Blood pressure effects of lower abdominal aortic surgery with particular reference to the use of morphine and droperidol in modifying the responses. Hypertensive responses during operations on the abdominal aorta are common. We have shown that hypertensive changes occur before cross-clamping of the aorta. We have also shown that droperidol modifies the hypertensive changes in operations on the lower aorta. Although further studies are required, our preliminary results show a higher rise of serum epinephrine and norepinephrine levels in one patient receiving morphine, as compared to one patient receiving the same dose per unit body weight of morphine plus droperidol. With maintenance of adequate pre-operative hydration and intra-operative fluid and blood replacement, declamping hypotension was transient in both groups of our patients.


The effect of total peripheral resistance on indicator-dilution curves from the central circulation of the dog. The influence of total peripheral resistance on the dispersion of indicator particles passing through the central circulation was studied in five dogs. The standard deviation of indocyanine green indicator-dilution curves provided a measure of dispersion. The peripheral vascular resistance of the dog was altered during the experiment by the intravenous administration of sodium nitroprusside and methoxamine. We calculated the area of each curve by fitting the first portion of the curve to a random walk function and the cardiac output and the total peripheral resistance were calculated in the usual manner. The mean, standard deviation and area of each of the curves were calculated from the least squares estimate of the random walk function. The relationship of the cardiac output, mean arterial pressure, and total peripheral resistance to the mean and the standard deviation were compared in a multiple stepwise linear regression. For each dog there was a definite linear relationship between total peripheral resistance and standard deviation (p less than 0.01). These results indicate that the dispersion of particles passing through the central circulation is directly related to the total peripheral resistance.


Lung water estimation using an external sensing catheter. A method for measuring pulmonary oedema (ETVL) is described using sodium ions as the intravascular indicator and heat as the extravascular indicator. The technique offers the advantage of virtually unlimited repeatability, relatively uncomplicated instrumentation and no blood loss. There is also the potential for differentiating between pulmonary oedema and pulmonary congestion and determining the efficacy of therapy. A sensing catheter to detect blood conductivity and temperature changes is required. Ideally the sensing catheter should be situated at a point just distal to the aortic valve. However, a commercially manufactured detecting catheter we had built proved troublesome to insert because of its relatively large diameter. Therefore, an external sensing catheter has been constructed to measure ETVL and the values obtained have been compared with those from an internal catheter. There was no difference between the internal and external catheter QT, but ETVL was consistently overestimated by 2.46 +/- 0.26 ml/kg. However, this value can be related to the internal value and can be used for clinical studies.


Foetal deterioration following thiopentone-nitrous oxide anaesthesia in the pregnant ewe. Six pregnant sheep were chronically prepared with indwelling catheters in maternal and foetal vessels and a flow probe around a maternal uterine artery. They were anaesthetized the following day with thiopentone and nitrous oxide (70 per cent)--oxygen (30 per cent) maintenance with tracheal intubation. Maternal uterine blood flow fell about 20 per cent following induction of anaesthesia. This resulted from uterine vasoconstriction which, in turn, probably resulted from maternal catecholamine release during light anaesthesia. The foetus in utero developed a mixed metabolic and respiratory acidosis and a fall in oxygen saturation. The possibility that light maternal anaesthesia increases rather than decreases neonatal depression is discussed.


Urine catecholamine excretion after large doses of fentanyl, fentanyl and diazepam and fentanyl, diazepam and pancuronium. The effects of fentanyl (0.5 mg/kg iv), fentanyl with diazepam (1 mg/kg iv) and fentanyl, diazepam and pancuronium (0.1 mg/kg iv) on heart rate (HR), mean arterial blood pressure (BP), cardiac output (QT), urine flow rate and urine epinephrine and norepinephrine excretion were determined in nine dogs. Fentanyl did not significantly change QT or BP but did reduce HR and urine flow rate (P less than 0.05). Urine epinephrine and norepinephrine excretion rates were signicantly increased by fentanyl (P less than 0.05). Diazepam caused no significant further changes in QT, BP or HR 30 minutes after administration, but urine epinephrine and norepinephrine excretion rates were reduced to control (pre-fentanyl) levels. Addition of pancuronium after fentanyl and diazepam increased urine flow rate to pre-fentanyl levels and elevated QT, BP and HR above controls but produced no significant change in urine epinephrine or norepinephrine excretion. These data suggest that fentanyl increases catecholamine blood levels and imply that the latter may be one mechanism by which cardiovascular dynamics are maintained stable during fentanyl anaesthesia. Our findings also demonstrate that cardiovascular stimulation after pancuronium is not associated with increased urinary catecholamine excretion.


Pulmonary lavage in the treatment of alveolar proteinosis. Alveolar proteinosis is a rare disease characterized by the accumulation of a proteinoseous material inside the alveolus. Various forms of treatment have been tried without much success. Pulmonary lavage has been found effective in bringing relief to patients as well as objective amelioration. This presentation describes the technique used and the problems encountered.


The effect of assisted ventilation on arterial carbon dioxide. The effect on the arterial carbon dioxide tension of assisting spontaneously breathing patients under halothane anaesthesia has been studied. A reduction in PaCO2 is usually seen, but it was reduced below 40 torr in only 30 per cent of cases, while a rise was recorded in 20 per cent.


The use of serum cholinesterase in succinylcholine apnoea. Fifteen patients demonstrating unexpected prolonged apnoea lasting several hours after succinylcholine have been treated by a new preparation of human serum cholinesterase. Adequate spontaneous respiration was re-established in an average period of ten minutes after the injection. In 12 patients biochemical genetic examinations confirmed the presence of an atypical serum cholinesterase. In three patients none of the usual variants were found. It is therefore supposed that other unknown variants of serum cholinesterase exist which cannot hydrolyze succinylcholine. The use of serum cholinesterase in succinylcholine apnoea provided considerable relief to both patient and anaesthetist.


An evaluation of the anaesthetic techniques used in an outpatient unit. Our follow-up of 250 gynaecology patients and 100 dental patients who had received anaesthesia for elective outpatient surgical procedures indicates: (1) The practice of outpatient anaesthesia in proper facilities with proper selection of patients appears to be safe. (2) There is widespread patient acceptance of surgery and anaesthesia on an outpatient basis. (3) Complications are frequent but minor. (4) Many of the complications may be minimized: (i) Adequate depth of anaesthesia preferably with a volatile agent will do away with awareness during operation. (ii) Methoxyflurane should be avoided to minimize late arousal. Volatile agents such as enflurane or halothane would seem to be preferable to intravenous agents. (iii) Post-fasciculation pain could be minimized by avoiding succinylcholine for short procedures like D & C and using adequate depth instead. For dental procedures requiring tracheal intubation, one could perhaps use non-depolarizing muscle relaxants, like pancuronium, with reversal at the end of the procedure. (5) Nausea, vomiting, dizziness and headache are complications that occur very frequently in all series reported and this is an area where more research is indicated.


The Bartlett-Edwards incentive spirometer: a preliminary assessment of its use in the prevention of atelectasis after cardio-pulmonary bypass. A preliminary assessment of the effectiveness of the Bartlett-Edwards incentive spirometer was made using it as part of the post-operative treatment of 34 patients after open-heart surgery. Its effects on atelectasis were assessed by measuring vital capacity, arterial oxygen tensions, clinical signs and radiological changes. It was found that vital capacity fell after surgery to 41.5 per cent of the pre-operative level, but this rose after the use of the incentive spirometer by an average of 15.5 per cent. Arterial oxygen tensions were unaltered by the use of the incentive spirometer. Temperature, pulse and respirations were of little help in the early diagnosis of atelectasis, but all patients who were still pyrexic four days after operation showed radiological evidence of atelectasis. The incidence of atelectasis demonstrated radiologically was total 84 per cent and plate 69 per cent, sub-segmental 34 per cent and segmental 9 per cent. The implications of these findings are discussed.


The action of diazoxide analogs on aortic reactivity in normotensive and spontaneously hypertensive rats. Diazoxide and four related 1,2,4-benzothiadiazines reduced the maximum responses to noradrenaline (NA) and KCl in aortic rings from normotensive and spontaneously hypertensive rats (SHR). These agents were more active against NA than against KCl responses, and more effective against either NA or KCl responses in aortae from hypertensive animals. Further studies with V (3-cyclopentenyl-6,7-dichloro-1,2,4-benzothiadiazine-1,1-dioxide), the most active member of the series, showed it to be significantly more effective against NA responses; when calcium was added to calcium-free solutions containing eith NA or KCl, V was more effective in inhibiting either response in SHR aortae and was about 100 times more effective in inhibiting the calcium-NA responses. These data are consistent with previous findings that vascular smooth muscle from normotensive and SHR differs in sensitivity to diazoxide. However, the site(s) at which this difference is exhibited remains to be elucidated.


