The chemical synthesis of E. coli tRNA(Lys) anticodon loop fragment and its analogues

E. coli tRNA(Lys) anticodon loop fragment (Umnm5s2UUUt6A) 1 and its analogues 2-6 were synthesized by the classical phosphotriester approach in solution. The preparation of suitably protected derivatives of N6-threonylcarbamoyladenosine 18 is also described.     

Synthesis of willardiine and 6-azawillardiine analogs: pharmacological characterization on cloned homomeric human AMPA and kainate receptor subtypes

Both willardiine and azawillardiine analogs (18-28) have been reported to be potent and selective agonists for either AMPA or kainate receptors. We report here the novel synthesis and pharmacological characterization of a range of willardiine (18-23) and 6-azawillardiine (24-28) analogs on cells individually expressing human homomeric hGluR1, hGluR2, hGluR4, or hGluR5 receptors. Reaction of the sodium salts of substituted uracils (7-12) or 6-azauracils (13-16) with (S)-3-[(tert-butoxycarbonyl)amino]oxetan-2-one (17) in dry DMF, subsequent deprotection in TFA, and purification by ion-exchange chromatography gave mainly the willardiine analog in which alkylation took place on N1 of the uracil ring. We have investigated the subtype selectivity of these compounds by examining their binding affinity for homomeric hGluR1, -2, -4, or -5 (and hGluR6 in the case of 5-iodowillardiine (22)). From this study we have demonstrated that 22 has high affinity for hGluR5 and, compared to kainate, displays excellent selectivity for this receptor over both the AMPA receptor subtypes and the homomeric kainate receptor, hGluR6. 5-Fluorowillardiine (19) has higher affinity than AMPA for both homomeric hGluR1 and hGluR2 and compared to AMPA displays greater selectivity for AMPA receptor subtypes over the kainate receptor, hGluR5. Some structural features required for optimal activity at homomeric AMPA or kainate receptor subtypes have also been identified. It would appear that quite large lipophilic substituents at the 5-position of the uracil ring not only are accommodated by hGluR5 receptors but also lead to enhanced affinity for these receptors. In contrast to this, for optimal binding affinity to hGluR1, -2, or -4, smaller, electron-withdrawing substituents are required. For optimal activity at hGluR4 receptors a 6-aza-substituted willardiine is favored. The subtype-selective compounds described here are likely to be useful tools to probe the distribution and the physiological roles of the various glutamate receptor subunits in the central nervous system.     

Risperidone versus haloperidol: I. Meta-analysis of efficacy and safety

Stepwise solution syntheses are described of the homo-oligomers Z-(Thr)n-NHCH3 (n=1-4, I1-4), Z-?[Gal(Ac)4beta]Thr?n-NHCH3(n=1-5, II1-5) and Z-[(Galbeta)Thr]n-NHCH3 (n=1-5, III1-5). Members of the III1-5 series were obtained by de-acetylation of the corresponding oligomers of the II1-5 series. The conformational preferences of the terminally protected homo-peptides of the three series were investigated by FT-IR absorption spectroscopy both in the solid state and in CDCl3 solution, at various concentrations. Proton NMR measurements in CDCl3 and in DMSO-d6 were also carried out and the effect of temperature variation on the chemical shifts of amide protons was determined in DMSO-d6 (range 298-335 K) and in CDCl3 (range 298-320K). CD spectra were recorded in water and in TFE. Solubility problems prevented measurements in CDCl3 solution for Z-(Thr)4-NHCH3 and for the entire III1-5 series. The existence of unordered structures in the carbohydrate-free oligomers and of more or less extended, organized structures in the glycosylated derivatives is indicated by the NMR and IR measurements. The sugar moieties apparently show a structure-inducing effect on the peptide chain.     

