M. Kudla - Academia.edu (original) (raw)
Papers by M. Kudla
Transplantation Proceedings, 2016
Background. The aim of our study was to evaluate the rate of surgical complications, patient outc... more Background. The aim of our study was to evaluate the rate of surgical complications, patient outcomes, and impact on graft function in renal transplant recipients in whom cholecystectomy for acute cholecystitis was performed. Methods. We reviewed data on transplant patients from January 1, 2006, to December 31, 2013. The subgroup of patients who required subsequent cholecystectomy for acute cholecystitis was assessed, and their data were further analyzed. Results. Thirty-one patients who underwent cholecystectomy for acute cholecystitis after renal transplantation were included in the study. Clinical signs such as pain in the right upper quadrant, temperature >38 C, and elevation in bilirubin levels occurred in 20 (64.5%), 8 (25.8%), and 3 (9.7%) patients, respectively. Ultrasound signs of acute cholecystitis were present in 27 patients (87.1%). In terms of laboratory values, white blood cell counts >10 Â 10 9 / L occurred in 17 patients (54.8%), and C-reactive protein levels >40 mg/L were reported in 21 patients (67.7%). The conversion rate to open surgery was 32.3% (10 patients). In 13 cases, acalculous cholecystitis was present (41.9%). The average serum creatinine level 1 year after cholecystectomy had no statistically significant differences. One patient required temporary dialysis during the postoperative period (with subsequent graft recovery), and 1 graft was lost. Conclusions. Acute cholecystitis in kidney transplant recipients is a serious complication, with frequent difficulties related to evaluation and diagnosis. Because clinical signs could be very mild compared with severity of gallbladder affliction, there is little room if any for conservative treatment in these patients. We have not noticed adverse impact of acute cholecystitis on 1-year graft function.
Rozhledy v chirurgii : měsíčník Československé chirurgické společnosti, 2009
Transplantation of the small intestine is a standard treatment method in patients with small inte... more Transplantation of the small intestine is a standard treatment method in patients with small intestinal failures. The aim of this study was to master the surgical technique, optimalize immunosuppression regimes, diagnose acute cellular graft rejection based on cellular and humoral indicators. The authors performed a total of 43 transplantation procedures in pigs. The first, surgical part of the experiment was aimed at mastering two principal methods of vascular anastomosis- firstly, connecting the graft with mesenteric vessels (Group n1 = 18) and secondly, connecting the graft with the aorta and the inferior vena cava (Group n2 = 25). The second part of the experiment included assessment of rejection changes in various immunosuppression regimes. Only animals who did not die because of a technical failure of the procedure or due to internal reasons (n = 24) were assessed. The study animals were assigned to four groups (A (n = 3)--autotransplantation, without immunosuppresion; B (n = ...
Bratislavské lekárske listy, 2009
The technical aspects of a procedure are most important for the outcome of the experiment. This s... more The technical aspects of a procedure are most important for the outcome of the experiment. This study was designed to compare two techniques suitable for graft revascularization. The first technique, where the animal is both donor and the recipient, consists of connecting the grafts' vascular anastomoses to the mesenteric vessel bed. In the second technique, one animal is the graft donor and the other is the recipient, with revascularization to the central vessel bed (subrenal inferior vena cava and aorta). Techniques of restoring digestive tract continuity and creation of diagnostic "chimney ileostomy" were identical in both groups. All experimental animals were monitored clinically regularly basis as per protocol (weight, temperature, stoma appearance, output and nature of stools). Blood and biopsy samples were obtained on days 0, 3, 5, 7, 10, 20, and 30. Overall, 43 transplant procedures were performed. The first group included 18 transplants and 66.7% animals had v...
Bratislavské lekárske listy, 2007
Clinical success of small bowel transplantation depends on quality of the preservation small bowe... more Clinical success of small bowel transplantation depends on quality of the preservation small bowel graft which is notoriously sensitive to ischemia. There is still no general agreement as to which segment of the small bowel is preferred (jejunum or ileum) for clinical use. In our study, using a light microscopy and concentrations of tissue serotonin-positive cells, we tried to identify a part of the human intestine, which is more resistant to preservation injury sustained by HTK preservation solution with 1-24 hr of cold ischemia. Statistical analysis of both parameters did not reveal any significant differences between the jejunum and ileum. According to our data, there is no difference between jejunal and ileal grafts in susceptibility to ischemic injury due to cold ischemia within 24 hours when using HTK preservation solution. A significant difference was observed in histological pictures only after 12-hour of cold ischemia in both groups (jejunum and ileum) (Fig. 2, Ref. 11). Fu...
