Evidence of the existence of a soluble mediator of cold preservation injury.

A study was designed to determine whether soluble mediators of injury are released during cold preservation. A first set of livers consisting of three groups was stored in cold Euro-Collins solution. These were a control group stored for 10 min (group 1), an experimental group stored for 16 hr (group 2), and an "antiprotease" group to which a cocktail of antiproteases had been added, which was also stored for 16 hr (group 3). The preservation solution in these livers was washed out at the end of preservation, and this effluent was concentrated and infused into a second set of livers that were all cold-stored for 4 hr. Then, the second-set livers were either perfused-fixed at 4 degrees C with universal fixative or reperfused at 37 degrees C for 180 min in the isolated perfused rat liver (IPRL). Morphometric assessment of sinusoidal lining cells (SLC) on light and electron microscopy showed an increased degree of microcirculatory injury in livers preserved with concentrates from livers of the experimental group. On light microscopy, only 2.2 +/- 0.4% (mean +/- SD) of the SLC had a normal flattened morphology compared with 11.9 +/- 2.0% in the control group, and 10.7 +/- 2.3% of the SLC appeared completely detached from the underlying hepatocytes compared with 2.6 +/- 0.8% in the control group, the differences being statistically significant (P < 0.05). This injury was prevented by the addition of antiproteases to EC solution. Similar results were obtained in the IPRL model, in which a number of typical changes related to cold preservation injury were noted in livers preserved with concentrates from the experimental group. Compared with controls, livers preserved with concentrates from the experimental group had early and significant alterations in markers of microcirculatory injury, including a reduction in portal flow and an increase in creatinine kinase-BB isoenzyme release, followed by an increase in perfusate transaminases, LDH, and a decrease in bile production. Again the injuries were largely prevented by the addition of antiproteases. There were no differences among groups in the degree of white cell and platelet adherence during reperfusion. Experiments using UW solution showed similar results, indicating that the soluble mediator(s) is not specific for a particular preservation solution. These observations are consistent with the hypothesis that soluble mediators are produced during the hypothermic period, and are responsible for a significant part of cold preservation injury, and that proteolytic reactions are involved in this type of injury.

Innominate artery interposition graft simplifies the portal venous drainage method of pancreas transplantation.

Pancreas transplantation utilizing portal venous and enteric exocrine drainage has potential benefits over the standard systemic venous and bladder exocrine drainage method. Unfortunately, technical difficulties are often experienced with the arterial anastomosis after the portal venous anastomosis is completed. We have found that the addition of an innominate artery interposition graft has greatly simplified the procedure.

Correlation of donor nutritional status with sinusoidal lining cell viability and liver function in the rat.

We have demonstrated that the sinusoidal lining cell injury sustained by rat liver allografts during hypothermic storage is a critical determinant of graft viability. The present study was designed to examine the effect of donor nutritional status on hepatic microcirculation and graft function. Rat livers from four nutritional groups (group I, fasted; group II, fed; group III, intraperitoneal glucose; and group IV, fed plus intraperitoneal glucose) were excised and stored for 24 hr in Marshall's isotonic citrate solution. Then the livers were perfused under anoxic conditions with trypan blue. The percentage of nonviable SLC in each group was 26.7 +/- 8.1, 24.9 +/- 7.9, 17.6 +/- 6.9, and 5.9 +/- 1.9 in groups I, II, III, and IV respectively; i.e., there was a significant improvement in SLC viability with nutritional repletion in group IV. Electron microscopy was performed on livers from groups I and IV following 30-hr preservation in University of Wisconsin solution and after 16-hr preservation in Marshall's isotonic citrate solution. Biopsies were taken at the end of storage and after 1 hr of reperfusion at 37 degrees C. At the end of preservation group IV livers contained glycogen and had much more normal liver ultrastructure than group I livers. After reperfusion there was partial recovery of normal SLC morphology in both groups and depletion of glycogen in group IV. Liver function was studied on the isolated perfused rat liver system at 37 degrees C following 30-hr storage in UW solution. Transaminase release into the perfusate was significantly lower in nutritionally repleted livers than in livers from fasted animals. A significant reduction in perfusate platelet count occurred only in livers from fasted animals. The results show that nutritional repletion can reduce the injury of cold preservation to both hepatocytes and endothelial cells and improve liver function in the postpreservation period.

