A novel ERK-dependent signaling process that regulates interleukin-2 expression in a late phase of T cell activation.

Engagement of the T cell antigen receptor (TCR) rapidly induces multiple signal transduction pathways, including ERK activation. Here, we report a critical role for ERK at a late stage of T cell activation. Inhibition of the ERK pathway 2-6 h after the start of TCR stimulation significantly impaired interleukin-2 (IL-2) production, whereas the same treatment during the first 2 h had no effect.

[A comparison between physicians' interpretation and a CAD system's cancer detection by using a mammogram database in a physicians' self-learning course].

We have been developing automated detection algorithms for masses and clustered microcalcifications in a mammography computer-aided diagnosis (CAD) system. In this study, we investigated the potential of our CAD system by comparing 579 physicians' interpretation results with that of the CAD system's cancer detection for 100 mammograms (21 malignant and 29 benign cases) employed in a physicians' self-learning course. As a result, our CAD system detected 7 out of 8 malignant lesions whose physicians' averaged sensitivity was less than 60%.

Quantification of the isomerization of Asp residue in recombinant human alpha A-crystallin by reversed-phase HPLC.

A method for determining the isomerization of Asp residues in proteins is described and demonstrated by quantifying the isomerization of Asp(151) in recombinant human alphaA-crystallin. First, four types of dodecapeptide fragment ((146)IQTGLD(151)ATHAER(157)) in which the Asp residue was either L-Asp, D-Asp, L-isoAsp or D-isoAsp were synthesized, and RP-HPLC conditions were established for their separation. Next, the Asp(151)-containing peptide fragments isolated from the tryptic hydrolysate of recombinant alphaA-crystallin were analyzed under these conditions.

Detection and monitoring of ongoing aseismic slip in the Tokai region, central Japan.

Analysis of global positioning system data shows that the rate of crustal deformations in the Tokai region of Japan, a seismic gap area, changed over the past 18 months. Kalman filtering analysis shows aseismic slip on the plate boundary in the western Tokai region centered on Lake Hamana, adjacent to the anticipated Tokai earthquake source area. The cumulative moment magnitude reaches 6.

Isolation of cathepsin B inhibitory peptides, Cabin-A1 and -A2, from a tryptic and chymotryptic hydrolysate of human serum albumin.

Two novel peptides that inhibit cathepsin B were isolated from a tryptic and chymotryptic hydrolysate of human serum albumin, and designated as Cabin-A1 and -A2. Cabin-A1 and -A2 were purified by reversed-phase HPLC and identified as Ser-Leu-His-Thr-Leu-Phe and Phe-Gln-Asn-Ala-Leu, respectively. These peptides correspond to f(65-70) and f(403-407) of human serum albumin.

Up-regulation of CD13/aminopeptidase N induced by phorbol ester is involved in redox regulation and tumor necrosis factor alpha production in HL-60 cells.

Here, we demonstrate the possible involvement of oxidative stress in altered CD13/aminopeptidase N (APN) expression during myeloid cell differentiation induced by TPA. In flow cytometrical analysis, CD13/APN protein was constitutively expressed in HL-60 cells. When the cells were treated with TPA, CD13/APN expression was up-regulated with increased intracellular peroxides and a morphological change into macrophage-like cells.

Synthesis of tag introducible (3-trifluoromethyl)phenyldiazirine based photoreactive phenylalanine.

An efficient synthesis of tag introducible (3-trifluoromethyl)phenyldiazirine based phenylalanine derivatives is described. Alkylation of a chiral glycine equivalent with a spacer containing (3-trifluoromethyl)phenyldiazirinyl bromides enables us to create photoreactive L-phenylalanine derivatives. After introduction of biotin at the spacer, the biotinylated and photoreactive amino acid was applied for L-amino acid oxidase and incorporated into a substrate binding site.

Isoserine-based biotinylated photoaffinity probes that interact with penicillin-binding protein 1b.

The photolytic decomposition of trifunctional carbene generating photoaffinity probes in methanolic solution was studied, a cleavage reaction with butylamine in water, the conjugation with a ligand (moenomycin), and experiments that demonstrate that the fully armed probes interact with penicillin-binding protein 1b.

N-acetyl-L-cysteine suppresses constitutive expression of CD11a/LFA-1alpha protein in myeloid lineage.

We investigated the possible involvement of redox regulation in constitutive expression of CD11a/LFA-1alpha, a leukocyte integrin alpha subunit, in myeloid cells using antioxidants. In unstimulated HL-60 cells, CD11a/LFA-1alpha was highly expressed, however, no expression of CD11b and CD11c proteins was detected. N-acetyl-L-cysteine (NAC) markedly down-regulated CD11a/LFA-1alpha expression in a dose-dependent manner.

Improvement in the properties of 3-phenyl-3-trifluoromethyldiazirine based photoreactive bis-glucose probes for GLUT4 following substitution on the phenyl ring.

We have developed two novel 3-phenyl-3-trifluoromethyldiazirinyl bis-glucose derivatives to investigate the properties of the adipocyte glucose transporter GLUT4. These compounds were substituted by electron-withdrawing (iodo and nitro) groups on the aromatic ring of 3-phenyl-3-trifluoromethyldiazirine photophore and were found to be more photosensitive than compounds without such substituents. The compounds were used as inhibitors of insulin-stimulated glucose transport activity in order to assess half-maximal inhibition or relative affinity values for GLUT4.

Protein domain of chicken alpha(1)-acid glycoprotein is responsible for chiral recognition.