Transplantation Proceedings, 2013
The diagnosis of sepsis is difficult in immunocompromised patients owing to their modified respon... more The diagnosis of sepsis is difficult in immunocompromised patients owing to their modified response to infection. Our experiment in minipigs was designed to compare responses to sepsis between experimental groups of septic minipigs with and without immunosuppression. Minipigs with identical baseline parameters were randomized into 3 groups: Sepsis (n = 10); immunosuppression (n = 11), including cyclosporine, methylprednisolone, and mycophenolate mofetil treatment before surgery, and a sham group (n = 6). Sepsis was induced by cecal ligation and puncture (CLP). We recorded selected clinical and laboratory parameters up to 24 hours postoperatively. All CLP animals developed septic shock with a febrile response, tachycardia, and hypotension requiring noradrenaline administration. The hemodynamic responses to sepsis in septic groups with and without immunosuppression were similar. Noradrenaline infusion was started on average later in the immunosuppression than in the group without immunosuppression; however, the difference was not significant. The kinetics of the plasma levels of most selected cytokines and C-reactive protein were similar in both septic groups. At 10 hours after surgery, the immunosuppression group showed significantly lower interleukin (IL)-6 levels compared with the sepsis group. At 19, 22, and 25 hours after surgery immunosuppressed animals displayed significantly greater increases in IL-10 levels compared with the cohort without immunosuppression. CLP is a simple, reproducible model of sepsis in minipigs. All CLP animals developed sepsis within 24 hours on average. Significant differences in IL-6 and IL-10 plasma levels were recorded between septic animals with versus without immunosuppression.
Journal of Chromatography B, 2008
Unconjugated bilirubin (UCB) exhibits potent antioxidant and cytoprotective properties, but cause... more Unconjugated bilirubin (UCB) exhibits potent antioxidant and cytoprotective properties, but causes apoptosis and cytotoxicity at pathologically elevated concentrations. Accurate measurement of UCB concentrations in cells, fluids and tissues is needed to evaluate its role in redox regulation, prevention of atherosclerotic and malignant diseases, and bilirubin encephalopathy. In the present study, we developed and validated a highly sensitive method for tissue UCB determinations. UCB was extracted from rat organs with chloroform/methanol/hexane at pH 6.2 and then partitioned into a minute volume of alkaline buffer that was subjected to HPLC using an octyl reverse phase (RP) column. Addition of mesobilirubin as an internal standard corrected for losses of UCB during extraction. Recoveries averaged 75 ± 5%. The detection limit was 10 pmol UCB/g wet tissue. Variance was ±2.5%. When used to measure UCB concentrations in tissues of jaundiced Gunn rats, this procedure yielded UCB levels directly comparable to published methods, and accurately determined very low tissue bilirubin concentrations (≤40 pmol UCB/g tissue) in non-jaundiced rats.
International Journal of Cancer, 2007
Statins are widely used for the treatment of hypercholesterolemia. However, their inhibitory acti... more Statins are widely used for the treatment of hypercholesterolemia. However, their inhibitory action on HMG-CoA reductase also results in the depletion of intermediate biosynthetic products, which importantly contribute to cell proliferation. The aim of the present study was to compare the effects of the individual commercially available statins on experimental pancreatic cancer. The in vitro effects of individual statins (pravastatin, atorvastatin, simvastatin, lovastatin, cerivastatin, rosuvastatin and fluvastatin) on the viability of human pancreatic cancer were evaluated in CAPAN-2, BxPc-3 and MiaPaCa-2 cell lines. The in vivo experiments were performed on nude mice xenotransplanted with CAPAN-2 cells. The mice received oral treatments either with a placebo, or with the statins mentioned earlier in a daily dose corresponding to a hypocholesterolemic dose in humans. The effect of these statins on the intracellular Ras protein, trafficking in MiaPaCa-2 transfected cells, was also investigated. Substantial differences in the tumor-suppressive effects of all statins were detected in both in vitro and in vivo experiments. While simvastatin exerted the highest tumor-suppressive effects in vitro, rosuvastatin (p = 0.002), cerivastatin (p = 0.002) and fluvastatin (p = 0.009) were the most potent compounds in an animal model. All statins (except pravastatin) inhibited intracellular Ras protein translocation. In summary, substantial tumor-suppressive effects of various statins on the progression of experimental pancreatic adenocarcinoma were demonstrated, with marked differences among individual statins. These results support greatly the potential of statins for the chemoadjuvant treatment of pancreatic cancer.
