Response to exogenous cholecystokinin of six human gastrointestinal cancers xenografted in nude mice.

Gastrointestinal hormones regulate growth of cancers as well as normal tissues. We investigated whether long-term cholecystokinin (CCK) administration might affect growth or metabolism of human tumors xenografted in nude mice. In each experiment, approximately 20 nude mice bearing subcutaneous xenografts of the particular cancer line being studied were used. Half received CCK and half received saline solution intraperitoneally twice daily for 14 days. Tumor volume and body weight were measured every 3 days. If the tumors produced marker substances, these were measured in nude mouse serum and also in the xenografts. Tumor growth was significantly retarded by CCK in two of the six cancers studied. In each case, DNA, RNA, and protein reflected tumor volumes. In one of these tumors (SLU 077), serum carcinoembryonic antigen (CEA) levels paralleled the tumor volumes. In another tumor (SLU 132), serum CEA levels and tumor immunolabeling for CEA and pancreatic oncofetal antigen increased in response to CCK administration, whereas tumor volumes did not. These findings suggest that exogenous highdose CCK altered the growth and metabolism in two of six human cancers studied.

Effect of chronically administered cholecystokinin on the nude mouse.

The nude mouse has been used to evaluate the effect of cholecystokinin (CCK) on xenografted tissues, but little is known about long-term actions of cholecystokinin on native organs in this animal. We investigated the impact of chronically administered synthetic cholecystokinin octapeptide on the nude mouse. Six groups of eight animals each received intraperitoneal injections twice daily for 14 days with diluent or a 4-log range of cholecystokinin. Overall health, behavior, and body weight were unaffected by this treatment. Among the seven organs examined at necropsy, pancreas alone showed a dose-related increase in weight. Pancreatic DNA content decreased with increasing dosages of CCK-8, while RNA content exhibited a biphasic response to CCK-8. The only histological abnormality occurred in the pancreas and was confined to the higher doses. These data indicate for the first time the action of CCK on the non-tumor-bearing nude mouse. Unlike other animal models, the nude mouse responds to cholecystokinin administration with pancreatic hypoplasia and hypertrophy, which is accompanied by pancreatitis at higher doses.

Effect of chronically administered pentagastrin on the nude mouse.

The nude mouse has been used to evaluate the effect of gastrin on xenografted tissues, but little is known about long-term actions of gastrin on native organs in this species. We investigated the impact of chronically administered synthetic pentagastrin on the nude mouse. Six groups of mice (eight animals each) received intraperitoneal injections twice daily for 14 days with saline or pentagastrin at 0.5, 5, 50, 500, or 5,000 micrograms/kg. Behavior, overall health, and body weight were unaffected by this treatment. Of the seven organs examined at necropsy, only the pancreas showed a weight gain in response to pentagastrin treatment, and this occurred only at the highest dose. Total DNA content of the pancreas decreased in a dose-related manner, indicating hypoplasia, whereas pancreatic RNA content increased, indicating hypertrophy. No effect on the stomach was observed. This work indicates that the nude mouse is less sensitive than other species to visceral growth regulation by pentagastrin, and that toxicity is low.

Effect of chronically-administered secretin on the nude mouse.

Secretin plays an important role in the growth regulation of certain cancers in vitro. The nude mouse is a suitable model for evaluation of the effects of this hormone on tumor xenografts in vivo, but little is known about long-term actions of secretin in this species. We investigated the impact of chronically administered synthetic porcine secretin in the nude mouse. Six groups of mice (eight animals each) received twice-daily intraperitoneal injections of saline or secretin at 0.5, 5, 50, 500, or 5,000 micrograms/kg for 14 days. Body weight and general health were unaffected by exogenous secretin, and no apparent behavioral effects were observed. Seven abdominal organs were examined at necropsy and all were histologically normal. The only organ that showed a weight change was the pancreas (13% decrease at the highest secretin dose). This was accompanied by decreases in DNA and RNA content, indicating pancreatic hypoplasia. Secretin administration caused changes in DNA and/or RNA content (but not protein content or weight) in liver, small bowel, cecum, and large bowel. No effect of secretin on stomach or kidney was observed. Our work demonstrates the safety of frequent injections of pharmacologic doses of secretin in this frail animal and suggests that the nude mouse is an appropriate model for the in vivo study of tumor growth regulation by secretin.

Diltiazem as an adjunct to cold blood potassium cardioplegia: a clinical assessment of dose and prospective randomization.

