Comparison of taurine biosynthesis ability between juveniles of Japanese flounder and common carp.

This study was conducted to investigate taurine deficiency and the ability of taurine biosynthesis in both juvenile Japanese flounder (JF) and juvenile common carp (CC) in vivo using low taurine level diets. Three different taurine level diets were prepared by the supplementation of taurine to the basal composition (JF--0, 0.5 and 1.5% in JF; CC--0, 1, 3% in CC). The final average body weight and feed efficiency of JF fed the JF - 1.5% was significantly higher than those of fish fed on the JF--0%. On the other hand, no significant difference was observed in CC fed with CC--0, 1, and 3% diets. The taurine retention rate was negative in the case of JF-fed with the taurine-free supplement (JF--0%). On the other hand, the taurine retention rate was about 280% in the case of CC-fed with the taurine-free supplement (CC--0%). These findings indicate that while taurine is essential for growth of JF, it is not essential for the growth of CC.

Nitrogen balance and ileal amino acid digestibility in growing pigs fed diets reduced in protein concentration.

Two studies were conducted to assess the effect of dietary protein reduction on N utilization, N excretion, and AA digestibility in growing pigs. The objective was to determine whether pigs fed diets with a reduced CP concentration could maintain the same N retention as pigs fed an adequate diet. The second objective was to test whether reducing dietary CP concentration decreases AA digestibility. In each study, six barrows were allotted to one of six dietary treatments in a Latin square design. Treatments consisted of four corn-soybean meal-based diets containing 15, 12, 9, and 6% CP, a casein-based diet containing 15% CP, and a protein-free diet. Crystalline AA were included in the 12, 9, and 6% CP diets. The indispensable:dispensable AA ratio was maintained at 45:55 with the addition of L-glutamic acid to the 9 and 6% CP diets. The casein-based and protein-free diets were used to determine endogenous total tract N and ileal AA losses. In the first study, total N losses and N absorbed decreased linearly (P < 0.001) as dietary CP concentration decreased from 15 to 6%. Both a linear (P < 0.001) and a quadratic (P < 0.05) decrease in N retention were found with decreasing dietary CP concentration. Nitrogen retained as a percentage of intake and absorbed increased (P < 0.001) as dietary CP concentration was reduced from 15 to 6%. In the second study, six barrows were surgically fitted with a T-cannula at the terminal ileum to determine ileal AA digestibility. For all dispensable and most indispensable AA, apparent and standardized ileal digestibility increased linearly (P 0.01, and for arginine, P < 0.05) as dietary CP concentration decreased. These results indicate that dietary CP concentration can be decreased from 15 to 12% with crystalline AA supplementation to meet an ideal AA profile without adversely affecting N retention, and that decreasing dietary CP concentration from 15 to 6% increases both dispensable and indispensable AA ileal digestibility.

Normalization of low biotinidase activity in a child with biotin deficiency after biotin supplementation.

We report findings in a Japanese boy with severe skin rash attributable to biotin deficiency. The patient had an intracranial malformation and developed biotin deficiency due to tube feeding with a single formula for over one year. Results of urinary organic acid analysis were consistent with multiple carboxylase deficiency, and low biotinidase activity was also observed. After biotin supplementation, the skin rash improved and biotinidase activity normalized. We speculate that biotin is one regulating factor in the biosynthesis of biotinidase.

Effects of eicosapentaenoic acid on blood pressure, cell membrane fatty acids, and intracellular sodium concentration in essential hypertension.