Acta Veterinaria Brno, 2010
An experiment was carried out on pigs during preparation of a clinical project of intestinal tran... more An experiment was carried out on pigs during preparation of a clinical project of intestinal transplantation. The objective of this study was to find the best surgical technique of vascular and intestinal anastomosis in different experimental settings (animals with and without immunosuppression) which could have a major impact on everyday veterinary practice. Transplantation was performed in 43 pigs. In the surgical part of the experiment we examined the most suitable surgical technique of vascular anastomosis. A running continuous single-layer seromuscular suture was used for all types of intestinal anastomoses. With regard to vascular anastomosis, the animals were divided into two groups. In group 1 (n = 18) one animal was both donor and recipient of the intestinal graft. Anastomoses were constructed to the mesenteric vessel bed. In group 2 (n = 25), one animal was the graft donor and another was the graft recipient, with revascularization to the central vessel bed. In the second ...
Rozhledy v chirurgii : měsíčník Československé chirurgické společnosti, 2007
The small intestine transplantations represent a logical alternative to final total parenteral nu... more The small intestine transplantations represent a logical alternative to final total parenteral nutrition in patients with chronic intestinal failures. It is considered a life- saving procedure in patients with intestinal failure, where standard treatment procedures cannot be further implemented. Perfect harvesting technique is very important for succesful clinical small bowel transplantation. The authors studied the surgical view of the small intestinal transplant harvesting and monitored differences in ischemic injuries to jejunum and ileum depending on duration of the cold ischemia time. The study is one of the inital works in the planned intestinal transplantation clinical programme.
Journal of Chromatography B, 2008
Unconjugated bilirubin (UCB) exhibits potent antioxidant and cytoprotective properties, but cause... more Unconjugated bilirubin (UCB) exhibits potent antioxidant and cytoprotective properties, but causes apoptosis and cytotoxicity at pathologically elevated concentrations. Accurate measurement of UCB concentrations in cells, fluids and tissues is needed to evaluate its role in redox regulation, prevention of atherosclerotic and malignant diseases, and bilirubin encephalopathy. In the present study, we developed and validated a highly sensitive method for tissue UCB determinations. UCB was extracted from rat organs with chloroform/methanol/hexane at pH 6.2 and then partitioned into a minute volume of alkaline buffer that was subjected to HPLC using an octyl reverse phase (RP) column. Addition of mesobilirubin as an internal standard corrected for losses of UCB during extraction. Recoveries averaged 75 ± 5%. The detection limit was 10 pmol UCB/g wet tissue. Variance was ±2.5%. When used to measure UCB concentrations in tissues of jaundiced Gunn rats, this procedure yielded UCB levels directly comparable to published methods, and accurately determined very low tissue bilirubin concentrations (≤40 pmol UCB/g tissue) in non-jaundiced rats.
International Journal of Cancer, 2007
Statins are widely used for the treatment of hypercholesterolemia. However, their inhibitory acti... more Statins are widely used for the treatment of hypercholesterolemia. However, their inhibitory action on HMG-CoA reductase also results in the depletion of intermediate biosynthetic products, which importantly contribute to cell proliferation. The aim of the present study was to compare the effects of the individual commercially available statins on experimental pancreatic cancer. The in vitro effects of individual statins (pravastatin, atorvastatin, simvastatin, lovastatin, cerivastatin, rosuvastatin and fluvastatin) on the viability of human pancreatic cancer were evaluated in CAPAN-2, BxPc-3 and MiaPaCa-2 cell lines. The in vivo experiments were performed on nude mice xenotransplanted with CAPAN-2 cells. The mice received oral treatments either with a placebo, or with the statins mentioned earlier in a daily dose corresponding to a hypocholesterolemic dose in humans. The effect of these statins on the intracellular Ras protein, trafficking in MiaPaCa-2 transfected cells, was also investigated. Substantial differences in the tumor-suppressive effects of all statins were detected in both in vitro and in vivo experiments. While simvastatin exerted the highest tumor-suppressive effects in vitro, rosuvastatin (p = 0.002), cerivastatin (p = 0.002) and fluvastatin (p = 0.009) were the most potent compounds in an animal model. All statins (except pravastatin) inhibited intracellular Ras protein translocation. In summary, substantial tumor-suppressive effects of various statins on the progression of experimental pancreatic adenocarcinoma were demonstrated, with marked differences among individual statins. These results support greatly the potential of statins for the chemoadjuvant treatment of pancreatic cancer.