Ovoglycoprotein from chicken egg whites (OGCHI) has been used as a chiral selector to separate drug enantiomers. However, neither the amino acid sequence of OGCHI nor the responsible part for the chiral recognition (protein domain or sugar moiety) has yet to be determined. First, we isolated a cDNA clone encoding OGCHI, and clarified the amino acid sequence of OGCHI, which consists of 203 amino acids including a predictable signal peptide of 20 amino acids.

An epidemiological study on relationship between the hours of sleep and life style factors in Japanese factory workers.

To prevent "life style-related diseases", it is necessary to evaluate not only the factors directly related to sleep but also the relationship between sleep and other life style-related factors (such as smoking, alcohol drinking, food habits, and exercise routines). There have been no extensive studies conducted on these relationships. A survey was conducted on 2,000 employees of a large plant over a 6-year period to provide data that would allow one to analyze correlation between hours of sleep and other life style factors, such as smoking, alcohol drinking, dietary habit, and exercise.

Filamin A-interacting protein (FILIP) regulates cortical cell migration out of the ventricular zone.

Precisely regulated radial migration out of the ventricular zone is essential for corticogenesis. Here, we identify a mechanism that can tether ventricular zone cells in situ. FILIP interacts with Filamin A, an indispensable actin-binding protein that is required for cell motility, and induces its degradation in COS-7 cells.

Isolation of novel peptides, cabin-1, -2, -3, and -4, that inhibit cathepsin B from a thermolysin digest of human plasma.

Four novel peptides that inhibit cathepsin B, designated as Cabin-1, -2, -3, and -4, were isolated from a thermolysin digest of human plasma. After gel filtration and cation-exchange chromatography, the peptide mixture was purified by reverse-phase HPLC to isolate Cabin-1, -2, -3, and 4, with the amino acid sequences LGPVTQE, VLQSSGLYS, VVSVLT, and LVYDAY, respectively. These peptides correspond to f(64-70) of human apolipoprotein A-I for Cabin-1, f(56-64) and f(185-190) of the human immunoglobulin G gamma chain for Cabin-2 and -3, and f(66-71) of human transferrin for Cabin-4.

Photoaffinity labeling in drug discovery and developments: chemical gateway for entering proteomic frontier.

One of the major events occurring at biological interfaces is the specific recognition of bioactive ligands by their receptor proteins. The elucidation of interacting partners is an immediate entrance into the discovery of medicinal leads. The method of photoaffinity labeling enables the direct probing of target protein through a covalent bond introduced between a ligand and its specific receptor.

Genetic evidence for Shc requirement in TCR-induced c-Rel nuclear translocation and IL-2 expression.

Shc, a prototypic adapter molecule, has been implicated in T cell receptor (TCR) signal transduction, but its role has not been identified clearly. Here we report that Shc is essential for TCR-induced IL-2 production but is dispensable for CD69 or CD25 expression. Engagement of TCR in mutant Jurkat T cells lacking Shc fails to produce IL-2 because of impaired mitogen-activated protein kinase activation.

Increased plasma levels of the soluble form of Fas ligand in patients with acute myocardial infarction and unstable angina pectoris.

Objectives: To examine whether the Fas/Fas ligand system is involved in the pathogenesis of acute myocardial infarction (AMI), we measured the levels of the soluble form of the Fas ligand (sFasL) in the plasma of patients with AMI and stable or unstable angina pectoris (AP).

Background: The Fas ligand (FasL) is rapidly cleaved off by a metalloproteinase from the cell membrane to become a soluble form as a cytokine. Fas is expressed in most cells, including cardiomyocytes, whereas FasL is mainly expressed in inflammatory cells such as macrophages, which are greatly accumulated in unstable plaque.

Epidemiological study on improving the QOL and oral conditions of the aged--Part 2: Relationship between tooth loss and lifestyle factors for adults men.

Oral health in early- and mid-adulthood is essential for the improvement of one's QOL, this study was investigated to include an epidemiological analysis of the relationship between tooth loss and life style, such as smoking, regular exercise, and the food habits of approximately 2,000 employees. Compared with the group with mild or no periodontal disease (CPI of 0, 1, or 2), the frequency of tooth loss in the group with advanced periodontal disease (CPI of 4) was 2.00 times (odds ratio, 2.

Epidemiological study on improving the QOL and oral conditions of the aged--Part 1: The relationship between the status of tooth preservation and QOL.

In part 1 of this epidemiological study, a survey was conducted for all senior citizens aged 70 and over who resided in a mountainous village in the mid-section of Hyogo Prefecture. It focused on the relationship among the number of existing teeth, life environment, health status, and activities of daily living; and the correlation between oral status and QOL was analyzed. The daily activities of individuals were compared between those having one or more teeth and others who were totally edentulous.

Development of an automated method for detecting mammographic masses with a partial loss of region.

Recently, we have been developing several automated algorithms for detecting masses on mammograms. For our algorithm, we devised an adaptive thresholding technique for detecting masses, but our system failed to detect masses with a partial loss of region that were located on the edge of the film. This is a common issue in all of the algorithms developed so far by other groups.

Decreased expression of Bcl-x protein during hepatocarcinogenesis induced exogenously and endogenously in rats.

Dysregulations of apoptosis have been widely recognized as important events in multi-stage carcinogenesis. Bcl-x, a member of the Bcl-2 family, is known to act as a regulator of apoptosis. The present study was conducted to assess the role of altered Bcl-x protein expression in exogenous and endogenous hepatocarcinogenesis in rats.

Versatile synthesis of phenoxydiazirine-based fatty acid analogues and photoreactive galactosylceramide.

A versatile synthesis of diazirine-based photoreactive fatty acid analogues is reported. The key step is phenoxy alkylation of diazirine with halo alkyl acid esters. The conditions described will be acceptable for the synthesis of various alkyl-length derivatives.