Expression of macrophage migration inhibitory factor in corneal wound healing in rats.

Purpose: The study was conducted to evaluate the expression of macrophage migration inhibitory factor (MIF) during penetrating corneal injury.

Method: A penetrating linear incision (2 mm) was made in the center of the right cornea with a razor blade. The expression of MIF in the lacerated eye and in the contralateral eye was examined by immunohistochemistry at 3, 6, 24, 48, and 72 hours after injury.

Impaired contractile response to beta adrenoceptor stimulation in diabetic rat hearts: alterations in beta adrenoceptors-G protein-adenylate cyclase system and phospholamban phosphorylation.

The aim of this study was to explore the cellular mechanisms underlying the impaired contractile response to beta adrenoceptor stimulation in diabetic hearts. Chronic diabetes was induced in rats by a streptozotocin injection. Four to six weeks later, papillary muscles isolated from diabetic hearts exhibited marked reductions in the positive inotropic responses to isoproterenol, norepinephrine and epinephrine.

Expression of macrophage migration inhibitory factor in rat retina and its immunohistochemical localization.

Macrophage migration inhibitory factor (MIF) plays an important regulatory role for the T-cell activation induced by mitogenic or antigenic stimuli. We examined expression of MIF in rat retina. Reverse transcription-polymerase chain reaction analysis of a retinal tissue homogenate revealed that MIF mRNA was constitutively expressed.

Identification of macrophage migration inhibitory factor in adipose tissue and its induction by tumor necrosis factor-alpha.

Macrophage migration inhibitory factor (MIF) has been rediscovered as a proinflammatory cytokine, pituitary hormone, and glucocorticoid-induced immunoregulator. A survey of tissue distribution revealed that MIF expression is not limited to T lymphocytes, but exists in several other tissues; however, its presence in adipose tissue has never been investigated. In this study, we examined the expression of MIF in adipose tissue using the rat epididymal fat pad and murine 3T3-L1 adipocytes.

Macrophage migration inhibitory factor in the human ovary: presence in the follicular fluids and production by granulosa cells.

Cytokines play an important role in ovarian function. We unexpectedly found high expression of macrophage migration inhibitory factor (MIF) mRNA in human ovarian tissues. Hence, we examined the presence of MIF in the follicular fluid because the follicular microenvironment is important for oocyte fecundity.

Theoretical study on the interaction between dopamine and its receptor by ab initio molecular orbital calculation.

Dopamine has been implicated in the function of a diverse set of central nervous system and peripheral functions. We theoretically evaluated the chemistry of interaction between dopamine and its receptor using ab initio molecular orbital calculation. First, we calculated the total energy of dopamine on either a protonated or deprotonated molecule at the meta- or para-position hydroxy group of the catechol ring, and then evaluated the hydrogen bond effect in these hydroxy groups.

Escherichia coli positive regulator OmpR has a large loop structure at the putative RNA polymerase interaction site.

The C-terminal DNA-binding domain of OmpR, a positive regulator involved in osmoregulation expression of the ompF and ompC genes in Escherichia coli, has a helix-turn-helix variant motif. The 'turn' region, consisting of 11 residues, forms an RNA polymerase contact site.

Identification and immunohistochemical localization of macrophage migration inhibitory factor in human kidney.

Macrophage migration inhibitory factor (MIF) was the first lymphokine identified in activated T-lymphocytes. MIF can induce proinflammatory cytokines, such as interleukin-1 and tumor necrosis factor-alpha. In this study, we identified MIF expression in a tissue specimen of a normal portion of a nephrectomized human kidney by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot analysis.

Effect of vitamin E on expression of cyclooxygenase-2 in lipopolysaccharide-stimulated rat macrophages.

To clarify the role of vitamin E (alpha-tocopherol) for the induction of cyclooxygenase-2 (COX-2) in rat macrophages stimulated by lipopolysaccharide (LPS), vitamin E-enriched macrophages were prepared by intraperitoneal injection of vitamin E for 6 days at a rate of 5 mg per day. The production of PGE2 was increased in dose- and time-dependent manners by addition of LPS in both control and vitamin E-enriched peritoneal macrophages. The maximum effect of LPS was observed in 12 h at concentration of 5 micrograms/ml.