Transplantation Proceedings, 2013
The diagnosis of sepsis is difficult in immunocompromised patients owing to their modified respon... more The diagnosis of sepsis is difficult in immunocompromised patients owing to their modified response to infection. Our experiment in minipigs was designed to compare responses to sepsis between experimental groups of septic minipigs with and without immunosuppression. Minipigs with identical baseline parameters were randomized into 3 groups: Sepsis (n = 10); immunosuppression (n = 11), including cyclosporine, methylprednisolone, and mycophenolate mofetil treatment before surgery, and a sham group (n = 6). Sepsis was induced by cecal ligation and puncture (CLP). We recorded selected clinical and laboratory parameters up to 24 hours postoperatively. All CLP animals developed septic shock with a febrile response, tachycardia, and hypotension requiring noradrenaline administration. The hemodynamic responses to sepsis in septic groups with and without immunosuppression were similar. Noradrenaline infusion was started on average later in the immunosuppression than in the group without immunosuppression; however, the difference was not significant. The kinetics of the plasma levels of most selected cytokines and C-reactive protein were similar in both septic groups. At 10 hours after surgery, the immunosuppression group showed significantly lower interleukin (IL)-6 levels compared with the sepsis group. At 19, 22, and 25 hours after surgery immunosuppressed animals displayed significantly greater increases in IL-10 levels compared with the cohort without immunosuppression. CLP is a simple, reproducible model of sepsis in minipigs. All CLP animals developed sepsis within 24 hours on average. Significant differences in IL-6 and IL-10 plasma levels were recorded between septic animals with versus without immunosuppression.
Transplantation Proceedings, 2016
Background. The aim of our study was to evaluate the rate of surgical complications, patient outc... more Background. The aim of our study was to evaluate the rate of surgical complications, patient outcomes, and impact on graft function in renal transplant recipients in whom cholecystectomy for acute cholecystitis was performed. Methods. We reviewed data on transplant patients from January 1, 2006, to December 31, 2013. The subgroup of patients who required subsequent cholecystectomy for acute cholecystitis was assessed, and their data were further analyzed. Results. Thirty-one patients who underwent cholecystectomy for acute cholecystitis after renal transplantation were included in the study. Clinical signs such as pain in the right upper quadrant, temperature >38 C, and elevation in bilirubin levels occurred in 20 (64.5%), 8 (25.8%), and 3 (9.7%) patients, respectively. Ultrasound signs of acute cholecystitis were present in 27 patients (87.1%). In terms of laboratory values, white blood cell counts >10 Â 10 9 / L occurred in 17 patients (54.8%), and C-reactive protein levels >40 mg/L were reported in 21 patients (67.7%). The conversion rate to open surgery was 32.3% (10 patients). In 13 cases, acalculous cholecystitis was present (41.9%). The average serum creatinine level 1 year after cholecystectomy had no statistically significant differences. One patient required temporary dialysis during the postoperative period (with subsequent graft recovery), and 1 graft was lost. Conclusions. Acute cholecystitis in kidney transplant recipients is a serious complication, with frequent difficulties related to evaluation and diagnosis. Because clinical signs could be very mild compared with severity of gallbladder affliction, there is little room if any for conservative treatment in these patients. We have not noticed adverse impact of acute cholecystitis on 1-year graft function.
