Current understanding of the role of tyrosine kinase 2 signaling in immune responses

Immune system is a complex network that clears pathogens,toxic substrates,and cancer cells.Distinguishing self-antigens from non-self-antigens is critical for the immune cell-mediated response against foreign antigens.The innate immune system elicits an early-phase response to various stimuli,whereas the adaptive immune response is tailored to previously encountered antigens.During immune responses,B cells differentiate into antibody-secreting cells,while na?ve T cells differentiate into functionally specific effector cells[T helper 1(Th1),Th2,Th17,and regulatory T cells].However,enhanced or prolonged immune responses can result in autoimmune disorders,which are characterized by lymphocytemediated immune responses against self-antigens.Signal transduction of cytokines,which regulate the inflammatory cascades,is dependent on the members of the Janus family of protein kinases.Tyrosine kinase 2(Tyk2)is associated with receptor subunits of immune-related cytokines,such as type I interferon,interleukin(IL)-6,IL-10,IL-12,and IL-23.Clinical studies on the therapeutic effects and the underlying mechanisms of Tyk2 inhibitors in autoimmune or chronic inflammatory diseases are currently ongoing.This review summarizes the findings of studies examining the role of Tyk2 in immune and/or inflammatory responses using Tyk2-deficient cells and mice.     

Single amino acid substitutions in hydrophobic cores at a head-coiled coil junction region of cohesin facilitate its release of DNA during anaphase

Cohesin holds sister chromatids together and is cleaved by separase/Cut1 to release DNA during the transition from mitotic metaphase to anaphase. The cohesin complex consists of heterodimeric structural maintenance of chromosomes (SMC) subunits (Psm1 and Psm3), which possess a head and a hinge, separated by long coiled coils. Non-SMC subunits (Rad21, Psc3 and Mis4) bind to the SMC heads. Kleisin/Rad21''s N-terminal domain (Rad21-NTD) interacts with Psm3''s head-coiled coil junction (Psm3-HCJ). Spontaneous mutations that rescued the cleavage defects in temperature-sensitive (ts) separase mutants were identified in the interaction interface, but the underlying mechanism is yet to be understood. Here, we performed site-directed random mutagenesis to introduce single amino acid substitutions in Psm3-HCJ and Rad21-NTD, and then identified 300 mutations that rescued the cohesin-releasing defects in a separase ts mutant. Mutational analysis indicated that the amino acids involved in hydrophobic cores (which may be in close contact) in Psm3-HCJ and Rad21-NTD are hotspots, since 80 mutations (approx. 27%) were mapped in these locations. Properties of these substitutions indicate that they destabilize the interaction between the Psm3 head and Rad21-NTD. Thus, they may facilitate sister chromatid separation in a cleavage-independent way through cohesin structural re-arrangement.     

Economic assessment of batch biodiesel production processes using homogeneous and heterogeneous alkali catalysts

An economic feasibility study on four batch processes for the production of biodiesel ranging from 1452 tonnes/year (5000 l/day) to 14,520 tonnes/year (50,000 l/day) is conducted. The four processes assessed are the (1) KOH-W process, characterized by a homogeneous KOH catalyst and hot water purification process; (2) KOH-D process, characterized by a homogeneous KOH catalyst and vacuum FAME distillation process; (3) CaO-W process, characterized by a heterogeneous CaO catalyst and hot water purification process; and (4) CaO-D process, characterized by a heterogeneous CaO catalyst and vacuum FAME distillation process. The costs of the waste cooking oil, fixed costs, and manufacturing costs for producing 7260 tonnes/year (25,000 l/day) of biodiesel by means of the four processes are estimated to be $248–256, $194–232, and $584–641 per tonne of biodiesel, respectively. Among the four processes, the manufacturing costs involved in the CaO-W process are the lowest, in the range from 1452 tonnes/year to 14,520 tonnes/year.     

Ultrastructural changes and proliferation of pituicytes in mouse posterior lobe during water deprivation and rehydration

Ultrastructural changes and proliferation of pituicytes during water deprivation and rehydration were studied in the posterior lobe of C57BL/Tw mice. Deprivation for 3 days brought about a significant increase in the number of electron-dense bodies (lysosomes) in pituicyte perikarya and their processes. The frequency of pituicytes enclosing neurosecretory axons in their cytoplasm significantly decreased as compared with that of the controls. 12-hour rehydration following deprivation for 3 days induced extensive development of rough endoplasmic reticulum and Golgi apparatus and an increase in frequency of neurosecretory axons enwrapped by pituicyte cytoplasm. However, at 2 days of rehydration the morphology of pituicytes was no more different from that of the controls. Mitotic figures of pituicytes were not encountered throughout the deprivation period of 6 days, but rehydration for 12 h and 1, 2, or 3 days following deprivation for 3 or 6 days was effective in eliciting an increase in mitotic activity. The present results indicate that pituicytes in the mouse posterior lobe are intimately related with the secretory mechanism of neurosecretory material from the neurosecretory axons and that the proliferation of pituicytes is stimulated in conditions of reaccumulation of neurosecretory material.     