Identification of macrophage migration inhibitory factor in murine neonatal calvariae and osteoblasts.

Bone resorption and formation are dynamic processes that occur in both normal and injured bone tissues. Regulation of these processes is mediated at the local level by cytokines and growth factors. Macrophage migration inhibitory factor (MIF) is one of the proinflammatory cytokines that activates macrophages and regulates production of other cytokines, such as tumour necrosis factor-alpha and interleukin-1.

Identification of macrophage migration inhibitory factor in human leukemia HL-60 cells and its induction by lipopolysaccharide.

Cytokines play an important role in inflammation and immunity. In this study, we examined the expression and presence of human macrophage migration inhibitory factor (MIF) in human myelomonocytic leukemia cell line, HL-60 by reverse transcription-polymerase chain reaction (RT-PCR) and immunological methods (Western blot analysis and immunohistochemistry), respectively. The RT-PCR showed that MIF mRNA was constitutively expressed, and the expression was further induced by lipopolysaccharide (LPS) stimulation.

Detection and immunolocalization of macrophage migration inhibitory factor in rat iris and ciliary epithelium.

To elucidate the role of macrophage migration inhibitory factor (MIF) in ocular inflammation, we examined the localization of MIF in the normal anterior uveal tract of rats. Immunohistochemistry using an anti-MIF antibody revealed that MIF was present in non-pigment epithelial cells of the ciliary body and the epithelial cells of the iris. Western blot analysis of these tissues showed a single band specific for MIF protein.

Evidence for the presence of macrophage migration inhibitory factor in murine reproductive organs and early embryos.

Cytokines appear to play a critical role in the establishment of early pregnancy. In this study, we examined the mRNA expression of murine macrophage migration inhibitory factor (MIF), one of the cytokines activating macrophages, in the murine reproductive system by Northern blot analysis. It revealed that MIF mRNA was expressed in the ovary, the oviduct and the uterus during the pre-implantation period and all stages of the estrus cycle.

Crystal structure of macrophage migration inhibitory factor from human lymphocyte at 2.1 A resolution.

The three-dimensional structure of the macrophage migration inhibitory factor (MIF) from human lymphocytes has been determined by X-ray crystallography at 2.1 A resolution. The structure was solved by a molecular replacement technique using the coordinates of rat MIF.

Identification of macrophage migration inhibitory factor in synovial membranes of loosened total joint replacement.

Endosteal bone resorption often occurs around loosened total joint replacements. In the process of the loosening, macrophages play an important role by releasing cytokines such as interleukin-1, tumor necrosis factor-alpha and prostaglandin E2. In this study, we investigated the involvement of macrophage migration inhibitory factor (MIF) in the pathological state of the loosening of a total hip replacement.

Identification and immunohistochemical localization of macrophage migration inhibitory factor in human cornea.

We identified macrophage migration inhibitory factor (MIF) mRNA expression in human cornea, and demonstrated its immunohistological localization. Reverse transcription-polymerase chain reaction analysis revealed that MIF mRNA was expressed in both the corneal epithelial and endothelial cells. Immunohistochemical study using the polyclonal antibody prepared from immunizing a rabbit with human recombinant MIF showed that MIF was present in the basal cells of corneal epithelium and endothelial cells.

Degradation of T-kininogen by cathepsin D and matrix metalloproteinases.

Cathepsin D, matrix metalloproteinase (MMP)-2, MMP-3 (stromelysin), and MMP-9 were isolated from rat granulomatous tissues. HT1080 human fibrosarcoma cells and rheumatoid synovial cell CM. At acidic conditions, cathepsin D cleaved T-kininogen into small peptides and released Met-T-kinin-Leu (kinin precursor), but failed to release kinin.

