Expression of uncoupling protein 1 in bovine muscle cells.

Uncoupling protein 1 (Ucp1) is predominantly expressed in brown/beige adipocytes in mammals. Although myogenic cells have been suggested to commit to a brown adipocyte lineage through the induction of Prdm16 expression, Prdm16 is also expressed in skeletal muscle. Thus, we examined expression of Ucp1 in bovine myogenic cells. Considering that Ucp1 is a principle molecule that induces energy expenditure in brown/beige adipocytes, expression of Ucp1 is not preferable in beef cattle because of potential decrease in energy (fattening) efficiency. The RT-PCR analyses revealed the expression of Ucp1 in the skeletal muscle of cattle; expression levels were markedly lower than those in the brown fat of calves. Immunohistochemical analyses showed that Ucp1 surrounded muscle fibers, but not adipocytes residing in skeletal muscle. Myosatellite cells cultured in myogenic medium showed an increase in the expression levels of myogenic regulatory factors ( < 0.05), while those in cells cultured in adipogenic medium were decreased ( < 0.05). The Ucp1 expression was also detected in myosatellite cells; expression levels were greater in cells after myogenic culture for 12 d than in those after myogenic culture for 6 d ( < 0.05) and were decreased when cells were cultured in adipogenic medium ( < 0.05). The Prdm16 expression was not affected by culture conditions, suggesting that the expression of Ucp1 is not regulated by that of Prdm16. The results of the present study provide an insight into the unexpected expression of Ucp1 in bovine skeletal muscle, which suggests the necessity for further studies on Ucp1-mediated energy expenditure in bovine skeletal muscle.

Ultrafast electronic state conversion at room temperature utilizing hidden state in cuprate ladder system.

Photo-control of material properties on femto- (10(-15)) and pico- (10(-12)) second timescales at room temperature has been a long-sought goal of materials science. Here we demonstrate a unique ultrafast conversion between the metallic and insulating state and the emergence of a hidden insulating state by tuning the carrier coherence in a wide temperature range in the two-leg ladder superconductor Sr(14-x)Ca(x)Cu24O41 through femtosecond time-resolved reflection spectroscopy. We also propose a theoretical scenario that can explain the experimental results. The calculations indicate that the holes injected by the ultrashort light reduce the coherence among the inherent hole pairs and result in suppression of conductivity, which is opposite to the conventional photocarrier-doping mechanism. By using trains of ultrashort laser pulses, we successively tune the carrier coherence to within 1 picosecond. Control of hole-pair coherence is shown to be a realistic strategy for tuning the electronic state on ultrafast timescales at room temperature.

Relationship between the Peripheral Lymphocyte Response to Mycophenolic Acid in vitro and the Level of ATP in Peripheral CD4+ Lymphocytes before and after Renal Transplantation.

OBJECTIVE: The lymphocyte immunosuppressant sensitivity test has been used to predict the pharmacodynamics of immunosuppressive drugs for the purpose of preventing acute rejection and infection after renal transplantation. On the other hand, measuring the ATP levels in peripheral CD4+ lymphocytes is also able to monitor the risks of rejection and infection in transplant recipients. In the present study, we examined the relationship between the mycophenolic acid pharmacodynamics and the ATP levels in peripheral lymphocytes before and after renal transplantation. METHODS: We examined both the pharmacological efficacy of mycophenolic acid and the lymphocyte ATP levels before and 2, 4 and 6 weeks after the operation in 20 renal transplant recipients. The drug's pharmacological efficacy was evaluated by the 50% inhibitory concentration of the drug against the in vitro proliferation of peripheral blood lymphocytes activated by T cell mitogen. The ATP levels in peripheral CD4+ lymphocytes were measured by the Immuknow assay kit. The relationships between the mycophenolic acid pharmacodynamics and ATP levels in peripheral lymphocytes were examined in these recipients. RESULTS: The immunosuppressive effects of mycophenolic acid against mitogen-activated lymphocyte proliferation were significantly and positively correlated with the lymphocyte ATP levels, but only at 6 weeks after transplantation. The relationship was not significant before or at 2 or 4 weeks after the operation. CONCLUSION: Our present data raised the possibility that evaluating the pharmacological efficacy of mycophenolic acid pre-transplantation and monitoring the ATP level 6 weeks after transplantation can predict the risk of rejection and/or infection in renal transplant recipients.

