Circadian factors BMAL1 and RORα control HIF-1α transcriptional activity in nucleus pulposus cells: implications in maintenance of intervertebral disc health.

BMAL1 and RORα are major regulators of the circadian molecular oscillator. Since previous work in other cell types has shown cross talk between circadian rhythm genes and hypoxic signaling, we investigated the role of BMAL1 and RORα in controlling HIF-1-dependent transcriptional responses in NP cells that exist in the physiologically hypoxic intervertebral disc. HIF-1-dependent HRE reporter activity was further promoted by co-transfection with either BMAL1 or RORα. In addition, stable silencing of BMAL1 or inhibition of RORα activity resulted in decreased HRE activation. Inhibition of RORα also modulated HIF1α-TAD activity. Interestingly, immunoprecipitation studies showed no evidence of BMAL1, CLOCK or RORα binding to HIF-1α in NP cells. Noteworthy, stable silencing of BMAL1 as well as inhibition of RORα decreased expression of select HIF-1 target genes including VEGF, PFKFB3 and Eno1. To delineate if BMAL1 plays a role in maintenance of disc health, we studied the spinal phenotype of BMAL1-null mice. The lumbar discs of null mice evidenced decreased height, and several parameters associated with vertebral trabecular bone quality were also affected in nulls. In addition, null animals showed a higher ratio of cells to matrix in NP tissue and hyperplasia of the annulus fibrosus. Taken together, our results indicate that BMAL1 and RORα form a regulatory loop in the NP and control HIF-1 activity without direct interaction. Importantly, activities of these circadian rhythm molecules may play a role in the adaptation of NP cells to their unique niche

Rare case of single left coronary artery in a Japanese cadaver

A single left coronary artery with a single orifice in the left aortic sinus was observed during anatomical practice in an 81-year-old male Japanese cadaver. The single left coronary artery bifurcated into the anterior interventricular branch (IVa) and circumflex (CXa) branches. The IVa descended into the anterior interventricular sulcus to supply the apex of the heart, leaving a branch that traversed the upper part of the infundibulum to supply the anterior upper region of the right ventricle. The CXa curved leftward in the atrioventricular sulcus to reach the posterior surface, after which it continued to emerge into the anterior surface. The vascular running pattern showed that CXa directly supplied blood to the upper right ventricle (but not the conus branch), with three branches connected to the apex. The atrial arteries showed no anomalous distribution patterns. These findings are useful during surgical procedures, including cardiac catheterization

ニワトリの精巣内プロジェスチン標的細胞の局在 オートラジオグラフによる検討

オートラジオグラフィーによって, 25-70日齢の未熟鶏および120日齢の成鶏の精巣の細胞成分におけるプロジェスチン(P)の局在と濃度を調査した.精巣の凍結切片標本を放射能で標識した合成P (3H-Promegestoneまたは3H-R5020)溶液中で培養した後, 常法通りオートラジオグラフィーを行い, 組織または細胞上の黒化銀粒子数を算定して, 放射活性の強さまたはP濃度の指標とした.その結果, 1)精細管上皮と間質細胞に放射活性の特異的集中が見られた.2)放射活性の強さは動物の日齢によって変動した.25, 70および120日齢で高く, 50日齢で低かった.3)細胞内のP濃度は動物をあらかじめエストロジェンで処置しておくと増強した.4)放射活性の強さは間質細胞よりも精細管上皮で高かった.しかし, 凍結切片標本では高放射活性を示す細胞の同定は困難であった.以上の結果から, 成長期のニワトリ精巣の精細管壁と間質にPレセプター含有細胞が存在することが示唆されるUsing an autoradiographic technique, the localization and concentration of progestin (P) in cell constituents of testes from the immature 25-to-70-day-old and mature 120-day-old chickens were investigated. After incubation of the frozen tissue sections with 3H-Promegestone (R5020), a synthetic P, specific radioactivity, was found in the seminiferous epithelium and interstitium in all the animals in the presence or in the absence of various non-radioactive steroids, and the intensity of radioactivity differed with age: high in 25-, 70-and 120-day-old chickens, and low in 50-day-old chickens. Estrogen treatment enhanced the intensity of radioactivity in the cells at all ages. The concentration of radioactivity was higher in the seminiferous epithelium than in the interstitial cells. In the frozen tissue sections, it was not possible to identify the kind of cell taking up the high amount of radioactivity. These results indicate that the testes from chicken during their growing stages contain cell constituents that specifically bind P

Effects of Chemotherapy and Radiotherapy on Spermatogenesis: The Role of Testicular Immunology

Substantial improvements in cancer treatment have resulted in longer survival and increased quality of life in cancer survivors with minimized long-term toxicity. However, infertility and gonadal dysfunction continue to be recognized as adverse effects of anticancer therapy. In particular, alkylating agents and irradiation induce testicular damage that results in prolonged azoospermia. Although damage to and recovery of spermatogenesis after cancer treatment have been extensively studied, there is little information regarding the role of differences in testicular immunology in cancer treatment-induced male infertility. In this review, we briefly summarize available rodent and human data on immunological differences in chemotherapy or radiotherapy

In-situ Real-Time Monitoring of Volatile Organic Compound Exposure and Heart Rate Variability for Patients with Multiple Chemical Sensitivity

