Precise Spectroscopy of the 3n and 3p Systems via the

To search for low-energy resonant structures in isospin T=3/2 three-body systems, we have performed the experiments ^{3}H(t,^{3}He)3n and ^{3}He(^{3}He,t)3p at intermediate energies. For the 3n experiment, we have newly developed a thick Ti-^{3}H target that has the largest tritium thickness among targets of this type ever made. The 3n experiment for the first time covered the momentum-transfer region as low as 15 MeV/c, which provides ideal conditions for producing fragile systems. However, in the excitation-energy spectra we obtained, we did not observe any distinct peak structures. This is in sharp contrast to tetraneutron spectra. The distributions of the 3n and 3p spectra are found to be similar, except for the displacement in energy due to Coulomb repulsion. Comparisons with theoretical calculations suggest that three-body correlations exist in the 3n and 3p systems, although not enough to produce a resonant peak.

Polyphenols and fatty acids responsible for anti-cyanobacterial allelopathic effects of submerged macrophyte Myriophyllum spicatum.

Myriophyllum spicatum is known to inhibit the growth of cyanobacteria such as Microcystis aeruginosa by releasing anti-cyanobacterial allelochemicals. The allelochemicals possibly responsible for the inhibition include five polyphenols and three fatty acids, but the extent to which these are indeed responsible for the anti-cyanobacterial effects is unclear. The goal of this research was to determine the contribution of these compounds to the allelopathic effect of M. spicatum on M. aeruginosa. We first collected information on the release rates of these compounds and then added the compounds to a cyanobacterial medium on the basis of their release rates so as to simulate their excretion by M. spicatum. Addition of the polyphenols and fatty acids inhibited the growth of M. aeruginosa, and the interaction of the polyphenols and fatty acids was additive. The EC50 of a polyphenol and fatty acid mixture was compared with that of M. spicatum itself as previously determined in a mixed culture system in which M. spicatum and M. aeruginosa were incubated. The former was about 1.9 times higher than that of the latter, the implication being that the inhibitory effect of the polyphenols and fatty acids contributed about 53% of the allelopathic effect of M. spicatum. This paper is the first to describe allelochemicals that account for a half of the anti-cyanobacterial allelopathic effect of a macrophyte.

Reduction of Alzheimer's disease amyloid-ß in plasma by hemodialysis and its relation to cognitive functions.

BACKGROUND/AIMS: Rapid removal of plasma amyloid-ß (Aß) by blood purification may serve as a peripheral Aß sink from the brain for Alzheimer's disease therapy. We investigated the reduction of plasma Aß during hemodialysis and cognitive states. METHODS: Aß concentrations and Mini-Mental State Examinations (MMSE) were investigated in 37 hemodialysis patients (68.9 ± 4.1 years). RESULTS: The dialyzers effectively removed Aß(1-40) and Aß(1-42), 63.9 ± 14.4 and 51.6 ± 17.0% at 4 h dialysis, resulting in the reduction of Aßs in whole-body circulation by 51.1 ± 8.9 and 32.7 ± 12.0%, respectively. Although the plasma Aßs before dialysis (750.8 ± 171.3 pg/ml for Aß(1-40)) were higher than those reported for Alzheimer's disease patients, the cognitive states of hemodialysis patients were relatively normal, especially of longer dialysis vintages. CONCLUSIONS: Dialyzers effectively reduced Aßs in whole-body circulation. Repeated rapid decrease of plasma Aßs might maintain cognitive state.

Interleukin 2 inhibits in vitro growth of human T cell lines carrying retrovirus.

Four human T cell lines, TL-Mor, TL-Su, TL-TerI, and TL-OmI, carrying human T cell leukemia virus (HTLV), were established previously. TL-Mor, TL-Su, and TL-TerI were derived from interleukin 2 (IL-2)-dependent parental cell lines cloned from peripheral blood leukocytes (PBL) of three healthy HTLV carriers, while TL-OmI was directly established from PBL of a patient with adult T cell leukemia. These four TL cell lines grow autonomously without IL-2. When they were cultured in the presence of IL-2, their growth was inhibited after a few days. This growth inhibition depended on the dose of IL-2, and the effective dose significantly promoted growth of their parental IL-2-dependent cell lines. The growth inhibition is demonstrated to be due to specific binding of IL-2 to receptors on the TL cells.