Synthesis and in vitro antitumor activity of some amino-deoxy 7-hexofuranosylpyrrolo[2,3-d]pyrimidines

7-(6-amino-6-deoxy-beta-D-glucofuranosyl)-5-cyanopyrrolo[2,3 -d]pyrimidine (22) and 7-(3-amino-methyl-3-deoxy-beta-D-allofuranosyl)-5- cyanopyrrolo[2,3-d]pyrimidine (28) were synthesized by sequentially coupling silylated 4-amino-6-bromo-5- cyanopyrrolo[2,3-d]pyrimidine with the corresponding protected sugars 9 and 17, followed by deblocking and catalytic hydrogenation. Conversion of the 5-nitrile in 22 and 28 into a carboxamide gave the corresponding sangivamycin derivatives 23 and 29. Whereas 5'-aminomethyl nucleosides 22 and 23 inhibited the growth of four different human tumor cell lines at microM concentrations, the 3'-aminomethyl analogs 28 and 29 were much less active against these cells.     

Synthesis of 2"-oxidized derivatives of 5-deoxy-5-epi-5-fluoro-dibekacin and -arbekacin, and study on structure-chemical shift relationships of urethane(or amide)-type NH protons in synthetic intermediates

Three 2"-modified dibekacin-analogs have been prepared as potential compounds active against resistant bacteria producing 2"-O-phosphotransferases; one is 5-deoxy-5,2"-diepi-5-fluorodibekacin (9) prepared from a suitably protected 2"-O-triflyl derivative through the 2" 3"-cyclic carbamate, and the others are 2"-oxo derivatives (12 and 22, both as the hydrate) of 5-deoxy-5-epi-5-fluoro-dibekacin and -arbekacin prepared through oxidation at C-2" of suitably protected derivatives. Relationships between the t-butoxycarbonyl(= Boc)-NH-shifts of per-N-Boc synthetic intermediates and their structures were studied. It was found that the shifts, measured in pyridine-d5 at 80 degrees C, which spread over a close range (delta 6-7 ppm), are sensitively influenced by nearby and surrounding groups around the BocNH group in respect of electron-withdrawing character, hydrogen bonding (BocNH ... acceptor), and also solvent effects (BocNH ... NC5H5).     

Ketomethylenebestatin: synthesis and aminopeptidase inhibition

The synthesis of (6R,5S,2RS)-6-amino-5-hydroxy-2-isobutyl-4-oxo-7- phenylheptanoic acid (9), a carbaanalogue of the aminopeptidase (AP) inhibitor bestatin (1) is described. This synthesis was carried out by a malonic ester alkylation with the suitably protected halomethyl ketone of (2S,3R)-AHPBA*), followed by a second alkylation with isobutyl bromide of the resulting 4-ketodiester, and subsequent decarboxylation and deprotection. The inhibitory potencies of the 1:1 diastereomeric mixture 9 against AP-B, AP-M and Leu-AP were approximately 10-fold lower than those of bestatin.     

Synthesis and biological properties of 5-o-carboranyl-1-(2-deoxy-2-fluoro-beta-D-arabinofuranosyl)uracil

A novel 5-o-carboranyl-containing nucleoside, 5-o-carboranyl-1-(2-deoxy-2-fluoro-beta-D-arabinofuranosyl)uracil (6, CFAU), was synthesized as a potential intracellular neutron capture agent. This compound was prepared in five steps starting from 5-iodo-1-(2-deoxy-2-fluoro-beta-D-arabinofuranosyl)uracil (1). The desired carboranyl derivative was obtained by addition of decaborane [as the bis(propionitrile) adduct] to the protected acetylenic nucleoside precursor followed by debenzoylation. The synthesis of CFAU was also performed by glycosylation of the suitably protected 5-o-carboranyluracil with the appropriate 2-fluoroarabinosyl derivative. This compound was evaluated for its cytotoxicity in human lymphocytes, monkey cells, and rat and human gliomas cells, as well as for antiviral activity against human immunodeficiency virus and herpes simplex virus type 1. Its biological activity was compared to 5-o-carboranyl-1-(2-deoxyribofuranosyl)uracil in these cell culture systems, human bone marrow cells, and mice. The results obtained to date suggest that CFAU has suitable characteristics as a sensitizer for boron neutron capture therapy.     