This study was designed to clarify the effects of orally administered eicosapentaenoic acid (EPA) on blood pressure, intracellular sodium content, and cell membrane fatty acid composition in patients with essential hypertension. After a 4-week run-in period, a study group of 17 male patients was assigned to an 8-week treatment with EPA (2.7 g/day) or placebo in a randomized, double-blind fashion with a crossover at week 4. Systolic blood pressure (SBP) was lower after treatment with EPA than after treatment with placebo (152.9+/-17.3 vs. 162.6+/-20.6 mmHg; p<0.01), while diastolic blood pressure was not statistically different. Compared with the placebo treatment, EPA supplementation resulted in a decrease in intraerythrocyte sodium content (R-Na; 11.17+/-0.63 vs. 10.44+/-1.28 nmol/l cells; p<0.05) accompanied by an increase (p<0.001) in erythrocyte membrane EPA content. The increase in membrane EPA content was related to the decrease in SBP (r=-0.52, p<0.05) and the decrease in R-Na (r=-0.57, p<0.02) during EPA treatment. The decrease in R-Na correlated positively with the decrease in SBP (r=0.54, p<0.05), and correlated negatively with the change in Na+-K+ ATPase activity (r= -0.59, p<0.02). However, the change in Na+-K+ ATPase activity did not directly correlate with the change in membrane EPA content. In conclusion, oral EPA supplementation increased membrane EPA content and reduced SBP in patients with essential hypertension. Based on the association between the increase in membrane EPA content and the decrease in intracellular sodium concentration, EPA may lower blood pressure by altering the activities of the membrane sodium transport systems.

[A case of central retinal artery occlusion after anterior posterior fusion of the lumbar spine].

We report a case of central retinal artery occlusion after anterior-posterior fusion of the lumbar spine. The patient suddenly lost his vision of the right eye in the ICU just after the end of long procedure for anterior-posterior fusion of the lumbar spine. The patient was diagnosed as having central retinal artery occlusion, and treated successfully with treatments including immediate administration of urokinase and PGE1, stellate ganglion block, and hyperbaric oxygen therapy. The patient was discharged from the hospital on the 54th postoperative day with adequate vision to drive a car. Central retinal artery occlusion is a rare but very serious complication during and after supine surgery with prone position. It is very important for us to be aware of its possible occurrence. We have to diagnose and treat, as soon as possible, the vision loss after the spine surgery.

Roles of carbohydrates on Cry j 1, the major allergen of Japanese cedar pollen, in specific T-cell responses.

BACKGROUND: Carbohydrates expressed on allergens are known to be important for allergenicity. However, little is known about whether the carbohydrates drive the T(H)2 response. OBJECTIVE: We sought to determine a role for carbohydrates expressed on Cry j 1, which is the major allergen of Cryptomeria japonica pollen and causes the most prevalent pollinosis in Japan, in in vitro cellular responses. METHODS: Carbohydrates on Cry j 1 were destroyed by periodate-oxidation under mild conditions. Proliferative responses and cytokine productions against native, periodate-treated, and mock-treated Cry j 1 were compared in peripheral blood mononuclear cells, Cry j 1-specific T-cell lines, and clones from patients with Japanese cedar pollinosis. RESULTS: We found that peripheral blood mononuclear cells from patients with Japanese cedar pollinosis displayed a significant decrease in proliferation and IL-5 production in response to periodate-treated Cry j 1 in comparison with native and mock-treated Cry j 1. Decreased proliferative responses against periodate-treated Cry j 1 were also seen in polyclonal T-cell lines, and the responses showed a heterogeneity. In addition, Cry j 1-specific CD4+ T-cell clones also displayed a significant decrease in proliferation and IL-4 and IL-5 production-but not IFN-gamma production-in comparison with the control antigens. However, most of the clones showed decreased but positive proliferative responses against periodate-treated Cry j 1. Blockade of the mannose receptor had no effect on cellular responses. CONCLUSION: The results suggest that carbohydrates on Cry j 1 play a major role in promoting Cry j 1-specific T(H)2 response in vitro, though they are not major targets as T-cell epitopes.

Progression of T cell lineage restriction in the earliest subpopulation of murine adult thymus visualized by the expression of lck proximal promoter activity.