The effects of enzymatic digestion on the elastic properties of isolated human cerebral arteries. Measurement of volume, pressure, and length were made on eight segments of human cerebral arteries perfused with chymotrypsin (CT) (EC 3.4.21.1) solution (pH = 7.8) for no more than 19 h, and on nine arterial segments perfused with combined enzyme (CT, trypsin (EC 3.4.21.4), elastase (EC 3.4.21.11)) solutions (pH = 7.8) for no more than 4 h. Circumferential tension-strain (and absolute radius) curves were obtained through the Law of Laplace (tension = pressure X radius). Initial and final elastances (tension/strain) were calculated after 0, 0.5, 1.0, 2.0, and 4.0 h of perfusion under the combined enzyme category, and after 0, 0.5, 1.0, 2.0, 4.0, 6.0, and 19.0 h of perfusion with CT. The initial elastance showed a significant increase (0.02 less than p less than 0.05) after about 6 h of perfusion. Increases in the final elastance became significant only after prolonged periods of perfusion with CT. Histological studies using light and scanning electron microscopy confirmed the removal of the elastic lamina as well as portions of the medial region. Fragmentation of the internal elastic lamina did not appear to affect the distensibility of major cerebral arteries in the 50- to 80-year-old age group.


Influence of infused prolactin on hormone binding to tissue slices. The influence of circulating prolactin on the binding of labeled hormone to tissue slices in vitro was investigated in rats bearing dimethylbenzanthracene-induced mammary tumors. Prolactin was infused into four animals over four 30-min periods to give serum levels ranging from 20 to 2000 ng/ml. Serial biopsies of liver and tumor were taken during infusion, and binding of labeled prolactin and insulin measured in tissue slices. Whereas insulin binding remained constant, prolactin binding was significantly depressed in both tissues when samples were obtained at serum prolactin levels greater than 300 ng/ml. This decrease was due to diminished numbers of available binding sites rather than a change in binding-site affinity for prolactin. The data indicate that under the conditions of this experiment, endogenous circulating prolactin may interfere with in vitro measurements of hormone uptake when serum levels excee 300 ng/ml.


Effects of reserpine on water, cation, and norepinephrine contents of cardiovascular tissues of normotensive dogs. Six adult male mongrel dogs were treated orally with reserpine (0.03 mg/kg) for 6 weeks, after which time the arterial tissue content of water, cations (sodium, potassium, magnesium, and calcium), and norepinephrine was determined and compared with those of six untreated dogs. Reserpine administration resulted in sustained decreases in the mean arterial blood pressure and heart rate, and the arterial tissues of the treated dogs contained less potassium, calcium, and magnesium, the last-named being affected most, but the sodium remained the same. Arterial norepinephrine content was significantly decreased. There was a slight but significant increase in plasma magnesium, without any noticeable changes in other plasma cations. The reduction in the arterial magnesium by reserpine may help to regulate vascular tone and (or) vascular reactivity.


Effects of estrogen and progesterone on pulsatile discharges of luteinizing hormone in the ovariectomized rat. The effects of both acute and chronic systemic administration of estrogen and progesterone on pulsatile discharges of luteinizing hormone were observed in ovariectomized rats. Intravenous injection of 10 microgram of 17beta-estradiol resulted in a gradual decrease in plasma luteinizing hormone (LH) concentrations but did not completely block the pulsatile discharges. Subcutaneous implantation of Silastic capsules containing 17beta-estradiol caused similar decreases; plasma LH concentrations decreased for the first 6 h but periodic increases continued, with nearly complete suppression of such pulsatile discharges by 24 h. Such inhibition was maintained for the 14-day course of the experiments. Progesterone alone, whether administered by Silastic capsule or by daily subcutaneous injection in oil, was without apparent effect on either plasma LH concentrations or temporal patterns of the hormone. Further, progesterone combined with 17beta-estradiol was little, if any, more effective in this regard than estradiol alone. These results indicate that estrogen is an effective feedback signal inhibiting pulsatile release of LH in the rat and that progesterone alone has little, if any, influence in this regard.


Can nonadrenergic inhibitory varicosities be identified structurally. The distribution of vesicles in axon varicosities was examined in rabbit jejunum and opossum esophagus. Circular muscle cells of both rabbit jejunum and opossum esophagus responded to stimulation of intrinsic nerves with inhibitory junction potential in nearly all cases. Inhibitory junction potentials were rare and small in longitudinal muscle cells of rabbit jejunum. There were few axon varicosities with a predominance of large opaque vesicles or large dense-cored vesicle in any muscle layer of rabbit or opossum or in the plexuses of rabbit jejunum. There were almost as high a proportion of such varicosities in longitudinal as in circular muscle of rabbit jejunum, and there were almost none in circular muscle of opossum esophagus. The distribution of vesicles in varicosities was found often to be heterogeneous and could account for the observed occurrence of a small proportion of varicosities with a predominance of large vesicles. It is suggested that there is no structural distinction in vertebrates between axons which are cholinergic and axons which are nonadrenergic inhibitory.


Oxygen-tension-dependent pulmonary vascular responses to vasoactive agents. There have been recent indications that oxygen may nonspecifically oppose pulmonary vasoconstriction induced by a few vasoactive agents. Therefore, we examined the effect of four inspired oxygen tensions on the pulmonary vascular responses to exogenous prostaglandin F2alpha (PGF2alpha), serotonin (5-HT), 2-methylhistamine (2-MeH)(an H1-receptor agonist), histamine (after H2-receptor blockade with metiamide), and prostaglandin E1 (PGE1) in anesthetized dogs. An oxygen tension dependency on the pulmonary vascular responses to these vasoactive agents was observed, with each agent exhibiting maximal responses at different ranges of oxygen tension. PGF2alpha and PGE1 were most effective during hypoxia, while 5-HT, histamine, and 2-MeH produced maximal responses during normoxia. A comparison of dose-response curves for PGF2alpha during breathing of two inspired oxygen tensions indicated a decreased sensitivity, but not decreased reactivity, with the higher oxygen tension. The action of variable oxygen tensions on pulmonary vascular responsiveness to vasoactive agents suggests another role for oxygen in the control of the pulmonary circulation. It is not clear if oxygen acts non-specifically on the vascular smooth muscle, or if it alters the metabolic mechanisms of vasoactive agent action.


Myocardial lesions in rats fed rapeseed oil. I. Influence of strain of rats. The influence of strain of rat on the development of myocardial lesions was investigated in an experiment which included two factors: strain (Wistar, Sherman, Chester Beatty (Hooded), and Sprague-Dawley from two sources designated Sprague-Dawley A, Sprague-Dawley C) and diet (5% corn oil, 20% corn oil, and 20% Brassica napus var. Zephyr rapeseed oil). Groups of 30 rats, housed 2 per cage, from each of three different strains of rats and groups of 10 rats, housed 2 per cage, from one strain of rats (Hooded) were fed semisynthetic diets containing the test oils for 16 weeks on an ad libitum basis. Rats of the Hooded strain consumed considerably less feed and grew at a significantly (P less than 0.01) slower rate than did rats of the Sprague-Dawley C strain, which in turn ate less and grew significantly (P less than 0.01) slower than rats of the other three strains. No marked differences were observed in the fatty acid composition of total cardiac  lipids among strains. Rats from all strains except Hooded fed the diet containing 20% Zephyr RSO had a significantly (P less than 0.001) higher incidence of myocardial lesions than rats fed the 20% corn oil diet which in turn had a significantly (P less than 0.001) higher incidence than rats fed 5% corn oil. Similarly, significantly (P less than 0.05) more Sprague-Dawley C rats had myocardial lesions than Sprague-Dawley A rats regardless of diet. None of the Hooded rats fed the diet containing 20% Zephyr RSO developed myocardial lesions whereas all other strains fed this diet developed a high incidence of myocardial lesions.


Myocardial lesions in rats fed rapeseed oil. II. Effects of castration. The effects of castration on the incidence of myocardial lesions in rats were investigated in an experiment which included two factors: sex (entire males, castrated males, entire females and castrated (ovariohysterectomized) females) and diets (5% corn oil, 20% corn oil, and 20% Brassica napus var. Zephyr rapeseed oil). For 16 weeks, each of the 12 groups of 30 Sprague-Dawley rats, housed 2 per cage, were fed ad libitum the test oils incorporated in a semisynthetic diet. At each weighing, the mean body weights for each diet were highest in entire males followed by castrated males, castrated females, and entire females with all differences significant (P less than 0.05). The results indicated that castration did not influence cardiac fatty acid composition. The incidence of myocardial lesions in entire and castrated females and in castrated males was similar while significantly more entire males developed lesions (P less than 0.001). Rats fed a diet containing 20% Zephyr rapeseed oil showed a significantly (P less than 0.001) higher incidence of heart lesions than did rats fed diets containing 5% or 20% corn oil. Similarly, significantly (P less than 0.05) more rats fed the 20% corn oil diet had lesions than rats fed the 5% corn oil diet. The involvement of androgens in the formation of myocardial lesions is suggested, since castration significantly lowered the incidence in males but not in females.


