Effects of medium-chain fatty acids and tributyrin supplementation in milk replacers on growth performance, blood metabolites, and hormone concentrations in Holstein dairy calves.

This study aimed to evaluate the effects of triglycerides containing medium-chain fatty acids (MCT) and tributyrin (TB) supplementation in a milk replacer (MR) on growth performance, plasma metabolites, and hormone concentrations in dairy calves. Sixty-three Holstein heifer calves (body weight at 8 d of age, 41.1 ± 2.91 kg; mean ± SD) were randomly assigned to 1 of 4 experimental MR (28% crude protein and 18% fat): (1) containing 3.2% C8:0 and 2.8% C10:0 (in fat basis) without TB supplementation (CONT; n = 15), (2) containing 6.7% C8:0 and 6.4% C10:0 without TB supplementation (MCT; n = 16), (3) containing 3.2% C8:0 and 2.8% C10:0 with 0.6% (dry matter basis) TB supplementation (CONT+TB; n = 16), (4) containing 6.7% C8:0 and 6.4% C10:0 with 0.6% TB supplementation (MCT+TB; n = 16). The MR were offered at 600 g/d (powder basis) from 8 to 14 d, up to 1,300 g/d from 15 to 21 d, 1,400 g/d from 22 to 49 d, down to 700 g/d from 50 to 56 d, 600 g/d from 57 to 63 d, and weaned at 64 d of age. All calves were fed calf starter, chopped hay, and water ad libitum. The data were analyzed using a 2-way ANOVA via the fit model procedure of JMP Pro 16 (SAS Institute Inc.). Medium-chain fatty acid supplementation did not affect the total dry matter intake. However, calves that were fed MCT had greater feed efficiency (gain/feed) before weaning (0.74 ± 0.098 vs. 0.71 ± 0.010 kg/kg) compared with non-MCT calves. The MCT calves also had a lower incidence of diarrhea compared with non-MCT calves during 23 to 49 d of age and the weaning period (50 to 63 d of age; 9.2% vs. 18.5% and 10.5% vs. 17.2%, respectively). Calves fed with TB had a greater total dry matter intake during postweaning (3,465 vs. 3,232 g/d). Calves fed TB also had greater body weight during the weaning (90.7 ± 0.97 vs. 87.9 ± 1.01 kg) and postweaning period (116.5 ± 1.47 vs. 112.1 ± 1.50 kg) compared with that of non-TB calves. The plasma metabolites and hormone concentrations were not affected by MCT or TB. These results suggest that MCT and TB supplementation in the MR may improve the growth performance and gut health of dairy calves.

Evaluation of the suppressive actions of glucosamine on the interleukin-1beta-mediated activation of synoviocytes.

OBJECTIVE: Recently, we found that administration of glucosamine to adjuvant arthritis, a model for rheumatoid arthritis, suppressed the progression of arthritis in rats. To clarify its anti-inflammatory mechanism, we evaluated the actions of glucosamine on the activation of synoviocytes in vitro. MATERIALS AND METHODS: Synoviocytes isolated from human synovial tissues were stimulated with interleukin (IL)-1beta in the presence of 0.01-1 mM glucosamine. IL-8 and prostaglandin (PG) E(2) were measured by ELISA, and nitric oxide was quantitated by Griess assay. IL-8 mRNA was detected by RT-PCR. Furthermore, the effect of glucosamine on the phosphorylation of p38 mitogen-activated protein kinase (MAPK) and the binding of [(125)I] IL-1beta to its receptors were examined using a primary human synovial cell line (CSABI- 479). RESULTS: Glucosamine significantly suppressed the IL-1beta-induced IL-8 production as well as its mRNA expression (p < 0.05) at 1 mM. Furthermore, glucosamine (1 mM) inhibited the IL-1beta-induced nitric oxide and PGE(2) production (p < 0.05). Moreover, glucosamine suppressed the IL-1beta-induced phosphorylation of p38 MAPK (p < 0.05 at >0.1 mM) and the IL-1beta-binding to its receptors (p < 0.05 at 1 mM). CONCLUSIONS: These observations suggest that glucosamine can suppress the IL-1beta-mediated activation of synoviocytes (such as IL-8-, nitric oxide- and PGE(2)-production, and phosphorylation of p38 MAPK), thereby possibly exhibiting antiinflammatory actions in arthritis.

