Expression of mouse osteocalcin transcripts, OG1 and OG2, is differently regulated in bone tissues and osteoblast cultures.

Osteocalcin is a noncollagenous protein that is abundant in mineralized bone matrix. Mice have a gene cluster of osteocalcin that consists of OG1, OG2, and ORG. We established a new method to directly analyze the expression levels of OG1, OG2, and ORG mRNAs relative to total osteocalcin mRNA. They were amplified as 371-bp fragments by reverse transcription-polymerase chain reaction (RT-PCR) at the same time using common primers, digested with ApaLI, and separated in a polyacrylamide gel. ApaLI digestion did not affect the mobility of the OG1-derived 371-bp fragment, whereas both 371-bp fragments, derived from OG2 and ORG, were digested into 350 bp. Total RNA prepared from mouse bone was then subjected to RT-PCR followed by ApaLI digestion. OG1 and OG2 mRNAs were found to be expressed at ratios of 80%-86% and 14%-20%, respectively, to the total osteocalcin mRNA in mouse bone. The ratios were almost constant in various bones in vivo, independent of the animal's genetic background, age, or gender, or different parts of bone. RT-PCR using specific primers revealed that mouse bone tissues strongly expressed osteocalcin mRNA derived from OG1 and OG2, but not ORG. In contrast, cells cultured in vitro showed different expression ratios of osteocalcin mRNA: 53%-65% for OG1 and 35%-47% for OG2 to the total osteocalcin mRNA in the osteoblast cell line and primary osteoblasts in culture even though they formed many mineralized bone nodules. Similar results were obtained in both KS483 osteoblasts and C2C12 myoblasts, when they were cultured with bone morphogenetic protein-2 (BMP-2) to induce osteocalcin mRNA. Taken together, these findings indicate that OG1 is the predominant transcript among the three osteocalcin genes in mouse bone in vivo. It is also suggested that the expression of OG1 and OG2 is regulated differently in bone tissues and osteoblast cultures.

Essential oil constituents from Japanese and Indian Curcuma aromatica rhizomes.

The chemical compositions of the essential oils from rhizomes of Curcuma aromatica cultivated in Japan and India were analyzed by GC and GC/MS. These oils were extremely different. The major constituents in both oils from Japanese samples were curdione, germacrone, 1,8-cineole, (45,5S)-germacrone-4,5-epoxide, beta-elemene, and linalool, whereas those in the oil from Indian were beta-curcumene, ar-curcumene, xanthorrhizol, germacrone, camphor, and curzerenone.

Hepatic oxygen supply, energy charge, and histological findings in dogs with portal vein arterialization.

Hepatic oxygen supply, energy charge (EC), and histology were examined comparatively in dogs with portal vein anastomosis (PVA group), and PA in addition to PVA (PA group). The PVA group showed a lower level of hepatic oxygen supply than those of the PA group throughout the experimental period, and also showed decreases of adenosine triphosphate (ATP) and EC level after blood perfusion. In contrast, the oxygen supply and consumption were stable in the PA group. A temporary fall of ATP level was followed by recovery to the preperfusion level in the PA group. Histological examination indicated the collapse of hepatic cords with granular and vacuolar degeneration in only the PVA group. These findings suggested that PA, when supplemented to PVA, is an available technique for preventing hepatic failure caused by ischemic conditions.

UDP-GT involvement in the enhancement of cell proliferation in thyroid follicular cell proliferative lesions in rats treated with thiourea and vitamin A.

The mechanisms underlying enhanced cell proliferation in thyroid proliferative lesions of rats simultaneously treated with large amounts of vitamin A (VA) and thiourea (TU) were investigated. Male F344 animals were initiated with N-bis(2-hydroxypropyl)nitrosamine (2800 mg/kg body weight, single s.c. injection). Starting 1 week later, groups received water containing 0.2% TU (TU group), diet containing 0.1% VA (VA group), both 0.2% TU and 0.1% VA (TU + VA group) or tap water/basal diet without supplement (control group) for 10 weeks. The serum levels of triiodothyronine (T3) and thyroxine (T4) were decreased and the thyroid stimulating hormone (TSH) levels were elevated in the TU and TU + VA groups, with the degree of change being significantly greater in the combined treatment group. The induction of P450 isoenzymes by TU was not enhanced by VA supplementation, but uridine diphosphate glucuronosyltransferase (UDP-GT) activity in the liver was significantly increased in the TU + VA group compared to the TU group. Thyroid weights were increased in both the TU and TU + VA groups, this being more pronounced with VA supplementation. Thyroid follicular cell hyperplasias and neoplasias were induced to similar extents in both TU treated groups, but their cell proliferation appeared to be increased by the VA supplementation. The results of the present study suggest that enhanced cell proliferation is due to increased TSH stimulation, resulting from the decrease in serum T3/T4 levels brought about by induction of liver UDP-GT activity with the combined action of TU + VA as well as inhibition by TU of thyroid hormone synthesis in the thyroid.

Eosinophilic neuronal inclusions in the thalamus of ageing B6C3F1 mice.

The age-related incidence of thalamic neuronal inclusions in the brains of aged B6C3F1 mice and their histopathological and ultrastructural features were studied. Round to oval or rod-shaped eosinophilic inclusions, which were frequently observed in the neurons of the thalamic area, were stained positively with phosphotungstic acid haematoxylin. The inclusions were detected first at 32 weeks of age and the incidence and severity were higher in older animals, all of the mice being affected after 58 weeks of age. Ultrastructurally, the inclusions appeared as sheaves of parallel osmiophilic filaments in the perikaryon of the neurons in the thalamus. Thus thalamic neuronal inclusions in mice seem to be age-related, but their significance remains unclear.

Spontaneous globoid mineralization in the cerebellum of rats.

Cerebellar globoid mineralization in two rats was examined by light and electron microscopy, and by X-ray microanalysis. The mineralization was round to oval in shape; it varied in size and was positive for the periodic acid-Schiff and von Kossa reactions. Ultrastructurally, a concentric lamellar structure was prominent in moderately electron-dense depositions. Elemental analysis revealed the presence of large amounts of calcium and phosphorus, and small amounts of zinc, potassium and aluminum.

Hyperbaric oxygen therapy in experimentally induced acute cerebral ischemia.

Effects of hyperbaric oxygen (HBO) on acute cerebral ischemia were studied in spontaneously hypertensive rats, which had the carotid artery bilaterally ligated. The animals were exposed to HBO (100% 02 at 2 ATA) for 30 min at 1 or 3 h after carotid ligation (treated group). Survival time and brain tissue metabolites were measured after HBO in these animals and compared with ischemic animals without HBO exposure (nontreated group). The animals treated at 3 h after ligation survived longer (6.5 +/- 0.7 h) than did nontreated ones (4.3 +/- 0.2 h) (P less than 0.05). The cerebral lactate increased much less in these treated animals (24.60 +/- 1.67 mM/kg) than in nontreated ones (31.78 +/- 1.68 mM/kg) (P less than 0.05). Cerebral ATP levels tended to decrease less in the former (0.66 +/- 0.17 mM/kg) than in the latter (0.59 +/- 0.07 mM/kg). When HBO started at 1 h after carotid ligation, however, there were no significant differences of survival time or brain metabolites between treated and nontreated groups of animals. The present results indicate that HBO administered at 3 h after brain ischemia prevents further increase in cerebral lactate and produces a slight but significant increase in survival time.