Marko Lempinen

Serum Phospholipase A2 in Human Acute Pancreatitis

The main problem in the diagnosis of acute pancreatitis is the early detection of the fulminant forms. So far, there is no single laboratory test that affords an unequivocal measure of the severity of the disease. We have studied the serum phospholipase A2 concentrations of 66 consecutive patients with acute or chronic relapsing pancreatitis. Fifty-two patients had a mild spontaneously healing form of the disease, whereas eight patients developed a pseudocyst or an abscess. Six patients had operatively established haemorrhagic pancreatitis. The serum phospholipase A2 concentrations were significantly higher in the patients with haemorrhagic pancreatitis than in those with a spontaneously healing disease. Moreover, in the patients with a milder form of the disease the serum phospholipase A2 concentrations correlated with prognostic signs suggested by others. The results suggest that the assaying of serum phospholipase A2 might be a valuable tool in the diagnosis and follow-up study of patients with acute pancreatitis.

Chlorpromazine Treatment of Experimental Acute Fulminant Pancreatitis in Pigs

Acute pancreatitis was induced in 20 piglets. In the controls (10 piglets) no specific treatment was given, while in the experimental group the animals were treated with chlorpromazine, a potent phospholipase A2 inhibitor in vitro. Blood samples were taken from all animals for the assay of amylase, lipase, phospholipase A2, calcium, blood glucose and arterial blood gases both before the experiment and at various time intervals after its commencement. At autopsy, tissue specimens were taken from the pancreas, liver, lungs, heart and kidneys for histological studies. In the untreated animals there was a continuous rise in the phospholipase A2 levels, while the level decreased to zero on the second day in the treated animals (p less than 0.01, Students t test). Moreover, in the untreated animals there was a sharp drop in the arterial pO2, while this remained unchanged in the treated animals (p less than 0.1, Students t test). The mortality was significantly less (p less than 0.01, chi-square test) in the chlorpromazine-treated animals than in the controls. Serum phospholipase A2 levels correlated with the changes in pulmonary function and mortality. The results support the view that phospholipase A2 is of importance in the pathophysiology of acute pancreatitis.

High frequency of gastroduodenal cytomegalovirus infection in liver transplant patients.

The prevalence and significance of cytomegalovirus (CMV) detected in biopsy specimens from the gastroduodenal mucosa of liver transplant patients, patients with chronic or acute liver failure and immunocompetent patients with dyspeptic symptoms were evaluated. 80 liver transplant patients with upper gastrointestinal symptoms, 132 patients with chronic and 25 with acute liver failure, and 33 immunocompetent, dyspeptic patients underwent oesophagogastroduodenoscopies, with biopsies from the duodenum and stomach. CMV was demonstrated by immunohistochemistry in frozen sections, using a monoclonal antibody against CMV‐specific antigens (pp65 matrix protein), and in paraffin sections by a monoclonal antibody against delayed early protein (p52). 71% of the liver transplant patients, 45% of the patients with chronic liver disease, 20% with acute liver failure, and 45% of the immunocompetent, dyspeptic patients had CMV‐positive findings in the gastroduodenal mucosa (liver transplant patients vs other groups, p<0.01). Histopathological findings in CMV‐positive samples were focal inflammation, including increased inflammation of the lamina propria, infiltrating leukocytes intra‐epithelially, regenerative changes in the epithelial cells and inclusion bodies. In conclusion, CMV‐positive cells and inclusions are often found in the gastroduodenal mucosa of liver transplant patients, as well as in patients suffering from chronic liver disease or even in otherwise healthy patients with dyspeptic symptoms.