Rozhledy v chirurgii : měsíčník Československé chirurgické společnosti, 2009
Transplantation of the small intestine is a standard treatment method in patients with small inte... more Transplantation of the small intestine is a standard treatment method in patients with small intestinal failures. The aim of this study was to master the surgical technique, optimalize immunosuppression regimes, diagnose acute cellular graft rejection based on cellular and humoral indicators. The authors performed a total of 43 transplantation procedures in pigs. The first, surgical part of the experiment was aimed at mastering two principal methods of vascular anastomosis- firstly, connecting the graft with mesenteric vessels (Group n1 = 18) and secondly, connecting the graft with the aorta and the inferior vena cava (Group n2 = 25). The second part of the experiment included assessment of rejection changes in various immunosuppression regimes. Only animals who did not die because of a technical failure of the procedure or due to internal reasons (n = 24) were assessed. The study animals were assigned to four groups (A (n = 3)--autotransplantation, without immunosuppresion; B (n = ...
Bratislavské lekárske listy, 2009
The technical aspects of a procedure are most important for the outcome of the experiment. This s... more The technical aspects of a procedure are most important for the outcome of the experiment. This study was designed to compare two techniques suitable for graft revascularization. The first technique, where the animal is both donor and the recipient, consists of connecting the grafts' vascular anastomoses to the mesenteric vessel bed. In the second technique, one animal is the graft donor and the other is the recipient, with revascularization to the central vessel bed (subrenal inferior vena cava and aorta). Techniques of restoring digestive tract continuity and creation of diagnostic "chimney ileostomy" were identical in both groups. All experimental animals were monitored clinically regularly basis as per protocol (weight, temperature, stoma appearance, output and nature of stools). Blood and biopsy samples were obtained on days 0, 3, 5, 7, 10, 20, and 30. Overall, 43 transplant procedures were performed. The first group included 18 transplants and 66.7% animals had v...
Bratislavské lekárske listy, 2007
Clinical success of small bowel transplantation depends on quality of the preservation small bowe... more Clinical success of small bowel transplantation depends on quality of the preservation small bowel graft which is notoriously sensitive to ischemia. There is still no general agreement as to which segment of the small bowel is preferred (jejunum or ileum) for clinical use. In our study, using a light microscopy and concentrations of tissue serotonin-positive cells, we tried to identify a part of the human intestine, which is more resistant to preservation injury sustained by HTK preservation solution with 1-24 hr of cold ischemia. Statistical analysis of both parameters did not reveal any significant differences between the jejunum and ileum. According to our data, there is no difference between jejunal and ileal grafts in susceptibility to ischemic injury due to cold ischemia within 24 hours when using HTK preservation solution. A significant difference was observed in histological pictures only after 12-hour of cold ischemia in both groups (jejunum and ileum) (Fig. 2, Ref. 11). Fu...
Transplantation Proceedings, 2013
The diagnosis of sepsis is difficult in immunocompromised patients owing to their modified respon... more The diagnosis of sepsis is difficult in immunocompromised patients owing to their modified response to infection. Our experiment in minipigs was designed to compare responses to sepsis between experimental groups of septic minipigs with and without immunosuppression. Minipigs with identical baseline parameters were randomized into 3 groups: Sepsis (n = 10); immunosuppression (n = 11), including cyclosporine, methylprednisolone, and mycophenolate mofetil treatment before surgery, and a sham group (n = 6). Sepsis was induced by cecal ligation and puncture (CLP). We recorded selected clinical and laboratory parameters up to 24 hours postoperatively. All CLP animals developed septic shock with a febrile response, tachycardia, and hypotension requiring noradrenaline administration. The hemodynamic responses to sepsis in septic groups with and without immunosuppression were similar. Noradrenaline infusion was started on average later in the immunosuppression than in the group without immunosuppression; however, the difference was not significant. The kinetics of the plasma levels of most selected cytokines and C-reactive protein were similar in both septic groups. At 10 hours after surgery, the immunosuppression group showed significantly lower interleukin (IL)-6 levels compared with the sepsis group. At 19, 22, and 25 hours after surgery immunosuppressed animals displayed significantly greater increases in IL-10 levels compared with the cohort without immunosuppression. CLP is a simple, reproducible model of sepsis in minipigs. All CLP animals developed sepsis within 24 hours on average. Significant differences in IL-6 and IL-10 plasma levels were recorded between septic animals with versus without immunosuppression.