Preventive actions of a high dose of glucosamine on adjuvant arthritis in rats.

OBJECTIVE: Glucosamine, a naturally occurring amino monosaccharide has been used to treat or prevent osteoarthritis in humans. In this study, we evaluated the effect of glucosamine on rat adjuvant arthritis, a model of rheumatoid arthritis. MATERIALS AND METHODS: Adjuvant arthritis was induced in male Wistar rats by injection of Freund's complete adjuvant (FCA) into the right hind paw, and 300 mg/kg of glucosamine, an extra-dose compared with a regular dose for osteoarthritis patients (1.5 g/day, approximately 25 mg/kg), was orally administered once a day to the arthritic rats for 22 days. RESULTS: Glucosamine significantly suppressed the increase in arthritis score (p < 0.05) after day 10 of adjuvant injection, and inhibited the swelling of FCA-injected right and -uninjected left hind paws (p < 0.01) after day 18. In addition, histopathological examination of the arthritic joints revealed that glucosamine suppressed synovial hyperplasia, cartilage destruction and inflammatory cell infiltration. Furthermore, glucosamine reduced the production of nitric oxide and prostaglandin E(2) in plasma (p < 0.05). CONCLUSIONS: These observations suggest that glucosamine is able to suppress the progression of adjuvant arthritis in rats. Glucosamine may be expected as a novel anti-inflammatory agent for treatment of rheumatoid arthritis.

Combined radiation and cytochrome CYP4B1/4-ipomeanol gene therapy using the EGR1 promoter.

BACKGROUND: Cytochrome p450 isozyme CYP4B1 converts the inert prodrug 4-ipomeanol (4-IM) into toxic alkylating metabolites. Induction of cytotoxicity by 4-IM combined with ionizing radiation (IR) in cells transfected with a fusion protein of rabbit cytochrome CYP4B1 under control of the radiation inducible EGR1 promoter was investigated. The capability of activated 4-IM to sensitize cells to IR was also assessed. MATERIALS AND METHODS: Survival fractions of cells, determined by MTT assays, stably transfected with EGR1-CYP4B1 were compared with that of cells transfected with a control plasmid after IR followed by 4-IM. Radiosensitization was tested by comparing clonogenic survival curves of cells transfected with the CYP4B1 cassette under a CMV promoter instead of EGR-1, irradiated with or without 4-IM. RESULTS: MTT assays for cytotoxicity indicated a decrease in relative survival fractions (survival with 4-IM/survival without 4-IM) of the EGR1-CYP4B1 transfected cells with increasing radiation dosage, but not of control cells. Clonogenic assays revealed decreased survival fractions with increasing radiation doses (CYP4B1 transfected and control cells) and 4-IM concentrations (CYP4B1 transfected cells), but showed no significant differences in slope of survival curves with 4-IM. CONCLUSION: The results indicate IR potentiates the cytotoxic activity of the EGR1-CYP4B1/4-IM transgene system, but activated 4-IM does not sensitize cells to IR. Thus, the EGR1-CYP4B1/4-IM system is a viable radiation-gene therapy system that may allow for improved spatial and temporal control of cytotoxicity by therapeutic radiation fields.

Different effects of St John's wort on the pharmacokinetics of simvastatin and pravastatin.

OBJECTIVE: St John's Wort, a widely used herbal product, is an inducer of CYP3A4 and it decreases blood concentrations of CYP3A4 substrates. The effects of St John's Wort on the pharmacokinetics of 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitors simvastatin (an inactive lactone pro-drug) and pravastatin were determined in this study. METHODS: Sixteen healthy male subjects (n = 8 in group 1 and n = 8 in group 2) took a St John's Wort caplet (300 mg) or matching placebo three times a day for 14 days in a double-blind, crossover study. On day 14, a single oral dose of 10 mg simvastatin and 20 mg pravastatin was given to subjects in group 1 and group 2, respectively. Blood samples were obtained during a 24-hour period after the administration of each drug. RESULTS: Repeated St John's Wort treatment tended to lower plasma simvastatin concentration and significantly (P <.05) lowered concentrations of simvastatin hydroxy acid, its active metabolite. The peak concentration in plasma (ratio, 0.72 of placebo) of simvastatin hydroxy acid tended to be decreased and its area under the plasma concentration-time curve between time zero and 24 hours after administration (ratio, 0.48 of placebo) was significantly decreased (P <.05) by St John's Wort. On the other hand, St John's Wort did not influence plasma pravastatin concentration. No significant differences were observed in the elimination half-life of simvastatin or pravastatin between the placebo and St John's Wort trials. CONCLUSIONS: This study showed that St John's Wort decreases plasma concentrations of simvastatin but not of pravastatin. Because simvastatin is extensively metabolized by CYP3A4 in the intestinal wall and liver, which are induced by St John's Wort, it is likely that this interaction is partly caused by the enhancement of the CYP3A4-mediated first-pass metabolism of simvastatin in the small intestine and liver.