Xylocaine Treatment in Experimental Pancreatitis in Pigs

Experimental haemorrhagic pancreatitis was induced in 12 piglets by infusing Nataurocholate trypsin into the pancreatic duct with simultaneous intravenous secretin stimulation. Within some minutes after the infusion all animals developed severe pancreatitis accompanied by the production of bloody ascites. Unless given specific treatment the pigs died within 24 h. Of the animals treated with xylocaine infusion (50 microgram/kg/min for 24 h) one died within 24 h, one during the second day, and four lived for over a week, at which time they were killed. Although xylocaine treatment signficantly improved the survival of the animals, it did not seem to influence the local damage of the pancreatic tissue. Xylocaine has been shown to inhibit phospholipase-A in vitro. It is possible that xylocaine also acts in vivo by inhibiting phospholipase-A, thus preventing lethal tissue damages at an early stage of pancreatitis.

Prognostic value of serum MMP-8, -9 and TIMP-1 in patients with hepatocellular carcinoma

Abstract Aim. Prediction of prognosis in hepatocellular carcinoma (HCC) is difficult. The aim of this study was to evaluate the prognostic value of serum MMP-8, -9, -13, and TIMP-1 in patients with HCC. Methods. Pre-treatment serum samples from 134 patients with HCC were retrospectively analyzed. The serum concentration of MMP-8 was analyzed with immunofluorometric assay (IFMA), and those of MMP-9, MMP-13, and TIMP-1 were determined by enzyme-linked immunosorbent assays (ELISA). Clinical data were retrieved from patient records and survival data obtained from Statistics Finland. Results. The overall cumulative disease-specific survival was 69% at 1 year, 50% at 2 years, and 33% at 5 years. Kaplan–Meier overall survival analysis showed that patients with low concentrations of serum MMP-8 or TIMP-1 had a statistically significantly better overall survival than patients with high concentrations of serum MMP-8 or TIMP-1 (P = 0.013 and P = 0.003). Interestingly, the overall survival in patients with high MMP-9/TIMP-1 ratio was statistically significantly better than in those patients with low MMP-9/TIMP-1 ratio (P = 0.004). Conclusion. Our results suggest that serum MMP-8, TIMP-1, and the ratio of MMP-9/TIMP-1 might be useful adjuncts as predictors of prognosis in patients with HCC.

Prognostic significance of tumor-associated trypsin inhibitor (TATI) and human chorionic gonadotropin-β (hCGβ) in patients with hepatocellular carcinoma

Abstract Aim. Hepatocellular carcinoma (HCC) is the sixth most common cancer and the third most frequent cause of cancer death worldwide. The aim of this study was to evaluate the prognostic value of serum tumor-associated trypsin inhibitor (TATI) and the free β subunit of human chorionic gonadotropin (hCGβ) in patients with HCC. Methods. The serum concentrations of TATI and hCGβ were determined by time-resolved immunofluorometric assays (IFMA) in pretreatment serum samples from 144 patients with HCC. Clinical data were retrieved from patient records and survival data obtained from Statistics Finland. Results. The overall cumulative disease-specific survival was 69% at 1 year, 50% at 2 years and 33% at 5 years. Disease-specific median survival time was 26 months. The overall survival in patients with low serum concentrations of TATI or hCGβ was statistically significantly better than in patients with elevated concentrations (p = 0.003 and 0.003, respectively). In multivariate analysis, both serum TATI and serum hCGβ were independent prognostic markers. Conclusion. The results imply that elevated serum concentrations of TATI and hCGβ are predictors of adverse prognosis in patients with HCC and appear to be useful adjuncts in predicting prognosis in patients with HCC.

HHV-6B is frequently found in the gastrointestinal tract in kidney transplantation patients.

In immunosuppressed patients human herpesvirus 6 (HHV‐6) reactivations are common. The aim of the study was to determine to which extent HHV‐6 can be found in the gastrointestinal tract in kidney transplant recipients and in patients on chronic dialysis. The HHV‐6 and cytomegalovirus (CMV) examinations were performed on gastro duodenal and colon biopsy specimens obtained from 81 kidney transplant recipients and on 46 chronic dialysis patients. The HHV‐6 and CMV were demonstrated by immunohistochemistry detecting both HHV‐6A and HHV‐6B, and CMV‐specific antigens. The HHV‐6B‐positive cells, were found in gastroduodenal biopsy specimens from 34% of the transplant recipients and 28% of the patients on chronic dialysis, CMV‐positive cells were found in specimens from 53% of the transplant recipients and 28% of the patients on chronic dialysis. The HHV‐6B positive cells were found in the colonic mucosa specimens from 36% of the transplant recipients and 22% of the patients on chronic dialysis, CMV‐positive cells were found in specimens from 36% of the transplant recipients and 17% of the patients on chronic dialysis. The HHV‐6B positive cells were found equally often in the gastroduodenal as in the colorectal mucosa. The HHV‐6B positive cells as well as CMV positive cells were simultaneously found in every fifth of transplant recipients.

