Dendritic cell vaccination in human melanoma: relationships between clinical effects and vaccine parameters

Eleven years have passed since the start of the first trial of dendritic cell (DC) vaccination for melanoma. A review of 54 trials was performed to evaluate the relationship between clinical effects and vaccine parameters. Significant differences were found between use of immature and mature DCs with regard to progressive disease (PD), between stage III and IV for clinical response, between use and non-use of adjuvants with regard to stable disease (SD) in treatment with tumor/tumor lysate-pulsed DCs, between positive and negative delayed-type hypersensitivity (DTH) for PD, and between increased and unchanged interferon (IFN)-γ-secreting T cells for clinical response. These results are consistent with the partial efficacy of vaccination with mature DCs in early stage melanoma and the partial correlation of efficacy with positive DTH and increased IFN-γ-secreting T cells. DC vaccination alone had a limited clinical effect and a modified regimen is needed to enhance antigen-specific cytotoxic T cells and decrease immunosuppression.     

Effects of athletic strength and endurance exercise training in young humans on plasma endothelin-1 concentration and arterial distensibility

Strength exercise training induces a decrease in arterial distensibility, whereas endurance exercise training causes an increase in arterial distensibility. Endothelin-1 (ET-1), which is produced by vascular endothelial cells, has potent vasoconstrictor and proliferative activity on vascular smooth muscle cells. We hypothesized that endogenous ET-1 participates in alteration of arterial distensibility by different exercise training types (i.e., strength and endurance exercise training). The purpose of the present study was to investigate plasma ET-1 concentration and arterial distensibility in strength- and endurance-trained athletes. Subjects were male strength-trained athletes (discus, hammer, or javelin throwers; 22.2 years; SA), male endurance-trained athletes (long- or middle-distance runners; 20.7 years; EA), and sedentary healthy men (20.6 years; sedentary control, SC). Maximum hand-grip strength was markedly greater in SA compared with EA and SC (55.3 vs. 41.1 vs. 40.5 kg, P < 0.05). Maximum oxygen uptake was markedly greater in EA than in SA and SC (60.9 vs. 43.1 vs. 43.6 ml/kg/min, P < 0.05). Arterial pulse wave velocity (PWV), which is an index of arterial distensibility, was significantly higher in SA than in EA and SC (688 vs. 529 vs. 601 cm/sec, P < 0.05). In EA, PWV was significantly lower in comparison to that in SC (P < 0.05). Thus arterial distensibility was lower in SA than in EA and SC and higher in EA than in SC. Plasma ET-1 concentration was significantly higher in SA compared with EA and SC (1.64 vs. 1.12 vs. 1.24 pg/ml, P < 0.05). Plasma ET-1 concentration tended to be lower in EA than in SC. These results suggest that the difference in plasma ET-1 level may participate in the mechanism underlying different adaptation of arterial distensibility between strength- and endurance-trained athletes.     

The Plasmodium vivax homolog of the ookinete adhesive micronemal protein, CTRP

The Plasmodium circumsporozoite protein/thrombospondin-related anonymous protein-related protein (CTRP) is expressed at the mosquito midgut ookinete stage and is considered to be a transmission-blocking vaccine candidate. CTRP is composed of multiple von Willebrand factor A (vWA) and thrombospondin type 1 domains in the extracellular portion of the molecule, and a short acidic cytoplasmic domain that interacts with the actomyosin machinery. As a means to predict functionally relevant domains within CTRP we determined the nucleotide sequences of CTRP from the Plasmodium vivax Sall and the Plasmodium yoelii 17XL strains and characterized the conservation of domain architectures and motifs across Plasmodium genera. Sequence alignments indicate that the CTRP 1st to 4th vWA domains exhibit greater conservation, and thereby are perhaps functionally more important than the 5th and 6th domains. This point should be considered for the development of a transmission-blocking vaccine that includes CTRP recombinant subunit. To complement previous cellular studies on CTRP, we further determined the expression and cellular localization of CTRP protein in P. vivax and P. yoelii.     

Correlation between [U-14C]glucose metabolism and function in perfused cat brain

In brain perfusion experiments conducted with blood containing [U-14C]glucose the relative specific activity (RSA) of blood glucose carbon incorporated in brain intermediate metabolites was measured. It was demonstrated that the so-called metabolic pattern of Geiger is not constant, but it bears a close relation to the function of the brain. The results of the study may be summarized briefly as follows. (1) In a group (A) of cats with a high level of brain function, the RSA of lactic acid was 75 per cent; that of glutamic acid 80 per cent; aspartic acid 75 per cent; glutamine 61 per cent; GABA 43 per cent; and respiratory CO2 55 per cent. It was observed that the major part of the carbon of amino acids, such as glutamic acid and aspartic acid, which are directly associated with the tricarboxylic acid cycle are derived from blood glucose. (2) In a group (B) showing a low level of brain function, the RSA of each amino acid was considerably lowered. The RSA of glutamic acid and aspartic acid was about 50 per cent and that of respiratory CO2 was 27 per cent. (3) In a group (C) with a still lower level of brain function, each amino acid as well as the respiratory CO2 had still lower RSA values. (4) The metabolic pattern of Geiger corresponds to values obtained during low functional activity of the brain in our experiment.     