The proximal promoter of lck directs gene expression exclusively in T cells. To investigate the developmental regulation of the lck proximal promoter activity and its relationship to T cell lineage commitment, a green fluorescence protein (GFP) transgenic (Tg) mouse in which the GFP expression is under the control of the proximal promoter of lck was created. In the adult GFP-Tg mice, >90% of CD4(+)CD8(+) and CD4(+)CD8(-) thymocytes, and the majority of CD4(-)CD8(+) and CD4(-)CD8(-) [double-negative (DN)] thymocytes were highly positive for GFP. Slightly lower but substantial levels of expression of GFP was also observed in mature splenic T cells. No GFP(+) cells was detected in non-T lineage subsets, including mature and immature B cells, CD5(+) B cells, and NK cells, indicating a preserved tissue specificity of the promoter. The earliest GFP(+) cells detected were found in the CD44(+)CD25(-) DN thymocyte subpopulation. The developmental potential of GFP(-) and GFP(+) cells in the CD44(+)CD25(-) DN fraction was examined using in vitro culture systems. The generation of substantial numbers of alphabeta and gammadelta T cells as well as NK cells was demonstrated from both GFP(-) and GFP(+) cells. However, no development of B cells or dendritic cells was detected from GFP(+) CD44(+)CD25(-) DN thymocytes. These results suggest that the progenitors expressing lck proximal promoter activity in the CD44(+)CD25(-) DN thymocyte subset have lost most of the progenitor potential for the B and dendritic cell lineage. Thus, progression of T cell lineage restriction in the earliest thymic population can be visualized by lck proximal promoter activity, suggesting a potential role of Lck in the T cell lineage commitment.

Transcription factor BACH1 is recruited to the nucleus by its novel alternative spliced isoform.

The transcription factor Bach1 is a member of a novel family of broad complex, tramtrack, bric-a-brac/poxvirus and zinc finger (BTB/POZ) basic region leucine zipper factors. Bach1 forms a heterodimer with MafK, a member of the small Maf protein family (MafF, MafG, and MafK), which recognizes the NF-E2/Maf recognition element, a cis-regulatory motif containing a 12-O-tetradecanoylphorbol-13-acetate-responsive element. Here we describe the gene structure of human BACH1, including a newly identified promoter and an alternatively RNA-spliced truncated form of BACH1, designated BACH1t, abundantly transcribed in human testis. The alternate splicing originated from the usage of a novel exon located 5.6 kilobase pairs downstream of the exon encoding the leucine zipper domain, and produced a protein that contained the conserved BTB/POZ, Cap'n collar, and basic region domains, but lacked the leucine zipper domain essential for NF-E2/Maf recognition element binding. Subcellular localization studies using green fluorescent protein as a reporter showed that full-length BACH1 localized to the cytoplasm, whereas BACH1t accumulated in the nucleus. Interestingly, coexpression of BACH1 and BACH1t demonstrated interaction between the molecules and the induction of nuclear import of BACH1. These results suggested that BACH1t recruits BACH1 to the nucleus through BTB domain-mediated interaction.

Purification, identification, and cDNA cloning of Jun a 2, the second major allergen of mountain cedar pollen.

The second major allergen of Juniperus ashei (mountain cedar) pollen, Jun a 2, has been purified and its cDNA cloned. The purified protein has a molecular mass of 43 kDa and its N-terminal 9-residue amino acid sequence is highly homologous to those of Cry j 2 and Cha o 2, the second major allergen of Cryptomeria japonica and Chamaecyparis obtusa pollen, respectively. cDNA clones encoding Jun a 2 were isolated after PCR based amplification, and their nucleotide sequences were determined. The cDNA contains an open reading frame of 507 amino acid residues, and encodes a putative 54-residue signal sequence and a 453-residue intermediate, which releases a C-terminal fragment upon maturation. Three possible N-linked glycosylation sites and 20 cystein-residues are found in the deduced amino acid sequence. The amino acid sequence of Jun a 2 shows 70.7 and 82.0% identity with those of Cry j 2 and Cha o 2, respectively. Immunological observations that IgE antibodies in sera of Japanese pollinosis patients bind not only to Cry j 2 and Cha o 2 but also to Jun a 2 strongly suggest that Jun a 2 is an allergen of mountain cedar pollen, and that allergenic epitopes of these three allergens are similar.

Isoproterenol-induced myocardial injury resulting in altered S100A4 and S100A11 protein expression in the rat.

S-100 proteins (S100) are characterized by calcium-binding ability with two structural EF hands. Several S100 are expressed in cardiomyocytes and thought to play a crucial role in calcium signaling. To examine whether the expression of S100 is a response to detectable myocardial damage or regeneration, we investigated, immunohistochemically, the expression of S100A4 and S100A11 in the isoproterenol (ISP)-treated rat heart. Definite expression of S100A4 and S100A11 was demonstrated in normal cardiomyocytes, and their staining patterns were enhanced in the ISP-treated rat heart, suggesting the possible involvement of S1-A4 and S100A11 in ISP-induced myocardial damage.