Effect of cold preservation on lymphocyte adherence in the perfused rat liver.

A study was designed to determine if cold preservation induces an increase in lymphocyte adherence to liver sinusoids on reperfusion. Rat livers were stored at 1 degree C in University of Wisconsin solution for 45 min, 8 hr, or 30 hr, and then reperfused for 90 min at 37 degrees C in an isolated perfused rat liver apparatus. Just prior to reperfusion, isogeneic rat lymphocytes prepared on a Ficoll-Paque gradient were added to the perfusate. In some studies lymphocytes were labeled with a fluorescent lipophilic membrane marker. There was no change in the number of circulating lymphocytes in an anhepatic circuit. When livers were present in the circuit, lymphocytes were lost from the perfusate into the liver in all studies, with the most rapid decrease occurring within 10 min of reperfusion. The length of preservation had a marked and statistically significant effect on the rate of disappearance of lymphocytes from the perfusate. Reduction by 50% of the number of lymphocytes infused did not affect the results when expressed as percent lymphocytes remaining in perfusate. To exclude the possibility that the loss of lymphocytes into the liver was due to a damaged subpopulation of lymphocytes, two livers stored 3 for 45 min were put into the circuit in sequence. The percent reduction in cells due to exposure to a second liver was not significantly different from that observed when cells were exposed only to a single liver. Histological studies showed fluorescence-labeled lymphocytes adherent in sinusoids, and the number of labeled cells was directly related to the length of preservation. Cold preservation induces an increase in lymphocyte adherence in the reperfused liver, which might be important in graft malfunction and rejection.

Proposed classification of complications of surgery with examples of utility in cholecystectomy.

Lack of uniform reporting of negative outcomes makes interpretation of surgical literature difficult. We attempt to define and classify negative outcomes by differentiating complications, sequelae, and failures. Complications and sequelae result from procedures, adding new problems to the underlying disease. However, complications are unexpected events not intrinsic to the procedure, whereas sequelae are inherent to the procedure. Failures are events in which the purpose of the procedure is not fulfilled. We propose a classification of complications based on four grades: Grade I complications are alterations from the ideal postoperative course, non-life-threatening, and with no lasting disability. Complications of this grade necessitate only bedside procedures and do not significantly extend hospital stay. Grade II complications are potentially life-threatening but without residual disability. Within grade II complications a subdivision is made according to the requirement for invasive procedures. Grade III complications are those with residual disability, including organ resection or persistence of life-threatening conditions. Finally, grade IV complications are deaths as a result of complications. To illustrate the relevance of the classification, we reviewed 650 cases of elective cholecystectomy. Risk factors for development of complications were determined, and the classification was also used to analyze the value of a modified APACHE II as a preoperative prognostic score. Both supported the relevance of the proposed classification. The advantages of such a classification are (1) increased uniformity in reporting results, (2) the ability to compare results of two distinct time periods in a single center, (3) the ability to compare results of surgery between different centers, (4) the ability to compare results of surgical versus nonsurgical measures, (5) the ability to perform adequate metaanalysis, (6) the ability to identify objective preoperative risk factors, and (7) the ability to establish preoperative prognostic scores.

Risk factors in elective laparoscopic cholecystectomy for conversion to open cholecystectomy.

BACKGROUND: Most cholecystectomies can be performed using a laparoscopic approach. However, 3 to 10 percent of laparoscopic cholecystectomies (LC) must be converted to open cholecystectomies (OC) and preoperative factors that predict risk for conversion are still not defined. STUDY DESIGN: Preoperative and intraoperative data were collected and analyzed from 628 patients who were scheduled for elective LC by two surgeons in an academic institution. Logistic regression was performed on data from two groups of patients: LC completed, 596 patients (95 percent) and LC converted, 32 patients (5 percent). RESULTS: Elective LC was accomplished with no common bile duct injuries, low morbidity rate (7.3 percent), and zero mortality rate. Both patient and surgeon factors predicted conversion from LC to OC. Older patients (65 years of age or older, (p < 0.01), males (p < 0.01), and patients with multiple attacks (ten or more) of biliary colic (p < 0.01), or a documented history of acute cholecystitis (p < 0.01) had a greater risk for conversion. Both surgeons had higher rates of conversion (p < 0.05) during the learning phase (fewer than 50 LC) of their experience. CONCLUSIONS: Risk factors for conversion may be predicted and awareness of these factors should help in the selection of the appropriate procedure for patients and in selection of cases for resident training.