Power of recent multiple comparison procedures as applied to a complete set of planned orthogonal contrasts.

Four recent multiple comparison strategies (Y. Hochberg, 1988; S. Holm, 1979; G. Hommel, 1988; and J. P. Shaffer, 1986), as well as a stagewise protected procedure proposed herein, were compared as tests of significance for a complete set of planned orthogonal contrasts. The Holm, Shaffer, and stagewise protected procedures were further modified using an adjustment suggested by B. S. Holland and M. D. Copenhaver (1987, 1988). The 2 F-protected methods and 3 unprotected methods all maintain experimentwise error rate at or below α. Simulation results indicate that the relative power of the 5 procedures depends on the configuration of the treatment effects contained in all contrasts. Specifically, the unprotected tests favor configurations with relatively small amounts of variability that are due to treatment effects, but the protected tests have more power in cases with a relatively large amount of treatment variability. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Supplemental protein plus ruminally protected methionine and lysine for primiparous beef cattle consuming annual rye hay

Primiparous beef cows (n = 35 Bos taurus, average initial BW of 498 kg) were allotted to treatments in a split-plot designed experiment to determine the effects of supplemental ruminally protected amino acids on cow and calf productivity and metabolic changes in the cows. Cows were fed chopped annual rye hay at 1.5% of BW. The following treatments were used: 1) .8 kg soybean hulls, 1.4 kg ground corn, .6 kg soybean meal (CON); 2) 1.4 kg ground corn, 1.4 kg soybean meal (PRO); 3) PRO plus ruminally protected methionine and lysine (supplied 5 and 10 g, respectively; PRO1); and 4) PRO with twice the level of ruminally protected amino acids in PRO1 (PRO2). Cow weight gain was not different (P > .26) among treatments and averaged 1.2 kg/d for the 45 + 6 d before parturition. After parturition, cow weight gain did not differ (P > .47) between CON and PRO treatments, but it decreased quadratically (P < .01) with increasing level of ruminally protected amino acids. Total milk yield, protein, and fat (4 h) were greater (P < .05) for cows consuming PRO supplements than for CON, whereas CON cattle tended (P = .11) to lose less body condition. Total milk protein showed a quadratic increase (P < .05) in response to level of ruminally protected amino acids that was the inverse of the quadratic response noted for cow weight gain. Serum urea N concentration was greater (P = .07) for cattle consuming additional protein. Metabolic hormones were not affected (P > .18) by dietary treatment, but they responded (P < .05) to changes in physiological state. Supplements with additional protein supported greater (P = .0001) milk urea N concentration and output. Milk urea N concentration increased (P < .05) and milk IGF-I decreased (P < .05) as the lactation period progressed. The measurement of CON and PRO diets revealed that supplements with additional soybean meal had greater (P < .05) DM and N degradation; the extent of forage DM and NDF degradation was similar (P > .05) among treatments. Production shifted away from body weight gain to increased milk protein production when daily supplementation of ruminally protected methionine and lysine increased from 5 and 10 to 10 and 20 g, respectively. This shift in production was not reflective of changes in the metabolic regulators measured.     

Inactivation of S-adenosyl-L-homocysteine hydrolase and antiviral activity with 5',5',6',6'-tetradehydro-6'-deoxy-6'-halohomoadenosine analogues (4'-haloacetylene analogues derived from adenosine)