Altered cochlear fibrocytes in a mouse model of DFN3 nonsyndromic deafness.

DFN3, an X chromosome-linked nonsyndromic mixed deafness, is caused by mutations in the BRN-4 gene, which encodes a POU transcription factor. Brn-4-deficient mice were created and found to exhibit profound deafness. No gross morphological changes were observed in the conductive ossicles or cochlea, although there was a dramatic reduction in endocochlear potential. Electron microscopy revealed severe ultrastructural alterations in cochlear spiral ligament fibrocytes. The findings suggest that these fibrocytes, which are mesenchymal in origin and for which a role in potassium ion homeostasis has been postulated, may play a critical role in auditory function.

Higher survival rates of chronic hemodialysis patients on anti-hypertensive drugs.

BACKGROUND: The effects of anti-hypertensive drugs on survival have not been examined in a large cohort of hemodialysis (HD) patients. METHODS: We examined the relationship between blood pressure, anti-hypertensive drug therapy, and survival using the nationwide HD registry of the Japanese Society for Dialysis Therapy. Outcomes were confirmed using the coded ID numbers of the 2005 and 2006 registries. Logistic analyses were performed to determine the effect of anti-hypertensive drug therapy on survival. RESULTS: A total of 163,668 patients (50.6% men; 31.5% with diabetes mellitus; mean age 63.6 years) on HD 3 times a week in 2005 were studied. Mean (SD) levels of systolic and diastolic blood pressure were 153.4 (24.1) and 78.7 (13.7) mm Hg, respectively, before the HD session. Two-thirds of the HD patients were prescribed anti-hypertensive drugs and the numbers of anti-hypertensive medications were: 1 in 26.8%, 2 in 24.4%, and 3 or more in 14.5% of the total patients. The 1-year mortality rate was 6.6% overall: 8.5% in patients not prescribed anti-hypertensive drugs and 5.6% among those prescribed anti-hypertensive drugs. The odds ratio (95% confidence interval) for the 1-year mortality rate was 0.724 (0.681-0.770, p < 0.0001) for patients prescribed anti-hypertensive drugs, after adjusting for age, sex, diabetes mellitus, body mass index, HD duration, serum albumin, and systolic blood pressure. CONCLUSION: Survival was better in patients prescribed anti-hypertensive drugs, particularly renin-angiotensin system inhibitors, than in those not prescribed anti-hypertensive drugs. The causality on this association remained to be determined and prospective studies on blood pressure target levels and the effects of anti-hypertensive drug class in HD patients are warranted.

Evaluation of ethanol washing on dioxins-polluted soil and sediment based on adsorption relationships.

Multi-stage ethanol washing on dioxins-polluted soil and sediment were performed. The results indicated the existence of limit washing concentration (LWC), where no more dioxins were removed from the soil or the sediment by further washing. In each stage, dioxins concentration in the soil, sediment and ethanol could be described satisfactory by the Freundlich equation. The Freundlich capacity factor, K(ef) correlated with the LWC which was estimated to be ca. 1000 pmol g(-1) in the case of soil, and about 150 pmol g(-1) in the case of sediment. Organic contents in the soil and sediment affected the Freundlich intensity parameter, n(-1) but not K(f). A model, which enables the calculation of removal efficiency of PCDD/DFs at each stage using K(f), n, and initial PCDD/DFs concentration, is presented.

Kidney Transplantation, Cardiovascular Risk, and Long-Term Dialysis in Japan.

BACKGROUND: The waiting time for deceased-donor kidney-only transplantations in Japan is long. Herein, we assessed the effect of length of dialysis on the outcomes of these patients. METHODS: We divided patients into 2 groups based on length of dialysis (Group A, <15 years, and Group B, ≥15 years), and compared the background and outcomes after kidney transplantation. RESULTS: Group A included 210 patients and Group B included 35 patients. In Group B, 20% of transplants were from living donors. Patient age (P = .017) and the hepatitis C infection rate (P = .018) were significantly higher in Group B, whereas hypertension (P = .011), diabetes (P = .041), and ABO-incompatibility rates (P = .015) were significantly higher in Group A. The 5- and 10-year survival rates were 97.0% and 95.4%, respectively, in Group A and 97.1% and 97.1%, respectively, in Group B. The 5- and 10-year graft survival rates were 95.4% and 84.8%, respectively, in Group A and 97.1% and 73.1%, respectively, in Group B. There were no significant differences between the groups in patient survival (P = .74) and graft survival (P = .72). The 5- and 10-year cardiovascular event-free survival rates were 95.9% and 92.4%, respectively, in Group A and 88.6% and 76.8%, respectively, in Group B. Cardiovascular event-free survival was significantly higher in Group A (P = .038). Cox stepwise multivariate analysis indicated that length of dialysis was a significant predictor of cardiovascular events (hazard risk, 1.007; range, 1.001-1.012; P = .012). CONCLUSION: The prognosis after kidney transplantation is promising even after a long length of dialysis, although evaluation of the cardiovascular risk is needed in these cases.