Suppression of protein kinase C signaling by the novel isoform for bovine PGF(2alpha) receptor.

A cDNA clone for a novel isoform of prostaglandin (PG) F(2alpha) receptor (FP) was isolated from the cDNA pool of the bovine corpus luteum. The sequence analysis revealed that the new FP isoform (FP(a)) encodes a 295-amino acid protein carrying a specific 28-amino acid sequence from the middle of transmembrane segment VI to the carboxyl terminus. Because only one copy gene has been identified for FP, FP(a) was generated by alternative mRNA splicing at the middle of the VI transmembrane region, resulting in the lack of a VII transmembrane segment and an intracellular carboxyl tail. The RT-PCR analysis for FP and FP(a) indicated that both mRNAs are expressed similarly during the estrous cycle and pregnancy. The PGF(2alpha) stimulation drastically enhanced protein kinase C (PKC) activity in the COS-7 cell transfected with FP, whereas no PKC activation was detected in FP(a)-transfected cells. Cotransfection of an excess amount of FP(a) markedly reduced FP-mediated PKC activity, suggesting that the novel FP isoform might play a role as a negative regulator to attenuate normal FP function.

p55CDC/hCDC20 is associated with BUBR1 and may be a downstream target of the spindle checkpoint kinase.

Eukaryotic cells have evolved a mechanism that delays the progression of mitosis until condensed chromosomes are properly positioned on the mitotic spindle. We have been studying genes that regulated the spindle checkpoint in human cells. Enforced expression of human BUBR1, but not a BUBR1 mutant allele, enhances accumulation of mitotic cells. Yeast two-hybrid system and GST-pulldown analyses show that p55CDC/hCdc20, a protein known to link spindle checkpoint components such as MAD2 to anaphase promoting complex (APC), interacts with BUBR1. In addition, p55CDC is capable of pulling down BUBR1 in sf-9 cells infected with both p55CDC and His6-BUBR1 recombinant baculoviruses but not in the cells infected with p55CDC baculoviruses or with the baculoviral vector alone. Moreover, immunoprecipitation followed by Western blot analyses confirmed that native p55CDC is associated with BUBR1 in HeLa cells. Spindle checkpoint activation by nocodazole treatment enhances the association between p55CDC and His6-BUBR1. In nocodazole-arrested mitotic cells, both CDC16 and hyperphosphorylated CDC27, two APC components, preferentially associate with His6-BUBR1 resins, but not the control resins. Furthermore, BUBR1 phosphorylates p55CDC in vitro, and the phosphorylation of p55CDC by BUBR1 appears to be correlated with spindle checkpoint activation. Together, our studies strongly suggest that BUBR1 may target APC via p55CDC.

Synergistic effects of thearubigin and genistein on human prostate tumor cell (PC-3) growth via cell cycle arrest.

There is evidence that habitual consumption of green tea by Japanese men is correlated with a reduction in cancers, including prostate; soybean isoflavones are also associated with increased protection. The present study compared the anti-proliferative effect of black tea (Camellia sinensis) polyphenol, thearubigin (TR), alone or combined with the isoflavone genistein, on human prostate (PC-3) carcinoma cells. TR administered alone did not result in any alteration of cell growth. When combined with genistein, however, TR significantly inhibited cell growth and induced a G2/M phase cell cycle arrest in a dose dependent manner. These findings indicate the potential use of combined phytochemicals to provide protection against prostate cancer.

The effect of diet on blood vitamin K status and urinary mineral excretion assessed by a food questionnaire.

