• Korean Journal of Veterinary Research
• /
• v.34 no.2
• /
• pp.275-286
• /
• 1994

Plasma protein which has been known as one of nonspecific immunostimulators was added to feedstuff to examine its effect on the enhancement of cellular immune response in porcine immune system. A total of 40 piglets, 20 male and 20 female each, were fed for 30 days with or without plasma protein. The peripheral blood were collected and analyzed for the investigation of leukocyte subpopulations and their activities by using a panel of monoclonal antibodies specific to porcine leukocyte differentiation antigens and flow cytometry. The results obtained as follows. 1. Total weight gain, daily feed intake and feed conversion rate for 10 days were significantly improved to 56%, 20% and 22% in the piglets fed plasma protein, respectively. 2. A significant increase in N (null or non T/non B) cells was also noticed. Leukocyte proportion from piglets fed plasma protein was 20.2-24.7%, otherwise that from piglets fed without plasma protein was 12.3-13.4%, respectively. 3. A significant increase in the proportion of B cells and cells expressing poCD1 was not found in piglets fed plasma protein. 4. Reaction with monoclonal antibodies specific to adhesion molecules, poCD11a, poCD11b, poCD44 and poCD45A and poCD45B, has shown that leukocyte subpopulation from piglets fed plasma protein did not significantly higher than that from piglets fed without plasma protein. 5. Total proportion of granulocytes and monocytes was about 50% in both group and the proportion after treated with Hypaque/Ficoll was 2.7% and 5.8% in each group, respectively.

• Journal of Microbiology and Biotechnology
• /
• v.28 no.5
• /
• pp.707-717
• /
• 2018

Leukocytes are reportedly the first line of the innate immune defense and essential for the control of common bacterial infections. Therefore, in this work, the antibacterial activity of crocodile leukocyte extract against Propionibacterium acnes was evaluated, and we also characterized the related activity of skin infection. The leukocyte extract showed the minimum inhibitory concentration to be $100{\mu}g/ml$ to P. acnes. SEM imaging demonstrated that the leukocyte extract adversely affected P. acnes cell permeability in a concentration-dependent manner. Furthermore, the crocodile leukocyte extract could significantly reduce proinflammatory markers and decrease inflammatory signs in infected mouse ears. The crude leukocyte extract was further purified using FPLC and RP-HPLC. The resulting fraction F5 was indicated as the anti-acne peptide-containing fraction. The molecular mass of the peptide contained in F5 was calculated to be 4,790.5 Da. N-Terminal sequencing revealed the amino acid sequence as GPEPVPAIYQ, which displays similarities to immunoglobulin A and leucine-rich repeat neuronal protein. This is the first reported amino acid sequence of a crocodile leukocyte extract that possesses anti-acne activity. To attempt to use it in a prototype cosmetic, an anti-acne gel containing crude crocodile leukocyte extract was formulated, resulting in seven gel formulations (G1, G2, G3, G4, G5, G6, and G7). The formulations G5, G6, and G7 exhibited 2-fold higher anti-acne activity than G1-G4. Investigation of accelerating stability studies of anti-acne gel formulations G5, G6, and G7 demonstrated that a low storage temperature ($4^{\circ}C$) is suitable for maintaining the physical properties and biological activity of the anti-acne gel products.