Effect of 6-hydroxydopamine and alpha-methyltyrosine on brown adipose tissue of infant rats. A single-dose administration of 6-hydroxydopamine (50 mg/kg body weight, sc) to infant rats resulted in an enlargement, higher fresh weight, markedly elevated lipid content, and higher total protein content of their interscapular and cervical brown adipose tissue. The protein kinase (EC 2.7.1.37) activation ratio in the tissue was decreased as was the phosphoenolpyruvate carboxykinase (EC 4.1.1.32) activity. Fatty acid synthetase, on the other hand, showed an increased activity. These changes commenced as soon as 24 h after the administration of the drug, were fully developed 2-4 days later, and persisted for at least 14 days. The results are in line with the assumption that 6-hydroxydopamine administration causes chemical sympathectomy of brown adipose tissue. This is further supported by the fact that treatment with alpha-methyltyrosine, which is known to competively inhibit norepinephrine systhesis, results in similar changes in brown fat of infant rats. Hence it seems that 6-hydroxydopamine administration offers a simple and inexpensive experimental model for studies of the role norepinephrine-mediated sympathetic nervous system in brown tissue function and development.


Superior colliculus cell responses to to electrical stimulation of the retina. Superior colliculus cell discharges in response to electrical stimulation of the retina were investigated in rabbit. In contrast with the responses at the optic tract level both polarities evoked discharges of equal latencies suggesting a convergence of ON- and OFF-centre retinal ganglion cells upon one collicular unit. Three typical patterns of responses could be distinguished. Thus, 40% of cells reacted with a burst, 47% with a prolonged discharge, and 5% responded by a transient inhibition. The responses of the remaining cells could not be classified.


Does indomethacin antagonize the effects of peptides and other agents on the coronary circulation of rabbit isolated hearts. The coronary vasodilator effects of substance P, bradykinin, eledoisin, adenosine, and nitroglycerin have been tested in rabbit isolated hearts perfused with physiological medium. It has been found that substance P is the most potent vasodilator and that the peptides are 10-100 times as active as adenosine and nitroglycerin. The effect of bradykinin is reduced by 50% in the presence of indomethacin (2.8-5 X 10(-6) M) while the actions of the other vasodilator are not influenced. At such concentrations, indomethacin is inactive on its own and blocks the effects of small and intermediate doses of arachidonic acid, the prostaglandin precursor. It suggested that edogenous prostaglandins do not play any major role in mediating the coronary vasodilation produced by several peptide and nonpeptide agents, but may account for about 50% of the effect of bradykinin.


Further identification of PGE1 metabolites formed in the cat lung. Tritium-labeled compounds appearing in the venous effluent of isolated blood perfused cat lungs after infusion of [3H]prostaglandin E1 (PGE1) appear to be unchanged [3H]PGE1 and 13,14-dihydro-15-keto-PGE1. No 15-keto-PGE1 could be detected in the venous effluent.


Further studies on the formation of choline sulfate by bacteria. Cell extracts of Pseudomonas C12B synthesized choline sulfate (COS) from SO42-, choline chloride, and ATP. However, most of the COS-forming activity was found in culture medium supernatants of this bacterium, and that which remained with the cells was cell wall-associated. Enzyme release was independent of the carbon and (or) sulfur source used for growth and was not suppressed by increasing the divalent cation concentration of the medium. The COS-synthesizing system was inhbited in vitro by L-cysteine (greater than or equal to 10(-3) mM), SO42- (greater than 0.1 mM), and choline chloride (greater than 0.1 M). L-Cysteine (0.1-5.0 mM) did not repress the synthesis of enzymes present in the system. COS formation from SO42- in vitro was increased 2.8-fold by 10 mM adenosine 5'-phosphosulfate (APS) and 5-fold by 1 mM 3'-phosphoadenosine,5'-phosphosulfate (PAPS) during a 4-h incubation period. APS (10 mM) also inhibited the incorporation of 35SO42- into COS. Culture supernatants incubated with Na235SO4 produced two 35S-labelled metabolites having electrophoretic mobilities similar to those exhibited by authentic APS and PAPS. The synthesis of these metabolites was also inhibited in vitro by unlabelled APS and by L-cysteine.


Ultrastructural localization of succinate dehydrogenase in a self-parasitic isolate of Saprolegnia megasperma. The enzyme succinate dehydrogenase (SDH, succinate: (acceptor) oxidoreductase, EC 1.3.99.1) was localized by the combined techniques of cytochemistry and electrom microscopy in the hyphae of a self-parasitizing isolate of Saprolegnia megasperma Coker. The enzyme was localized in the mitochondrial membranes; its activity was inhibited by malonate. Electron-dense deposits, whose formation was not prevented by the addition of malonate, appeared outsided of the hyphal cell walls. No evidence was found at the ultrastructural level within the vegetative hyphae for any abnormalities which could be linked to the phenomeonon of self-parasitism.


The relative ability of four rubella antigens to elicit blast cell transformation of lymphocytes from immune individuals. The ability of crude, semi-purified, and purified rubella antigens to elicit a specific and significant blast cell transformation of lymphocytes from immune individuals was investigated in 25 seronegative and 25 seropositive young adults. The type of preparation and the purity of the antigens were critical. Titration of the antigens by complement fixation or hemagglutination was of little value for selecting the best antigen which was a whole virion-purified antigen.


ATP and acetylene-reducing activity of a sulfate-reducing bacterium. A sulfate-reducing bacterium, a strain of Desulfovibrio vulgaris, when lactate or pyruvate was supplied as an electron and energy source, reduced acetylene only in the presence of sulfate. Acetylene reduction started after a lag of 1 h. H2 which acts as an electron and energy source for sulfate reduction was unable to reduce acetylene even in the presence of sulfate. It was suggested, from the measurements of the amounts of adenine nucleotides of the bacterium incubated under various conditions, that the bacterium is able to reduce acetylene at a high ATP level or a high ATP/ADP ratio.


Effects of temperature and antibiotics on appressorium formation in spores of Colletotrichum lagenarium. Effects of temperature (32 degrees C), cycloheximide, and blasticidin-S on spore germination and appressorium formation of Colletotrichum lagenarium were investigated. Temperature treatment at 32 degrees C, given just before the emergence of the germ tube 4 h after incubation at 24 degrees C, significantly inhibited appressorium formation. Cycloheximide (1 ppm) or blasticidin-S (7 ppm) appeared to have reversed the effect of 32 degrees C treatment by producing appressoria in 30% of the germinated spores.


Simultaneous presence of antiviral activity and its degrader in Bacillus extracts. Extracts of uninfected Bacillus species ATCC 27877 cells contained both an antiviral activity and a degrader for this activity. The antiviral activity was stable to 90 degrees C, caused a rapid inhibition of phage adsorption, and resulted in a gradual loss of phage infectivity. When extracts were incubated at 37 degrees C, the degrader inactivated the antiviral activity and the degrader was retained. When the extracts were heated at 90 degrees C, the degrader was inactivated and the antiviral activity was retained. A comparison was made of these two activities in extracts from cells of various incubation ages. The apparent association of maximal antiviral activity with prespore cells seemed due to the high content of degrader in vegetative and stationary cells. Both antiviral activity and its degrader appeared to be characteristics of vegetative cells.


Numerical taxonomy and ecology of oligotrophic bacteria isolated from the estuarine environment. Slow-growing bacteria, isolated on nutrient-rich and nutrient-limited media, from Chesapeake Bay water and sediment samples, were examined for 119 biochemical, cultural, morphological, nutritional, and physiological characters. Those bacteria which grow on low nutrient media, termed oligotrophs, a total of 162 strains, were subjected to taxonomic analysis, as a preliminary step in determining their ecological significance. The data for all strains included in the study were examined by computer and the simple matching (S SM) and Jaccard (SJ) coefficients calculated. Clustering was achieved by the unweighted average-linkage method. From sorted similarity matrices and dendrograms, 148 strains, 90% of the total, were recovered in 24 phenetic groups defined at the 80 to 85% similarity level. Only 12 phena could be presumptively identified and these included representatives of Alcaligenes, Corynebacterium, Hyphomicrobium, Hyphomonas polymorpha, Listeria, Nocardia marina, Pedomicrobium, Planococcus citreus, Sphaerotilus, Streptothrix, and Streptomyces. Of the remaining organisms, 10% were unidentified sheathed bacteria. It is concluded that slow-growing bacteria are distributed throughout the estuarine environment and can account for a large proportion of the colonies observed on media after prolonged periods of incubation. The oligotrophic bacteria appear to predominate in areas where the concentration of available nutrients is low and are more characteristic of non-eutrophic aquatic systems.


Occurrence of indole in cells of extremely halophilic bacteria. The neutral lipids of several halophilic bacteria contain a non-isoprenoid compound which was identified by its chromatographic and spectroscopic properties as indole.


Rapid solid-phase radioassay for staphylococcal protein A. Solid-phase radioassay was applied to the determination of protein A after release from staphylococcal cells with lysostaphin. Assays can be completed in 60-90 min.