Early laparoscopic cholecystectomy for acute cholecystitis: a safe procedure.

Acute cholecystitis is increasingly managed by laparoscopic cholecystectomy. Some reports have shown conversion and complication rates that are increased in comparison to elective laparoscopic cholecystectomy. This study reviews the combined experience of two hospitals where the intention was to perform early laparoscopic cholecystectomy for acute cholecystitis. A total of 152 cases of laparoscopic cholecystectomy for acute cholecystitis (evidence of acute inflammation clinically and pathologically) were identified. Conversion to open cholecystectomy was required in 14 cases (9%) in the total series. Laparoscopic cholecystectomy was performed within 2 days of admission in 76% (115 of 152) of patients. Conversion was significantly less likely in patients undergoing laparoscopic cholecystectomy within 2 days of admission (4 of 115) compared to those undergoing surgery beyond 2 days (10 of 37; P<0.0001). Eleven patients (7%) had postoperative complications; however, there were no cases of injury to the biliary system and no perioperative deaths. This series shows that laparoscopic cholecystectomy can be performed safely in patients with acute cholecystitis and suggests that early laparoscopic cholecystectomy is preferable to delaying surgery. Although the conversion rate to open surgery is higher than for elective cholecystectomy, the majority of patients (91%) still derive the well-recognized benefits of laparoscopic cholecystectomy. Early laparoscopic cholecystectomy is an acceptable approach to acute cholecystitis for the experienced laparoscopic surgeon.

Familial segregation in the occurrence and severity of periampullary neoplasms in familial adenomatous polyposis.

BACKGROUND: Familial adenomatous polyposis (FAP) patients often develop periampullary adenomas that may progress to periampullary cancer, a common cause of death in this population. The risk of periampullary cancer in FAP is unclear, and variables that predict the occurrence and severity of periampullary tumors are not well understood. The specific aim of this study was to determine whether the risk of periampullary neoplasia segregates in specific FAP families. MATERIALS AND METHODS: A total of 144 FAP patients from 74 families were either screened by gastroduodenoscopy (n = 132) or information was obtained from surgical or autopsy reports (n = 12). The severity of periampullary neoplasia was recorded for each patient and graded based on maximum polyp size and histology. Linear regression was used to determine the significance of a number of variables with respect to periampullary neoplasia. A blood sample was available from at least one member of 50 unrelated families and used to detect germline mutations in codons 686 through 1693 of the adenomatous polyposis coli (APC) gene. RESULTS: Statistically significant familial segregation was found for the incidence and severity of periampullary neoplasia (P < 0.02). Age was also a statistically significant variable (P < 0.01). No correlation was observed between specific APC germline mutations and periampullary polyp frequency and severity. CONCLUSIONS: The occurrence and severity of periampullary neoplasms in patients with FAP segregates in families. This familial association may be related to as yet unidentified modifier genes or perhaps common environmental factors. These results should prove useful in developing upper gastrointestinal screening protocols for FAP patients at risk for periampullary neoplasia.

The dynamics of glutathione species and ophthalmate concentrations in plasma from the VX2 rabbit model of secondary liver tumors.

PURPOSE: Available tumor markers have low sensitivity/specificity for the diagnosis of liver tumors. The present study was designed to evaluate the oxidoreductive status of the liver as surrogates of tumor subsistence and growth. METHODS: Glutathione species (GSH:GSSG), ophthalmate (OA) concentrations, and their turnover were measured in plasma of rabbits (n = 6) in their healthy state and in the state of tumor growth after implantation of the VX2 carcinoma in their liver. Tumors were allowed to grow for a period of 14 days when rabbits were sacrificed. Livers were removed and cysteine concentration was measured in liver tissue. RESULTS: Tumor growth was found in 100% of the rabbits. Concentration and labeling of GSH/GSSG were similar in experimental animals before and after tumor implantation and to sham animals. In contrast, OA concentration increased significantly in experimental animals after tumor implantation when compared to same animals prior to tumor implantation and to sham animals (P < .05). The concentration of cysteine, a precursor of GSH, was found to be significantly lower in the liver tissue adjacent to the tumor (P < .05). CONCLUSION: Disturbances in the oxidoreductive state of livers appear to be a surrogate of early tumor growth.