Shear stress enhances glutathione peroxidase expression in endothelial cells.

Hemodynamic forces have profound effects on vasculature. Laminar shear stress upregulates superoxide dismutase (SOD) expression in endothelial cells. SOD converts superoxide anion to H(2)O(2), which, however, promotes atherosclerosis. Therefore, defense against H(2)O(2) may be crucial in reducing oxidative stress. Since glutathione peroxidase (GPx-1) reduces H(2)O(2) to H(2)O, the regulation of GPx-1 expression by mechanical stress was examined. Cultured bovine aortic endothelial cells (BAECs) were subjected to laminar shear stress and stretch force. Shear stress upregulated GPx-1 mRNA expression in a time- and force-dependent manner in BAECs, whereas stretch force was without effect. Furthermore, shear stress increased GPx activity. L-NAME, an inhibitor of nitric oxide synthase, did not affect shear stress-induced GPx-1 mRNA expression. The ability of laminar shear stress to induce GPx-1 expression in endothelial cells may be an important mechanism whereby shear stress protects vascular cells against oxidative stress.

Effect of L-arginine administration on myocardial thallium-201 perfusion during exercise in patients with angina pectoris and normal coronary angiograms.

OBJECTIVES: We tested the hypothesis that an exogenous supplement of L-arginine could alleviate coronary perfusion abnormality during exercise in patients with angina pectoris and normal coronary arteries. METHODS AND RESULTS: Twelve patients underwent exercise thallium-201 scintigraphy without medication (control) and after intravenous administration of L-arginine. Exercise time was prolonged in the L-arginine study compared with the control (482 s vs 540 s, P < .05). TI-201 extent score was improved in the L-arginine study (0.33 vs 0.26, P < .05), and the severity score was also improved (23.7 vs 16.9, P < .05). In 7 of the 12 patients whose TI-201 redistribution disappeared in the L-arginine study, the percent increase in serum L-citrulline concentration during exercise was larger than that of the remaining 5 patients (18% vs 0.9%, P < .01). The percent reduction in epicardial coronary diameter in response to acetylcholine was also greater in the former group (28.3% vs 11.1%, P < .05). CONCLUSION: Exogenous L-arginine improved myocardial perfusion during exercise in a subset of patients with angina pectoris and normal coronary arteries, probably by increasing production of nitric oxide.

Hypothermia with acute renal failure in a patient suffering from diabetic nephropathy and malnutrition.

We report a rare case of hypothermia with acute renal failure in a patient suffering from diabetic nephropathy. A 71-year-old male who had been receiving insulin therapy for the treatment of diabetes mellitus complicated with advanced diabetic nephropathy since 1998 was malnourished with an extremely decreased muscle mass. Without any prolonged exposure to excessively low external temperatures or hypothyroidism, pituitary insufficiency, adrenal insufficiency, sepsis, hypoglycemia, and diabetic ketoacidosis, acute hypothermia appeared together with an aggravation of diabetic nephropathy. His skin temperature fell to below measurable levels and his rectal temperature fell to 30.0 degrees C. His consciousness was drowsy and the hypothermia was not accompanied by shivering. Skeletal muscle is known to play an important role as a center of heat production and shivering thermogenesis in skeletal muscle mainly operates on acute cold stress. Therefore, in this case, hypothermia may have occurred because the shivering thermogenesis could not fully act on the acute cold stress due to the dramatically reduced muscle mass. We should always keep in mind that older, malnourished diabetic patients can easily suffer from impairments of the thermoregulatory system.

Oral taurine supplementation prevents the development of ethanol-induced hypertension in rats.