Hydrophobicity determined by a fluorescence probe method and its correlation with surface properties of proteins.

The fluorescence method of Sklar et al. (Sklar, L.A., Hudson, B.S. and Simoni, R.D. (1977) Biochemistry 16, 5100-5108) using cis-parinaric acid as a probe was applied to determine the effective hydrophobicity of proteins. The initial slope (S0) of fluorescence intensity vs. protein concentration plot was used as an index of the protein hydrophobicity. A good correlation was observed for S0 of native proteins, denatured proteins and surfactant-bound proteins with an effective hydrophobicity determined by the hydrophobic partition method. The effective hydrophobicity determined fluorometrically showed significant correlations with interfacial tension and emulsifying activity of the proteins studied. The fluorescence technique using cis-parinaric acid is useful for determination of the effective hydrophobicity, as the procedure is much simpler and quicker than hydrophobic chromatography and hydrophobic partition.

The relationship between hydrophobicity and interfacial tension of proteins.

Charge-free hydrophobic gels of Hjerten et al. (Hjerten, S., Rosengren, J. and Pahlman, S. (1974) J. Chromatogr. 101, 281--288) were used for hydrophobic affinity chromatography. The effective hydrophobicity of proteins was expressed as their retention volumes from columns of butylepoxy- and hexylepoxy-Sepharose 4B. The effective hydrophobicity was also estimated by a partition method of Shanbhag and Axelsson ((1975) Eur. J. Biochem. 60, 17--22) from the partition coefficients of proteins between two phases, poly (ethylene glycol) and dextran. The former contained a hydrophobic ligand, palmitate. A close correlation was observed between the hydrophobicities determined by the two methods. However, no significant relationship was observed between these effective hydrophobicities and the average hydrophobicity of Bigelow ((1967) J. Theoret. Biol. 16, 187--211) that was calculated from the total amino acid composition of each protein. The interfacial tensions at the 0.2% protein/corn oil interface revealed negative correlations with the effective hydrophobicities determined by both methods indicating lower interfacial tensions with more hydrophobic proteins.

The Mfd protein of Bacillus subtilis 168 is involved in both transcription-coupled DNA repair and DNA recombination.

Inactivation of Bacillus subtilis orf1177 in an otherwise Rec+ strain reduced genetic exchange and DNA repair. When the mutation was transferred into a set of recombination-deficient and repair-deficient strains, the DNA repair and recombination ability of the double or triple mutant strains was drastically reduced. B. subtilis Orf1177 protein shares substantial homology with the Escherichia coli Mdf, RecG and UvrB proteins. In vivo analysis of UV-induced mutations suggests that Orf1177 is necessary for strand-specific DNA repair, as is the case for the E. coli MFD protein. Therefore, orf1177 and Orf1177 were termed mfd gene and Mfd protein, respectively. The purified Mfd protein has a native molecular mass of 140 kDa (expected molecular mass 133 kDa). The Mfd protein is a sequence-independent DNA binding protein with weak ATPase activity. The Mfd protein was able to displace in vitro B. subtilis or E. coli RNA polymerase stalled at a lesion. Therefore, Mfd protein appears to target the transcribed strand for repair by recognizing a stalled RNA polymerase and dissociating it from the DNA. In addition, the strong recombination-deficient phenotype of mfd- rec- strains suggest that Mfd protein is involved in homologous DNA recombination.

Induction of differentiation and enhancement of vincristine sensitivity of human erythroleukemia HEL cells by vesnarinone, a positive inotropic agent.