In-situ real-time monitoring of volatile organic compound (VOC) exposure and heart rate variability (HRV) were conducted for eight multiple chemical sensitivity (MCS) patients using a VOC monitor, a Holter monitor, and a time-activity questionnaire for 24 h to identify the relationship between VOC exposure, biological effects, and subjective symptoms in actual life. The results revealed no significantly different parameters for averaged values such as VOC concentration, HF (high frequency), and LF (low frequency) to HF ratio compared with previous data from healthy subjects (Int. J. Environ. Res. Public Health 2010, 7, 4127–4138). Significant negative correlations for four subjects were observed between HF and amounts of VOC change. These results suggest that some patients show inhibition of parasympathetic activities along with VOC exposure as observed in healthy subjects. Comparing the parameters during subjective symptoms and normal condition, VOC concentration and/or VOC change were high except for one subject. HF values were low for five subjects during subjective symptoms. Examining the time-series data for VOC exposure and HF of each subject showed that the subjective symptoms, VOC exposure, and HF seemed well related in some symptoms. Based on these characteristics, prevention measures of symptoms for each subject may be proposed

Association of Odor Thresholds and Responses in Cerebral Blood Flow of the Prefrontal Area during Olfactory Stimulation in Patients with Multiple Chemical Sensitivity.

Multiple chemical sensitivity (MCS) is a disorder characterized by nonspecific and recurrent symptoms from various organ systems associated with exposure to low levels of chemicals. Patients with MCS process odors differently than controls do. Previously, we suggested that this odor processing was associated with increased regional cerebral blood flow (rCBF) in the prefrontal area during olfactory stimulation using near-infrared spectroscopic (NIRS) imaging. The aim of this study was to investigate the association of odor thresholds and changes in rCBF during olfactory stimulation at odor threshold levels in patients with MCS. We investigated changes in the prefrontal area using NIRS imaging and a T&T olfactometer during olfactory stimulation with two different odorants (sweet and fecal) at three concentrations (zero, odor recognition threshold, and normal perceived odor level) in 10 patients with MCS and six controls. The T&T olfactometer threshold test and subjective assessment of irritating and hedonic odors were also performed. The results indicated that the scores for both unpleasant and pungent odors were significantly higher for those for sweet odors at the normal perceived level in patients with MCS than in controls. The brain responses at the recognition threshold (fecal odor) and normal perceived levels (sweet and fecal odors) were stronger in patients with MCS than in controls. However, significant differences in the odor detection and recognition thresholds and odor intensity score between the two groups were not observed. These brain responses may involve cognitive and memory processing systems during past exposure to chemicals. Further research regarding the cognitive features of sensory perception and memory due to past exposure to chemicals and their associations with MCS symptoms is needed

GRP78 suppresses lipid peroxidation and promotes cellular antioxidant levels in glial cells following hydrogen peroxide exposure.

Oxidative stress, caused by the over production of reactive oxygen species (ROS), has been shown to contribute to cell damage associated with neurotrauma and neurodegenerative diseases. ROS mediates cell damage either through direct oxidation of lipids, proteins and DNA or by acting as signaling molecules to trigger cellular apoptotic pathways. The 78 kDa glucose-regulated protein (GRP78) is an ER chaperone that has been suggested to protect cells against ROS-induced damage. However, the protective mechanism of GRP78 remains unclear. In this study, we used C6 glioma cells transiently overexpressing GRP78 to investigate the protective effect of GRP78 against oxidative stress (hydrogen peroxide)-induced injury. Our results showed that the overexpression of GRP78 significantly protected cells from ROS-induced cell damage when compared to non-GRP78 overexpressing cells, which was most likely due to GRP78-overexpressing cells having higher levels of glutathione (GSH) andquinone oxidoreductase 1 (NQO1), two antioxidants that protect cells against oxidative stress. Although hydrogen peroxide treatment increased lipid peroxidation in non-GRP78 overexpressing cells, this increase was significantly reduced in GRP78-overexpressing cells. Overall, these results indicate that GRP78 plays an important role in protecting glial cells against oxidative stress via regulating the expression of GSH and NQO1

Effects of Dexmedetomidine on the Localization of α2A-Adrenergic and Imidazoline Receptors in Mouse Testis

Dexmedetomidine (DEX) used for sedation was reported to have organ-protecting effects in ischemia–reperfusion injury model animals. However, no testicular cell-protecting effect was observed with DEX treatment. The effects of DEX on a normal testis in vivo have not been reported. Therefore, DEX was administered to mice for 14 days to investigate the reproductive toxicology of DEX on the testis and the localization of DEX-responsive receptors. The testes, pituitary glands, and serum were examined and analyzed using real-time PCR, immunofluorescence staining, and liquid chromatography–mass spectrometry. In the testis, α2A-adrenergic receptors were observed in the cytoplasm of Leydig cells, while imidazoline receptors were observed in germ cells and Leydig cell cytoplasm. The levels of luteinizing hormone and follicle-stimulating hormone mRNA in the pituitary gland significantly temporarily decreased. Serum DEX could not be detected 26 h after DEX administration. DEX administration did not affect serum testosterone levels, some testicular mRNA related to spermatogenesis, and oxidative stress factors. Therefore, although DEX receptors are present in the testis, DEX is metabolized relatively quickly, and DEX administration has no damaging effects on the testis. This study is the first in vivo report about the effects of DEX administration on the testis