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1.
Congenit Anom (Kyoto) ; 53(1): 46-8, 2013 Mar.
Article in English | MEDLINE | ID: mdl-23480357

ABSTRACT

Japan Association for Laboratory Animal Medicine (JALAM) recommends humane handling of rat fetuses. However, it is a challenge to accept proposed euthanizing methods such as cervical dislocation, decapitation and/or intracardiac injection of potassium chloride, because these methods would damage fetal specimens for skeletal and visceral examinations in developmental toxicity studies. The present study aimed at seeking better methodologies for fetal euthanasia and anesthesia. We were unable to accomplish fetal euthanasia directly, but instead, we could euthanize fetuses under pain-controlled anesthesia. It is recommended that hypothermia by immersion in cold physiological saline is an appropriate method for anesthesia. Moreover, we recommend that the anesthetized fetuses should be euthanized immediately by removal of the vital organs or immersion in appropriate fixatives.


Subject(s)
Anesthesia/methods , Euthanasia , Teratogens/toxicity , Animals , Female , Fetus/drug effects , Fetus/physiology , Heart Rate , Male , Rats , Rats, Sprague-Dawley
2.
Thromb Res ; 128(3): 283-92, 2011 Sep.
Article in English | MEDLINE | ID: mdl-21624645

ABSTRACT

Neutrophil elastase released from activated neutrophils contributes in combating bacterial infection. While chronic inflammation results in anemia and decreased bone marrow activities, little is known about the effect of neutrophil elastase on hematological cell growth in severe inflammatory states. Here, we demonstrated that α1-antitrypsin, a physiological inhibitor of neutrophil elastase, functions as a regulator for cell growth by neutralizing neutrophil elastase activity in lipopolysaccharide-primed hematological cells. HL-60 cells were resistant to neutrophil elastase, as they also expressed α1-antitrypsin. The growth of HL-60 cells transduced with a LentiLox-short hairpin α1-antitrypsin vector was significantly suppressed by neutrophil elastase or lipopolysaccharide. When CD34(+) progenitor cells were differentiated towards a granulocytic lineage, they concomitantly expressed neutrophil elastase and α1-antitrypsin and prevented neutrophil elastase-induced growth inhibition. These results suggest that granulocytes might protect themselves from neutrophil elastase-induced cellular damage by efficiently neutralizing its activity through the simultaneous secretion of endogenous α1-antitrypsin.


Subject(s)
Hematopoietic System/enzymology , Leukocyte Elastase/antagonists & inhibitors , Leukocyte Elastase/metabolism , alpha 1-Antitrypsin/metabolism , Cell Growth Processes/physiology , HL-60 Cells , Hematopoietic System/cytology , Humans , Immunohistochemistry , K562 Cells , Lipopolysaccharides/pharmacology , Megakaryocytes/drug effects , Megakaryocytes/metabolism
3.
Thromb Res ; 127(4): 349-55, 2011 Apr.
Article in English | MEDLINE | ID: mdl-21236472

ABSTRACT

An alternative pathway for fibrinolysis that comprises leukocyte elastase and its interaction with the plasminogen activator-plasmin system has been suggested. Plasma levels of cross-linked fibrin degradation product by leukocyte elastase (e-XDP) were significantly increased in patients with sepsis induced disseminated intravascular coagulation (DIC) compared with healthy subjects (18.6±19.9 vs 0.58±0.47U/mL, p<0.001). Twenty seven unique spots were identified from e-XDP dominant patients by immune-purification and two-dimensional difference gel electrophoresis, and they contained fibrinogen Bß-chain derived fragments Bß Asp-164, Ser-200, Gln-301, Ala-354, Ile-484 and γ-chain derivatives γ Val-274 at their amino-termini by acquired and processed tandem mass spectrometer. The Sequential Organ Failure Assessment Scores in patients with e-XDPs levels 3-10U/mL were significantly lower than those with e-XDPs levels -3U/mL, 10-30U/mL, and 30- U/mL. The adjusted odds for 28-day mortality rate in patients with e-XDP levels less than 3U/mL (hazard ratio, 4.432; 95% CI, 1.557-12.615 [p=0.005]) were significantly higher than those in patients with e-XDP levels of 3-10U/mL. These data suggest that leukocyte elastase might contribute to the degradation of cross-linked fibrin in sepsis-induced DIC.


Subject(s)
Disseminated Intravascular Coagulation/blood , Fibrin/metabolism , Fibrinolysin/metabolism , Fibrinolysis , Leukocyte Elastase/metabolism , Sepsis/complications , Aged , Disseminated Intravascular Coagulation/diagnosis , Disseminated Intravascular Coagulation/etiology , Disseminated Intravascular Coagulation/metabolism , Female , Fibrin Fibrinogen Degradation Products/metabolism , Humans , Male , Middle Aged , Prognosis
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