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4.  Time-dependent gene expression and immunohistochemical analysis of the injured anterior cruciate ligament 
Bone & Joint Research  2012;1(10):238-244.
This study aimed to investigate time-dependent gene expression of injured human anterior cruciate ligament (ACL), and to evaluate the histological changes of the ACL remnant in terms of cellular characterisation.
Injured human ACL tissues were harvested from 105 patients undergoing primary ACL reconstruction and divided into four phases based on the period from injury to surgery. Phase I was < three weeks, phase II was three to eight weeks, phase III was eight to 20 weeks, and phase IV was ≥ 21 weeks. Gene expressions of these tissues were analysed in each phase by quantitative real-time polymerase chain reaction using selected markers (collagen types 1 and 3, biglycan, decorin, α-smooth muscle actin, IL-6, TGF-β1, MMP-1, MMP-2 and TIMP-1). Immunohistochemical staining was also performed using primary antibodies against CD68, CD55, Stat3 and phosphorylated-Stat3 (P-Stat3).
Expression of IL-6 was mainly seen in phases I, II and III, collagen type 1 in phase II, MMP-1, 2 in phase III, and decorin, TGF-β1 and α-smooth muscle actin in phase IV. Histologically, degradation and scar formation were seen in the ACL remnant after phase III. The numbers of CD55 and P-Stat3 positive cells were elevated from phase II to phase III.
Elevated cell numbers including P-Stat3 positive cells were not related to collagens but to MMPs’ expressions.
PMCID: PMC3626253  PMID: 23610654
Anterior cruciate ligament; ACL; Synovial fibroblast-like cell; Matrix metalloproteinases (MMPs); Stat3; Gene expression; Immunohistochemical analysis
5.  Role of K+ ATP channels and adenosine in the regulation of coronary blood flow during exercise with normal and restricted coronary blood flow. 
Journal of Clinical Investigation  1996;97(4):996-1009.
Regulation of coronary vasomotor tone during exercise is incompletely understood. We investigated the contributions of K+ ATP channels and adenosine to the coronary vasodilation that occurs during exercise in the normal heart and in the presence of a coronary artery stenosis. Dogs that were chronically instrumented with a Doppler flow probe, hydraulic occluder, and indwelling catheter on the left anterior descending coronary artery were exercised on a treadmill to produce heart rates of approximately 200 beats/min. By graded inflation of the occluder to produce a wide range of coronary stenosis severities, we determined the coronary pressure-flow relation. K+ atp channel blockade with intracoronary glibenclamide (10-50 microgram/kg per min) decreased coronary blood flow during exercise at coronary pressures within and below the autoregulatory range, indicating that coronary K+ ATP channel activation is critical for producing coronary vasodilation with either normal arterial inflow or when flow is restricted by a coronary artery stenosis. Adenosine receptor blockade with intravenous 8-phenyltheophylline (5 mg/kg) had no effect on coronary flow at pressures within the autoregulatory range but decreased flow at pressures < 55 mmHg. In contrast, in the presence of K+ ATP channel blockade, the addition of adenosine receptor blockade further decreased coronary flow even at coronary pressures in the autoregulatory range, indicating increased importance of the vasodilator influence of endogenous adenosine during exercise when K+ atp channels are blocked. Intracoronary adenosine (50 microgram/kg per min) increased coronary flow at perfusion pressures both within and below the autoregulatory range. In contrast, selective K+ ATP channel activation with intracoronary pinacidil (0.2-5.0 microgram/kg per min) increased flow at normal but not at lower coronary pressures (< 55 mmHg). This finding demonstrates that not all K+ ATP channels are activated during exercise at pressures in the autoregulatory range, but that most K+ ATP channels are recruited as pressures approach the lower end of the autoregulatory plateau. Thus, K+ ATP channels and endogenous adenosine play a synergistic role in maintaining vasodilation during exercise in normal hearts and distal to a coronary artery stenosis that results in myocardial hypoperfusion during exercise.