Identification of macrophage migration inhibitory factor (MIF) in human skin and its immmunohistochemical localization.

The presence and tissue localization of macrophage migration inhibitory factor (MIF) in human skin were examined. Reverse transcription-polymerase chain reaction analysis revealed that MIF mRNA was expressed in both surgically obtained normal human epidermis and primary cultured human keratinocytes. The expression of MIF was further confirmed by Western blot analysis, which demonstrated a single band at about 12.

Crystal structure of the macrophage migration inhibitory factor from rat liver.

The tertiary structure of the macrophage migration inhibitory factor (MIF) from rat liver (12,300 Mr) is presented at 2.2 A resolution. Each monomer consists of two beta/alpha/beta motifs aligned in quasi two-fold symmetry, comprising a domain consisting of a four-stranded mixed beta-sheet and two antiparallel alpha-helices.

The role of macrophage migration inhibitory factor in pregnancy and development of murine embryos.

Cytokines appear to play a critical role in the establishment of pregnancy. In this study, we measured serum and amniotic fluid concentrations of macrophage migration inhibitory factor (MIF) on the maternal hosts in murine pregnancy by enzyme-linked immunosorbent assay. The MIF concentrations in the murine maternal circulation in pregnancy ranged from 10 to 200 ng/ml and the level of MIF was lowest on Day 3 of pregnancy.

Evidence for the production of platelet-activating factor by murine embryos and its putative role in the maternal physiology.

Thrombocytopenia has been shown to be an initial maternal response to conception. We investigated a potent bioactive lipid, platelet-activating factor (PAF), as one of the candidates derived from embryos to induce the decrease of platelet counts in maternal blood. First, we examined the potential of murine embryos to liberate PAF.

Crystallization of rat liver macrophage migration inhibitory factor for MAD analysis.

The recombinant macrophage migration inhibitory factor (MIF) of a rat liver was crystallized using the hanging-drop vapor diffusion method. We also crystallized a selenomethionyl rat MIF under similar conditions for X-ray structure analysis using the multiwavelength anomalous diffraction method. Furthermore, two kinds of selenomethionyl rat MIF mutants, in which one or two of three methionine residues were replaced by alanine [(Met101Ala), (Met2Ala and Met101Ala)], were constructed and crystallized.

The C-terminal region, Arg201-Gln209, of glutathione S-transferase P contributes to stability of the active-site conformation.

The C-terminal region of rat glutathione S-transferase P (GST-P) was deleted by either carboxypeptidase (CPase) A and B or site-specific truncation to evaluate the role of the region in the catalytic mechanism. The C-terminal sequence from the 201st to 209th amino-acid residues is Arg-Pro-Ile-Asn-Gly-Asn-Gly-Lys-Gln. When seven of the C-terminal amino-acid residues from the C-terminus were removed by the CPases, the catalytic activity decreased in parallel with the amino-acid removal, amounting to less than 5% of that of the wild-type GST-P.

Identification of the electrophilic substrate-binding site of glutathione S-transferase P by photoaffinity labeling.

We determined the electrophilic substrate-binding site of rat glutathione S-transferase P (GST-P) by photoaffinity labeling using the photosensitive compound S-[2-(2-fluoro-4-nitrophenoxy)ethyl]glutathione. This photosensitive glutathione analogue inhibited the catalytic activity in a competitive manner against both glutathione and 1-chloro-2,4-dinitrobenzene, a putative electrophilic substrate. The enzyme kinetics indicated that the photoactivatable glutathione analogue was specifically bound at the active site, which consisted of glutathione-binding (G-site) and the electrophilic substrate-binding (H-site) regions.

The pathophysiological roles of heterogeneous eosinophils in allergic rhinitis caused by house dust mites.

We examined the biological characteristics of normodense and hypodense eosinophils prepared from the peripheral blood of the patients with allergic rhinitis caused by house dust mites by measuring leukotriene C4 (LTC4), platelet-activating factor (PAF), and superoxide anions. The normodense (density: 1.100-1.