Treatment of a protected 9-(5, 6-dideoxy-beta-D-ribo-hex-5-ynofuranosyl)adenine derivative with silver nitrate and N-iodosuccinimide (NIS) and deprotection gave the 6'-iodo acetylenic nucleoside analogue 3c. Halogenation of 3-O-benzoyl-5,6-dideoxy-1, 2-O-isopropylidene-alpha-D-ribo-hex-5-enofuranose gave 6-halo acetylenic sugars that were converted to anomeric 1,2-di-O-acetyl derivatives and coupled with 6-N-benzoyladenine. These intermediates were deprotected to give the 6'-chloro 3a, 6'-bromo 3b, and 6'-iodo 3c acetylenic nucleoside analogues. Iodo compound 3c appears to inactivate S-adenosyl-L-homocysteine hydrolase by a type I ("cofactor depletion") mechanism since complete reduction of enzyme-bound NAD+ to NADH was observed and no release of adenine or iodide ion was detected. In contrast, incubation of the enzyme with the chloro 3a or bromo 3b analogues resulted in release of Cl- or Br- and Ade, as well as partial reduction of E-NAD+ to E-NADH. Compounds 3a, 3b, and 3c were inhibitory to replication of vaccinia virus, vesicular stomatitis virus, parainfluenza-3 virus, and reovirus-1 (3a < 3b < 3c, in order of increasing activity). The antiviral effects appear to correlate with type I mechanism-based inhibition of S-adenosyl-L-homocysteine hydrolase. Mechanistic considerations are discussed.     

First total synthesis of ganglioside GT1a alpha

A first total synthesis of ganglioside GT1a alpha (IV3III6II3)Neu5AcGg4Cer) is described. The suitably protected sialyl alpha-(2-->6) ganglioside was glycosylated with the phenylthioglycoside of sialic acid in the presence of N-iodosuccinimide (NIS)-trimethylsilyl trifluoromethanesulfonate (TMSOTf), followed by further glycosylation with the methyl thioglycoside promoted by dimethyl(methylthio)sulfonium triflate (DMTST), to give the heptasaccharide. The oligosaccharide obtained was converted into the title ganglioside by the introduction of ceramide and then complete deprotection.     

Antioxidative action of diterpenoids from Podocarpus nagi

Diterpenoids, totarol (1), totaradiol (2), 19-hydroxytotarol (3), totaral (4), 4 beta-carboxy-19-nortotarol (5), sugiol (6), isolated from Podocarpus nagi, were evaluated as antioxidants. Microsomal lipid peroxidation induced by Fe(III)-ADP/NADPH and mitochondrial lipid peroxidation induced by Fe(III)-ADP/ NADH were inhibited by these terpenoids. They inhibited linoleic acid autoxidation but not generation of superoxide anion. Totarol (1) protected mitochondrial respiratory enzyme activities against NADPH induced oxidative injury. Totarane diterpenes from P. nagi were shown to be effective to protect biological systems and function against various oxidative stresses.     

When dreaming is believing: The (motivated) interpretation of dreams.

This research investigated laypeople's interpretation of their dreams. Participants from both Eastern and Western cultures believed that dreams contain hidden truths (Study 1) and considered dreams to provide more meaningful information about the world than similar waking thoughts (Studies 2 and 3). The meaningfulness attributed to specific dreams, however, was moderated by the extent to which the content of those dreams accorded with participants' preexisting beliefs--from the theories they endorsed to attitudes toward acquaintances, relationships with friends, and faith in God (Studies 3-6). Finally, dream content influenced judgment: Participants reported greater affection for a friend after considering a dream in which a friend protected rather than betrayed them (Study 5) and were equally reluctant to fly after dreaming or learning of a plane crash (Studies 2 and 3). Together, these results suggest that people engage in motivated interpretation of their dreams and that these interpretations impact their everyday lives. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Human immunodeficiency virus 1 (HIV-1)-specific reverse transcriptase (RT) inhibitors may suppress the replication of specific drug-resistant (E138K)RT HIV-1 mutants or select for highly resistant (Y181C-->C181I)RT HIV-1 mutants