To assess Vitamin K (VK) status by food questionnaire, healthy young males (32) and females (9) were given a food list of 50 VK rich foods (identified in the 4th edition standard tables of food composition in Japan). After checking the food names and approximate amount eaten for 7 days, early morning blood and urine samples were collected. Prothrombin and hepaplastin was tested and plasma protein induced by VK absence factor II (PIVKA-II), osteocalcin, and calcium, phosphorous and magnesium in plasma and urine were determined. Participants who have a habit of eating natto, a traditional Japanese food very rich in Vitamin K, VK were excluded, and lowest and highest VK consumers were compared (males; lowest 7 vs. highest 7, females; 3 vs. 3). Plasma PIVKA-II levels, and urinary calcium and magnesium excretion of the lowest was significantly higher, but urinary phosphorus was lower, than that of the highest in females. In the natto eaters, daily mean VK intakes and hepaplastin test results of natto eaters were significantly higher, but urinary calcium excretions were lower than that of non natto eaters in males. These results suggest that Daily VK intake estimated from a questionnaire, is well related to real VK status, and also that natto is a good dietary source of vitamin K.

Study of the cause of the temperature rise at the muscle-bone interface during ultrasound hyperthermia.

Undesirable temperature rise at the muscle-bone interface has been one of the major problems during ultrasound hyperthermia treatment. In this study, we examined by both computer calculation and phantom experiment the cause of this problem. Ultrasound penetrates a bone in two different waveforms, longitudinal and transversal. The transmission coefficient of these two waves vary greatly with the incident angle. From both theoretical and experimental results, the incident angle dependency of the interface heat was confirmed. When the incident angle is less than the critical angle of the longitudinal wave, the main cause of the temperature elevation is the absorption of the longitudinal wave in the bone. When the incident angle is larger than the critical angle of the longitudinal wave, the transversal wave becomes the major cause of the heat generation. At the incident angles larger than the critical angle of the transversal wave, no temperature rise is produced by the absorption of the ultrasound at the bone; the incident longitudinal wave, strengthened by the reflected wave, is absorbed in the muscle just in front of the bone. The heat generated in the muscle is transported to the interface so that the temperature of the interface and bone increases slightly.

Novel membrane transporter OCTN1 mediates multispecific, bidirectional, and pH-dependent transport of organic cations.

In the present study, functional characteristics of organic cation transporter (OCTN)1, which was cloned as the pH-dependent tetraethylammonium (TEA) transporter when expressed in mammalian human embryonic kidney (HEK)293 cells, were further investigated using Xenopus oocytes as well as HEK293 cells as gene expression systems. When OCTN1-derived complementary RNA was injected into Xenopus oocytes, pH-dependent transport of [14C]TEA was observed as the same in HEK293 cells. In contrast, a replacement of sodium ions with potassium ions in the surrounding medium did not cause any change in [14C]TEA uptake in Xenopus oocytes expressed with OCTN1. In addition, when OCTN1 was expressed in HEK293 cells, efflux of TEA from the cells was pH dependent, with an accelerated rate at acidic external medium pH. Accordingly, membrane potential or sodium ions are suggested to have no influence on [14C]TEA transport and the transport activity of OCTN1 is directly affected by pH itself. Furthermore, addition of the unlabeled TEA in external medium enhanced the efflux of preloaded [14C]TEA. These observations suggest that OCTN1 is a pH-dependent and bidirectional TEA transporter. OCTN1-mediated [14C]TEA uptake was inhibited by various organic cations such as cimetidine, procainamide, pyrilamine, quinidine, quinine, and verapamil. In addition, uptakes of cationic compounds such as [3H]pyrilamine, [3H]quinidine, and [3H]verapamil and zwitterionic L-[3H]carnitine were increased by expression of OCTN1 in Xenopus oocytes. Accordingly, OCTN1 was functionally demonstrated to be a multispecific and pH-dependent organic cation transporter, which presumably functions as a proton/organic cation antiporter at the renal apical membrane and other tissues.

[Spinal cord stimulation therapy at an early stage for unresponsive patients with hypoxic encephalopathy].