Diltiazem was evaluated as an adjunct to cold blood potassium cardioplegia in 63 patients undergoing elective coronary artery bypass grafting. The dual-phase study compared incrementally increasing doses (50, 100, and 150 micrograms/kg) of diltiazem using a single-blind, randomized schedule with an equivalent volume of placebo added to each of three infusions of cold (10 degrees C +/- 2 degrees C) blood containing potassium chloride at 25 mEq/L for the initial infusion (400 ml) and at 12 mEq/L for the next two infusions (300 ml each). Observations included a number of operative variables, creatine kinase (CK)-MB curves, two-dimensional echocardiography, and pulsed Doppler sonography before operation and on postoperative days 1 and 5. Pulmonary artery thermistor catheter responses were observed for 16 hours postoperatively, as were left ventricular micromanometer-tipped catheter responses in 7 patients. As the dose of diltiazem was increased, there was increasing time to atrioventricular node refunction (23.6 to 62.0 minutes). Diltiazem at 100 micrograms/kg (D-100) resulted in a significantly lower peak CK-MB activity than its placebo. Peak - dp/dt increased in treated patients and decreased in patients given the placebo. The cardiac index in D-100 patients was greater on the first postoperative day than preoperatively. The stroke index returned to the control level by the fifth postoperative day in D-50 and D-100 patients only, and it remained depressed in placebo patients. Although few benefits were realized from the addition of diltiazem to cold blood potassium cardioplegia, there was dose-related prolongation of the atrioventricular node recovery time, which required cardiac pacing and thus was associated with its attendant risks.

Reperfusion arrhythmias during coronary reperfusion therapy in man. Clinical and angiographic correlations.

We hypothesized that patients suffering acute myocardial infarction who have reperfusion arrhythmias (RPA) during intracoronary streptokinase infusion (ICSK) would have different clinical and angiographic characteristics and a larger infarction size than those who achieved reperfusion without RPA. Of 35 patients who received intracoronary streptokinase, 27 had successful reperfusion documented by angiography. Successful reperfusion was accompanied by RPA in eight patients and no RPA in 19 patients. RPA included episodes of ventricular tachycardia in one, idioventricular rhythm in four, junctional bradycardia in one, or AV block in two patients which occurred at the time of reperfusion. Myocardial infarction size was calculated using creatine kinase-MB (CK-MB) isoenzyme time activity curves using standard methods. The mean CK-MB g equivalents (eq) for those with RPA was 71 +/- 25 (+/- 1 SD) and for those with no RPA was 45 +/- 24 (p less than .04). In patients with RPA, ejection fraction rose 5 +/- 14 percentage points before discharge, but fell 10 +/- 13 points in those with RPA (p less than .03). There was no difference between groups in total dose of streptokinase, final degree of stenosis of the affected vessel, reocclusion rate, or time from onset of symptoms to reperfusion. We conclude that patients suffering acute myocardial infarction who have RPA during ICSK in most cases have a larger infarction site or a more "stunned myocardium," as indicated by greater CK-MB release and fall in ejection fraction which is not due to increased time of ischemia.

Quantitation of CK-MB release: diagnostic utility in coronary artery bypass grafting.

The quantitative release of creatine kinase (CK-MB) into the circulation of 97 patients receiving between three and five distal aorto-coronary bypass grafts was used to quantitate the minimal operative myocardial injury and to determine the diagnostic utility of this measurement in the detection of perioperative myocardial infarction. Independently read electrocardiography (ECG) was used to define infarction. The +/- SD confidence range for traumatized heart tissue based on 88 patients without infarction was 0-40.5 g equivalents. Six patients with perioperative myocardial infarction had values significantly above this range. Three patients with indeterminate ECG both released CK-MB significantly above the reference range and were clearly abnormal from a clinical standpoint.

Effect of chronically administered somatostatin on the nude mouse.

We investigated the impact of chronically administered synthetic cyclic somatostatin-14 on the nude mouse. Four groups of seven animals each received intraperitoneal injections twice daily for 14 days with diluent or 5, 50, or 500 mcg/kg somatostatin in hydrolyzed gelatin. Overall health and body weight were unaffected by this treatment. Among the seven organs examined at necropsy, kidney alone showed a dose-related gain in weight with increasing somatostatin dose. Renal RNA content also increased in response to somatostatin treatment, but liver DNA content and small bowel DNA content and protein content decreased. This work indicates that somatostatin affects metabolism of gastrointestinal tissues and provides evidence that renal growth can be regulated by somatostatin. Whether similar effects occur at a physiological somatostatin level is unknown, but the current findings suggest new therapeutic possibilities for this hormone.