Disturbances in the glutathione/ophthalmate redox buffer system in the woodchuck model of hepatitis virus-induced hepatocellular carcinoma.

Purpose. The incidence of liver tumors is rising in USA. The purpose of this study was to evaluate liver oxido-reductive status in the presence of chronic liver disease and hepatocellular carcinoma (HCC). Methods. Glutathione species and ophthalmate (OA) concentrations were measured by LC-MS in processed plasma and red blood cells (RBC) from infected Woodchuck with hepatitis virus (WHV). Blood samples were obtained from: (i) infected animals with tumors (WHV+/HCC+), (ii) infected animals without tumors (WHV+/HCC-) and (iii) healthy animals (WHC-/HCC-). Results. The concentration of reduced glutathione (GSH) and the ratio GSH/GSG were lower in plasma from WHV+/HCC+ animals when compared to WHV+/HCC- and WHV-/HCC- (P < 0.01). In contrast, the concentration of oxidized glutathione (GSSG) was found to be higher in plasma from WHV+/HCC+ animals when compared to WHV+/HCC- and WHV-/HCC- (P < 0.01). The Glutathione species and its ratio from the RBC compartment were similar among all groups. OA concentration in both plasma and RBC was significantly higher from WHV+/HCC+ when compared to WHV+/HCC- and WHV-/HCC- (P < 0.01). Conclusions. Disturbances of the glutathione redox buffer system and higher concentrations of OA were found in the WCV+/HCC+ animal model. The role of these compounds as biomarkers of early tumor development in patients with end stage liver disease remains to be determined.

Complications of laparoscopic cholecystectomy.

Laparoscopic cholecystectomy is an extremely safe procedure. The most common serious complications are bile-duct injury and injury to vessels or bowel secondary to the insertion of trocars. These complications may be avoided with appropriate precautions. Indirect complications of surgery, such as pulmonary and cardiac complications, appear less common than with open cholecystectomy.

Effect of deoxycholate on immunoglobulin G concentration in bile: studies in humans and pigs.

Because an increase in biliary deoxycholate levels seems to be a risk factor for cholesterol gallstone formation, we determined the relationship between deoxycholate levels and levels of the pronucleating protein, immunoglobulin G (Ig) in human gallbladder bile. Patients with cholesterol gallstones had a higher concentration of biliary IgG compared with a pigmented stone group and control patients. This was associated with the simultaneous presence of two conditions in the cholesterol stone group, supersaturated bile and a high deoxycholate/cholate ratio. The other patient groups met only one of the two conditions. Next, animal studies were performed to determine if model biles mimicking the two conditions could affect IgG secretion by the gallbladder. Gallbladders were exposed in vivo and then in an Ussing chamber to model biles. The voltage clamp technique was used to monitor functional integrity of the preparation. Three different model biles were tested: (1) taurodeoxycholate (TDC), 80%; taurocholate (TC), 20%; and cholesterol saturation index (CSI), 1.2; (2) TDC, 20%; TC, 80%; and CSI, 1.2; and (3) TDC, 80%; TC, 20%; and CSI, 0.6. IgG concentrations became significantly higher in group 1 than in the other two groups. The concentration of mucous glycoprotein was also significantly greater in group 1 when compared with group 2. Plasma cells were increased in number in mucosal and submucosal layers in group 1. We conclude that cholesterol supersaturated model bile with high content of TDC induces gallbladder epithelial alterations, which increase the luminal concentration of IgG and mucous glycoprotein.

Diffusion of substances into human cholesterol gallstones.