Recently, spinal cord stimulation (SCS) has been used for the treatment of patients in prolonged coma. However, the results of SCS in unresponsive patients with hypoxic encephalopathy at the chronic stage have not been satisfactory. Considering these circumstances, we began SCS from one month after the onset of hypoxic encephalopathy and evaluated its effect. Twelve patients (5 males and 7 females) with hypoxic encephalopathy, ranging in age from 7 to 72 years, were treated with SCS. The causes of hypoxia were acute cardiac failure in 4, automobile exhaust gas poisoning in 2, and asthma, pneumothorax, anaphylaxis, asphyxia, drowning and hypotension during aortic surgery in one patient each. One month after the onset, an electrode for electrical stimulation was implanted in the epidural space at the C2-C4 level under general anesthesia. The spinal cord was stimulated for 8 hours each day, starting on the day after implantation, and was continued for 3 months. Magnetic resonance imaging (MRI), cerebral blood flow (CBF) measurement using xenon-computed tomography (Xe-CT), and measurement of auditory evoked potential (AEP) and somatosensory evoked potential (SEP) were carried out 3 weeks after the onset for presurgical evaluation. Among the 12 patients, 7 (58%) showed clinical improvement, beginning within two weeks after starting stimulation. They were able to communicate with others and to express their emotion. However, disturbance of writing, picture drawing and calculation were not improved by stimulation. From presurgical evaluation, cases in which SCS therapy was effective had the following features: 1) No hemorrhagic infarction in the basal ganglia was demonstrable by MRI. 2) Mean hemispheric CBF measured by the Xe-CT method exceeded 25 ml/100 g per min. 3) The mean increase in hemispheric CBF 20 min after acetazolamide administration exceeded 5 ml/100 g per min. 4) An N20 peak was evident on the median nerve SEP, SCS appears to be an effective supplementary for unresponsive patients with hypoxic encephalopathy at the subacute stage, in addition to rehabilitation and drug therapy.

Mineral concentrations in the blood of male smokers and non-smoking males and females measured with fluoro-X-ray analyzer.

For clarification of the effects of smoking on mineral absorption in red blood cells (RBC) and serum, their concentrations at these sites were examined in ten male smokers and 12 male and 10 female non-smokers by fluoro-X-ray analysis. K in serum was high, and P, Ca and Fe low, compared to RBC. Mg and S in serum and RBC were essentially the same. S, Mg, P and K in RBC of smokers were higher than in male non-smokers, and Ca was lower. S and Ca in serum of smokers were significantly lower than in male non-smokers. P in smokers was higher than in non-smokers. P in RBC may possibly activate the reflux of Mg into RBC and may suppress that of Ca. In smokers, the correlation coefficient (gamma) between Fe and Ca was r = -0.68 in RBC, and r = 0.76 in serum. Also gamma between P and Mg was r = 0.4 in RBC and r = -0.48 in serum. Thus Fe in RBC may suppress the reflux of Ca. Mg and Ca in serum of females were significantly higher than in male non-smokers. Ca in RBC of females was significantly higher than in male non-smokers, and P, K and Fe were significantly lower. The product of Ca and P in RBC of females was lower than in male non-smokers, while in serum it was higher. gamma between P and Ca, and between Fe and Ca in RBC of females were negative, while they were positive in male non-smokers. The correlations in female RBC had the same pattern as in male smokers.

[Balloon-occluded ethanol ablation therapy: (BEAT) for hepatocellular carcinoma].

To obtain simultaneous embolization of feeding arteries and portal veins in the hepatocellular carcinoma, we performed a new treatment, balloon occluded ethanol ablation therapy (BEAT), in 7 patients. A Balloon catheter was inserted into the hepatic vein draining the tumor bearing segment of the liver. During occlusion of the hepatic vein by balloon catheter, an absolute ethanol-Lipiodol emulsion was injected through a microcatheter or a microballoon catheter placed in the feeding artery. There were no major side effects. Histological examination of the specimen taken from one case showed complete necrosis of the tumor and accumulation of Lipiodol within the portal veins of the surrounding tissue.

Functional oxytocin receptors in bovine granulosa cells.