Preoperative irradiation in operable cancer of the rectum: report of the Toronto trial. The authors have studied the value of preoperative radiotherapy in 125 patients with cancer of the rectum. The patients were divided into two groups, control (no irradiation) and experimental (irradiation), and according to Dukes' pathological stage. The crude survival indicated no difference between the two groups, but analysis by stage and group demonstrated that for patients with a Dukes' stage C lesion, 500 rads of cobalt-60 gamma  irradiation a few hours before excision of the lesion approximately doubles the probability of survival at 5 years. The prospect of surgical cure is three times higher in cancers arising above the peritoneal reflection than in those arising below it. If the practice of abdominoperineal resection is to continue the authors recommend that preoperative irradiation become a routine procedure.


Canine pancreatic exocrine secretory response to low- and high-fat elemental diets. Elemental or chemically defined diets are used in a variety of gastrointestinal disorders to sustain enteral nutrition, avoid the potential danger of central venous hyperalimentation and reduce intestinal and pancreatic secretions. The two diets available commercially are used interchangeably despite important differences in composition. The present study compared pancreatic exocrine secretory responses to these two diets in dogs. Infusion of high- and low-fat elemental diets resulted in less enzymatic output than a standard blenderized diet. Pancreatic volume and bicarbonate content were similar with the low-fat elemental diet and the blenderized diet, but volume was increased by 30% and bicarbonate increased nearly twofold during infusion of the high-fat elemental diet. The results support the hypothesis that a low-fat elemental diet is preferable for intragastric feeding when the therapeutic goal is to minimize pancreatic secretion.


Dysphagia lusoria: anterior approach for definitive treatment. An aberrant right subclavian artery causing symptoms is rare in adults. Several surgical approaches have been advocated. On the basis of results obtained in two patients, the authors recommended an anterior approach, either a transternal bilateral thoracotomy or a median sternotomy, that affords excellent exposure for flush ligation and division of the right subclavian artery at the aorta, and reconstruction on the ascending aorta.


Grey-scale ultrasound in the diagnosis of abdominal mass lesions. With the introduction of grey-scale imaging, the diagnostic capability of ultrasound has been greatly increased. Previously, ultrasound images were limited to a bistable presentation that did not distinguish between high- and low-level echoes. As a result the images could be interpreted by only a few experienced ultrasonographers. Grey-scale imaging with the use of a television scan converter allows both the strong and the weaker echoes to be displayed, so that both the outline and the internal texture of normal abdominal organs and lesions can be visualized on a single scan. Abnormalities are now more readily identified and their relation to normal organs is more easily appreciated so that the scans can be interpreted not only by the ultrasonographer but also by the clinician. Several cases are presented in which grey-scale ultrasound was used.


Comparison of epicardial and endocardial pacemaker insertion. The authors compared the results of 100 pacemaker insertion procedures: 50 endocardial and 50 epicardial. The late pacing threshold and the postoperative pain and pulmonary complications were higher in the epicardial group. Although dislodgement occurred in 28% of cases of insertion through the endocardial route, the authors concluded that this route was preferable.


Spontaneous spinal extradural hematoma.

Spontaneous spinal extradural hematoma. Spontaneous spinal extradural hematoma is an uncommon but reversible cause of acute paraplegia. Only 44 cases have been reported in the English literature. This paper adds two cases and analyzes the data in previous case reports to identify prognostic factors. Patients with a history of trauma or an incomplete myelographic block have a good prognosis. In most patients no cause of bleeding is found at operation.


Early and late results of resection of abdominal aortic aneurysms. Resection of the abdominal aortic aneurysm is being performed with decreasing operative mortality and morbidity. Among 190 patients undergoing this procedure at the Ottawa Civic Hospital between 1970 and 1975, 53 (28%) had a ruptured aneurysm and 137 (72%), a nonruptured aneurysm. Mean age of the patients was 66.2 years. Concomitant disease was frequent, 73% of patients having two or more associated diseases; the average number of associated diseases per patient was 2.25. Operative mortality in the group with ruptured aneurysms was 51%, and in the group with nonruptured aneurysms, 4%. Postoperative morbidity was 85% among those with a ruptured aneurysm, 67% among those with imminent rupture before operation and 34% among the others with a nonruptured aneurysm. Graft complications occurred in 15% of those with a ruptured aneurysm and 9% of those with a nonruptured aneurysm. Among survivors of the operation 73% and 81% of those with a ruptured and a nonruptured aneurysm, respectively, are known to be alive. In both groups causes of late death included infection or thrombosis of the graft and mesenteric thrombosis, as well as causes unrelated to the operation. Surgical management of the abdominal aortic aneurysm is advocated in all but patients at poor risk for operation who have asymptomatic aneurysms less than 6 cm in diameter.


Pericarditis as a manifestation of essential thrombocythemia. A 58-year-old woman presented with pericarditis and pericardial effusion. Investigation revealed that she was suffering from a thrombocythemic myeloproliferative disorder; she died of massive pulmonary embolism 10 days after admission. Histologic study verified epicardial and pericardial trilineage hematopoiesis. Pericarditis is an unusual feature of essential thrombocythemia and it may occur in direct relation to the abnormal cellular proliferation.


Hairy cell leukemia. Enzyme histochemical characterization, with special reference to splenic stromal changes. Seven spleens and two peripheral blood specimens from eight patients with hairy cell leukemia were examined with enzyme cytochemical and histochemical methods. Hairy cells consistently exhibited acid phosphatase and tartrate-resistant acid phosphatase. However, nonspecific esterases characteristic of monocytes and histiocytes were consistently absent or very weak. beta-glucuronidase and cytoplasmic membrane-bound ATPase were positive in four cases, suggesting a possible relationship to the B-lymphocytic series. Fundamental splenic changes were accumulation of hairy cells and benign macrophages within the pulp cords, with resulting extreme expansion of the cords. Abnormally well developed ellipsoids were identified around the sheathed arteries within the cords. Sinuses, specifically delineated with the NASDA reaction, were atrophic and often destroyed. No cytogeneologic relationship was found between sinus endothelial cells and hairy cells. The pulp cords are the primary site of involvement of the spleen in hairy cell leukemia. A simultaneous proliferation of neoplastic cells, histiocytes and reticulum fibers accounts for the splenomegaly and clinical hypersplenism characteristic of the disease.


HLA antigens in malignant melanoma.

HLA antigens in malignant melanoma. HLA-A, -B, -C and -D typing was performed on 54 Norwegian patients with microscopically verified malignant melanoma. A slight,  but non-significant increase in the frequency of HLA-B27 and LD 108 was demonstrated among the patients compared with healthy Norwegian controls.


Prolonged disease-free survival in bronchogenic carcinoma associated with HLA-Aw19 and HLA-B5. A two-year prospective study. Seventy patients with squamous carcinoma or adenocarcinoma of the lung were HLA typed at the time of their diagnosis and initial therapy. No abnormal HLA antigen frequencies were found. However, the possession of HLA-Aw19 or HLA-B5 was significantly correlated with two year disease-free survival. Twelve of 21 patients (57%) with either Aw 19 or B5 were disease-free at two years. In contrast, only six of 48 patients (13%) not having either antigen were disease-free (p less than .001). One patient was lost to follow-up. It is possible that Aw 19 and/or B5 confer resistance to progression of bronchogenic carcinoma on their possessors.


Immunosuppression and human cancer: role of prostaglandins. Prostaglandins, unsaturated fatty acid derivatives with diversified pharmacologic activity, have been implicated in the pathophysiology of many diseases. Prostaglandin E (PGE) levels were measured by radioimmunoassay in the plasma of 41 normocalcemic patients with various stages of malignancies. Delayed hypersensitivity was assessed by a battery of six recall skin test antigens (ST) and by Dinitrochlorobenzene (DNCB) sensitization and challenge. Twenty-five patients with one or more positive skin tests had a mean PGE level of 87+/-8 pg/ml, whereas 16 patients with negative ST had a mean PGE level of 96+/-12 pg/ml. Twenty-one DNCB negative patients had a mean PGE level of 98+/-12 pg/ml and eight totally anergic patients had a mean PGE of 96+/-12 pg/ml. All PGE values were within the normal range and there was no statistical difference between the four groups. (p less than 0.1). We concluded that circulating PGE does not correlate with the non-specific immunosuppression seen in cancer patients.


Immunomorphological lymph node changes in patients with operable bronchogenic squamous cell carcinoma. Histological changes in tracheobronchial and bronchopulmonary lymph nodes of patients with operable (i.e. stages T1/2, N0/1, M0) bronchogenic squamous cell carcinoma were examined histometrically. Out of 29 patients, 10 survived less than 12 months and 19 lived longer than two years after surgery. Quantitative methods were employed for measuring structural changes in lymph node sections at the histological and cellular level. The following parameters reflecting both cellular and humoral immune responses correlated with survival: relative numbers of large lymphoid cells and mitotic figures in the paracortical ("thymus-dependent") area; volume of the follicular (predominantly B-cell) cortex; and volume of germinal centers. Pronounced accumulation of histiocytes or dust-loaded macrophages in the paracortex and presence of "empty" lymph sinusoids (i.e. without sinuhistiocytosis) correlated inversely with survival. In addition to the theoretical importance and considerations of a documented immune response, such data could help in identifying high risk groups within the same stage of bronchogenic carcinoma of a given type.