• Journal of Chest Surgery
• /
• v.33 no.3
• /
• pp.213-220
• /
• 2000

Background: Adhesion of leukocytes to myocardium or vascular endothelium has been known as an importation initial step in the ischemia-reperfusion injury which may affect the cardiac function. Therefore, leukocyte-depleted reperfusion may inhibit ischemia-reperfusion induced functional and ultrastructural deterioration. In this study, we quantified the time-dependent expression of the vascular cell adhesion molecule-1(VCMA-1) on piglet myocardium and demonstrated its relation to functional recovery using isolated piglet heart perfusion model. Material and Method: Neonatal(1 to 3 day old) piglet heart was harvested with 4$^{\circ}C$ University of Wisconsin solution (UWS) and presrved in the same solution for 12 hours. Ex vivo model of an isolated working neonatal piglet heart perfusion consisting of membrane oxygenator and roller-pump was used (Fig. 1). Hearts were grouped into leukocyte-non-depleted (group A, n=8) and leukocyte-depleted group(group B, n=8). In group B, hearts were reperfused with leukocyte-depleted blood using a leukocyte filter (Sepacell R, Asahi Medical, Japan). Segments of right atrium were taken before and after 1, 2, 3, and 4 hours of reperfusion for the evaluation of expression of VCAM-1. The intensity of immunohistochyemical satining of the VCAM-1 on the myocardium were graded semiquantitatively (0 to 4). For the evaluation of myocardial stroke work indices were calculated as well at the same time-points. Result: Mean expressins of VCAM-1 on the myocardium at 0, 1, 2, 3, adn 4 hours of reperfusion were 0.63, 1.44, 1.64, 2.65, and 3.34 in group A, while 0.56, 1.40, 1.50, 1.88 and 2.14 in group B (Fig. 3). Mean stroke work indices at 0.5, 1, 2, 3, and 4 hours after reperfusion were 1.35$\times$104, 1.32$\times$104, 1.14$\times$104, 0.81$\times$104, 0.68$\times$104 erg/gm in group A, while 1.40$\times$104, 1.43$\times$104, 1.43$\times$104, 1.28$\times$104, and 1.12$\times$104 erg/em in group B(Fig. 4). Conclusion : In this study, we demonstrated that leukocyte-depletion attenuated the expression of VCAM-1 during reperfusion and the time-dependent functional deterioration of the myocardium was well correlated with the degree of VCAM-1 expression.

• Journal of Chest Surgery
• /
• v.38 no.12 s.257
• /
• pp.807-814
• /
• 2005

Background: Cardiopulmonary bypass is an essential process to maintain circulation for saving life during the cardiac surgery, But it is a process in which systemic inflammation was evoked inevitably because of the exposure of blood to foreign surface. The injuries to distal organs during the cardiopulmonary bypass were resulted from systemic inflammation and the disturbances of micro-circulations in the organs. We designed this study to research the effects of leukocyte depletion from pump-oxygenator priming solution on the systemic inflammation, and the micro-circulation of gastric mucosa that is suggested by the gastric mucosal $CO_{2}$ partial pressure and acidity. Material and Method: The dogs were divided into three groups according to the different pump-oxygenator priming solutions; non-hemic crystalloid solution; leukocyte-depleted homologous blood; and non leukocyte-depleted homo-logous blood. Each priming solution group contained five dogs. In all three groups, 2 hours of cardiopulmonary bypass, and 4 consecutive hours of general anesthesia was maintained on the mechanical ventilation. Each dog was evaluated for the gastric mucosal pH, $CO_{2}$ partial Pressure, arterial pH, $CO_{2}$ partial pressure, the exhaled air $CO_{2}$ partial pressure and the level of IL-8 on before the cardiopulmonary bypass, 1 hour after the cardiopulmonary bypass, 2 hours after the cardiopulmonary bypass, 2 hours after the restoration of normal circulation, and 4 hours after the restoration of normal circulation after the cardiopulmonary bypass. The levels of IL-8 were measured with ELISA (enzyme linked immunosorbent assay) technique. Result: 1. There were significant differences of gastric mucosal $CO_{2}$ partial pressure between the leukocyte-depleted homologous blood group and other two groups(vs non leukocyte-depleted homologous blood group; P=0.02, vs non-hemic crystalloid solution group; P=0,01). 2. The gastric mucosal pH of leukocyte-depleted homologous blood group was significantly different from non leukocyte-depleted homologous blood group (p=0.01). 3. The levels of IL-8, which examine the systemic inflammation, showed signi- ficantly better results in leukocyte-depleted homologous blood group and non-hemic crystalloid solution group than non leukocyte-depleted homologous blood group (p=0.01, 0.01). Conclusion: Based upon these results, we concluded that the leukocyte depletion from the pump-oxygenator priming solution has a beneficial effects in reducing systemic inflammation and the preserving of gastric mucosal micro-circulation.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
• /
• v.29 no.5
• /
• pp.301-314
• /
• 2003

