High activity enzyme microcrystal multilayer films.
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Catalase↗
Biomedical subjects
W Jin
Publications and source records attributed to W Jin.
Requirement for transforming growth factor beta1 in controlling T cell apoptosis.
Transforming growth factor (TGF)-beta1, a potent immunoregulatory molecule, was found to control the life and death decisions of T lymphocytes. Both thymic and peripheral T cell apoptosis was increased in mice lacking TGF-beta1 (TGF-beta1(-/-)) compared with wild-type littermates. Engagement of the T cell receptor enhanced this aberrant T cell apoptosis, as did signaling through either the death receptor Fas or the tumor necrosis factor alpha receptor in peripheral T cells. Strikingly, TGF-beta was localized within the mitochondria of normal T cells, and the absence of TGF-beta1 resulted in disruption of mitochondrial membrane potential (Deltapsi(m)), which marks the point of no return in a cell condemned to die. This TGF-beta-dependent regulation of viability appears dissociable from the TGF-beta1 membrane receptor-Smad3 signaling pathway, but associated with a mitochondrial antiapoptotic protein Bcl-XL. Thus, TGF-beta1 may protect T cells at multiple sites in the death pathway, particularly by maintaining the essential integrity of mitochondria. These findings may have broad implications not only for T cell selection and death in immune responses and in the generation of tolerance, but also for defining the mechanisms of programmed cell death in general.
Animals↗
"Opening" the ferritin pore for iron release by mutation of conserved amino acids at interhelix and loop sites.
Ferritin concentrates, stores, and detoxifies iron in most organisms. The iron is a solid, ferric oxide mineral (< or =4500 Fe) inside the protein shell. Eight pores are formed by subunit trimers of the 24 subunit protein. A role for the protein in controlling reduction and dissolution of the iron mineral was suggested in preliminary experiments [Takagi et al. (1998) J. Biol. Chem. 273, 18685-18688] with a proline/leucine substitution near the pore. Localized pore disorder in frog L134P crystals coincided with enhanced iron exit, triggered by reduction. In this report, nine additional substitutions of conserved amino acids near L134 were studied for effects on iron release. Alterations of a conserved hydrophobic pair, a conserved ion pair, and a loop at the ferritin pores all increased iron exit (3-30-fold). Protein assembly was unchanged, except for a slight decrease in volume (measured by gel filtration); ferroxidase activity was still in the millisecond range, but a small decrease indicates slight alteration of the channel from the pore to the oxidation site. The sensitivity of reductive iron exit rates to changes in conserved residues near the ferritin pores, associated with localized unfolding, suggests that the structure around the ferritin pores is a target for regulated protein unfolding and iron release.
Amino Acid Motifs↗
Restriction fragment length dimorphism-PCR method for the detection of extended-spectrum beta-lactamases unrelated to TEM- and SHV-types.
The diagnostic ability of the restriction fragment length dimorphism-polymerase chain reaction (RFLD-PCR) method was evaluated. Seven primer pairs, newly designed from 44 beta-lactamase genes encoding extended-spectrum beta-lactamases not related to TEM- and SHV-types, were used to differentiate OXA-2, FOX-3, CMY-3, IMP-1, and IMI-1 beta-lactamases. The RFLD-PCR was carried out successfully, and these genes were differentiated by the sizes of their PCR products and by the difference in restriction fragment length when each amplicon was digested with a unique restriction enzyme. This discriminatory detection of the genes was confirmed by sequencing the PCR products.
Bacterial Proteins↗
Immunohistochemical localization of interleukin-2 and its receptor subunits alpha, beta and gamma in the main olfactory bulb of the rat.
Endogenous interleukin-2 (IL-2) was found in the adult rat brain, however, it has not been reported whether this cytokine is present in the olfactory bulb. Immunohistochemical techniques were used to examine the cellular localization of IL-2 and its receptor subunits in the main olfactory bulb of the rat. Strong IL-2 immunoreactivity was localized in glial cells, specifically in the olfactory nerve layer, glomerular layer and external plexiform layer. IL-2 mRNA was detected in the olfactory bulb by RT-PCR. All three IL-2 receptor subunits also showed distinct laminar distributions. The IL-2Ralpha and IL-2Rbeta immunoreactivity was found both in neurons and glial cells, whereas IL-2Rgamma imunoreactivity was found in glial cells, and thus resembled IL-2 immunostaining. The present results demonstrated a wide distribution of IL-2 and its receptor subunits in the main olfactory bulb of the rat, suggesting that IL-2 might play a role in the olfactory function through autocrine or paracrine pathways. The exclusive high expression of IL-2 in glial cells in distinct laminar structures, where neuron-glia interactions are closely associated with olfactory nerve regeneration, imply that IL-2 might be involved in the process of nerve regeneration in the olfactory bulb.