Mutant HIV-1 that expresses a Glu138-->Lys substitution in its RT [(E138K)RT] is resistant to the HIV-1-specific RT inhibitor 2',5'-bis-O-(tert-butyldimethylsilyl)-3'-spiro-5"-(4"-amino-1",2"- oxathiole-2",2"-dioxide)pyrimidine (TSAO). However, cell cultures infected with this mutant were completely protected against virus-mediated destruction by micromolar concentrations of the HIV-1-specific RT inhibitors tetrahydroimidazo[4,5,1-jk][1,4]benzodiazepin-2(1H)-one and -thione (TIBO), nevirapine, and bis(heteroaryl)piperazine (BHAP). In contrast, cells infected with a virus mutant that expresses a Tyr181-->Cys substitution in its RT [(Y181C)RT] were not protected by nevirapine and TIBO and were only temporarily protected by BHAP. HIV-1 mutant that emerged under the latter conditions contained a Cys181-->Ile substitution in their RT [(LC181I)RT]. This mutant proved highly resistant to all HIV-1-specific RT inhibitors tested, except for several 1-(2-hydroxyethoxymethyl)-6-(phenylthio)thymine (HEPT) derivatives. When recombinant (C181I)RT was evaluated for susceptibility to the HIV-1-specific RT inhibitors, it was resistant to all inhibitors except the HEPT compounds. Since a (Y181F)RT HIV mutant strain was isolated from cells infected with (Y181C)RT HIV-1 and treated with BHAP, we postulate that the Ile codon was derived from a Cys-->Phe transversion mutation (TGT-->TTT), followed by a Phe-->Ile transversion mutation (TTT-->ATT).     

The marital repercussions of behavior therapy for agoraphobia.

Examined the effects of intensive behavior therapy on the husbands and marriages (mean length of 12.7 yrs) of 32 females with long-standing agoraphobia (mean length 9.5 yrs). Pre- and posttreatment assessments were made of Ss and their husbands to measure fear, hostility, experience, and husband's attitude. Further assessments were made at 3-, 6-, and 12-mo follow-up. In one pattern of marital interaction, couples had increased marital conflict and exposed problems relating to sex roles and wives' autonomy following rapid improvement in wives' phobias, assertiveness, and social confidence. 12 husbands developed psychological symptoms in relation to these problems, which also impeded wives' progress in the 1st 6 mo of follow-up. One year after treatment, marital systems had largely adapted, and both partners were more autonomous within the marriage. In the 2nd pattern of interaction, partners were protected from personal conflict in the 1st 6 mo followed by a mutual adherence to sex-role stereotypes. Husbands reported a marked decrease in psychological symptoms. In the last 6 mo of follow-up, conflict over sex-role stereotypes emerged, and the extent of the patient's continuing symptomatic improvement was determined mainly by the couples' success in resolving these conflicts. (21 ref) (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

单轨列车用镁合金与铝合金在NaCl溶液中的电偶腐蚀行为研究

采用浸泡失重、SEM、XRD等方法研究了AZ31镁合金与A6N01S-T5铝合金偶接后在3.5%NaCl溶液中电偶腐蚀行为。结果表明,浸泡腐蚀试验时间越长,偶对中AZ31镁合金、A6N01S-T5铝合金的腐蚀失重量越大,但是平均腐蚀速率则呈下降趋势。在偶对中AZ31镁合金作为阳极腐蚀加速,A6N01S-T5铝合金作为阴极,与镁合金接触的部分得到保护,而暴露在溶液中的部分发生了严重的阴极腐蚀。     

DNA-protein interaction sites in differentiating cells. I. Two-dimensional mapping of modulated sites

We have previously shown that DNA-protein attachment sites form during the induction of hematopoietic cell differentiation. Affinity phase-partitioning studies of DNA/protein complexes demonstrated that the DNA involved is not randomly distributed throughout the genome. The object of this study was to use filter binding followed by two-dimensional (2D) polyacrylamide gel electrophoresis (using a neutral 6% gel in the first dimension and a denaturing gradient gel in the second dimension) to gain insight into changes in DNA-protein interactions during induced granulocytic and monocytic differentiation of HL60 cells. Nitrocellulose filter-binding enriched samples for protein-associated DNA sufficiently to change the pattern of DNA spots on 2D gels. The patterns of spots obtained was reasonably reproducible between experiments and highly reproducible within experiments. Gels obtained from cells induced to differentiate by either phorbol ester or all-trans retinoic acid (RA) showed identical patterns for the majority of spots but changes in a small proportion of spots with respect to uninduced controls. Both intensification and reduction/disappearance of spots was observed, demonstrating the existence of both invariant and variant DNA/protein attachment sites during the early stages of hematopoietic cell differentiation. Previous studies have implicated DNA topoisomerase II in chromatin structural changes that are necessary for induction of granulocytic differentiation. We therefore examined the filter-binding DNA preparation by 5'-exonuclease digestion (since topoisomerase II is known to bind covalently to the 5'termini on either side of its cleavage sites). The filter-associated DNA exhibited increased 5' exonuclease protection (with respect to filter flow-through DNA), and the degree of protection increased significantly with exposure to phorbol ester and less markedly with retinoic acid. However, since not all filter DNA was 5' protected, it remains unresolved whether the specific differentiation-associated DNA-protein interactions revealed here involve DNA topoisomerase II or some other protein.     