The purposes of this retrospective study were to assess the change of serum parameters in oral and maxillofacial surgery patients after operation and to determine what laboratory parameters on treatment periods were associated with the recovery of systemic condition. For purposes of assessing systemic nutritional status, several serum parameters were chosen. The sample patients were randomsubjects extracted from three category patient groups- oral cancer, odontogenic abscess, facial bone fracture based on treated patients at department of oral and maxillofacial surgery in Pusan National University Hospital from September 1, 1998, to September 1, 2002. Each groups were consisted with 10 patients. Each patient chart was examined and blood sample parameters were reviewed with clinical signs, symptoms and vital sign at preoperative day, postoperative 1 day, postoperative 1 week. Several parameters were analyzed statistically for extraction of mean values and differences between the periods groups. The findings of serum parameters of cancer, abscess and fracture groups were as follows: 1. In cancer patients, Hb, MCV, albumin, cholesterol, LDH, AST, ALT, neutrophil, platelet, leukocyte, Na, K, Cl, BUN, creatinine were analyzed. Values of Hb, albumin, AST, neutrophil, leukocyte, Cl showed significantly differences according to periods. 2. In abscess patients, CRP, ESR, leukocyte, body temperature, neutrophil were analyzed. Values of CRP, leukocyte, body temperature, neutrophil showed significanlty differences according to periods. 3. In fracture patients, same parameters with cancer patient's were chosen. Values of platelet, Cl only showed significantly differences according to periods. 4. In cancer patients, data regarding correlation was analyzed statistically as Pearson's value. A positive correlation was found between Hb and albumin, K, Na(P<0.05). A positive correlation was also found between neutrophil and leukocyte(P<0.05). Positive correlations were found between cholesterol and ALT, LDH and platelet, creatinine both, Platelet and BUN, Na and K(P<0.01). 5. In abscess patients, Peason's correlation values were analyzed on parameters. A positive correlation was found only between CRP and neutrophil(P<0.05). 6. In fracture patients, The correlations of parameters also were statistically analyzed. Positive correlations were found between MCV and K, albumin and LDH, AST and three parameters of creatinine, Na, Cl, K and neutrophil, neutrophil and three parameters of leukocyte, BUN, K(P<0.05). Positive correlations were found between LDH and AST, ALT and AST, creatinine both(P<0.01). This retrospective clinical study showed the CRP levels only on abscess patients may be useful in determination of clinical infected status, but the levels of other parameters on cancer, fracture patients did not showed significant values as diagnostic aids for clinical status.

• Korean Journal of Veterinary Research
• /
• v.39 no.2
• /
• pp.247-256
• /
• 1999