Animals↗
A novel reductive dimerization/oxidative dehydrogenation of aldimines mediated by lanthanoid metals.
A lanthanoid metal-mediated novel reductive dimerization/oxidative dehydrogenation of a variety of aldimines has been achieved. Aromatic aldimines (1) were dimerized in the presence of 0.5 mol of ytterbium metal (Yb) and 1-naphthaldehyde (1-NpCHO) to give the corresponding vicinal diimines (2) in good to high yields. Samarium metal (Sm) or samarium(II) diiodide (SmI2) gives unsatisfied yields under the same reaction conditions. As an oxidant, 1-NpCHO gives the best result. In addition, HMPA plays an important role as a cosolvent in this reaction. Reaction of various aldimines with Yb metal is discussed in detail. Moreover, it has been found that the electron-donating substitutents on the benzene ring promote the reaction and that the electron-withdrawing substitutents retard the reaction. The reaction of deuterated N-benzylideneaniline with Yb metal confirmed that transformation of the hydride occurred from aldimines to the oxidant aldehyde. The mechanism of the reaction was discussed.
Journal Article↗
Monitoring human serum transferrin by capillary zone electrophoresis with end-column amperometric detection.
Capillary zone electrophoresis was employed for the determination of human serum transferrin using end-column amperometric detection with a carbon fiber microelectrode at a constant potential of 1.9 V vs. saturated calomel electrode (SCE). The optimum conditions of separation and detection are 7.5 x 10(-4) mol/L Tris-3.44 x 10(-4) mol/L HCl for the buffer solution, 20 kV for the separation voltage, 5 kV and 10 s for the injection voltage and the injection time, respectively. The limit of detection is 6.7 x 10(-8) mol/L or 440 amol (S/N = 2). The relative standard deviations are 0.67% for the migration time and 1.5% for the electrophoretic peak current. The method was applied to the determination of transferrin in human serum. The recovery is between 93-104%.
Adult↗
Monitoring diclofenac sodium in single human erythrocytes introduced by electroporation using capillary zone electrophoresis with electrochemical detection.
A method for determination of the drug diclofenac sodium introduced into individual human erythrocytes by electroporation using capillary zone electrophoresis with electrochemical detection at a carbon fiber array microelectrode was developed. In this method, the whole cell was injected into the separation capillary by electromigration. Cell lysis was accomplished by injecting a plug of the separation buffer (1.25 x 10(-2) mol/L Na2B4O7-3.13 x 10(-3) mol/L NaOH). The optimum conditions of separation and detection were 20 kV for the separation voltage and 1.0 V for the detection potential. The concentration of diclofenac sodium in the single cells was quantified by a calibration curve. The mean concentration of diclofenac sodium introduced into the cell was 4.21 micromol/L. The relative standard deviation of the concentration of diclofenac sodium introduced into ten cells is 10%.
Anti-Inflammatory Agents, Non-Steroidal↗
Direct electrochemical determination of pyruvate in human sweat by capillary zone electrophoresis.
Capillary zone electrophoresis was employed for the determination of pyruvate in human sweat using electrochemical detection with a carbon fiber microdisk bundle electrode at a constant potential of 1.60 V vs. saturated calomel electrode. The optimum separation conditions are 3.6 x 10(-3) mol/L Na2HPO4-1.4 x 10(-3) mol/L NaH2PO4 (pH 7.2) for the buffer solution, and 18 kV for the separation voltage. The limits of detection of pyruvate are 8.0 x 10(-6) mol/L or 24 fmol (S/N = 3) for the injection voltage of 6 kV and the injection time of 10 s. The relative standard deviation is 2.0% for the migration time and 5.7% for the electrophoretic peak current. The method was applied to determining pyruvate in human sweat.
Electrochemistry↗
Determination of free intracellular amino acids in single mouse peritoneal macrophages after naphthalene-2,3-dicarboxaldehyde derivatization by capillary zone electrophoresis with electrochemical detection.
A method is described for the direct identification and quantification of amino acids in individual mouse peritoneal macrophages by capillary zone electrophoresis with electrochemical detection after on-column derivatization with naphthalene-2,3-dicarboxaldehyde (NDA) and CN-. In this method, individual macrophages and then the lysing/ derivatizing buffer are injected into the front end of the separation capillary by electromigration with the aid of an inverted microscope. The front end of the separation capillary is used as a chamber to lyse the macrophage and derivatize its contents, which minimizes dilution of amino acids of a single macrophage during derivatization. Six amino acids (serine, alanine, taurine, glycine, glutamic acid, and aspartic acid) in single mouse peritoneal macrophages have been identified. Quantitation has been accomplished through the use of calibration curves, where the concentration ratios of these standard amino acids are similar to the concentration ratios of amino acids in macrophages. Cellular levels of the amino acids in these cells range from 0.27 +/- 0.20 fmol/ cell for alanine to 6.4 +/- 4.6 fmol/cell for taurine.