We examined the effect of vesnarinone, an oral cardiotonic, on the growth and differentiation of human myeloid leukemia cells. Vesnarinone alone markedly induced erythroid differentiation of HEL cells. All-trans-retinoic acid also induced erythroid differentiation of the cells, and the differentiation was greatly enhanced by combined treatment with vesnarinone and retinoic acid. HEL cells are highly resistant to some anticancer drugs, including vincristine, but treatment with vesnarinone greatly increased the sensitivity of HEL cells to vincristine. Enhancement of vincristine sensitivity by vesnarinone was not as significant for other leukemia cells. Expression of P-glycoprotein in HEL cells was effectively inhibited by vesnarinone, suggesting that the restoration of vincristine sensitivity is associated with decrease of P-glycoprotein expression in HEL cells. The plasma level of vesnarinone required to induce differentiation of leukemia cells is 30 micrograms/mL, which could be achieved with oral administration. These results suggest that vesnarinone should be useful in differentiation therapy for some types of myelogenous leukemia.

Systematic sequencing of the 180 kilobase region of the Bacillus subtilis chromosome containing the replication origin.

We have determined a 180 kb contiguous sequence in the replication origin region of the Bacillus subtilis chromosome. Open reading frames (ORF) in this region were unambiguously identified from the determined sequence, using criteria characteristic for the B. subtilis gene structure, i.e., starting with an ATG, GTG or TTG codon preceded by sequences complementary to the 3' end of the 16S rRNA. Four rRNA gene sets, 7 individual tRNA genes and 1 scRNA gene were identified, occupying 20 kb in total. In the remaining 160 kb region, 158 ORFs were identified, suggesting that 1 ORF is coded on average by 1 kb of DNA of the B. subtilis genome. Among the 158 ORFs, the functions of 48 ORFs were assigned and those of 11 ORFs are suggested through significant similarities to known proteins present in data banks. However, the functions of more than half of the ORFs (63%) remain to be determined.

Sequence analysis of the 36-kb region between gntZ and trnY genes of Bacillus subtilis genome.

Within the framework of an international Bacillus subtilis genome sequencing project, we have determined a 36-kb sequence covering the region between the gntZ and trnY genes. In addition to five genes sequenced and characterized previously, 27 putative protein coding sequences (open reading frame; ORF) were identified. A homology search for the newly identified ORFs revealed that six of them had similarities to known proteins. It is notable that new ORFs belonging to response-regulator aspartate phosphatase (Rap) and its regulator (Phr) families, and response regulator and sensory kinase families of two-component signal transduction systems have been identified. Furthermore, we found that some 180-bp non-coding sequence, that might be an remnant of an ancient IS element, is preserved in at least five loci of the B. subtilis genome.

Sequence analysis of the groESL-cotA region of the Bacillus subtilis genome, containing the restriction/modification system genes.

We have determined a 35-kb sequence of the groESL-gutR-cotA (45 degrees-52 degrees) region of the Bacillus subtilis genome. In addition to the groESL, gutRB and cotA genes reported previously, we have newly identified 24 ORFs including gutA and fruC genes, encoding glucitol permease and fructokinase, respectively. The inherent restriction/modification system genes, hsdMR and hsdMM, were mapped between groESL and gutRB, and we have identified two open reading frames (ORFs) encoding 5-methylcytosine forming DNA methyl transferase and an operon probably encoding a restriction enzyme complex. The unusual genome structure of few ORFs and lower GC content around the restriction/modification genes strongly suggests that the region originated from a bacteriophage integrated during evolution.

Integration of woodchuck hepatitis virus (WHV) DNA at two chromosomal sites (Vk and gag-like) in a hepatocellular carcinoma.