Synthesis of the C-terminal region of opioid receptor like 1 in an SDS micelle by the native chemical ligation: effect of thiol additive and SDS concentration on ligation efficiency.

In the process of developing a method for the synthesis of membrane proteins, the conditions for native chemical ligation, namely, detergent concentration and the chemical characteristics of the thiol additive were investigated in detail. The C-terminal region of the opioid receptor like 1, ORL1(288-370), which contains C-terminal intracellular and transmembrane domains, was chosen as a model. The building blocks, ORL1(329-370) and ORL1(288-328)-SR-Gly-Arg(5)-Leu (-SR- : -SCH(2)CH(2)CO-) were most effectively condensed slightly below the critical micelle concentration of SDS and in the presence of mercaptoethanesulfonic acid as a thiol additive. The results showed that the concentration of SDS and the charge on the thiol additive are crucial factors for the effective synthesis of a membrane protein by native chemical ligation.     

Mating reaction in Saccharomyces cerevisiae

A diffusible sex-specific substance called substance-I (S-I) was isolated from culture filtrate of type strains of the yeast Saccharomyces cerevisiae. The isolated S-I, an oligopeptide, induced sexual cell agglutinability in inducible a type strains and enhanced the agglutinability in constitutive a type strains. The induction of sexual agglutinability was detected in 30 min and reached maximum in 90 min, when 0.2 g/ml of S-I was added to inducible a type cells. The a type-specific factor responsible for sexual cell agglutination, called a type agglutination factor (aAF), was shown to be produced during the induction or the enhancement of agglutinability of a type cells by S-I. The aAF produced in response to S-I was not different in the susceptibility to proteolytic enzymes and disulfide-cleaving agents from those produced constitutively in the absence of S-I.     

Widely Targeted Metabolomics Based on Large-Scale MS/MS Data for Elucidating Metabolite Accumulation Patterns in Plants

Metabolomics is an ‘omics’ approach that aims toanalyze all metabolites in a biological sample comprehensively.The detailed metabolite profiling of thousands of plant sampleshas great potential for directly elucidating plant metabolicprocesses. However, both a comprehensive analysis and a highthroughput are difficult to achieve at the same time due tothe wide diversity of metabolites in plants. Here, we have establisheda novel and practical metabolomics methodology for quantifyinghundreds of targeted metabolites in a high-throughput manner.Multiple reaction monitoring (MRM) using tandem quadrupole massspectrometry (TQMS), which monitors both the specific precursorions and product ions of each metabolite, is a standard techniquein targeted metabolomics, as it enables high sensitivity, reproducibilityand a broad dynamic range. In this study, we optimized the MRMconditions for specific compounds by performing automated flowinjection analyses with TQMS. Based on a total of 61,920 spectrafor 860 authentic compounds, the MRM conditions of 497 compoundswere successfully optimized. These were applied to high-throughputautomated analysis of biological samples using TQMS coupledwith ultra performance liquid chromatography (UPLC). By thisanalysis, approximately 100 metabolites were quantified in eachof 14 plant accessions from Brassicaceae, Gramineae and Fabaceae.A hierarchical cluster analysis based on the metabolite accumulationpatterns clearly showed differences among the plant families,and family-specific metabolites could be predicted using a batch-learningself-organizing map analysis. Thus, the automated widely targetedmetabolomics approach established here should pave the way forlarge-scale metabolite profiling and comparative metabolomics.     

Carbonic anhydrase inhibitors: cloning and sulfonamide inhibition studies of a carboxyterminal truncated alpha-carbonic anhydrase from Helicobacter pylori

A library of sulfonamides/sulfamates has been investigated for the inhibition of the carboxyterminal truncated form of the alpha-carbonic anhydrase (CA, EC 4.2.1.1) isolated from the gastric pathogen Helicobacter pylori (hpCA). This enzyme, incorporating 202 amino acid residues, showed a catalytic activity similar to that of the full length hpCA, with k(cat) of 2.35 x 10(5)s(-1) and k(cat)/K(M) of 1.56 x 10(7)M(-1)s(-1) at 25 degrees C and pH of 8.9, for the CO(2) hydration reaction. All types of activity for inhibition of the bacterial enzyme have been detected. Dorzolamide and simple 4-substituted benzenesulfonamides were weak hpCA inhibitors (inhibition constants, K(I)s, in the range of 830-4310 nM). Sulfanilamide, orthanilamide, some of their derivatives, and indisulam showed better activity (K(I)s in the range of 310-562 nM), whereas most of the clinically used CA inhibitors, such as methazolamide, ethoxzolamide, dichlorophenamide, brinzolamide, topiramate, zonisamide, etc., acted as medium potency hpCA inhibitors (K(I)s in the range of 124-287 nM). Some potent hpCA inhibitors were detected too (K(I)s in the range of 20-96 nM) such as acetazolamide, 4-amino-6-chloro-1,3-benzenedisulfonamide, 4-sulfanilyl-aminoethyl-benzenesulfonamide, and 4-(2-amino-pyrimidin-4-yl)-benzenesulfonamide. Most of the investigated derivatives acted as better inhibitors of the human isoform hCA II than as hpCA inhibitors. Since hpCA is essential for the survival of the pathogen in acid, its inhibition by compounds such as those investigated here might be used as a new pharmacologic tool in the management of drug resistant H. pylori.