Evaluation of the Beckman ASTRA enzyme system.

We have evaluated the ASTRA enzyme analyzer. Six enzymes can be analyzed individually or as part of a larger profile of chemistry tests. The system can analyze all six enzymes at a rate of 35-40 specimens per hour. Within-run and day-to-day precision were comparable to that of other enzyme analyzers. A useful feature of the ASTRA enzyme analyzer permits the automatic reassay of specimens having activities that exceed certain defined limits. A comparison of methods for the ASTRA enzyme analyzer with those used on the SMA 12/60 and the ABA-100 showed good correlation with minimal random error.

Assessment of the Corvac blood collection tube for drug specimen processing.

The Corvac integrated serum separator tube (TM Monoject Scientific) has been examined for use in processing specimens for drug analysis. This study was undertaken because the SST serum separator tube (Becton-Dickinson) was shown to cause decreases in drug levels of lidocaine, pentobarbital, and phenytoin, which were dependent upon blood volume and time of contact with the gel. Pools of donor blood were spiked with lidocaine or phenytoin and processed in Corvac tubes and red top tubes. Lidocaine levels were depressed by 5.2% when Corvac was used. Phenytoin levels were unchanged. The lidocaine depression was independent of blood volume, time of gel contact, or tube lot number. Paired patient specimens for quinidine, lidocaine, or theophylline analysis were obtained in Corvac and red top tubes. Lidocaine levels were depressed by 6.0% (p less than 0.01). Quinidine and theophylline levels were unchanged. All drug analyses were carried out using the Syva enzyme multiplied immunoassay technique. The Corvac tube has been shown to cause decreased levels of lidocaine but not of the other drugs tested. Corvac (silicone gel) results differ from SST (polyester gel) results because of the different construction of these tubes.

Intraoperative myocardial protection: a comparison of blood and asanguineous cardioplegia.

Cardiac arrest was achieved in 84 patients using asanguineous cardioplegia and in 97 patients using cold blood potassium cardioplegia. The patient groups were similar in age, sex ratio, and preoperative risk factors. Other than the cardioplegic solution used, the conduct of each operation was identical. There were no differences in mean total pump time (118 minutes for the asanguineous cardioplegia group versus 117 minutes for the cold blood cardioplegia group) or cross-clamp time (73.5 versus 70 minutes, respectively). However, the blood cardioplegia group had a greater number of distal anastomoses per patient (3.9 versus 3.7; p less than 0.05). Myocardial protection was assessed clinically and by serial electrocardiograms. Cellular integrity was determined by release of the myocardial isoenzyme of serum creatine kinase (CK-MB). Cellular morphology was studied in 6 randomly selected patients in each group by electron microscopic examination of left ventricular myocardial samples obtained before and after bypass. Three patients given blood cardioplegia and 5 given asanguineous cardioplegia required intraaortic balloon counterpulsation at termination of bypass. There were no ultrastructural changes in either group. Electrocardiographic changes (Minnesota code) occurred in 12 of 84 patients receiving asanguineous cardioplegia versus 12 of 97 patients receiving cold blood potassium cardioplegia. To maintain a satisfactory cardiac index (greater than 2.0 L/min/m2), 38 of 84 patients given asanguineous cardioplegia versus 25 of 97 patients given blood cardioplegia required inotropic support up to 24 hours postoperatively (p less than 0.05). Infarct size determined from CK-MB release was significantly greater (p less than 0.05) in patients given asanguineous cardioplegia (36.27 gm-equivalents) than in those given blood cardioplegia (26.7 gm-equivalents).(ABSTRACT TRUNCATED AT 250 WORDS)

Carcinoembryonic antigen in experimental rat gastrointestinal carcinoma.

Detection of gastric carcinoma in the experimental rat can be difficult. We investigated whether or not serological testing for carcinoembryonic antigen might aid in the diagnostic process. Twenty-five young adult male Wistar rats were studied; 15 were treated with the carcinogen N-methyl-N'-nitro-N-nitrosoguanidine (MNNG), and 10 received no treatment. A complete autopsy was done on the 22 animals (12 in MNNG group, 10 in control group) that completed the 37-week study. Serum obtained at autopsy was evaluated by radioimmunoassay for the presence of CEA-like immunoreactivity. Grossly normal stomach, as well as gastric and small bowel cancers, were also stained for CEA. Of the 12 autopsied animals in the MNNG group, eight had a total of 12 cancers (seven gastric and five proximal small bowel); in no animal was an elevated serum CEA level detected. Immunoreactive tissue CEA was demonstrable in normal stomach and in gastrointestinal cancers; the tumors usually stained less intensely than adjacent normal stomach and did not exhibit normal intracellular CEA distribution.

