elicited by MEHP principally injured mitochondrial function and induced the release of cytochrome c, thereby inducing apoptosis of spermatocytes and causing atrophy of the testis.

smoking and the risk of venous thromboembolism (VTE) is an as-yet-unsolved clinical question. 10,11 Because VTE events occur after censoring the data to evaluate deaths caused by cancer or CVD, data regarding VTE events have not been counted and thus excluded from analysis. However, when VTE events are analyzed as the only outcome, heavy smoking is associated with provoked VTE. 12 Prevalence of Tobacco SmokingThe estimated global prevalence of tobacco smoking in 2010 was 36.6% for men and 7.5% for women. 13 In Japan, smoking prevalence rates in 2017 were 29.4% for men and 7.2% for women. 14 Although the average smoking rate in Japan has declined by half during the past 30 years, the rate among middle-aged men remains high, at approximately 40%. 14 This fact indicates that a high incidence of smokingrelated CVDs will continue for several more decades in Japan.

Aerobically grown Euglena gracilis, a bleached mutant, shows a prompt synthesis of wax esters with the concomitant fall of the paramylon (a β‐1,3‐glucan) content upon exposure to anaerobiosis. Bringing the anaerobic cells back to aerobiosis causes the reverse conversions. The anaerobic wax ester formation is accompanied by a net synthesis of ATP. The transition between the fermentation and respiration occurs at 10−5–10−7 M of the O2 concentration.

The vitamin B(12) concentration of an algal health food, spirulina (Spirulina sp.) tablets, was determined by both Lactobacillus leichmannii ATCC 7830 microbiological and intrinsic factor chemiluminescence methods. The values determined with the microbiological method were approximately 6-9-fold greater in the spirulina tablets than the values determined with the chemiluminescence method. Although most of the vitamin B(12) determined with the microbiological method was derived from various vitamin B(12) substitutive compounds and/or inactive vitamin B(12) analogues, the spirulina contained a small amount of vitamin B(12) active in the binding of the intrinsic factor. Two intrinsic factor active vitamin B(12) analogues (major and minor) were purified from the spirulina tablets and partially characterized. The major (83%) and minor (17%) analogues were identified as pseudovitamin B(12) and vitamin B(12), respectively, as judged from data of TLC, reversed-phase HPLC, (1)H NMR spectroscopy, ultraviolet-visible spectroscopy, and biological activity using L. leichmannii as a test organism and the binding of vitamin B(12) to the intrinsic factor.