Soluble endoglin production is upregulated by oxysterols but not quenched by pravastatin in primary placental and endothelial cells.

INTRODUCTION: Preeclampsia is a serious pregnancy complication. Soluble endoglin (sEng) is released from the placenta and contributes to the maternal endothelial dysfunction seen in preeclampsia. Recently oxysterols, which activate the Liver X Receptor (LXR), have been implicated in producing sEng, by upregulating matrix metalloproteinase-14 (MMP14; cleaves endoglin to produce sEng) and down-regulating tissue inhibitor of metalloproteinase-3 (TIMP-3; inhibitor of MMP14). The functional experiments in that study were performed on JAR cells (human choriocarcinoma cell line) and placental explants. METHODS: We characterized LXR in severe preeclamptic placentas, and assessed whether oxysterols increase release of sEng from primary human umbilical vein endothelial cells (HUVECs), primary trophoblasts and placental explants. Given pravastatin is thought to block oxysterol production and inhibit the LXR, we examined whether pravastatin reduces sEng release. RESULTS: LXRα and ß were localized to the syncytiotrophoblast and villous tips and were significantly up-regulated in preeclamptic placenta. Oxysterols upregulated sEng production in HUVECs and placental explants although the increases were far more modest than that recently reported. Oxysterols did not upregulate sEng in primary trophoblasts. Furthermore, mRNA expression of MMP14 and TIMP-3 were not altered by oxysterols in any tissue. Surprisingly, pravastatin did not decrease oxysterol-induced upregulation of sEng. DISCUSSION: LXR is up-regulated in preeclamptic placenta. Oxysterols upregulate sEng production from human tissues, but the increase is modest, suggesting this may not be the main mechanism for the very significant elevations in sEng seen in preeclampsia. Pravastatin does not decrease sEng production. CONCLUSION: Oxysterols modestly up-regulate sEng production which is not quenched by pravastatin.

Effects of d-α-tocopherol and dietary energy on growth and health of preruminant dairy calves.

To observe the effects of supplemental dietary d-α-tocopherol in relation to dietary energy on growth and immune status in dairy calves, 32 newborn Holstein bull calves were assigned to 1 of 4 treatments for 5 wk in a 2 × 2 factorial, randomized complete block, split-plot design. Calves received moderate growth (MG) or low growth (LG) all-milk dietary treatments, formulated to support daily gains of 0.5 or 0.25 kg/d, respectively, per the dietary energy recommendation for milk-fed calves according to the National Research Council's Nutrient Requirements of Dairy Cattle. Calves in both groups were either injected i.m. with Vital E-A+D (injectable solution of vitamins E, A, and D) on d 1 and supplemented with Emcelle Tocopherol (micellized vitamin E) via milk daily (MG-S and LG-S), or were not supplemented (MG-C and LG-C) during the study period. Total weight gain of MG calves was greater than that of LG calves and tended to be greater in MG-S calves than in MG-C calves. Calves receiving vitamin supplementation demonstrated greater concentrations of plasma α-tocopherol, retinol, and 25-(OH)-vitamin D than did control calves, whereas MG calves demonstrated a lower concentration of plasma α-tocopherol than did LG calves. The apparent increased utilization of α-tocopherol by MG calves was accompanied by a rise in serum haptoglobin, a positive acute-phase protein and indicator of inflammation, especially in MG-C calves. Serum amyloid A, also a positive acute-phase protein, was not different among groups, but was elevated from baseline in all groups during wk 1 through 3. Plasma IgG1 concentrations were higher in MG-S and LG-S calves than in their nonsupplemented dietary counterparts, whereas plasma IgG2, IgA, and IgM concentrations were not different among groups. In summary, dietary supplementation of d-α-tocopherol improved plasma α-tocopherol status and tended to increase growth in calves fed for 0.5 kg of average daily gain. Vitamin supplementation ameliorated the rise of serum haptoglobin associated with acute inflammation in MG calves, and may have improved passive transfer of maternal antibody. These results indicate a role for α-tocopherol in prevention of proinflammatory state associated with greater dietary energy and onset of infectious disease.