A disease of young Holstein calves characterized by recurrent pneumonia, ulcerative and granulomatous stomatitis, enteritis with bacterial overgrowth, periodontitis, delayed wound healing, persistent neutrophilia and death at an early age had been originally described in 1983 and again in 1987. Most of these calves had stunted growth and a persistent, progressive neutrophilia (often exceeding 100,000/ml). By investigation of pedigrees, all of the affected calves have now been traced to a common sire and confirmed by polymerase chain reaction (PCR) diagnostic DNA testing to be homozygous carriers of a defective allele for bovine CD18. Neutrophils from these calves have several functional deficits and, most importantly, fail to adhere in a ${\beta}_2$-integrin dependent manner. The ${\beta}_2$-integrins represent a family of glycoproteins which participate in various leukocyte adhesion reactions during host defense. The presence or absence of ${\beta}_2$-integrin molecules can be demonstrated on the surface of neutrophils, monocytes and lymphocytes from normal or affected calves using specific monoclonal antibodies and flow cytometry, or by colloidal gold immunolabeling and scanning electron microscopy in backscatter mode. Deficiency of the ${\beta}_2$-integrins on all leukocyte types in Holstein calves is analogous to leukocyte adhesion deficiency (LAD) seen in humans. Neutrophils in bovine (BLAD) and human LAD patients are unable to adhere to the endothelial lining of the cardiovascular system thus interrupting egression of neutrophils into infected tissues. Other leukocytes, while still deficient in expression of the ${\beta}_2$-integrins, are still able to efficiently egress from the blood stream due to interactions of other adhesion molecules that are not as highly expressed on neutrophils. Both BLAD cattle and LAD children (who do not receive bone marrow transplants) often die at an early age as a result of the failure of neutrophils to extravasate into infected tissues. In 1991, Shuster, et $al^{27}$, identified two point mutations within the alleles encoding bovine CD18 in a Holstein calf afflicted with leukocyte adhesion deficiency. One mutation causes an aspartic acid to glycine substitution at amino acid 128 (D128G) in an extracellular region of this adhesion glycoprotein that is highly conserved (> 95% identity) between humans, cattle and mice. The other mutation is silent. Numerous calves with clinical symptoms of leukocyte adhesion deficiency have since been tested and all have been found homozygous for the D128G allele. In addition, calves homozygous far the D128G allele have been identified during widespread DNA testing in the United States. All cattle with the mutant allele are related to one bull, who through artificial insemination (A.I.), sired many calves in the 1950's and 1960's. The carrier frequency of the D128G CD18 allele among U.S. Holstein cattle had reached approximately 15% among active A.I. bulls and 8% among cows. By 1993, the organization of the dairy industry and the diagnostic test developed to genotype cattle, enabled virtually complete eradication of bovine leukocyte adhesion deficiency among current and future A.I. bulls.

• Applied Microscopy
• /
• v.5 no.1
• /
• pp.1-8
• /
• 1975

These studies were carried out the observation of polymorphonuclear leukocyte phagocytosis of Candida albicans in vitro and also detected to the cytoplasmic changes of polymorphonuclear leukocyte during phagocytosis by the method of electron microscopy. The results were summarized as follows: 1. In normal polymorphonuclear leukocyte, nuclear lobes showed a preponderance of dense, granular chromatin located peripherally. The cytoplasm of polymorphonuclear leukocyte was not extensive; the cytoplasmic matrix was moderate dense and of a granular appearance. Golgi complex and rough endoplasmic reticulum system was poorly developed. But a various type of granules were seen abundantly. 2. After 30 minutes of incubation, Candida albicans was completely engulfed. These had come to lie in the vacuole which was limited by the membrane. 3. After 90 minutes of incubation, the phagocytic vacuoles were larger, and many granules devoid of membranes were seen within them. Though the granules has lost their membrane after entering the vacuoles. 4. After 2 hours of incubation, the cytoplamsic components of polymorphonuclear leuko cytes were changed their original morphology.