Use of a permanent cell line extracts to study the tumor associated immune reactions in colorectal cancer patients by leucocyte migration inhibition test. Crude extracts of colorectal surgical tumors, either individual or pooled, and HT29 line cells were compared in leucocyte migration inhibition test. HT29 cell extract gave more positive reactions with leucocytes of colorectal cancer patients than the other ones, and less with control leucocytes. It was thus used for all the subsequent experiments. As a whole, we obtained 44% (30/68) of positive inhibitions in patients afflicted with a colorectal carcinoma, against 9% (3/31e in normal controls and 20% (8/40) in patients bearing a carcinoma from another organ. Among the latest patients, those having a gynecological carcinoma were rarely positive, contrasting with those suffering from a carcinoma from the aerodigestive organs. It seemed that the antigen(s) involved in the reaction had no organ specificity, but rather a broad tissue specificity.


Biological markers in breast carcinoma. II. Clinical correlations with human chorionic gonadotrophin. Serum hCG levels were measured in female patients with breast carcinoma and examined with respect to the clinical stage of disease, the clinical tumor burden, prognosis, and the organ sites of involvement. Elevated levels were observed in 65/134 (48.5%) patients with metastatic disease, 5/14 (35.7%) patients preoperatively, and in 9/33 (27.2%) N+ patients from one-six months postoperatively. The greatest proportion of elevated values (60.5%) was in patients with one metastatic site of involvement. Although no correlations were demonstrated between the preoperative or postoperative samples and subsequent disease recurrence, it was observed that 4/10 patients who did have a recurrence had preceding hCG elevations. In metastatic disease the level in 13 patients starting with values greater than 5.8 mIU/ml fell with the attainment of a response or rose with therapeutic failure. The hCG level rose from normal to greater than 6.1 mIU/ml in six additional patients that developed progressive disease. A normal hCG level was associated with a 94.5% response rate to combination chemotherapy whereas the response rate with levels greater than 5 mIU/ml was 71.4%. Only hepatic involvement was associated with a disproportionate incidence of elevations, being 63% compared to 42-47% for other sites. Monitoring hCG levels may be of prognostic use in patients being treated with selected chemotherapy programs for metastatic disease. It appears that further studies are warranted to ascertain if these results will extend to additional treatment approaches.


Biological markers in breast carcinoma. III. Clinical correlations with carcinoembryonic antigen. Plasma CEA levels were evaluated by radioimmunoassay in patients with breast carcinoma in relation to clinical-pathologic staging, clinical tumor burden, prognosis and organ sites of involvement. Elevated levels were observed in 83/117 (70.9%) patients with metastatic disease, 2/14 preoperative patients and in 3/39 one-six month postoperative patients. Preoperative levels were elevated in two patients; the levels fell to normal after operation. Changes of elevated CEA levels followed the clinical response to therapy in 22/22 metastatic disease patient-trials. The levels decreased with a response in 15 trials and rose with progressive disease or relapse in seven trials. The incidence of CEA elevations and quantitative CEA levels both rose with increasing clinical tumor burden from the postoperative state through the preoperative state to two or more organ sites of metastatic involvement. No relationship was demonstrable among limited samples between preoperative or postoperative CEA levels and prognosis; however, in metastatic disease, pretherapy CEA levels greater than 5 ng/ml were associated with low response rates and early therapeutic failure to chemotherapy. The highest frequency of elevated CEA levels was observed in patients with osseous involvement (79%) and the lowest frequency with skin (52%) and breast (50%) metastases. Liver and osseous disease were also associated with higher mean CEA levels than were other sites of metastatic involvement. CEA levels appear to be elevated in the majority of patients with metastatic disease and be of prognostic importance in metastatic disease. The level in patients with metastatic disease appears to reflect the therapy-associated tumor burden of the host, especially in patients with elevated levels.


Comparative effect of anaerobic coryneforms on a murine melanoma. Ten strains of anaerobic Corynebacteria were compared in their effect on survival of C57BL/6 mice, bearing subcutaneously inoculated B16 melanomas. The corynebacterial suspensions were injected intralesionally twice weekly for five injections. Significant permanent tumor regression was not obtained. Significant prolongation of survival was observed in mice treated with seven of the corynebacterial strains.


Low-dose Adriamycin remission maintenance therapy for pediatric solid tumors. A low-dose maintenance schedule of Adriamycin was evaluated in six children whose metastatic solid tumors regressed following toxic induction therapy with Adriamycin. Three of these children are now disease-free more than one year following discontinuation of therapy. Adriamycin can be given on a low-dose maintenance schedule free from alopecia, fever, stomatitis, myelosuppression and recognizable cardiomyopathy. Further studies of similar schedules are warranted.


Combined treatment modalities of rhabdomyosarcoma in children. Thirty-nine previously untreated children with rhabdomyosarcoma were managed by a coordinated program of surgery, radiation therapy, and chemotherapy during the years 1960 to 1973. The primary tumor was located in the head and neck (24), chest wall (1), abdomen (1), pelvis (10), and lower extremity (3). Radiation therapy consisted of tumor doses of 5000 to 6000 rads delivered in five to six weeks. Combination chemotherapy with actinomyctin-D, vincristine and cyclophosphamide was used after 1968. Seventeen of 25 cases (68%) treated after 1968 are alive two to five years following treatment. Only four of 14 cases (29%) who received less radical therapy before 1968 are alive. A relatively high incidence of local failure (23%) was noted in spite of adequate doses of radiotherapy. None of the four cases with metastatic disease at the time of diagnosis survived. Major complications were mainly noted in patients with orbital rhabdomyosarcoma. Correlation of absolute survival with dose of irradiation, primary site, extent of disease and histologic subtypes of the tumor are discussed.


Diagnostic planning using computer assisted decision-making for patients with Hodgkin's disease. A family of computer programs has been developed to help physicians effectively utilize available clinical information as in indicator of tumor extent for the staging of Hodgkin's disease patients. Decision analysis techniques were employed to suggest chronologic sequencing of selected, risk-associated tests, tailored to the individual patient, that minimize the risk of iatrogenic mortality. These quantitative techniques provide an objective measure of expected survival for a given plan of action. By using this information a physician and patient can balance expected survival vs morbidity of proposed staging procedures. Physicians can then focus on the interrelation between the inherent diagnostic uncertainty, therapy-induced expected survival, and morbid consequences of staging procedures. The use of these decision-making tools enables the physician to identify patient-specific presentations where the formation of an individual diagnostic plan could lead both to reduction in testing and to an increased survival rate.


Natural history of patients after abdominal-perineal resection. Implications for radiation therapy. The records of one hundred twenty-three consecutive abdominal perineal resections for rectosigmoid carcinoma were reviewed. One hundred sixteen cases were adenocarcinoma, and seven cases squamous cell carcinoma. Clinical outcome was documented in 111 cases. Overall, 42% survived five years or more. Ultimately 44 of the 123 patients (36%) were proven to have died with tumor and 19 patients (15%) developed clinical manifestations of recurrent pelvic tumor. Five of the 116 patients with adenocarcinoma (4%) died with evidence of pelvic disease only. Second primary tumors were observed in 22 patients (18%). The authors suggest preoperative irradiation to reduce local recurrence and possibly increase disease-free survival. Physicians following these patients should consider them at high risk for cancer in general.


Persistent carcinoma of the oropharynx and oral cavity re-treated by after-loading interstitial 192Ir implant. After-loading implantation techniques apparently offer an effective alternative in the management of persistent (or recurrent) oropharynx and oral cavity carcinoma postirradiation. A total of 29 patients with such lesions was treated from February 1974, to July 1975, in the LAC/USC Radiation Medicine Section, using after-loading 192Ir implantation techniques. These patients all had extensive persistent cancer after "full tolerance" irradiation (with or without previous surgery) and were referred for palliation or for possible salvage. Additional doses administered ranged from 5000 to 7000 rads in three to five days. Eighteen of the 29 patients (63%) have had complete local control for 18 to 36 months. These patients have maintained a relatively satisfactory palliative status. Ten of the 29 are dead. Follow-up periods are short, but early indications are that problem patients with persistent cancer after "full tolerance irradiation" can be salvaged, or at least satisfactorily palliated, with after-loading interstitial implant techniques. Clinical details, sites of involvement, implantation techniques, dosimetry methodology, reactions and complications will be described.