Probing the metal-insulator phase transition in the (DMEDO-EBDT)2PF6 single crystal by optical measurements.

The temperature and polarization dependence of the optical reflectivity  spectra of a quasi-one-dimensional 1/4-filled band system, (DMEDO-EBDT)(2)PF(6), have been investigated. We observed clear anisotropy in the electronic structures corresponding to the anisotropic transport properties. The appearance of a charge gap (E(g) > 0.1 eV) and transfer of the spectral weight accompanied by the metal-insulator phase transition were clearly observed. In addition, a split of the intramolecular vibrational modes was observed, which strongly suggested the existence of charge disproportionation in the low temperature phase. We also observed a photoinduced reflectivity change, which implied the occurrence of a photoinduced phase transition from the low temperature insulating phase to the high temperature metallic phase.

Modification of the analysis of parathyroid hormone-related protein in milk and concentrations of this protein in commercial milk and milk products in Japan.

Parathyroid hormone-related protein (PTHrP), which causes hypercalcemia associated with malignant tumors, is known to be present in milk. Gene expression of PTHrP in the mammary gland increases markedly during parturition and with the onset of lactation. Even when circulating PTHrP levels are extremely low or below the detection limit, milk PTHrP levels are remarkably high. Parathyroid hormone-related protein derived from the mammary gland is assumed to play a role in maintaining the maternal calcium homeostasis and calcium transport from blood to milk. In previous studies that determined the PTHrP concentrations in milk, the pretreatments and diluent composition were not standardized. Here, we investigated the effect of various pretreatment procedures and diluent constitutions and the consequent PTHrP concentrations in commercial milk and milk products in Japan. Significant differences were found in PTHrP concentrations in raw milk samples subjected to different combinations of pretreatments (mixing, centrifugation, acidification, and heating) and diluents (0pM standard solution of PTHrP, plasma treated with protease inhibitors, and original diluent). We measured the PTHrP concentrations in normal liquid milk, processed milk, milk drinks, formulated milk powders, and skim milk powder by using the appropriate combination of pretreatment (acidification) and diluent (plasma treated with protease inhibitors). The PTHrP concentration in normal liquid milk, processed milk, and skim milk powder was as high as that in raw milk (>5nM), whereas that in milk drinks differed considerably. The PTHrP concentration in infant formulas (<2nM) was lower than that in the other milk products. These results indicate that a certain amount of PTHrP is ingested when milk and milk products are consumed.

Ultrasonic propagation of a metallic domain in Pr0.5Ca0.5CoO3 undergoing a photoinduced insulator-metal transition.

Femtosecond optical response was investigated on a perovskite-type cobalt oxide, Pr0.5Ca0.5CoO3 during an insulator-metal (I-M) transition accompanied with the change in spin configuration. After photoirradiation at 30 K, the reflectivity showed a sudden and large increase with subsequent variation depending on the observed photon energy. An exact calculation of Maxwell's equations for the Pr0.5Ca0.5CoO3 after the photoirradiation enabled us to observe the ultrafast dynamics of I-M phase transition and the motion of the photonically created metallic domain at the velocity of ultrasonic wave.

Contact dermatitis in dairy cattle caused by calcium cyanamide.

Nine of 250 cows on a dairy farm initially developed severe dermatitis on parts of their bodies that touched the floor, and it then spread over their entire body. The cause was suspected to be calcium cyanamide, which had been added to the material spread on the floor to prevent environmental mastitis. Experimental exposure of the skin of a cow to calcium cyanamide induced the same type of contact dermatitis, and histopathological investigations showed that it caused irritant and allergic reactions. To identify the cause of the dermatitis, a patch test with calcium cyanamide and its breakdown products, cyanamide, urea and ammonium bicarbonate, was carried out on four cows. Three of them had a positive reaction to calcium cyanamide and cyanamide; delayed and amplified reactions suggesting an allergic response were observed.

Comparison of natural estrogen removal efficiency in the conventional activated sludge process and the oxidation ditch process.