PMCID: PMC507146  PMID: 8613554
7.  Chromium reduction in Pseudomonas putida. 
Reduction of hexavalent chromium (chromate) to less-toxic trivalent chromium was studied by using cell suspensions and cell-free supernatant fluids from Pseudomonas putida PRS2000. Chromate reductase activity was associated with soluble protein and not with the membrane fraction. The crude enzyme activity was heat labile and showed a Km of 40 microM CrO4(2-). Neither sulfate nor nitrate affected chromate reduction either in vitro or with intact cells.
PMCID: PMC184598  PMID: 2389940
8.  Endogenous Nocardia asteroides endophthalmitis in a patient with systemic lupus erythematosus. 
We report a case of endogenous Nocardia endophthalmitis in a patient with systemic lupus erythematosus (SLE). He developed a parafoveal lesion in the right fundus while on systemic corticosteroid and antibiotic treatment. Initially we suspected a fungal origin and treated him with antifungal drugs. The intraocular disease progressed without improvement and advanced to the vitreous cavity. Nocardia asteroides was found in a specimen obtained at pars plana vitrectomy and was also cultured from the same specimen. The intraocular infection was controlled by antibacterial drugs, though the visual acuity of the right eye was reduced to only light perception owing to heavy vitreous opacity and secondary cataract. This case is the first report of endogenous Nocardia endophthalmitis in Japan and also the first case of this disease reported from outside the United States of America.
PMCID: PMC1042157  PMID: 2198934
9.  Effects of a phagocytosis-stimulating factor derived from polymorphonuclear neutrophils on the functions of macrophages. 
Infection and Immunity  1987;55(8):1762-1766.
The effects of phagocytosis-stimulating factor (PSF) derived from polymorphonuclear neutrophils on macrophage functions were studied. PSF enhanced the initial rate of phagocytosis of serum-opsonized zymosan particles by macrophages, whereas it did not affect the phagocytosis of immunoglobulin G-sensitized and inert zymosan particles. Kinetic studies showed that PSF accelerated the ingestion step, but not the attachment step, of phagocytosis by macrophages. On the other hand, PSF did not affect the other macrophage functions such as O2- generation, chemotaxis, adherence, and enzyme release. These results suggest that PSF may specifically modulate the complement receptor function of macrophages. Immunoblot assay showed the absence of components in macrophage which reacted with purified antibodies against polymorphonuclear neutrophil-derived PSF, and an extract from phagocytosing macrophages had no phagocytosis-stimulating activity, indicating that the macrophages did not produce PSF-like substances.
PMCID: PMC260598  PMID: 3038750
10.  Identification of precursor of phagocytosis-stimulating factor in guinea pig polymorphonuclear neutrophils. 
Infection and Immunity  1985;50(2):500-505.
A precursor of phagocytosis-stimulating factor (PSF) was identified by immunoblot assay with purified anti-PSF antibodies. The purified anti-PSF antibodies recognized not only the PSF with a molecular weight of 16,000 (16K) but also the 36K protein with a pI of 6.5 in the granule fraction of polymorphonuclear neutrophils, indicating that this 36K protein has an antigenic determinant common to PSF. In addition, the appearance of the PSF during phagocytosis by polymorphonuclear neutrophils was closely correlated to the decrease of the 36K protein. These results suggest that the 36K protein is the precursor of PSF which is converted to biologically active PSF in the granules during phagocytosis.
PMCID: PMC261982  PMID: 4055031
11.  Purification and characterization of a phagocytosis-stimulating factor from phagocytosing polymorphonuclear neutrophils: comparison with granule basic proteins. 
Infection and Immunity  1985;48(3):799-805.