Ultrastructural studies on the morphogenesis of psammoma bodies in ovarian serous neoplasia. Transmission electron microscopic studies have been carried out on psammoma bodies in two benign and seven malignant papillary serous neoplasms of the ovary. Ultrastructurally, psammoma bodies are composed of microcrystals in all respects similar to calcium-phosphate apatite crystals of bone. The formation of psammoma bodies is initiated intracellularly, in both the neoplastic epithelial cells and stromal histiocytes. The initial seeding site of apatite crystals is served by lipid-rich intracellular vesicles. These structures are produced in association with autophagocytosis in the neoplastic epithelial cells and heterophagocytosis of extracellular lipidic material in the stromal histiocytes. Extracellular lipids presumably derive from dehiscent tumor tissue. The close relationship between larger intraepithelial calcific bodies and microfilaments suggests that the latter provide supportive matrix for further intracellular calcification. Large extracellular psammoma bodies result from fused calcific bodies which have been extruded from calcified cells. Mineralization of extracellular collagen fibers is not observed. The results provide supportive evidence to the concept that psammoma bodies in ovarian papillary serous neoplasms and probably in other neoplastic and non-neoplastic conditions are a consequence of dystrophic calcification associated with cellular degeneration.


Cystadenoma of the pancreas. An ultrastructural study. A case of cystadenoma of the pancreas was studied at the light (LM) and the ultrastructural (EM) levels. The observation made under LM that the flattened epithelial cells lining the cystic spaces were similar to the centroacinar cells of a normal pancreas was supported by EM findings. Numerous glycogen granules were demonstrated in the cells, a feature characteristic of fetal but not adult centroacinar cells. A few epithelial cells contained occasional membrane bound dense granules of unknown nature. This observation and the finding of normally appearing islets of Langerhans raise the possibility that this may be an example of a mixed insular-duct cystadenoma. However, if the islets of Langerhans were considered entrapped structures, this tumor should be labelled as a simple serous cystadenoma.


Gastric carcinoma. A pathobiological classification. Gastric carcinomas had various pathological features. Based on patterns of growth and invasiveness, however, they fell into two types; expanding type and infiltrative type. These types were readily recognizable histologically: expanding carcinomas grew en masse and by expansion, resulting in the formation of discrete tumor nodules, whereas in infiltrative carcinoma tumor cells invaded individuality. Both types showed varying degrees of cell maturation, but glands were much more common in expanding carcinoma. The difference in growth pattern was reflected partly by gross appearance of the tumors. These two types of carcinoma appeared to be different in their histogenetic origins. Intestinal metaplasia probably played a role in the development of expanding, but not infiltrative, carcinoma. There were differences also in the sex and age of the patients, survival rate, and epidemiological distribution. Thus, this classification provided a simple basis for evaluation of various aspects of gastric cancer.


Ectomesenchymoma. A malignant tumor of migratory neural crest (ectomesenchyme) remnants showing ganglionic, schwannian, melanocytic and rhabdomyoblastic differentiation. A case is reported in which a tumor containing elements of mature and immature ganglioneuroma, Schwannoma, clusters of apparently benign melanocytes and embryonal rhabdomyosarcoma presented as a right-sided facial mass in a six-month-old child. Different elements of the tumor were intimately intermingled without distinct borders. Embryogenesis of the tumor, which is thought to arise from remnants of migratory neural crest cells (ectomesenchyme), showing multidirectional phenotypic expression is discussed. It was felt that the lesion contributed additional evidence in support of pluripotentiality of the migratory, neural crest derivatives, and their susceptibility to mutagenic and oncogenic influences.


Primary clear cell thyroid carcinoma with squamous features. A clear cell neoplasm of the thyroid with indistinguishable light microscopic features and similar tinctorial reactions for lipid and glycogen to those of clear cell renal carcinoma is described. However, electron microscopic examination disclosed cytoplasmic tonofibrils and keratohyaline granules as well as suggestive transitions between the tumor cells and acinar epithelium. The former appear to represent helpful "markers" for the distinction of this type of thyroid cancer from parathyroid or metastatic renal clear cell tumors. There is some evidence that not all clear cell thyroid neoplasms possess such squamous features. Nevertheless, it is apparent that electron microscopy may be necessary for the pathological discrimination of primary and secondary clear cell neoplasms of the thyroid, a situation analogous to that for clear cell tumors of other sites such as the lung.


Circumscribed carcinoma of the uterine cervix, with marked lymphocytic infiltration. The present study was carried out to investigate a distinctive type of carcinoma of the uterine cervix categorized under the designation as circumscribed carcinoma with lymphocytic infiltration. Grossly this carcinoma is characterized by defined circumscription with a superficial ulceration. A microscropic feature characteristic of this tumor is the presence of a loose fibrillary stroma infiltrated densely and uniformly by lymphocytes throughout the tumor mass. The tumor is arranged in solid cords separated by a lymphoid stroma with evidence of minimal squamous differentiation. The authors picked up 709 patients with carcinoma of the uterine cervix who underwent radical hysterectomy and pelvic lymphadenectomy at the Cancer Institute Hospital, Tokyo, between 1956-1967, without preoperative radio- or chemotherapy. Among them, 39 or 5.5% were identified as this type of carcinoma. Patients with this types of carcinoma had a significantly better prognosis than those with other types of cervical carcinoma of the same stage (p less than 0.05). This favorable prognosis is probably due to the less regional node metastases found in the group. A proposal was made to segregate this particular type of carcinoma from other types of cervical carcinoma on the basis of its morphologic and prognostic distinctiveness.


Studies on adenovirus type 9-induced mammary fibroadenomas in rats and their malignant transformation. After inoculating newborn W/Fu rats with adenovirus type 9, 27 of 27 females developed mammary fibroadenomas with a latency period of 14-25 weeks. No tumors were observed after inoculation with adenovirus type 5 or in males with the type 9 inoculation. After persistence of the tumors for 3-14 months, malignant transformation of the stroma resulted in different types of sarcoma in three rats: fibrosarcoma, round-cell liposarcoma, osteosarcoma and malignant mesenchymoma. In another animal the stroma of a fibroadenoma was highly cellular, suggesting a transition into fibrosarcoma. Malignant transformation of the epithelial component was not observed. Tumor cells contained adenovirus type 9-specific T-antigen, and rats with transplanted tumors were immunized to T-antigen. Mammary fibroadenomas without signs of malignant transformation developed in eight of nine female rats inoculated with adenovirus type 9 at an adult age. Neonatal thymectomy and total body x-irradiation neither significantly shortened the induction time of adenovirus 9-induced fibroadenomas nor increased the frequency of malignant transformation in females. One lipoma and one highly differentiated liposarcoma, however, appeared in two male rats. The results provide an example of the progression of a virus-induced benign tumor into a malignant neoplasm.


The carcinogenic effect of cigarette smoke. The effect of cigarette smoke on human gastric mucosal cells in organ culture. The carcinogenic effect of cigarette smoke on certain organs is well established epidemiologically, but the evidence in relation to gastric carcinoma is inconclusive. The present study reports the effect of cigarette smoke condensate (CSC) on gastric mucosa grown in organ culture and its effect is compared with that of the known gastric carcinogen, N-methyl-N1-nitro-N-nitrosoguanidine (NG). Viability of the cells was assessed by the uptake of labelled glucose and glycine. It was found that, as gauged by lactate and beta-glucuronidase production into the ambient fluid, CSC produced a response typical of malignancy that did not differ from that with NG.


Metabolic epidemiology of colon cancer. Fecal bile acids and neutral sterols in colon cancer patients and patients with adenomatous polyps. Because of potential significance of bile acids and cholesterol metabolites in the pathogenesis of colon cancer, fecal neutral sterols, and bile acids were determined in patients with colon cancer, adenomatous polyps or other digestive diseases and American or Japanese controls. The fecal excretion of cholesterol, coprostanol, coprostanone, total bile acids, deoxycholic acid, lithocholic acid was higher in patients with colon cancer and patients with adenomatous polyps compared to normal American and Japanese controls as well as patients with other digestive diseases. Patients with other digestive diseases excreted comparable levels of fecal bile acids and cholesterol metabolites compared to normal American controls; Japanese controls excreted reduced levels compared to normal American controls. These findings suggest that possible interactions between bile acids and cholesterol metabolites and colonic epithelial cells may be relevant in colon carcinogenesis.


Cancer and malpractice claims.

Cancer and malpractice claims. An analysis of 36 closed malpractice claims related to the diagnosis of cancer was conducted at a time when both subjects are receiving particular attention. Awareness of cancer statistics and the medicolegal literature is necessary. Factors related to the allegation of delay as well as the psychological aspects of cancer are presented. Suggestions for claims prevention have been delineated.


Role of bacterial overgrowth in the malabsorption syndrome of primary small intestinal lymphoma in Iran. Malabsorption studies were performed on five Iranian patients with primary small intestinal lymphoma. The effect of oral tetracycline (1.0 g daily) was also studied in three of the above subjects. The results of breath tests (utilizing glycine-1-14C-cholic acid) were abnormal in all five subjects before the antibiotic treatment. Oral tetracycline had a striking effect towards normalizing the results of breath tests. Schilling tests (with intrinsic factor) improve in two patients and steatorrhea improved in all and there was significant weight gain. The antibiotic had no apparent effect on D-xylose or folate absorption tests. It is concluded that bacterial overgrowth in the small intestinal lumen is an important contributory factor to the malabsorption syndrome of this disease.