The small subunits of calcium dependent proteases with different calcium sensitivities are identical

The small subunits of two calcium dependent proteases from rabbit with different calcium sensitivities were isolated by high performance liquid chromatography (HPLC) and their properties were compared. The isolated subunits were indistinguishable on polyacrylamide gel electrophoresis and HPLC. Their amino acid compositions were identical, and the peptides obtained on their digestion with lysyl-endopeptidase showed identical peptide maps on HPLC. Furthermore, the amino acid compositions and partial amino acid sequences of the corresponding peptides purified by HPLC were the same. These results indicate that the two calcium protease isozymes possess the same small subunit.     

Acceptor proteins for (ADP-ribose)n in the HeLa S3 cell cycle

The acceptor proteins for (ADP-ribose)n were investigated by using nuclei or chromosomes isolated from specific phases of the cell cycle of HeLa S3 cells. Analysis of HMG proteins and histone H1 by acetic acid/urea polyacrylamide gel electrophoresis demonstrated that the (ADP-ribosyl)n-ation of HMG 14 and 17 and histone H1 increased by 12- and 5-fold, respectively, in the metaphase chromosomes as compared with that in the G1 phase cell nuclei. The degree of (ADP-ribosyl)n-ation of these proteins in the S phase cell nuclei was as low as that in G1 phase cell nuclei. In the G2 phase cell nuclei, the degrees of (ADP-ribosyl)n-ation of HMG 14 and 17 and histone H1 were about 5- and 2-fold greater, respectively, as compared with that in the G1 phase cell nuclei. The (ADP-ribosyl)n-ation of HMG 1 and 2 was constant through the cell cycle except for a slight decrease in the S phase. The data may imply that the (ADP-ribosyl)n-ation of HMG 14 and 17 and histone H1 is linked to chromatin structural changes in mitosis.     

Isolation and sequence analyses of cDNA clones for the large subunits of two isozymes of rabbit calcium-dependent protease

Two sets of cDNA clones were isolated from cDNA libraries prepared from poly(A+) RNA of rabbit lung and spleen by screening with the cDNA probe for the large subunit (80-kDa subunit) of chicken calcium-dependent protease (Ca2+-protease; Ohno, S., Emori, Y., Imajoh, S., Kawasaki, H., Kisaragi, M., and Suzuki, K. (1984) Nature 312, 566-570). The two sets of clones were identified as cDNA clones for two Ca2+-protease isozymes with high (mu-type) and low (m-type) calcium sensitivities from a comparison of the primary structures deduced from the nucleotide sequences with partial amino acid sequences from the two isozymes. The cDNA clones for the 80-kDa subunits of the mu- and m-type Ca2+-proteases contained, in total, about 1.5- and 2.2-kilobase cDNA inserts, respectively, which correspond roughly to the C-terminal halves of the coding regions and the entire 3'-noncoding regions. The two isozymes are encoded by two distinct mRNA species present in all the tissues examined, although the amount of mRNA significantly differs among the various tissues. Four E-F hand structures, typical calcium-binding structures in various calcium-binding proteins such as calmodulin, were detected in the C-terminal regions of both isozymes, as in the case of chicken Ca2+-protease. Comparison of the amino acid sequences of the two rabbit isozymes and the corresponding region of the chicken enzyme revealed marked homology, which indicates that these three enzymes have the same evolutionary origin. Furthermore, we suggest that the mu-type rabbit Ca2+-protease, rather than the m-type, is similar to chicken Ca2+-protease, which is regarded as an m-type enzyme in the C-terminal region. The evolution and molecular basis of the differences in calcium sensitivities of the Ca2+-proteases are discussed.     

Endogenous indole-3-acetic Acid in the stem of tobacco in relation to flower neoformation as measured by mass spectroscopic assay

The contents of free indole-3-acetic acid (IAA) and alkali-labile, conjugated IAA were measured in relation to a `floral gradient' present in epidermis and subepidermis tissues of flowering plants of Nicotiana tabacum by capillary gas-chromatographic spectrometric analysis by selected ion monitoring (GC-SIM-MS) using 2,4,5,6,7-penta deutero IA (²H₅-IAA) as an internal standard. In floral axes, floral branches and stems with floral branches, free IAA levels (dry weight) were 387, 253, and 417 nanograms, and bound IAA levels were 99, 1089, and 268 nanograms. In vegetative tissue of the first plus second internodes (measured from top), and of the 11th to 13th internodes, free IAA levels were 826 and 500 nanograms, and bound IAA levels were 1421 and 286 nanograms, respectively. Since flower-forming ability of excised cells from the epidermis and subepidermis shows a gradient in an in vitro system, but levels of IAA in these tissues do not, there thus appears to be no correlation between flower-forming ability (in vitro) and endogenous IAA levels (at the time of excision) in tobacco stem tissues.     

Purification and characterization of a calcium-activated neutral protease from rabbit skeletal muscle which requires calcium ions of microM order concentration

One form of calcium-activated neutral protease (CANP) highly sensitive to calcium ions was purified by column chromatographic procedures to homogeneity. The purified enzyme required microM order Ca2+ (mu CANP), and the half-maximum activity was attained at 50 microM Ca2+. The electrophoretic mobility in a non-denaturing buffer showed that this enzyme is less acidic than another CANP which required mM order Ca2+ (mCANP). On SDS-polyacrylamide gel electrophoresis, the enzyme separated into two components with molecular weights of 79,000 and 28,000, respectively. Of these, the former was slightly larger than the counterpart of mCANP (Mr 76,000). Thus, mu CANP cannot be derived from mCANP by limited autolysis.