The aim of this study was to identify the presence of functional oxytocin (OT) receptors on bovine granulosa cells. Freshly prepared bovine granulosa cells from small (3-5 mm in diameter) or preovulatory (mature) follicles were examined for OT receptors by a radioreceptor assay. Scatchard analysis revealed that both binding capacity and affinity in granulosa cells from small follicles were significantly higher than those in granulosa cells from mature follicles (p < 0.01). With use of a reverse transcriptase polymerase chain reaction analysis, expression of OT receptor mRNA was detected in granulosa cells obtained from both small and mature follicles. When the granulosa cells obtained from small follicles were cultured in Dulbecco's Modified Eagle's Medium and Ham's F-12 medium (1:1 [v:v]) with 10% calf serum up to 72 h, as the period of culture was prolonged, the concentration of OT receptor decreased with increases of progesterone and OT release in the medium. However, the binding affinity was not changed during culture for 72 h. When bovine follicular oocytes with cumulus oophorus were cultured for 24 h in tissue culture Medium-199 with 10% fetal calf serum and OT (0-10 nM), the percentages of oocytes reaching maximum cumulus expansion were significantly increased at 0.5, 1, and 10 nM OT, although nuclear maturation in oocytes surrounded by compact cumulus cells was not affected by the addition of OT. Coexposure with OT antagonist blocked the stimulatory effect of OT on cumulus expansion, confirming the specificity of the effect. Furthermore, anti-OT rabbit serum inhibited the percentages of oocytes with expanded cumulus compared to those supplemented with normal rabbit serum (p <0.05). The overall results indicate the presence of functional OT receptors in bovine granulosa cells and support the hypothesis that OT plays a role (or roles) in regulating the function of granulosa cells as an autocrine factor during follicular growth.

Allyl sulfides from garlic suppress the in vitro proliferation of human A549 lung tumor cells.

The present studies compared the antiproliferative effects of diallyl trisulfide (DATS) and diallyl disulfide (DADS) on cultured human neoplastic (A549) and nonneoplastic (MRC-5) lung cells. Addition of 10 microM DATS reduced A549 growth by 47%, whereas 10 microM DADS decreased growth by only 20%. DATS treatment (10 microM) did not alter MRC-5 cell growth. DATS (10 microM) caused a marked and progressive increase in intracellular Ca2+ in A549 cells during the first four hours after treatment. Intracellular Ca2+ in A549 cells exposed to DATS returned to near control levels within one hour after refeeding complete medium without DATS. Exposure to 1 microM DATS for 24 hours significantly induced apoptosis, as indicated by increased DNA fragmentation. The ability of DATS and DADS to suppress neoplastic growth is consistent with increasing evidence that several garlic components have anticarcinogenic and antitumorigenic properties.

Dietary rosemary suppresses 7,12-dimethylbenz(a)anthracene binding to rat mammary cell DNA.

Commercially available ground rosemary powder was examined for its ability to modify the in vivo binding of 7,12-dimethylbenz(a)anthracene (DMBA) metabolites to mammary cell DNA in 55-d-old rats fed diets containing varying quantities and types of lipids. Supplementing a casein-based diet containing 20% corn oil with 1 % rosemary for 2 wk reduced by 76% the occurrence of DMBA-induced DNA adducts occurring 24 h after treatment with 50 mg DMBA/kg body weight. A comparable reduction in DNA adducts (66%) occurred when 0.5% rosemary was added to a diet containing 20% corn oil, and the quantity of DMBA given was reduced to 25 mg/kg body weight. The reduction in the occurrence of adducts occurring 24 h after DMBA treatment caused by 0.5% dietary rosemary was greater (P < 0.05) when added to a diet containing 20% corn oil than when added to a diet containing 5% corn oil and 15% coconut oil. Rosemary, 1% but not 0.5%, reduced DMBA-induced DNA adducts when the diet contained 5% corn oil. These studies demonstrate that rosemary is effective in reducing the binding of DMBA metabolites to rat mammary cell DNA. Furthermore, the present studies demonstrate that the benefits of rosemary are dependent on the source and concentration of dietary lipids.

Comparison of the inhibitory action of synthetic capsaicin analogues with various NADH-ubiquinone oxidoreductases.