Crowding stress and violent injuries among behaviorally inhibited rhesus macaques.

Environmental stressors and stable individual differences in human behavior have both been implicated etiologically in injuries. Because stress-related injuries are difficult to study experimentally in humans, the authors examined injury incidence in a troop of 21 male and 15 female free-ranging rhesus macaques (Macaca mulatta) before, during, and after a 6-month group confinement. Individual differences in behavioral inhibition to novelty were assessed, using multiple, quantified observations of behavior by 3 independent raters during 3 previous years. Incidence and severity of medically attended injuries were ascertained from veterinary records over a 2-year study period. A 5-fold increase in the incidence of injuries was documented during confinement stress, and an interaction was found between the stressor and behavioral inhibition in the prediction of injury incidence. Highly inhibited animals had significantly higher injury rates during confinement, compared with their uninhibited peers, but equal or lower rates in the low stress periods that preceded and followed confinement. Inhibited individuals appeared to have been specifically targeted for violence during the group stressor but were protected under normative, more predictable conditions. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Conditional immunomodulation following training with cyclophosphamide.

In 5 experiments, paired-group rats received a conditional stimulus (CS) paired with the immunosuppressive drug cyclophosphamide (CY). In Experiments 1–3, the CS was saccharin (SAC). Consistent with previous reports, these rats acquired a SAC aversion. However, there was no evidence of conditional immunosuppression. Rather, when reexposed to SAC in conjunction with an antigenic challenge, paired-group rats evidenced hemagglutination antibody titers similar to those seen in rats that never received the immunosuppressant. That is, the usual effect of CY in compromising immunological functioning was attenuated or eliminated by the CY-paired flavor. The findings of Experiments 1–3 were confirmed in Experiments 4–5, which used nongustatory CS. Both audiovisual (noise and flashing-light) and pharmacological (pentobarbital) cues were also effective signals for CY injection. Following pairing with CY, these cues protected animals from the immunosuppressive effects of the drug. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Charcot spinal arthropathy in congenital insensitivity to pain

Regioselective radioiodination of N-trifluoroacetyl 3,4-dimethoxy-6-trifluoroacetoxymercurio-L-phenylalanine ethyl ester 1 under no-carrier-added condition gave 6-[125I]iodo protected L-DOPA with a labeling efficiency of more than 85%, and no-carrier-added 6-[125I]I-L-DOPA was obtained with a radio-chemical purity of over 95% after hydrolysis and chromatography. A nonradioactive standard of 6-iodo protected L-DOPA was synthesized by the iododemercuration of 6-mercuric trifluoroacetate protected L-DOPA 1 using I2 in chloroform. 6-[125I]I-L-DOPA showed high brain accumulation and rapid blood clearance in mice. The rat brain slice studies indicated high affinity of 6-[125I]I-L-DOPA for carrier-mediated and stereoselective active transport systems. The tissue homogenate analysis revealed that most of the accumulated radioactivity was intact 6-[125I]I-L-DOPA. Thus, 6-[123I]I-L-DOPA appears to be a suitable single photon emission computed tomography (SPECT) tracer for the selective measurement of cerebral L-amino acid transport, having no affinity for dopamine metabolism.