Journal of Chromatography B, 2008
Unconjugated bilirubin (UCB) exhibits potent antioxidant and cytoprotective properties, but cause... more Unconjugated bilirubin (UCB) exhibits potent antioxidant and cytoprotective properties, but causes apoptosis and cytotoxicity at pathologically elevated concentrations. Accurate measurement of UCB concentrations in cells, fluids and tissues is needed to evaluate its role in redox regulation, prevention of atherosclerotic and malignant diseases, and bilirubin encephalopathy. In the present study, we developed and validated a highly sensitive method for tissue UCB determinations. UCB was extracted from rat organs with chloroform/methanol/hexane at pH 6.2 and then partitioned into a minute volume of alkaline buffer that was subjected to HPLC using an octyl reverse phase (RP) column. Addition of mesobilirubin as an internal standard corrected for losses of UCB during extraction. Recoveries averaged 75 ± 5%. The detection limit was 10 pmol UCB/g wet tissue. Variance was ±2.5%. When used to measure UCB concentrations in tissues of jaundiced Gunn rats, this procedure yielded UCB levels directly comparable to published methods, and accurately determined very low tissue bilirubin concentrations (≤40 pmol UCB/g tissue) in non-jaundiced rats.
International Journal of Cancer, 2007
Statins are widely used for the treatment of hypercholesterolemia. However, their inhibitory acti... more Statins are widely used for the treatment of hypercholesterolemia. However, their inhibitory action on HMG-CoA reductase also results in the depletion of intermediate biosynthetic products, which importantly contribute to cell proliferation. The aim of the present study was to compare the effects of the individual commercially available statins on experimental pancreatic cancer. The in vitro effects of individual statins (pravastatin, atorvastatin, simvastatin, lovastatin, cerivastatin, rosuvastatin and fluvastatin) on the viability of human pancreatic cancer were evaluated in CAPAN-2, BxPc-3 and MiaPaCa-2 cell lines. The in vivo experiments were performed on nude mice xenotransplanted with CAPAN-2 cells. The mice received oral treatments either with a placebo, or with the statins mentioned earlier in a daily dose corresponding to a hypocholesterolemic dose in humans. The effect of these statins on the intracellular Ras protein, trafficking in MiaPaCa-2 transfected cells, was also investigated. Substantial differences in the tumor-suppressive effects of all statins were detected in both in vitro and in vivo experiments. While simvastatin exerted the highest tumor-suppressive effects in vitro, rosuvastatin (p = 0.002), cerivastatin (p = 0.002) and fluvastatin (p = 0.009) were the most potent compounds in an animal model. All statins (except pravastatin) inhibited intracellular Ras protein translocation. In summary, substantial tumor-suppressive effects of various statins on the progression of experimental pancreatic adenocarcinoma were demonstrated, with marked differences among individual statins. These results support greatly the potential of statins for the chemoadjuvant treatment of pancreatic cancer.
Acta Veterinaria Brno, 2010
An experiment was carried out on pigs during preparation of a clinical project of intestinal tran... more An experiment was carried out on pigs during preparation of a clinical project of intestinal transplantation. The objective of this study was to find the best surgical technique of vascular and intestinal anastomosis in different experimental settings (animals with and without immunosuppression) which could have a major impact on everyday veterinary practice. Transplantation was performed in 43 pigs. In the surgical part of the experiment we examined the most suitable surgical technique of vascular anastomosis. A running continuous single-layer seromuscular suture was used for all types of intestinal anastomoses. With regard to vascular anastomosis, the animals were divided into two groups. In group 1 (n = 18) one animal was both donor and recipient of the intestinal graft. Anastomoses were constructed to the mesenteric vessel bed. In group 2 (n = 25), one animal was the graft donor and another was the graft recipient, with revascularization to the central vessel bed. In the second ...
Rozhledy v chirurgii : měsíčník Československé chirurgické společnosti, 2007
The small intestine transplantations represent a logical alternative to final total parenteral nu... more The small intestine transplantations represent a logical alternative to final total parenteral nutrition in patients with chronic intestinal failures. It is considered a life- saving procedure in patients with intestinal failure, where standard treatment procedures cannot be further implemented. Perfect harvesting technique is very important for succesful clinical small bowel transplantation. The authors studied the surgical view of the small intestinal transplant harvesting and monitored differences in ischemic injuries to jejunum and ileum depending on duration of the cold ischemia time. The study is one of the inital works in the planned intestinal transplantation clinical programme.