BACKGROUND/AIMS: The possibility that substances penetrate gallstones and accumulate after stones have formed has not been examined. The specific aims of this study were to determine whether cholesterol gallstones are permeable and, if so, the effect of molecular weight on permeability. METHODS: Cholesterol gallstones from patients with multiple stones were collected during surgery and incubated in fluorescein solution or in solutions of fluoresceinated albumin or immunoglobulin (Ig) G. To determine egress from the stones, some stones were removed from the fluoresceinated solution after incubation and placed in bicarbonate buffer. The total area of the stone and the area of dye that had diffused into the calculi were calculated. To determine mass of penetrating IgG, stones were powdered after incubation, and IgG was measured by an enzyme-linked immunosorbent assay. RESULTS: All substances penetrated stones. Although all compounds tested diffused back out of the stones when they were replaced in buffer, proteins did so more slowly than fluorescein. CONCLUSIONS: Substances of different molecular weights can diffuse into and out of cholesterol gallstones. These findings must be taken into account when considering the role of substances contained in stones on stone formation and growth.

Laparoscopic versus open cholecystectomy: a matched study.

To determine the efficacy of laparoscopic cholecystectomy (LC) in the treatment of gallstone disease, all patients who underwent elective surgery for cholelithiasis during three consecutive periods (1989, 1990 and 1991) were studied. There were 121 patients in each period. All patients in the first period underwent open cholecystectomy (OC), whereas 70 (58%) patients underwent laparoscopic procedures in the second period (OC-LC). LC was the treatment of choice in the third period. Multiple factors, including sex, age, clinical and biochemical presentation of the disease and modified Apache II score were comparable among the three groups. The authors found significant differences in length of hospitalization (6.4 +/- 4.2 days in the OC group, 3.6 +/- 2.4 days in the OC-LC group and 2.4 +/- 1.7 days in the LC group, p < 0.01 when compared with the OC group) and return to work after surgery (5.8 +/- 2.8 weeks, 2.8 +/- 1.2 weeks and 1.3 +/- 1.8 weeks respectively, p < 0.01 when compared with the OC group). There was no significant difference in postoperative complications among the groups, but complications in the OC patients were more severe. Although operative time increased significantly after the introduction of LC, it returned to the range of OC after 36 procedures. Nine patients (5%) with LC required conversion to OC. Benefits of LC include a shorter hospital stay and a shorter recovery period. There were no deaths, very low morbidity, a substantial decrease in overall cost and a high degree of patient satisfaction with LC.

Multiple intrahepatic cholesterol stones.

The authors report a case of multiple intrahepatic cholesterol stones found in an asymptomatic patient who had undergone cholecystectomy 12 years before. Biochemical abnormalities and radiologic and pathologic findings are noted. The patient underwent liver resection with Roux-en-Y choledochojejunostomy and received ursodeoxycholic acid postoperatively. Recovery was uncomplicated, and the patient was well at 1-year follow-up. Intrahepatic cholesterol lithiasis is rare but can be diagnosed preoperatively. Treatment depends on the presence of complications and the distribution of the stones.

A method for sequential excision biopsies of rat liver in an isolated perfused system.

We describe a simple method of performing sequential excision biopsies during liver reperfusion in the isolated perfused rat liver. After hepatectomy, four ligatures (5.0 silk) tied with a slip knot are placed around the pedicles of: (1) the inferior and (2) the superior parts of the caudate lobe, as well as (3) the inferior and (4) the superior parts of the right lateral lobe. At the time of biopsy, the prepared 5.0 silk ties are tightened with sufficient force to occlude the vascular pedicle, preventing leakage of circulating perfusate. The procedure provides four biopsies of more than 350 mg each without alteration of perfusate transaminases and tissue ATP contents. The total tissue removed by this method comprises 20-25% of the whole liver weight.

Superior mesenteric vein thrombosis after the Whipple procedure: an aggressive, combined treatment approach.

It is now recognized that occlusion of the mesenteric veins not only may complicate a number of disease processes but may occur as a life-threatening complication after abdominal surgery. A 32-year-old woman had mesenteric venous thrombosis after resection of a duodenal inflammatory pseudotumour by pancreatoduodenectomy. She recovered fully after treatment, which consisted of thrombectomy, flushing with urokinase and intravenous administration of heparin. Papaverine infused for 4 days substantially improved bowel viability. Current concepts in mesenteric vein occlusion and the principles of clinical management are reviewed.

Changes in bilirubin pigments secreted in bile after liver transplantation.