Fatal thyroid carcinoma. Anaplastic transformation of adenocarcinoma. Prognosis of well-differentiated carcinoma of the thyroid gland is generally favorable, while that of anaplastic carcinoma, extremely poor. Well-differentiated carcinoma may sometimes be fatal; the most common underlying cause is considered to be due to anaplastic transformation of the original well-differentiated carcinoma to a less differentiated form. We studied 27 consecutive autopsy cases of fatal thyroid cancer treated at the Ito Hospital, Tokyo, during a five-year period, 1969-1973. We found uniform histological features of anaplastic carcinoma in 10 cases and of well differentiated carcinoma in four cases. In addition, co-existence of well-differentiated and anaplastic carcinomas was observed in nine cases and well differentiated and squamous cell carcinomas in four. Circumstantial evidence strongly suggests that malignant transformation is a part of the natural history of thyroid carcinoma, from well-differentiated carcinoma to less differentiated forms, either squamous cell or anaplastic carcinoma.


Lobular carcinoma in situ of the breast. An approach to rational treatment. A review of 3299 benign breast lesions was carried out. Lobular carcinoma in situ (LCIs) was discovered in 52 cases; and of these, 44 had been treated by biopsy alone. During the follow-up period averaging 15.9 years, 11 patients developed invasive breast cancer (IBC): 9 ipsilateral lesions, and 4 contralateral. This was about 12 times the frequency expected. The development of IBC had no relation to the patient's age, nor could any correlation be drawn between the time periods which elapsed from the time of the diagnosis of the in situ cancer to the development of IBC. The ipsilateral occurrence of IBC was not isgnificantly different from the contralateral. Careful lifelong follow-up seems to be the rational treatment for women whose breast biopsy shows LCIs.


Chronic mastopathy and breast cancer. A follow-up study. Over an average period of seven years 2,9000 cases of benign breast lesions diagnosed by biopsy between 1948 and 1973 in the Department of Pathology, Kaiser Foundation Hospital, Oakland, were followed for breast cancer development. When classified according to traditional diagnostic categories, the cancer incidence per 1,000 person-years varies between 2.7 and 7.9 and appears to be elevated in comparison to expectations obtained from the Third National Cancer Survey, San Francisco Bay Area. Two thousand four hundred biopsies were also scored by the Black-Chabou method. There is an upward trend in the breast cancer incidence as the atypia score rises, a finding which confirms conclusions from a retrospective case-control study by Black et al.


Low cancer incidence and mortality in Utah. Utah cancer mortality for the years 1950-1969 and morbidity for the years 1966-1970 are reported. Utah had 18% fewer cases of cancer than expected based on the Third National Cancer Survey, and 24% fewer cancer deaths than expected based on national mortality data. Cancer sites associated with cigarette smoking and alcohol use accounted for nearly half of these differences. Several major sites not strongly associated with smoking showed lower incidence and mortality than expected. These included pancreas, colon, rectum, female breast, uterine cervix and ovary. A marked excess occurrence above expectation was observed for cancer of the lip. Some possible explanations of these findings are discussed, including some of the unique aspects of the Utah population.


The prognostic significance of pelvic lymph node morphology in carcinoma of the uterine cervix. Pelvic lymph nodes from 80 patients with primary invasive cervical cancer undergoing radical hysterectomy and pelvic lymphadenectomy were evaluated morphologically. Six to 25 (mean 14) lymph nodes from each patient were examined histologically and classified as follows: lymphocyte preominant, germinal center predominant, unstimulated, or lymphocyte depleted. Tumors were classified according to their primary cell type: large cell non-keratinizing, keratinizing squamous cell, small cell, and adenocarcinoma. There was no direct relationship between lymph node morphology and patient age, lesion size, or tumor cell type. A lymphocyte predominant nodal pattern was associated with a statistically significant decrease in lymph node metastases, and tumor recurrence, and an increase in patient survival. In contrast, patients with a lymphocyte depleted nodal pattern had a high incidence of metastatic disease and tumor recurrence, and a decreased survival. Patients with lymph nodes showing an unstimulated or germinal center predominant pattern had an intermediate incidence of nodal metastases and tumor recurrence. These findings suggest that regional lymph nodal morphology may be of prognostic significance in patients with invasive carcinoma of the uterine cervix.


Effect of hyperthermia on malignant cells in vivo. A review and a hypothesis. The relevant literature is reviewed in an attempt to clarify the mechanism of heat-dependent tumor cell destruction in vivo. Malignant cells in vivo appear to be selectively destroyed by hyperthermia in the range of 41-43 degrees C. Heat evidently affects nuclear function, expressed by an inhibited RNA, DNA and protein synthesis and characteristic arrest or delay of cells in certain locations of the cell cycle. However, as these effects appear to be reversible and are observed in normal cells as well as malignant cells, they probably do not explain the hyperthermic induced selective in vivo destruction of malignant cells. Heat-induced cytoplasmic damage appears to be of more importance. Increased lysosomal activation is observed, and is further intensified by a relatively increased anaerobic glycolysis which develops selectively in tumor cells. A hypothesis is proposed and discussed which explains the marked and selective in vivo tumor cell destruction as a consequence of the enhancing effect on the cytoplasmic damage of certain environmental factors (e.g. increased acidity, hypoxia and insufficient nutrition.


Cronkhite-Canada syndrome. Case report.

Cronkhite-Canada syndrome. Case report. The case report of a 72-year-old man with the characteristic clinical and pathologic-anatomic picture of the Cronkhite-Canada syndrome is presented. The clinical features and the radiographic appearance of the gastrointestinal tract suggested Ménétrier's disease. Subtotal gastrectomy was performed and at the subsequent pathologic-anatomic examination, the diagnosis of Cronkhite-Canada syndrome was made. The patient slowly recovered over the following five months while being treated with the so-called astronaut diet (Vivasorb) supplemented with vitamins, iron and electrolytes. He died five months after surgery from intercurrent disease. The differential diagnosis of gastrointestinal polyposis with or without mucocutaneous pigmentation is briefly discussed. The histochemical characteristics of the epithelium in the gastric and colonic polyps was studied in some detail. The authors believe that the gastrointestinal polyps in the Cronkhite-Canada syndrome are regenerative and non-neoplastic in nature.


Primary clear cell carcinoma of the ovary. An analysis of 15 cases with review of the literature. The clinical and pathological findings in 15 primary "pure" clear cell carcinoma of the ovary included in the Magee-Womens Hospital Tumor Registry from 1947 through 1974 are discussed. The ages of the patients ranged from 35 to 75 years (mean 53). The most common clinical finding was an abdominal mass in 14 cases. At surgery, five patients had extraovarian spread, one accompanied by ascitis. The tumors varied in diameter from 4 cm to 17 cm (mean 10.5 cm); grossly, about one-third were cystic, one-third solid and one-third had combined cystic and solid areas. Microscopically, four basic patterns were seen: papillary, tubular, solid, and cystic, and in several an admixture of the above. Clear cells predominated in 48% of the cases and 23% displayed hobnail cells. Most patients were treated by TAH & BSO or variants thereof, some followed by radiation or chemotherapy. Four patients died with cancer within 35 months after diagnosis; two others died of unrelated causes three and 12 years later. The actuarial survival was computed as 55% at five years. Five patients were stages as IIb (FIGO classification); four died with tumor and the remaining patient is alive with recurrent tumor in the pelvis 18 months after the original diagnosis. Three patients with tumors displaying the cystic pattern are still alive; two with a predominantly papillary pattern and vascular permeation are dead. It is suggested that survival rates in these 15 cases may be related more to the extent of the lesion at the initial presentation than to the gross or microscopic pattern.


Role of nutrition in the etiology of breast cancer. International differences in breast cancer morbidity and mortality and studies on migrating populations point to the overriding importance of environmental factors in the etiology of breast cancer. Factors directly or indirectly associated with ovarian activity do not appear to explain international differences. Population correlation studies have indicated that much of the difference appears to be explicable on the basis of nutritional factors, particularly, high total fat intake. Animal experimental studies confirm the importance of high fat diet, possibly mediated through prolactin. Other indirect support for the nutritional hypothesis is supplied by changing incidence rates in Iceland, possibly correlated with changing nutritional practices and the association of breast cancer with weight and possibly also with height. The association with height (which would suggest  nutritional effects mediated through childhood diet) has not been supported by a study in Canada. Direct investigation of the association between nutrition and breast cancer is difficult because of problems in dietary methodology. Nevertheless, the results of a Canadian case-control study provide support for the importance of high fat intake. Further studies of this question are undoubtedly required, however, before specific recommendations on dietary modification can be made.