Pig-liver perfusion. Extracorporeal pig-liver perfusion in a patient with acute hepatic failure.

Three extracorporeal pig-liver perfusions on a patient, for the treatment of an acute hepatic failure, are described. The patient had massive postoperative necrosis of the liver, possibly due to halothane sensitization. There was an improvement in the patients condition after the perfusions, but she died a few days later of a pulmonary embolism. Comparison between the pre-perfusion biopsy and autopsy specimens showed that liver cell regeneration had occurred. Histological study of the pig livers from the first two perfusions showed an accumulation of granulocytes in the liver sinusoids. The third perfusion had to be stopped after 20 minutes, owing to an acute increase in flow resistance. In this case, the histological study revealed compression of the sinusoids by oedematous liver cells and congestion of the portal areas.

In Vitro Synthesis of Triglycerides and Cholesterol in Human Gallbladder Mucosa

Triglyceride and sterol synthesis was investigated in vitro in the gallbladder mucosa from control subjects and patients with acalculous cholesterolosis. The incorporation rate of 14C-acetate was 1.6 +/- 0.2 nmol/g/h into cholesterol (sum of squalene, methyl sterols, and cholesterol) and 5.7 +/- 0.8 nmol/g/h into triglycerides. The rates were significantly correlated with each other (r = 0.667). The conversion of 3H-mevalonate into cholesterol (49 +/- 10 nmol/g/h) and triglycerides (4.7 +/- 1.2 nmol/g/h) indicated a high activity in the postmevalonate cholesterol synthesis and an active shunt pathway of mevalonate metabolism. The synthesis rates of cholesterol, triglycerides, and sterol esters were closely interrelated, were unaltered in cholesterolosis, and were not correlated with the serum, biliary, and mucosal lipid concentrations. Thus, despite marked lipid accumulation the lipid synthesis proceeds effectively in the gallbladder mucosa with cholesterolosis.

Clostripain, the Missing Link in the Enzyme Blend for Efficient Human Islet Isolation

Background Effective digestive enzymes are crucial for successful islet isolation. Supplemental proteases are essential as they synergize with collagenase for effective pancreas digestion. The presence of tryptic-like activity has been implicated in efficient enzyme blends and the present study aimed to evaluate if addition of clostripain, an enzyme with tryptic-like activity, could improve efficacy of the islet isolation procedure. Methods Clostripain was added to the enzyme blend just before pancreas perfusion. Islets were isolated per standard method and numerous isolation parameters, islet quality control, and the number of isolations fulfilling standard transplantation criteria were evaluated. Two control organs per clostripain organ were chosen by blindly matching against body mass index, cold ischemia time, hemoglobin A1c, donor sex, and donor age. Results There were no differences in pancreas weight, dissection time, digestion time, harvest time, percent digested pancreas, or total pellet volume before islet purification between control or clostripain pancreases. Glucose-stimulated insulin release results were similar between groups. Total isolation islet equivalents, purified tissue volume and islet equivalents/g pancreas as well as fulfillment of transplantation criteria favored clostripain processed pancreases. Conclusions The addition of clostripain to the enzyme blend soundly improved islet yields and transplantation rates. It gently aided pancreas digestion and maintained proper islet functionality. The addition of clostripain to the enzyme blend has now been implemented into standard isolation protocols at the isolation centers in Uppsala and in Oslo.