The species of bile pigments secreted in T-tube fistula bile after liver transplantation were ascertained by high-performance liquid chromatography in 15 patients for 10 days after liver transplant. Nine glycosidic conjugates and unconjugated bilirubin were resolved by the analytical procedure. The principal pigments in bile and their proportions in normal patients were the following: bilirubin diglucuronide = 83.0% +/- 3.1% (S.D.); bilirubin monoglucuronide = 9.7% +/- 1.4% (S.D.); bilirubin monoglucuronide monoglucoside = 4.0% +/- 2.8% (S.D.); and bilirubin monoglucuronide monoxyloside = 1.5% +/- 1.8% (S.D.). All of the other possible glucuronide, glucose and xylose monoconjugates and diconjugates and unconjugated bilirubin were also found, but each was normally less than 1% of the total. In 13 of the 15 transplant patients, a significant depression in proportions of bilirubin diglucuronide and elevation in proportions of bilirubin monoglucuronide were found after the transplant, with an accompanying but generally small increase in the proportions of the minor conjugates. In two patients with rejection of the transplant, the changes were of larger magnitude, with improvement occurring only with recovery from the rejection. In one of these patients, kidney failure was present, and in addition to the diglucuronide and monoglucuronide conjugates, diglucoside and monoglucoside monoxyloside conjugates were found in plasma. The underlying metabolic abnormalities are not clear but likely reflect underlying abnormal intracellular cofactor levels for conjugation. Glycogen depletion with reduction of UDP-glucuronate levels or reduced UDP-glucuronate formation from UDP-glucose, secondary to elevation of UDP-xylose, could potentially account for the changes in pigment excretion.

Lymphocyte adherence in the reperfused rat liver: mechanisms and effects.

Leukocyte adhesion may play a central role in the pathogenesis of preservation-reperfusion injury to liver grafts. We previously showed that lymphocyte adhesion to sinusoids is dependent on the length of cold ischemia. In the present study we examined the mechanisms of lymphocyte adherence after harvesting combined with a short and a long preservation time. The effects of lymphocyte adherence on liver function were also examined. Rat livers were stored at 1 degrees C in University of Wisconsin solution for 45 min or 30 hr and then reperfused at 37 degrees C in the isolated perfused rat liver with isogeneic lymphocytes in an asanguineous perfusate. The role of reactive oxygen intermediates was investigated with allopurinol, a vitamin E analog and ascorbate or superoxide dismutase and catalase. For us to determine the role of Kupffer cells, Kupffer cell blockade was produced by gadolinium chloride. Leukotriene B4 effects were examined with the lipooxygenase inhibitor, nordihydroguaiaretic acid. We evaluated the possible presence of mechanical obstruction by studying flow rates and the circulation of red blood cells. We examined the role of adhesion molecules by pretreating lymphocytes with trypsin or neuraminidase and by exposing livers to arabinogalactan. We investigated the effects of lymphocyte adhesion on liver function by comparing perfusate liver enzymes in livers reperfused with and without lymphocytes, with trypsinized lymphocytes and with an increased number of lymphocytes. Allopurinol significantly reduced hypoxanthine degradation, and nordihydroguaiaretic acid inhibited leukotriene B4 release into the perfusate. The ability of gadolinium chloride to inhibit Kupffer cells was shown by colloid carbon uptake. In livers harvested and preserved for 45 min, lymphocytes decreased about 40% during reperfusion. In livers preserved for 30 hr, the reduction was significantly greater (about 80%). Lymphocyte adherence was lessened in livers preserved for 45 min by all three of the reactive oxygen intermediate protectants and by gadolinium chloride. In contrast, neither reactive oxygen intermediate protectants nor gadolinium chloride reduced adherence in livers preserved for 30 hr. Nordihydroguaiaretic acid had no effect in livers preserved for either 45 min or 30 hr. Portal flow in livers preserved for 45 min and 30 hr was similar, suggesting an absence of mechanical obstruction, and this finding was supported by a complete absence of red cell trapping. Trypsinization of lymphocytes and exposure of livers to arabinogalactan significantly lessened lymphocyte adherence in livers preserved for 30 hr but not in those preserved for 45 min.(ABSTRACT TRUNCATED AT 400 WORDS)