Taurine is known to lower blood pressure in essential hypertension and some experimental hypertensive models. Taurine has also been reported to activate aldehyde dehydrogenase and to inhibit the elevation of plasma acetaldehyde concentration after ethanol intake. Because acetaldehyde, the first metabolite of ethanol, is suspected to be responsible for many adverse effects of alcohol consumption, we examined the effect of taurine supplementation on ethanol-induced hypertension and abnormalities in the intracellular cation metabolism in Witar-Kyoto rats. In Study 1, systolic blood pressure and intraplatelet free calcium were significantly higher in rats who received 15% ethanol in drinking water than in control rats. Oral taurine supplementation (1% taurine and 15% ethanol in drinking water) completely prevented the development of ethanol-induced hypertension. Intraerythrocyte sodium and intraplatelet free calcium were significantly decreased in taurine-supplemented rats as compared with rats who received 15% ethanol only. In Study 2, hemoglobin-associated acetaldehyde (HbAA) was measured as a marker of protein-bound acetaldehyde. HbAA was significantly elevated in rats who received 5% ethanol in drinking water as compared with control rats. Taurine supplementation (1% taurine and 5% ethanol in drinking water) significantly decreased HbAA. Our findings suggest that the oral supplementation of taurine prevents ethanol-induced hypertension by decreasing protein bound acetaldehyde and altering the cation handling by the membrane.

Effects of transforming growth factor beta-1 and all-trans-retinoic acid on androgen-induced development of neonatal mouse bulbourethral glands in vitro.

Effects of transforming growth factor beta-1 (TGF-beta1) and all-trans-retinoic acid (All-trans-RA) on development of bulbourethral glands (BUGs) of neonatal mice were investigated in vitro. BUGs from 0-day-old male mice were cultured for 6 days in serum-free, chemically defined medium containing transferrin and bovine serum albumin, supplemented with 5alpha-dihydrotestosterone (DHT; 10-8 M) and insulin (10 microg/mL) alone or in combination. Prior to culture, BUGs from 0-day-old mice consisted of a simple epithelial rudiment encapsulated by mesenchyme. Epithelial growth and ductal branching occurred in BUGs cultured in medium containing DHT and insulin or DHT alone, but epithelial branching did not occur in BUGs cultured in the presence of insulin alone. Addition of TGF-beta1 at concentrations of > 5 ng/mL (0.2 x 10-9 M) to medium containing both insulin and DHT, inhibited the expected increase in overall size of BUGs, epithelial area and ductal branching in a dose-dependent manner. TGF-beta1 also decreased [3H]-thymidine labelling indices of both epithelium and mesenchyme. TGF-beta1 at 10 ng/mL elicited these inhibitory effects on BUGs cultured in medium containing DHT alone. Addition of All-trans-RA (10-8 to 10-6 M) to the medium containing DHT plus insulin, or DHT alone did not exert significant effects on either overall size of BUGs or epithelial growth and ductal branching. All-trans-RA at 10-6 M decreased the [3H]-thymidine labelling index of mesenchyme of BUGs cultured in medium with DHT plus insulin or DHT alone, but did not decrease the [3H]-thymidine labelling index of epithelium. The present results indicate that TGF-beta1 inhibits androgen-induced epithelial and mesenchymal growth as well as epithelial morphogenesis of BUGs from neonatal mice. Such an inhibitory effect of TGF-beta1 is not mimicked by All-trans-RA at physiological concentrations.

Immunohistochemical study of the receptors for retinoic acid in prostatic adenocarcinoma.

BACKGROUND: Retinoids play an important role in regulating cellular proliferation and differentiation. Although their effects are mediated by retinoic acid receptors (RARs) and retinoid X receptors (RXRs), limited information is available about the expression of RARs and RXRs in prostatic adenocarcinoma. We intended in this study to elucidate further the participation of those receptors in tumor progression of human prostate. MATERIALS AND METHODS: The immunohistochemical expression of RARs and RXRs proteins was studied using paraffin-embedded archival tissues obtained from patients with and without neoadjuvant hormonal therapies for prostatic adenocarcinoma. RESULTS: The immunoreactivity against RAR-alpha, RAR-gamma, RXR-alpha and RXR-gamma were detected in many, if not all, prostatic adenocarcinoma cells of the cases without and with neoadjuvant hormonal therapy, whereas less expression of both RAR-beta and RXR-beta were identified. Positively stained adenocarcinoma cells with RAR-beta and RXR-beta were found to be decreased in the cases with neoadjuvant therapy. In addition, the specimens showing positive immunoreaction of RAR-beta were limited to the cases of moderately- and poorly-differentiated but not well-differentiated, adenocarcinomas. CONCLUSION: We first demonstrated the expression of RARs and RXRs proteins in prostatic adenocarcinoma using subtype-specific antibodies. Immunohistochemistry using those antibodies might give an indication of susceptibility of the patients to retinoid therapy as a possible treatment of prostatic adenocarcinoma.

