AIM:To evaluate the prophylactic properties of integrin CD18-βA peptide in a murine model of abdominal polymicrobial peritonitis and sepsis.METHODS:Bacterial sepsis was induced in Institute of Cancer Research(ICR) mi...AIM:To evaluate the prophylactic properties of integrin CD18-βA peptide in a murine model of abdominal polymicrobial peritonitis and sepsis.METHODS:Bacterial sepsis was induced in Institute of Cancer Research(ICR) mice by cecal ligation and puncture(CLP) surgery.Inflicted mice were then injected with either sterile saline or CD18-βA peptide intraperitoneally at 2 h after surgery,and were sacrificed at 12 and 24 h after surgery.Blood samples were immediately collected,and analyzed for endotoxin activity and tumor necrosis factor(TNF)-α and interleukin(IL)-6.Lungs and liver were studied for CD45+ leukocyte and CD3 mRNA content.Pulmonary expression of intercellular adhesion molecule(ICAM)-1,vascular cell adhesion molecule(VCAM) and E-selectin was also determined.RESULTS:Intraperitoneal injection of CD18-βA peptide significantly suppressed circulating endotoxin activity(P<0.01) at 24 h,as well as serum levels of TNF-α(P<0.05 at 12 and 24 h) and IL-6(P<0.01 at 12 h,P<0.05 at 24 h) in CLP-inflicted mice.CD18-βA peptide also abrogated leukocyte infiltration into liver and lungs as unveiled by reduced CD45+ leukocyte and CD3 mRNA contents.Furthermore,the peptide significantly reduced pulmonary expression of VCAM(P<0.01 at 12 h,P<0.001 at 24 h),E-selectin(P<0.01 at 12 and 24 h),and ICAM-1(P<0.01 at 12 h,P<0.001 at 24 h).These actions of CD18-βA peptide collectively protected septic mice against lethality(P<0.01).CONCLUSION:CD18-βA peptide is a potent endotoxin antagonist that can protect surgical patients against sepsis-associated lethality.展开更多
The most common method of shipping cells between institutes and companies is sending them frozen, usually treated with anti-freeze solution (most commonly DMSO because it is less toxic than many alternatives), and the...The most common method of shipping cells between institutes and companies is sending them frozen, usually treated with anti-freeze solution (most commonly DMSO because it is less toxic than many alternatives), and then packaging them in dry ice for shipment. However many countries place restrictions on dry ice shipments. An alternative to shipping frozen cell vials is to send flasks of growing cells in media. This also has problems because cells in media have limited viability and the flasks can leak. Here we report on an alternative method for shipping viable cells at ambient temperature without dry ice or in media filled flasks. In this study we report on the development and properties of HemSol?. This is an inexpensive, eco-friendly and protects cell integrity at ambient temperature while maintaining viability. We have previously shown that HemSol? protects platelet and RBC function in cold storage and circulating tumor cells up to 6 days. Therefore we wanted to know if HemSol? could also be used to transport live cells. Since HemSol? is a liquid, we experimented with encasing the cells with HemSol? and gelatin so as to prevent dry ice shipment of cells and circumvent the shipping of cells in media. We performed mock shipping experiments where cells were stored in HemSol? gel kept at room temperature on a lab benchtop and cells stored in dry ice was also kept on lab benchtop for up to 2 days. After the mock shipping period, we analyzed cells for their functions. Our results show that cells in HemSol? gel have greater than 95% viability and restored biological functions in 2 hours, whereas, cells shipped in dry ice required more than 24 hours to recover and needed media change to remove the DMSO.展开更多
基金Supported by Research Grants Council of Hong Kong,National University of Singapore and NMRC
文摘AIM:To evaluate the prophylactic properties of integrin CD18-βA peptide in a murine model of abdominal polymicrobial peritonitis and sepsis.METHODS:Bacterial sepsis was induced in Institute of Cancer Research(ICR) mice by cecal ligation and puncture(CLP) surgery.Inflicted mice were then injected with either sterile saline or CD18-βA peptide intraperitoneally at 2 h after surgery,and were sacrificed at 12 and 24 h after surgery.Blood samples were immediately collected,and analyzed for endotoxin activity and tumor necrosis factor(TNF)-α and interleukin(IL)-6.Lungs and liver were studied for CD45+ leukocyte and CD3 mRNA content.Pulmonary expression of intercellular adhesion molecule(ICAM)-1,vascular cell adhesion molecule(VCAM) and E-selectin was also determined.RESULTS:Intraperitoneal injection of CD18-βA peptide significantly suppressed circulating endotoxin activity(P<0.01) at 24 h,as well as serum levels of TNF-α(P<0.05 at 12 and 24 h) and IL-6(P<0.01 at 12 h,P<0.05 at 24 h) in CLP-inflicted mice.CD18-βA peptide also abrogated leukocyte infiltration into liver and lungs as unveiled by reduced CD45+ leukocyte and CD3 mRNA contents.Furthermore,the peptide significantly reduced pulmonary expression of VCAM(P<0.01 at 12 h,P<0.001 at 24 h),E-selectin(P<0.01 at 12 and 24 h),and ICAM-1(P<0.01 at 12 h,P<0.001 at 24 h).These actions of CD18-βA peptide collectively protected septic mice against lethality(P<0.01).CONCLUSION:CD18-βA peptide is a potent endotoxin antagonist that can protect surgical patients against sepsis-associated lethality.
文摘The most common method of shipping cells between institutes and companies is sending them frozen, usually treated with anti-freeze solution (most commonly DMSO because it is less toxic than many alternatives), and then packaging them in dry ice for shipment. However many countries place restrictions on dry ice shipments. An alternative to shipping frozen cell vials is to send flasks of growing cells in media. This also has problems because cells in media have limited viability and the flasks can leak. Here we report on an alternative method for shipping viable cells at ambient temperature without dry ice or in media filled flasks. In this study we report on the development and properties of HemSol?. This is an inexpensive, eco-friendly and protects cell integrity at ambient temperature while maintaining viability. We have previously shown that HemSol? protects platelet and RBC function in cold storage and circulating tumor cells up to 6 days. Therefore we wanted to know if HemSol? could also be used to transport live cells. Since HemSol? is a liquid, we experimented with encasing the cells with HemSol? and gelatin so as to prevent dry ice shipment of cells and circumvent the shipping of cells in media. We performed mock shipping experiments where cells were stored in HemSol? gel kept at room temperature on a lab benchtop and cells stored in dry ice was also kept on lab benchtop for up to 2 days. After the mock shipping period, we analyzed cells for their functions. Our results show that cells in HemSol? gel have greater than 95% viability and restored biological functions in 2 hours, whereas, cells shipped in dry ice required more than 24 hours to recover and needed media change to remove the DMSO.