The presence of natural estrogens, 17beta-estradiol (E2), estrone (E1) and estriol (E3), as well as estrogenic activity in wastewater influents and secondary effluents were investigated in 20 full-scale wastewater treatment plants in Japan. In all of the influent samples, natural estrogens were detected at concentrations above the minimum limits of detection (0.5ng/L). The concentrations of natural estrogens detected in the effluent of oxidation ditch plants were generally lower than previously reported values. On the other hand, in the conventional activated sludge plants, increments of E1 during biological treatment were frequently observed although E2 and E3 were removed effectively in the process. The removal rates of natural estrogens or estrogenic activity show no observed statistical relationship with the solids retention time (SRT) and the hydraulic retention time (HRT). However, the plants with high SRT or HRT generally showed high and stable removal of both natural estrogens and estrogenic activity.

Optical control of two-photon excitation efficiency of alpha-perylene crystal by pulse shaping.

Optimized pulse shaping experiments were carried out on the control of two-photon excitation efficiency of an alpha-perylene crystal in the temperature region from 30 to 290 K. It was found that a pulse train with a pulse interval of 90 fs and an alternately reversing phase relation increased the excitation efficiency by a factor of 2 for the whole temperature region. The pulse shape characteristic for effective efficiency increase was reduced by double pulse experiments in which the dependence of the emission intensity on the pulse interval and relative phase between pulses were measured. The mechanism of the efficiency increase is briefly discussed using a sliding-window Fourier transform of the pulse shape.

Multiple basilar artery trunk aneurysms associated with fibromuscular dysplasia.

An association between fibromuscular dysplasia (FMD) and intracranial aneurysms has been proposed but the occurrence of multiple aneurysms on the trunk of basilar artery (BA) associated with FMD has not been previously reported.

The behavior of estrogenic substances in the biological treatment process of sewage.

A study was conducted for about one year on the fate and behavior of estrogens, namely 17beta-estradiol (E2), estrone (E1), and estriol (E3) in an activated sludge process of a pilot scale plant supplied with domestic sewage. A simultaneous analytical method for these three substances using LC-MS/MS was developed and applied to sewage samples. The average removal of E2 was 94.7%, while that of E3 was 96.9%. In contrast, the average removal of E1 was relatively low at 69.2% with a maximum concentration of 55.4 ng/L detected in the treated water. The theoretical values of estrogenic activity calculated from the concentrations of each natural estrogen in treated water were found to correlate with the values of estrogenic activity measured by a yeast estrogen screening assay. The effect of E2 on estrogenic activity in influent was found to be high, while that of E1 in treated water was considerably higher. In batch treatment tests on E2, E2 turned into E1 immediately after being charged. After three hours of aeration, the values of both E1 and E2 were around threshold limits. It was determined from this that E1 and E2 were substances that could be degraded by biological treatment. As the removal of E2 was found to be sufficiently high at times, optimization of operational conditions based on E1 removal should be important for reducing estrogenic activity in treated water.

Evaluation of estrogen-like activity on sewage treatment processes using recombinant yeast.

Four sewage treatment plants based on an activated sludge process and a pilot scale plant for advanced sewage treatment located in Japan were evaluated for removal of estrogenic substances using in vitro recombinant yeast assay and chemical analysis. The results indicated that 17beta-estradiol (E2) significantly contributed to estrogen-like activity analyzed by yeast assay especially in secondary treated effluents. On the other hand, batch study showed that estrogen-like activity of spiked E2 was easily decreased by an activated sludge treatment. This result suggested that E2 concentrations measured by enzyme immunoassay (EIA) were interpreted as false positives in effluents, and that unknown estrogenic substances other than E2 might have contributed to estrogen-like activity in the secondary treated effluents. Further, in the pilot scale study, advanced sewage treatment processes such as a biological aerated filtration (BAF) process, an advanced oxidation process (AOP), were effective for the removal of those estrogenic activities contributed by unknown estrogenic substances in sewage secondary treated effluent.

[A clinical study of psychopathology in systemic lupus erythematosus].