Molecular identification and characterization of novel membrane-bound metalloprotease, the soluble secreted form of which hydrolyzes a variety of vasoactive peptides.

One class of zinc metalloproteases, represented by neutral endopeptidase 24.11 and endothelin-converting enzyme, has been shown to be involved in proteolytic activation or inactivation of many regulatory peptides. Here, we report molecular cloning and characterization of a novel member of this type II membrane-bound metalloprotease family, termed soluble secreted endopeptidase (SEP). Alternative splicing results in the generation of another transcript, SEP(Delta), which lacks a 69-base pair nucleotide segment following the transmembrane helix. Both SEP and SEP(Delta) mRNA are detected in all mouse tissues examined. Transfection of an SEP cDNA expression construct resulted in the expression of the membrane-bound form of SEP in the early secretory pathway as well as the soluble secreted form of the enzyme in the culture medium. In contrast, transfection of the SEP(Delta) cDNA only results in the expression of the membrane-bound form. In vitro enzymological analysis of the recombinant soluble form of SEP demonstrated that it hydrolyzes a variety of vasoactive peptides, including endothelin-1, atrial natriuretic peptide, and angiotensin I. This activity of SEP was inhibited by phosphoramidon and the neutral endopeptidase 24.11 specific inhibitor thiorphan, but it was only partially inhibited by the endothelin-converting enzyme specific inhibitor FR901533. These findings suggest that SEP is a novel metalloprotease that possesses a broad substrate specificity and that it may be involved in the metabolism of biologically active peptides intracellulary as well as extracellularly.

Purification, identification, and cDNA cloning of Cha o 2, the second major allergen of Japanese cypress pollen.

The second major allergen of Chamaecyparis obtusa (Japanese cypress) pollen, Cha o 2, has been purified and its cDNA cloned. Of patients with pollinosis caused by C. obtusa, 82.5% produce IgE antibodies which react with purified Cha o 2. The purified protein has a molecular mass of 46 kDa and its 12 N-terminal amino acid sequence displays a high homology with that of Cry j 2, the second major allergen of Cryptomeria japonica pollen. cDNA clones coding for Cha o 2 have been isolated using Cry j 2 cDNA as a probe. Cha o 2 cDNA clones were sequenced and found to code a putative 50-residue signal sequence and a 464-residue mature protein with a molecular weight of 50 kDa. Two possible N-linked glycosylation sites were found in the sequence. The deduced amino acid sequence of Cha o 2 shows 74.3% identity with that of Cry j 2. In its primary structure, Cha o 2 shows significant identity with those of the polygalacturonases of avocado, tomato, and maize as well as Cry j 2.

Argatroban, an attractive anticoagulant, for left heart bypass with centrifugal pump for repair of traumatic aortic rupture.

Systemic heparinization often increases the risk of fatal bleeding from other injured organs in surgical repair of the aorta using extracorporeal circulation in patients with traumatic aortic rupture associated with multisystem injuries. We used an antithrombin agent, argatroban, as an alternative anticoagulant in left heart bypass with the Bio-Medicus centrifugal pump in 7 of 9 recent patients who underwent aortic repair using left heart bypass. All these patients survived without obvious evidence of systemic thromboembolization. Surgical treatments for other organ injuries were carried out in 3 patients concomitantly or immediately after aortic repairs without undue blood loss. Argatroban may have a complementary effect for preventing thrombus formation without aggravating bleeding tendency because of its monotarget specificity to thrombin. We believe intravenous administration (0.5 to 2 micrograms/kg/min) of argatroban is a safe anticoagulant for left heart bypass in repairs of traumatic aortic rupture associated with multiple organ injuries.