Phagocytosis-stimulating factor (PSF) was purified by copper chelate chromatography and characterized in comparison with basic proteins in the granule of polymorphonuclear neutrophils. By copper chelate chromatography, PSF was eluted at pH 3.7; whereas cationic protein, lysozyme, and lactoferrin were eluted at pH 5.6, 5.1, and 4.0, respectively. Purified PSF has an approximate molecular weight of 16,000 and an isoelectric point at 8.7, which differ from those of basic proteins, such as cationic protein, lysozyme, and lactoferrin. Anionic substances such as DNA and heparin did not influence the phagocytosis-stimulating activity of PSF, whereas that of the granule basic protein fraction from resting polymorphonuclear neutrophils was abolished. PSF had little bactericidal activity against Escherichia coli and Staphylococcus aureus, whereas the granule basic protein fraction from resting PMNs had strong bactericidal activity against E. coli and weak activity against S. aureus. These results indicate that PSF is a basic protein which is distinguishable from cationic protein, lysozyme, and lactoferrin.
PMCID: PMC261268  PMID: 3997249
12.  Effects of a phagocytosis-stimulating factor on the phagocytic process of polymorphonuclear neutrophils. 
Infection and Immunity  1982;38(3):825-833.
The effect of phagocytosis-stimulating factor (PSF) on phagocytosis was studied in detail. PSF did not affect the Fc receptor-mediated phagocytosis, whereas PSF enhanced the ingestion step, but not attachment step, of C3b receptor-mediated phagocytosis by polymorphonuclear neutrophils (PMNs), suggesting that PSF may specifically modulate the C3b receptor function of PMNs. PSF generated from guinea pig PMNs enhanced phagocytosis by rabbit PMNs, and rabbit PMN-produced PSF accelerated the phagocytosis by guinea pig PMNs, indicating that PSF is not specific for animal species. The effects of PSF on some PMN functions, such as O2- generation, chemotaxis, adherence, and enzyme release, were also studied. Only O2- generation from PMNs was significantly increased in the initial phase of phagocytosis, and this stimulation of O2- generation was completely parallel with the stimulation of phagocytosis by PSF. Resting PMNs hardly generated the superoxide anions by PSF treatment, suggesting that PSF does not affect the O2- forming system directly.
PMCID: PMC347822  PMID: 6295947
13.  Gene expression analysis of rheumatoid arthritis synovial lining regions by cDNA microarray combined with laser microdissection: up-regulation of inflammation-associated STAT1, IRF1, CXCL9, CXCL10, and CCL5 
The main histological change in rheumatoid arthritis (RA) is the villous proliferation of synovial lining cells, an important source of cytokines and chemokines, which are associated with inflammation. The aim of this study was to evaluate gene expression in the microdissected synovial lining cells of RA patients, using those of osteoarthritis (OA) patients as the control.
Samples were obtained during total joint replacement from 11 RA and five OA patients. Total RNA from the synovial lining cells was derived from selected specimens by laser microdissection (LMD) for subsequent cDNA microarray analysis. In addition, the expression of significant genes was confirmed immunohistochemically.
The 14 519 genes detected by cDNA microarray were used to compare gene expression levels in synovial lining cells from RA with those from OA patients. Cluster analysis indicated that RA cells, including low- and high-expression subgroups, and OA cells were stored in two main clusters. The molecular activity of RA was statistically consistent with its clinical and histological activity. Expression levels of signal transducer and activator of transcription 1 (STAT1), interferon regulatory factor 1 (IRF1), and the chemokines CXCL9, CXCL10, and CCL5 were statistically significantly higher in the synovium of RA than in that of OA. Immunohistochemically, the lining synovium of RA, but not that of OA, clearly expressed STAT1, IRF1, and chemokines, as was seen in microarray analysis combined with LMD.
Our findings indicate an important role for lining synovial cells in the inflammatory and proliferative processes of RA. Further understanding of the local signalling in structural components is important in rheumatology.
PMCID: PMC3400100  PMID: 22401175

Results 1-13 (13)