• Journal of Chest Surgery
• /
• v.32 no.11
• /
• pp.978-983
• /
• 1999

Background: Complement activation with transpulmonary leukocyte sequestration is considered a main mediator leading to ischemia-reperfusion lung(I-R) injury. We studied the role of leukocytes in the formation of I-R injury in ovine cardiopulmonary bypass(CPB) model with a membrane oxygenator. Material and Method: Five sheep were used. CPB circuitry consisted of a roller pump(American Optical Corp., Greenwich, CT, USA) and a membrane oxygenator(UNIVOX-IC, Bentley, Baxter Health Corp, Irvine, CA, USA). The CPB time was fixed at 120 min. Ten minutes after the start of CPB, total CPB was established. Thereafter a total CPB of 100 min was performed, followed by another 10 min of partial CPB. The CPB was discontinued and the animals were fully recovered. For measuring left and right atrial leukocyte counts, blood samples were taken before thoracotomy, 5 min and 109 in after the start of CPB, and 30 min and 120 min after weaning. C3a was measured before thoracotomy, 109 min after the start of CPB, and 30 min and 120 min after weaning. Plasma malondialdehyde(MDA) was checked before thoracotomy, 109 min after the start of CPB, and 30 min after weaning. One to two grams of lung tissue were taken for water content measurement before thoracotomy, 109 min after the start of CPB, and 30 min after weaning. Lung biopsy specimens were examined by light and electron microscopy. Result: Of 5 animals, 4 survived the experimental procedures. Of these, 3 animals survived on a long-term basis. No significant differences in transpulmonary gradients of leukocyte were found and no significant complement activation was expressed by C3a levels. MDA level did not show significant changes related to lung reperfusion despite an increase after the start of CPB. On both light and electron microscopic examinations, mild to moderate acute lung change was observed. Interstitial edema, leakage of erythrocytes into the alveolar space and endothelial cell swelling were the main findings. Water content of the lung showed a slight increase after the start of CPB, but there was no statistical significance. Conclusion: These findings indicate that ischemia-repersusion lung injury may not be from complement activation-leukocyte sequestration but from another source of oxygen free radicals related to CPB.

• Journal of fish pathology
• /
• v.33 no.1
• /
• pp.23-34
• /
• 2020

In recent years, global warming is causing dramatic environmental changes and deterioration, such as hypoxia, leading to reduced survival rate and growth performance of farmed aquatic animals. Hence, understanding systemic immuno-physiological changes in fish under environmental stress might be important to maximize aquaculture production. In this study, we investigated physiological changes in rainbow trout exposed to hypoxic stress by monitoring changes in blood chemistry, leukocyte population, and expression levels of related cytokine genes. Hematological and serological factors were evaluated in blood obtained from rainbow trout sampled at a dissolved level of 4.6 mg O₂ L^-1 and 2.1 mg O₂ L^-1. Blood and head kidney tissue obtained at each sampling time point were used to determine erythrocyte size, leukocyte population, and cytokine gene expression. The level of LDH and GPT in fish under progressive hypoxia were significantly increased in plasma. Likewise, the (Granulocyte + Macrophage)/lymphocyte ratio (%) of fish exposed to hypoxia was significantly lower than that in fish in the control group. Such changes might be due to the rapid movement of lymphocytes in fish exposed to acute hypoxia. In this study, significant up-regulation in expression levels of IL-1β and IL-6 gene appeared to be involved in the redistribution of leukocytes in rainbow trout. This is the first study to demonstrate the involvement of cytokines in leukocyte trafficking in fish exposed to hypoxia. It will help us understand systemic physiological changes and mechanisms involved in teleost under hypoxic stress.

• Biomedical Science Letters
• /
• v.6 no.4
• /
• pp.281-288
• /
• 2000

Leukocyte activation with cardiac surgery procedures produces various iuflammatory substances and involves in postoperative pathophysiology. The present study was carried out to elucidate changes in leukocyte myeloperoxidase level and effect on the heart and lung during cardiac operation. Total leukocyte and differential counts in peripheral blood, myeloperoxidase (MPO) and troponin-T concentratiens (TnT) in coronary sinus blood, and pulmonary vascular resistance (PVR) were measured at preoperative and postoperative period. The parameters were compared between sampling periods, and relationship was investigated between MPO and each variable. At the end of operation, there were leukocytosis with neutrophilia (p＜0.01), and increases of MPO and TnT concentrations (p＜0.05), but decrease in PVR (p＜0.05). MPO had a positive correlation to TnT, total leukocyte, neutrophil, or operative times (p＜0.05), whereas PVR had a negative relationship to total leukocyte or neutrophil counts (p＜0.05). These results indicate that cardiac surgery leads to elevated liberations of myeloperoxidase from neutrophils and may harmfully affect myocardium.