Thirty patients (24 inpatients and 6 outpatients) with a clinical diagnosis of SLE were examined between September 1, 1998 and August 1, 1999 in the rheumatology clinic of Jichi Medical School Hospital. All of these patients fulfilled the 1982 revised criteria of the American Rheumatism Association for the classification of SLE and had some psychiatric manifestations (psychiatric SLE; P-SLE group). Mean patient age was 38.6 +/- 13.0, and there were 5 males and 25 females. When classified into 5 subgroups according to the most prominent symptoms, the distribution was as follows: consciousness disturbance group: 6 (20%), schizophrenia-like group: 5 (16.7%), mood disorder group: 7 (23.3%), neurosis-like group: 10 (33.3%), and convulsive disorder group: 2 (6.7%). Among all 37 psychiatric episodes, symptoms appeared in 37.8% of cases during the acute phase of SLE (during onset or recurrence) and in 62.9% during the chronic phase (during remission). The profile of the P-SLE group showed that the psychiatric symptoms of the SLE patients were milder and more chronic than those described in previous reports. To begin to comprehend the psychopathology of SLE, we put forward the concept of "Psychiatric basal state" and "psychiatric conjugated state". The former is considered a direct reflection of the acute-phase SLE process on mental condition. It is defined clinically as psychiatric symptoms that parallel the activity of SLE and respond well to steroid therapy. The latter include all other psychiatric problems in which one cannot rule out the effects of pharmacological, somatic, personality, and environmental effects on psychiatric symptoms. Only 3 patients in the P-SLE group fulfilled the criteria for the "psychiatric basal state". All three patients belonged to the consciousness disturbance group, whose clinical features were defined as slight clouding of consciousness, so-called "Amentia" in the sense of the German terminology. The clinical profile of this state is: 1. the patients are young (about 16 years old), 2. the onset of psychiatric symptoms is within 5 years after the onset of SLE, 3. confusion and disorientation are the most characteristic features, and 4. the clinical course of this state is almost 2 months. The experience structure of the "psychiatric basal state" consists of: 1. difficulty in selecting and holding a topic in cognition, 2. confusion and emotional instability as the basal mood, and 3. primitive and floating forms of delusions and hallucinations. Using this concept of the "psychiatric basal state" as a clue, we can hypothesize the continuity of diverse psychiatric symptoms in SLE. The "proper process of SLE (Harada)" has a disintegrating effect on the "ego" and it allows various psychopathological phenomena to emerge in the experience field. Against this background, additional factors, such as secondary organ damage, personality structure, and social environment, induce organization of the "psychiatric conjugated state".

Utilization of intestinal triglyceride-rich lipoproteins in mammary gland of cows.

Elution profiles of total lipoproteins, apolipoprotein B (apoB) concentrations in lipoproteins, and plasma triglyceride (TG) levels were examined in early-, late-, and non-lactating cows. Additionally, arteriovenous (A-V) differences were also measured to elucidate the uptake of TG and apoB-containing lipoproteins in mammary gland. Non-lactating cows showed three major peaks corresponding to triglyceride-rich lipoprotein (TRL), low density lipoprotein (LDL), and high density lipoprotein (HDL) fraction, whereas both early- and late-lactating cows revealed two peaks corresponding to TRL and HDL. The peak area of TRL in early- and late-lactating cows were significantly (p < 0.05) smaller than that in non-lactating cows. The plasma TG levels and apoB-48 concentrations of TRL in early- and late-lactating cows were also significantly (p < 0.01) lower. Furthermore, early lactating cows showed significantly (p < 0.05) larger A-V differences in both plasma TG and apoB-48 concentration of TRL than those in late- and non-lactating cows. These results suggested that TG in exogenous (intestinal) TRL was utilized for milk fat synthesis in lactating mammary gland of cows by the receptor-mediated uptake.

Rare-type mutations of MMAC1 tumor suppressor gene in human glioma cell lines and their tumors of origin.