Journal of Chromatography B, 2008
Unconjugated bilirubin (UCB) exhibits potent antioxidant and cytoprotective properties, but cause... more Unconjugated bilirubin (UCB) exhibits potent antioxidant and cytoprotective properties, but causes apoptosis and cytotoxicity at pathologically elevated concentrations. Accurate measurement of UCB concentrations in cells, fluids and tissues is needed to evaluate its role in redox regulation, prevention of atherosclerotic and malignant diseases, and bilirubin encephalopathy. In the present study, we developed and validated a highly sensitive method for tissue UCB determinations. UCB was extracted from rat organs with chloroform/methanol/hexane at pH 6.2 and then partitioned into a minute volume of alkaline buffer that was subjected to HPLC using an octyl reverse phase (RP) column. Addition of mesobilirubin as an internal standard corrected for losses of UCB during extraction. Recoveries averaged 75 ± 5%. The detection limit was 10 pmol UCB/g wet tissue. Variance was ±2.5%. When used to measure UCB concentrations in tissues of jaundiced Gunn rats, this procedure yielded UCB levels directly comparable to published methods, and accurately determined very low tissue bilirubin concentrations (≤40 pmol UCB/g tissue) in non-jaundiced rats.
International Journal of Cancer, 2007
Statins are widely used for the treatment of hypercholesterolemia. However, their inhibitory acti... more Statins are widely used for the treatment of hypercholesterolemia. However, their inhibitory action on HMG-CoA reductase also results in the depletion of intermediate biosynthetic products, which importantly contribute to cell proliferation. The aim of the present study was to compare the effects of the individual commercially available statins on experimental pancreatic cancer. The in vitro effects of individual statins (pravastatin, atorvastatin, simvastatin, lovastatin, cerivastatin, rosuvastatin and fluvastatin) on the viability of human pancreatic cancer were evaluated in CAPAN-2, BxPc-3 and MiaPaCa-2 cell lines. The in vivo experiments were performed on nude mice xenotransplanted with CAPAN-2 cells. The mice received oral treatments either with a placebo, or with the statins mentioned earlier in a daily dose corresponding to a hypocholesterolemic dose in humans. The effect of these statins on the intracellular Ras protein, trafficking in MiaPaCa-2 transfected cells, was also investigated. Substantial differences in the tumor-suppressive effects of all statins were detected in both in vitro and in vivo experiments. While simvastatin exerted the highest tumor-suppressive effects in vitro, rosuvastatin (p = 0.002), cerivastatin (p = 0.002) and fluvastatin (p = 0.009) were the most potent compounds in an animal model. All statins (except pravastatin) inhibited intracellular Ras protein translocation. In summary, substantial tumor-suppressive effects of various statins on the progression of experimental pancreatic adenocarcinoma were demonstrated, with marked differences among individual statins. These results support greatly the potential of statins for the chemoadjuvant treatment of pancreatic cancer.
Transplantation Proceedings, 2013
The diagnosis of sepsis is difficult in immunocompromised patients owing to their modified respon... more The diagnosis of sepsis is difficult in immunocompromised patients owing to their modified response to infection. Our experiment in minipigs was designed to compare responses to sepsis between experimental groups of septic minipigs with and without immunosuppression. Minipigs with identical baseline parameters were randomized into 3 groups: Sepsis (n = 10); immunosuppression (n = 11), including cyclosporine, methylprednisolone, and mycophenolate mofetil treatment before surgery, and a sham group (n = 6). Sepsis was induced by cecal ligation and puncture (CLP). We recorded selected clinical and laboratory parameters up to 24 hours postoperatively. All CLP animals developed septic shock with a febrile response, tachycardia, and hypotension requiring noradrenaline administration. The hemodynamic responses to sepsis in septic groups with and without immunosuppression were similar. Noradrenaline infusion was started on average later in the immunosuppression than in the group without immunosuppression; however, the difference was not significant. The kinetics of the plasma levels of most selected cytokines and C-reactive protein were similar in both septic groups. At 10 hours after surgery, the immunosuppression group showed significantly lower interleukin (IL)-6 levels compared with the sepsis group. At 19, 22, and 25 hours after surgery immunosuppressed animals displayed significantly greater increases in IL-10 levels compared with the cohort without immunosuppression. CLP is a simple, reproducible model of sepsis in minipigs. All CLP animals developed sepsis within 24 hours on average. Significant differences in IL-6 and IL-10 plasma levels were recorded between septic animals with versus without immunosuppression.