Capsaicin is a new naturally occurring inhibitor of proton-pumping NADH-ubiquinone oxidoreductase (NDH-1), that competitively acts against ubiquinone. A series of capsaicin analogues was synthesized to examine the structural factors required for the inhibitory action and to probe the structural property of the ubiquinone catalytic site of various NADH-ubiquinone reductases, including non-proton-pumping enzyme (NDH-2), from bovine heart mitochondria, potato tuber (Solanum tuberosum, L) mitochondria and Escherichia coli (GR 19N) plasma membranes. Some synthetic capsaicins were fairly potent inhibitors of each of the three NDH-1 compared with the potent rotenone and piericidin A. Synthetic capsaicin analogues inhibited all three NDH-1 activities in a competitive manner against an exogenous quinone. The modification both of the substitution pattern and of the number of methoxy groups on the benzene ring, which may be superimposable on the quinone ring of ubiquinone, did not drastically affect the inhibitory potency. In addition, alteration of the position of dipolar amide bond unit in the molecule and chemical modifications of this unit did not change the inhibitory potency, particularly with bovine heart and potato tuber NDH-1. These results might be explained assuming that the ubiquinone catalytic site of NDH-1 is spacious enough to accommodate a variety of structurally different capsaicin analogues in a dissimilar manner. Regarding the moiety corresponding to the alkyl side chain, a rigid diphenyl ether structure was more inhibitory than a flexible alkyl chain. Structure-activity studies and molecular orbital calculations suggested that a bent form is the active conformation of capsaicin analogues. On the other hand, poor correlations between the inhibitory potencies determined with the three NDH-1 suggested that the structural similarity of the ubiquinone catalytic sites of these enzymes is rather poor. The sensitivity to the inhibition by synthetic capsaicins remarkably differed between NDH-1 and NDH-2, supporting the notion that the sensitivity against capsaicin inhibition correlates well with the presence of an energy coupling site in the enzyme (Yagi, T. (1990) Arch. Biochem. Biophys. 281, 305-311). It is noteworthy that several synthetic capsaicins discriminated between NDH-1 and NDH-2 much better than natural capsaicin.

Non-destructive elemental analysis of vertebral body trabecular bone using muonic X-rays.

Non-destructive elemental analysis with muonic X-rays was performed on human vertebral bone and lumbar torso phantoms. It can provide quantitative information on all elements in small deep-seated localized volumes. The experiment was carried out using the superconducting muon channel at TRIUMF in Vancouver, Canada and a lithium drifted germanium detector with an active area of 18.5 cm2. The muon channel produced backward-decayed negative muons with wide kinetic energy range from 0.5 to 54.2 MeV. The muon beam was collimated to a diameter of 18 mm. The number of incoming muons was about 4 x 10(6) approximately 5 x 10(7) per data point. In the measurements with human vertebral bones fixed with neutralized formaldehyde, the correlation coefficient between calcium content measured by muons and by atomic absorption analysis was 0.99 and the level of significance was 0.0003. In the measurements with lumbar torso phantoms, the correlation coefficient between calcium content measured by muons and by atomic absorption analysis was 0.99 and the level of significance was 0.02. The results suggest that elemental analysis in vertebral body trabecular bone using muonic X-rays closely correlates with measurements by atomic absorption analysis.

A longitudinal follow-up study of seasonal affective disorder.

OBJECTIVE: The purposes of this study were to evaluate the long-term stability of a seasonal pattern of recurrent depression, identify possible factors associated with alteration of the seasonal pattern, and determine whether atypical vegetative symptoms during early seasonal depressive episodes predict future seasonal relapses. METHOD: The subjects were 41 patients satisfying the criteria used in the Japanese multicenter study of seasonal affective disorder who were consistently treated at the same outpatient clinic. Their longitudinal courses were evaluated by using case records and the Schedule for Affective Disorder and Schizophrenia--Life-time Version; the mean follow-up period was 10.4 years. RESULTS: Nine subjects (22.0%) consistently showed a fall-winter pattern of recurrence throughout follow-up. Seventeen patients with an initial fall-winter pattern subsequently tended to shift seasons or show less seasonality. This alteration in pattern was possibly associated with antidepressant therapy or life events. Eleven patients with an initial diagnosis of nonseasonal affective disorder subsequently developed seasonal affective disorder; no specific factors were associated with this change. Atypical vegetative symptoms were significantly more common in patients with stable seasonal patterns of recurrence than in those who lost seasonality. CONCLUSIONS: Although seasonal affective disorder appeared to be altered by antidepressant treatment, the presence of a core group of patients with a consistent seasonal pattern of recurrent depression suggests the validity of seasonal affective disorder as a distinct subtype of recurrent affective illness. The findings also suggest that atypical vegetative symptoms during early seasonal depressive episodes predict the subsequent seasonality of depression.