A total of 10 glioma cell lines were examined to evaluate the status of the MMAC1 gene, a candidate tumor suppressor gene. Six cell lines showed mutations with presumed loss of heterozygosity and 1 cell line showed no mRNA expression. The 6 mutations consisted of 3 3-bp deletions (codons 17, 101 or 199), 1 missense mutation (codon 252) and 2 truncation mutations (1 nonsense mutation at codon 233 and 12-bp insertion at codon 241). Among them, the 3-bp deletions, which are a rare type of mutation in MMAC1 gene, were located in the N-terminal half (codons 1-212) of the coding region, which is considered important in MMAC1 function. The missense mutation was located unusually in the C-terminal half (codons 212-403), but it was in a small region in which some other reported missense mutations are clustered. Thus, these 4 mutations were suggested to have functional effects on the MMAC1 activity, like the other 2 mutations with predicted protein truncations. By sequence analysis of cDNA clones, we confirmed that all the mutations including these 4 rare ones were in the MMAC1 gene, not in the PTH2 pseudogene. In 2 cases, we also examined the primary glioma tissues from which the cell lines had been derived and found the same mutations as in the cell lines in both cases. This suggested that the mutations in these cell lines were derived from the primary glioma tissues, but not from artifacts arising during long-term in vitro cultivation.

Intracranial malignant lymphomas: clinicopathological study of 26 autopsy cases.

We examined 26 autopsy-proven cases of intracranial malignant lymphoma (IML) in immunocompetent patients to determine the extent of neoplastic involvement of the central nervous system (CNS) and to evaluate the effects of radiation on the tumor and brain tissue. All tumors were identified as diffuse non-Hodgkin's lymphomas of B-cell origin. In six patients who had not received radiotherapy, the clinical course of the disease was short and extensive infiltration of the tumor was seen. The remaining 20 patients were treated with radiotherapy and had a longer survival time. Leptomeningeal involvement was common, but extensive subarachnoid proliferation of the tumor was seen in only two cases. The posterior, but not anterior, lobe of the pituitary was involved in 5 of 22 cases, and choroid plexus involvement was seen in 4 of 21. Direct invasion of the tumor into the spinal cord, which tended to occur in patients with posterior fossa masses, was observed in 5 of 21 cases. Following irradiation, coagulation necrosis was frequently found in the invading zone as well as in the tumor mass, and degeneration of the white matter was also seen. We suggest that IML can extensively infiltrate into the CNS, including the posterior lobe of the pituitary and spinal cord, and that radiation injury to the brain appears to occur relatively easily in this disease.

Establishment of two glioma cell lines from two surgical specimens obtained at different times from the same individual.

We established two glioma cell lines from two surgical specimens obtained at different times from the same patient. One (No. 9R), which was derived from the recurrent tumor (glioblastoma, grade IV), proliferated more rapidly in vitro than the other (No. 9) from the primary tumor (slightly anaplastic astrocytoma, grade II-III). No. 9R showed heterotransplantability in nude mice, whereas No. 9 did not. These findings indicate that No. 9R has a more aggressive or malignant nature than No. 9. Both cell lines showed homozygous deletion of the representative tumor suppressor p16 and p15 genes, but no p53 gene alteration. However, examination of the overall mRNA expression profile using a commercially available cDNA-spotted membrane revealed much higher expression levels of several mRNAs, at least, in No. 9R than in No. 9, although the relationship between these mRNAs and the growth potentials remained unknown. These two cell lines, derived from the same individual, with different proliferating potentials may be useful for studies on the molecular bases of glioma malignancy and progression.

Comparison of sub-Tenon's anaesthesia by different delivery techniques in cataract surgery.

PURPOSE: To compare the analgesic effects of three different delivery techniques of sub-Tenon's anaesthesia in cataract surgery by assessing patients' response to the visceral stimulus. METHODS: A prospective, randomised study was conducted on 345 eyes of 345 patients undergoing phacoemulsification and posterior chamber intraocular lens implantation. They received anaesthetic infiltration into the sub-Tenon's space through a conjunctival incision (115 eyes), infiltration into the posterior sub-Tenon's space (retrobulbar space) through a conjunctival incision (114 eyes), or injection into the intra-Tenon's space (subconjunctival space) without making a conjunctival incision (116 eyes). Pain scores were recorded when the anterior chamber was irrigated with an acetylcholine chloride solution to achieve miosis after lens implantation. RESULTS: There were no significant differences in pain scores among the three groups (chi-squared test of homogeneity, p = 0.814). Approximately 10-20% of patients reported slight to severe pain at the time of acetylcholine administration. CONCLUSIONS: The three anaesthetic delivery methods of sub-Tenon's anaesthesia possess similar and reasonable analgesic effects in cataract surgery, but may not block visceral stimuli completely.