Alanine↗
Subspeciation of nickel disulfide by carbon paste electrode voltammetry catalyzed by Ni2+.
In subspeciation of sulfidic nickel, carbon paste electrode voltammetry was developed for the specific determination of Ni3S2 or NiS, but NiS2 was found to be unreactive. Ni2+ in pH 7.2 acetate solution was able to catalyze NiS2 to undergo redox reactions. The cyclic voltammogram showed two anodic peaks at -0.3 and +0.7 V and a cathodic peak at -0.6 V. The first anodic peak at -0.3 V and the cathodic peak were common among the three nickel sulfides, but the second anodic peak at +0.7 V was unique to NiS2. This peak current gave a linear dose response to NiS2 from 40 to 610 microg, with a correlation coefficient of 0.994, and a detection limit of 40 microg.
Carbon↗
TGF-beta released by apoptotic T cells contributes to an immunosuppressive milieu.
T cell apoptosis is critical to development and homeostasis of the immune system. The most salient feature of apoptosis is the lack of an attendant inflammatory response or tissue damage. Here, we present evidence that apoptotic T cells release TGF-beta, thereby contributing to an immunosuppressive milieu. Apoptotic T cells released not only latent but also bio-active TGF-beta. Nonetheless, TGF-beta transcription was not upregulated, suggesting release of existing rather than synthesis of new TGF-beta. Localized within the intracellular membrane-bound compartment, which includes mitochondria, TGF-beta was redistributed into the cytosol following loss of mitochondrial membrane potential. TGF-beta secreted from apoptotic T cells inhibited proinflammatory cytokine production by activated macrophages to foster immune suppression. These findings broaden the potential mechanisms whereby induction of immune tolerance or deficiency occurs through T cell deletion.
Animals↗
The contributions of sex, genotype and age to transcriptional variance in Drosophila melanogaster.
Here we present a statistically rigorous approach to quantifying microarray expression data that allows the relative effects of multiple classes of treatment to be compared and incorporates analytical methods that are common to quantitative genetics. From the magnitude of gene effects and contributions of variance components, we find that gene expression in adult flies is affected most strongly by sex, less so by genotype and only weakly by age (for 1- and 6-wk flies); in addition, sex x genotype interactions may be present for as much as 10% of the Drosophila transcriptome. This interpretation is compromised to some extent by statistical issues relating to power and experimental design. Nevertheless, we show that changes in expression as small as 1.2-fold can be highly significant. Genotypic contributions to transcriptional variance may be of a similar magnitude to those relating to some quantitative phenotypes and should be considered when assessing the significance of experimental treatments.
Aging↗
A novel mechanism of matrix metalloproteinase-9 gene expression implies a role for keratinization.
To investigate the pathophysiological role of matrix metalloproteinase (MMP)-9 in the skin, we analyzed MMP-9 expression from human keratinocytes in culture. MMP-9 and the terminal differentiation marker involucrin were co-localized in the same keratinocytes with a high concentration of Ca(2+), a potent stimulator of differentiation. We identified the novel KRE-M9 element, further downstream to the previously reported TPA responsive element in the MMP-9 promoter, and both of these two elements were shown to be important for MMP-9 transcription and Ca(2+) induction. The concomitant upregulation of MMP-9 and involucrin transcripts was probably due to the very similar gene regulatory elements, KRE-M9 and KRE-4, in their respective promoters. These results indicate a novel mechanism of transcriptional regulation for MMP-9 in the process of keratinization, implying the probable association of apoptosis and differentiation of keratinocytes in epidermal skin tissue.
Calcium↗
Regulation of follicle-stimulating hormone secretion by estradiol and dimeric inhibins in the infantile female rat.