Drug-protein binding interferences caused by the plasticizer TBEP.

Tris (2-butoxyethyl) phosphate (TBEP) is a plasticizer used in the rubber stoppers of evacuated blood collection tubes which disrupts the binding of basic drugs to the carrier protein alpha-1-acid glycoprotein (AAG). When blood drawn for therapeutic monitoring is exposed to TBEP, drug-protein binding is disrupted leading to increased free drug which is taken into red blood cells, resulting in decreased serum drug levels. Units of donor blood were spiked with quinidine or lidocaine and then exposed under various conditions to stoppers with and without TBEP. Total drug levels were determined by enzyme immunoassay as were free drug levels after equilibrium dialysis. AAG levels were determined by rate nephelometry. TBEP lowered quinidine levels by 11% at 8.20 mumol/L and by 32% at 4.38 mumol/L; lidocaine levels were reduced by 10% at 32.39 mumol/L and by 18% at 12.74 mumol/L. Exposure to TBEP increased free lidocaine from 66.7% to 80.0% of total lidocaine in the serum. The TBEP effect is diminished when the AAG level is low.

Macro lactate dehydrogenase (LD) due to protein binding of LD isoenzyme 1.

Abnormal lactate dehydrogenase (LD) isoenzyme patterns apparently due to protein binding of LD-1 have been observed in a patient with hepatoma. The abnormal patterns were observed within 30 h of death but were preceded by normal LD isoenzyme patterns. Heat treatment of the abnormal specimens followed by addition of control serum reproduced the abnormal pattern. This is consistent with immunoglobulin binding of LD. Results such as those observed in this case could serve to confound the interpretation of LD isoenzyme analyses. The diagnostic significance of these results is not clear.

Macro-creatine kinase as an interference in CK isoenzyme determinations.

Creatine kinase (CK) isoenzyme determinations have been performed on a series of specimens containing macro-CK, an atypical isoenzyme variant seen in approximately 1% of patients. These specimens were obtained from 2 different patients. Isoenzyme determinations were made using four different methods: electrophoresis (Corning ACI), column chromatography (Dupont ACA), immunoinhibition (ESKACHEM), Smith Kline), and immunoprecipitation (Isomune-CK, Roche). In specimens containing macro-CK, electrophoresis showed no false-positive CK-MB determinations, and immunoprecipitation showed 1 in 8 false-positives, while column chromatography and immunoinhibition each showed 4 of 4 false positives. When utilizing these methods, the possibility of false-positive CK-MB determinations must be taken into consideration as a possible explanation for positive CK-MB results which do not agree with the clinical assessment of the patient.

Creatine kinase (CK) isoenzyme activities in cardiac surgery.

Creatine kinase (CK) isoenzymes were determined on 15 patients undergoing open heart surgery with bypass. The study was performed to examine the relationship between the hypothermia under which the surgery is conducted and the appearance of CK isoenzymes in the serum both during and after surgery. All patients showed dramatic rises in total CK activity commencing during surgery. Myocardial CK (CK-MB or CK-2) was seen in fourteen patients and brain CK (CK-BB or CK-1) was seen in ten patients. Peak activities of CK-BB did not coincide with peak activities of CK-MB. The serum elevations of CK-BB in these patients appear to arise from a mechanism different from that responsible for the elevations of CK-MB, and it is assumed that the former is due to intermittent disruptions in the perfusion and/or oxygenation of tissues rich in CK-BB. CK-MB elevations appear to be due directly to the surgical intervention. Hypothermia alone does not in itself appear to be solely responsible for elevations of CK-BB although it can not be completely excluded from playing some role in its production.

The detection of creatine kinase isoenzyme CK1 in serum.

The brain isoenzyme of creatine kinase (CK) (EC 2.7.3.2) has been identified in the serum of 21 of 950 patients tested. Samples were received over a period of about 20 months. Analysis was performed by electrophoresis on agarose gel with isoenzyme bands visualized by fluorescence after enzyme coupled reactions producing NADH. Specialized substrate preparations were used to rule out the possibility that observed fluorescence was due to enzymatic activity other than that of CK isoenzymes. Patients who were hypothermic and/or had undergone cardiopulmonary resuscitation accounted for 19 of the 21 patients showing the brain isoenzyme. The inciting mechanism for the release of CK1, into the circulation is not clearly understood.