Integration of woodchuck hepatitis virus (WHV) DNA into the liver DNA of a woodchuck infected by the virus was investigated. Clonal viral integration was not detected three months before the appearance of four hepatocellular carcinomas (HCC). Integration of the viral DNA was detected in all four HCCs, of which one was chosen to determine the structure of the viral integration completely in a single tumor. The integration occurred in two sites. One part contains the viral DNA from the middle of the gene encoding surface antigen to two-thirds of the way through the gene encoding X protein (X) with no structural changes. The coding frame of the truncated X gene continues into the chromosomal sequence to make a possible fusion protein. The integration seems to have occurred by recombination within two direct repeats of the viral genome in one junction and by homologous recombination between viral DNA and chromosomal DNA in the other junction. The viral DNA is integrated into a spacer of the immunoglobulin L-chain Vk (IgVk) region without any chromosomal rearrangement accompanying the integration. The viral DNA at the second site has a complex structural rearrangement: part of the preS gene is duplicated and attached to the terminus of the gene encoding core antigen in a head-to-tail fashion, followed by three small fragments derived from other parts of the viral DNA. The integrated preS gene has its own 5' regulatory element and a coding frame consisting of the truncated preS gene, the other parts of viral DNA and the chromosomal sequence.(ABSTRACT TRUNCATED AT 250 WORDS)

Inverted terminal repeat sequence in the macronuclear DNA of Stylonychia pustulata.

The structure of macronuclear DNA of a hypotrichous ciliate, Stylonychia pustulata, was examined by both electron microscopy and nucleotide sequence analysis. The DNA in the macronucleus consists of small linear molecules with average length of about 3400 base pairs (bp). Most, if not all, of these DNA molecules have identical inverted terminal repeat sequence of 20 nucleotide residues. This sequence is 5'-CCCCAAAACCCC-AAAACCCC.

Identification of a specific receptor for interleukin-1 on rat bone marrow cells.

The interleukin-1 (IL-1) receptor on rat bone marrow cells was investigated using 125I-labeled IL-1 alpha and -1 beta. These radiolabeled ligands bound to the rat bone marrow cells in a specific and saturable manner with Kd values of 0.36 +/- 0.072 nM and 1.9 +/- 0.27 nM, respectively. In a competitive binding experiment, IL-1 alpha and -1 beta inhibited the binding of 125I-IL-1 alpha with Ki values of 0.35 +/- 0.041 nM and 2.9 +/- 1.0 nM. The binding of 125I-IL-1 beta was inhibited by IL-1 alpha and -1 beta with Ki values of 0.27 +/- 0.020 nM and 0.74 +/- 0.12 nM. In cross-linking experiments, 125I-IL-1 alpha was covalently incorporated into two proteins of 163 kDa and 63 kDa. These results suggested the presence of two binding proteins for IL-1 on the rat bone marrow cells.

Application of polymannosylated cystatin to surimi from roe-herring to prevent gel weakening.

A recombinant glycosylated cystatin with a polymannosyl chain was added to roe-herring surimi for preventing gel weakening due to autolysis during cooking. Proteolysis of myosin heavy chain in the surimi was effectively suppressed while cooking at 90 degrees C for 20 min after preincubation at 40 degrees C for 30 min. The glycosylation of cystatin improved the stability against heating as well as proteolysis by cathepsin D. This process markedly improved the texture of the cooked surimi gel with gel strength 2.5 times that of unglycosylated control cystatin.

Protective effect of lysozyme-galactomannan or lysozyme-palmitic acid conjugates against Edwardsiella tarda infection in carp, Cyprinus carpio L.

The protective effect of lysozyme-galactomannan or lysozyme-palmitic acid conjugates orally administered to carp, Cyprinus carpio L. was investigated using a virulent strain of Gram-negative Edwardsiella tarda isolated from an infected fish. Lysozyme-galactomannan conjugate was prepared through controlled Maillard reaction. Lysozyme-palmitic acid conjugate was prepared through base-catalyzed ester exchange using N-hydroxysuccinimide ester of palmitic acid. The conjugates provided substantial protection to carp infected with a Gram-negative bacteria fish pathogen E. tarda NG 8104. Lytic activities of lysozyme conjugates with galactomannan and palmitic acid were about 80 and 71% of native lysozyme using Micrococcus lysodeikticus as a substrate. Feeding with lysozyme conjugates, for 8 days, significantly enhanced fish protection against E. tarda infection. The survival rate was 30% for lysozyme-galactomannan conjugate treated fish and 20% for lysozyme-palmitic acid conjugate treated fish after 6 days cultivation while all control fish died within 3 days. On the other hand, a recovery rate of 40% after 6 days was observed in the fish group that were fed lysozyme-palmitic acid conjugate 3 and 2 h before and after E. tarda challenge, respectively, and for 6 consecutive days. The results of this work show the possibility of utilizing lysozyme conjugates with galactomannan or palmitic acid as a therapeutic for infection in fish.