Plasma and ovarian levels of the dimeric forms of inhibin and plasma estradiol-17beta were investigated and compared with changes in plasma gonadotropins from Postnatal Day (PND) 5 to PND 30 in the female rat. The inhibin subunit proteins were localized in follicular granulosa cells of the ovary. Plasma immunoreactive inhibin levels were low until PND 15 and increased thereafter. Plasma levels of inhibin B (alpha and beta(B) subunits) remained very low until PND 15 and then increased by approximately 24-fold. In contrast, plasma levels of inhibin A (alpha and beta(A) subunits) were relatively low and steady until PND 20, then increased by approximately 3-fold at PND 25. Changes in ovarian inhibin A and B levels closely resembled those in plasma levels. Plasma FSH levels were low at PND 10 but started to peak from PND 15 and remained high until PND 20, followed by a remarkable reduction at PNDs 25 and 30. This dramatic fall in FSH coincided with the rise of inhibin A. A significant inverse correlation was observed between plasma FSH and plasma inhibin A (r = -0.67, P < 0.0002), ovarian inhibin A (r = -0.48, P < 0.01), plasma inhibin B (r = -0.48, P < 0.05), and ovarian inhibin B (r = -0.54, P < 0.01). Plasma estradiol-17beta levels were elevated from PND 5 through PND 15, then fell sharply through PND 30. Plasma estradiol-17beta was significantly and positively (r = 0.75, P < 0.0002) correlated with plasma FSH. Plasma LH rose to higher levels at PND 15 and tended to be lower thereafter. The inhibin alpha, beta(A), and beta(B) subunits were localized to primary, secondary, and antral and large antral follicles, but the types of these immunopositive follicles varied with age. It appeared that, at PND 25 and afterward, all three subunits were mainly confined to large antral follicles in the ovary. We conclude that estradiol-17beta likely is the major candidate in stimulation of FSH secretion in the infantile female rat. We also conclude that inhibin regulation of pituitary FSH secretion through its negative feedback in the infantile female rat begins to operate after PND 20. We suggest that this negative feedback is achieved by increases in plasma levels of the two dimeric forms, and that inhibin A appears to be the major physiological regulator of FSH secretion at the initiation of this mechanism. We also conclude that large antral follicles in the ovary are the primary source of these bioactive inhibins that are secreted in large amounts into the circulation after PND 20.
Aging↗
CT findings of portal vein aneurysm caused by gastric adenocarcinoma invading the portal vein.
The known causes of acquired origin portal vein aneurysm are portal hypertension, pancreatitis and trauma. We describe the CT findings of an additional cause of acquired origin portal vein aneurysm, namely gastric adenocarcinoma invading the portal venous system.
Adenocarcinoma↗
Comparison of tuberculous and pyogenic epididymal abscesses: clinical, gray-scale sonographic, and color Doppler sonographic features.
OBJECTIVE: The purpose of this study was to evaluate the capability of clinical, gray-scale sonographic, and color Doppler sonographic features for differentiating tuberculous and pyogenic epididymal abscesses. MATERIALS AND METHODS: Retrospective analysis was performed in 10 cases of tuberculous epididymal abscess and in 13 cases of pyogenic epididymal abscess. The following clinical, gray-scale sonographic, and color Doppler sonographic features were analyzed: patient's age; duration of symptoms; scrotal tenderness; presence of sinus tract; concurrent tuberculosis in other organs; location, size, and echogenicity of the abscess; hyperechoic rim; testicular involvement; hydrocele; and blood flow in the epididymal lesion. RESULTS: Tuberculous epididymal abscess had a longer duration of symptoms (p = 0.0001) and a lower frequency of scrotal tenderness (p = 0.0048) than pyogenic epididymal abscess. The size of the abscess was larger in tuberculous epididymal abscess than in pyogenic epididymal abscess (p = 0.0002). The degree of blood flow in the peripheral portion of the abscess was lower in tuberculous epididymal abscess (p = 0.001). The patient's age, location and echogeninicity of the abscess, presence of sinus tract, hyperechoic rim, testicular involvement, and hydrocele did not differ between the tuberculous and pyogenic epididymal abscesses. CONCLUSION: Some clinical findings, gray-scale sonography, and color Doppler sonography were useful in differentiating tuberculous epididymal abscess from pyogenic epididymal abscess. The presence of long-term scrotal swelling without tenderness and a lower degree of blood flow in the peripheral portion of a large abscess are suggestive of tuberculous epididymal abscess.
Abscess↗
Gene characterization of sciellin (SCEL) and protein localization in vertebrate epithelia displaying barrier properties.
Sciellin is a precursor of the cornified envelope of mammalian stratified epithelia characterized by a central core of nonidentical repeats. We characterized the genomic structure of human sciellin and showed that each homologous repeat was encoded by one exon. We also characterized mouse sciellin and showed that mouse sciellin and human sciellin (HGMW-approved symbol SCEL) share a similar gene organization and protein expression pattern. This one exon/one repeat organization is unique among other cornified envelope precursors characterized by homologous repeats. We identified an alternatively spliced isoform of human sciellin, absent in mouse, characterized by an additional repeat at the beginning of the core domain. During embryonic development, the accumulation of sciellin transcript and the accumulation of sciellin protein in the epidermis correlated with the activation of markers of terminal differentiation in epidermis. Mouse sciellin was also identified in simple epithelia with barrier properties, lending further support to its importance in epithelial function.
Alternative Splicing↗