Present status of thermography, ultrasound and mammography in breast cancer detection. Various biophysical methods have been utilized in the diagnosis of breast cancer. To date the best results have been obtained with x-ray mammography. Ultrasound and thermography have great appeal as non-destructive techniques but, in the present state of development, are of limited use. The spatial resolution presently obtainable in ultrasonograms is inadequate for the detection of subclinical cancer and thermography is also of questionable reliability. While an overall true positive rate of 70% to 75% may be anticipated with thermography, the bulk of false negatives would seem to occur in those tumors most amenable to therapy, i.e., subclinical cancers. The "false positive" rate of thermography is also excessive, but would be acceptable for establishing a high risk group if true positive rates could be improved. At present thermography finds its greatest use as an adjunct to mammography and physical examination; it should not be used as the sole modality in a screening program. The efficacy of mammography can be readily demonstrated but the propriety of its use as a screening device has been questioned. This is primarily related to the possible carcinogenic effect of radiation at diagnostic levels. Although the carcinogenic effect is unproven, the dose in radiologic procedures should be kept to a minimum consistent with adequate images. The present state of the art would indicate that the risk, if any, is minimal as contrasted with the natural incidence of breast cancer and the results of early diagnosis and treatment.


Cell-mediated immunity and serum blocking factors in cancer patients during chemotherapy and immunotherapy. The leukocyte adherence inhibition test was used to monitor tumor-specific cell-mediated immunity in 15 patients who had a variety of malignant tumors and were undergoing chemotherapy alone or in combination with immunotherapy by Corynebacterium parvum. A rapid and prolonged loss of cell-mediated immunity in blood leukocytes was observed after treatment in all but one of the patients studied. Abolition of reactivity was due to the lack of production of the soluble lymphokine-like factor affecting leukocyte adherence to glass. A new phenomenon of adherence stimulation by antigen, also mediated by a soluble factor, was observed after treatment in some patients. A drop in titer or total abrogation of serum blocking factors occurred in six of six patients tested following chemotherapy or immunotherapy. The lowered levels of blocking activity persisted during treatment and, with the possible exception of one patient, were not correlated with clinical benefit.


N4-Behenoyl-1-beta-D-arabinofuranosylcytosine as a potential new antitumor agent. N4-Acyl-1-beta-D-arabinofuranosylcytosines, which are lipophilic antitumor analogs of 1-beta-D-arabinofuranosylcytosine, were dissolved by the use of a detergent, HCO-60, and the differences in the antitumor activities when the drugs were administered in the forms of solution or suspension were compared. N4-Stearoyl-1-beta-D-arabinofuranosylcytosine (NSC 201290), which was the most active compound when administered as an aqueous suspension, diminished in its activities after it had been dissolved into a clear solution, whereas N4-behenoyl-1-beta-D-arabinofuranosylcytosine (NSC 239336) exhibited activities superior to those of the parent compound 1-beta-D-arabinofuranosylcytosine when administered as a solution. Moreover, the high efficacy of this compound was long lasting in the host animal, regardless of the treatment schedules or the presence of the 1-beta-D-arabinofuranosylcytosine-inactivating enzyme, cytidine deaminase.


Identification of aldophosphamide as a metabolite of cyclophosphamide in vitro and in vivo in humans. Aldophosphamide (NSC 254), a putative key metabolite of cyclophosphamide, has now been isolated as a cyanohydrin derivative from an incubation mixture of cyclophosphamide with mouse liver microsomes in vitro and from the plasma of a cyclophosphamide-treated patient. The cyanohydrin has been shown to be identical with an authenic synthetic sample by mass spectrometry and combined gas chromatography-mass spectrometry.


Epidemiological features of canine renal neoplasms. A retrospective study of medical records from 13 veterinary university hospitals and clinics yielded 73 cases of confirmed primary neoplasms of the kidney and renal pelvis. Analysis of the 34 cases of renal carcinoma revealed no evidence of familial (breed) predisposition, but an excess risk was identified in males, especially during middle life. The morphological and epidemiological features of canine renal carcinoma are similar to those of renal carcinoma in humans. The dog may be a suitable model for further research into the causes of this cancer.


Variation of levels of plasma guanosine diphosphate L-fucose:beta-D-galactosyl alpha-2-L-fucosyltransferase in acute adult leukemia. We have measured plasma levels of an alpha-2-L-fucosyltransferase in 18 patients with acute adult leukemia at various clinical stages along with simultaneous bone marrow aspirations and biopsies. Patients in remission had significantly lower levels of this enzyme than did nonresponding or relapsing patients. Furthermore, plasma levels were correlated with percentage of marrow blast cells. This enzyme may serve as a biological marker for monitoring patients with acute leukemia receiving chemotherapy. It is particularly helpful as a guide to chemotherapy in patients on whom bone marrow aspirates are technically difficult to obtain.


Increased plasma corticosterone and decreased plasma thyroid-stimulating hormone levels in rats treated with vincristine. A single i.p. injection of vincristine (1000 microgram/kg) into rats increased the plasma concentration of corticosterone after a latent period of 4 hr; the effect lasted for 48 hr. The response was dose related with the threshold dose being 100 microgram/kg and the maximal effect occurring after 250 microgram/kg. Vincristine also increased plasma corticosterone levels in hypophysectomized rats, suggesting that the drug may have a direct action on the adrenal gland. The injection of 500 or 1000 microgram/kg also reduced the plasma concentration of thyroid-stimulating hormone. Twelve and 18 hr after the injection of vincristine (1000 microgram/kg), the plasma concentration of thyroid-stimulating hormone was reduced, whereas the hypothalamic content of norepinephrine, a neurotransmitter involved in the regulation of thyrotropin-releasing hormone-thyroid-stimulating hormone secretion, remained unchanged. The adrenocortical stimulation produced by vincristine may play some role in the antineoplastic effects of this drug.


Characterization and quantitative estimation of activated cyclophosphamide in blood and urine. An assay for the characterization and quantitative determination of activated cyclophosphamide has been described. The method is based on high reactivity of the kryptaldehyde group in 4-hydroxycyclophosphamide with mercapto compounds to yield 4-(S-R)mercaptocyclophosphamide derivatives. The activated cyclophosphamide can be converted quantitatively to 4-(S-benzyl)mercaptocyclophosphamide with benzyl mercaptan, and the mercapto derivative can be separated by thin-layer chromatography on silica gel with ethyl acetate:methyl ethyl ketone as solvent. Using this assay, significant levels of activated cyclophosphamide in the blood of mice and rats, as well as of humans, were found to be present after cyclophosphamide application. The characterization of 4-(S-benzyl)mercaptocyclophosphamide formed was confirmed by comparing the thin-layer chromatographic and mass spectrometric data on this derivative obtained from an authentic sample  of 4-hydroxycyclophosphamide.


Pharmacokinetics of vindesine and vincristine in humans. Vindesine, a new Phase 1 Vinca alkaloid congener, exhibited serum pharmacokinetic behavior in humans compatible with a three-compartment, open mammilary model. The kinetic parameters included: t1/2 alpha=3.24+/-1.14 min, t1/2beta=99.0+/-44.5 min, t1/2gamma=1213+/-493 min, Vc (Valpha)=4.81+/-2.12 liters, Vbeta=58.2+/-50.5 liters, Vgamma=598+/-294 liters. Vincristine, studied only for the first 4 hr, behaved like a two-compartment system, with values of t1/2 alpha=3.37+/-0.72 min, t1/2beta=155+/-18 min, Valpha=4.53+/-0.49 liters, and Vbeta=57.3+/-21.1 liters. Urine excretion data demonstrated that most drug elimination occurred within the first 24 hr and amounted to 13.2+/-5.9% for vindesine and 9.5+/-5.1% for vincristine.


Isolation and immunochemical and chemical characterization of carcinoembryonic antigen-like substances in colon lavages of healthy individuals. Material with carcinoembryonic antigen (CEA)-like activity, isolated from colon lavages of healthy individuals, has been chemically characterized. The CEA-like activity in these lavages was purified by gel filtration on Sepharose 6B and Sephadex G-200 and by affinity chromatography on concanavalin A linked to Sepharose. The purified material migrated in polyacrylamide-sodium dodecyl sulfate electrophoresis as a single diffuse band with mobility identical with that of tumor CEA. The material with CEA activity from tumor tissue and from normal colon lavages gave lines of identity in double diffusion experiments and had similar inhibition curves with esssentially the same specific activities in a radioimmune assay for tumor CEA. The amino acid and carbohydrate compositions were similar to that observed for CEA tumor preparations. Lysine was the single amino terminus. Methylation analysis demonstrated that the monosaccharide linkages were similar to those in CEA isolated from tumor tissue.


Antibody-dependent cell-mediated cytolysis of human colon carcinoma cells induced by specific antisera against carcinoembryonic antigen. Antibody-dependent lymphocyte cytotoxicity against human colon carcinoma cells grown in vitro was demonstrated with rabbit anti-carcinoembryonic antigen (CEA) antisera and normal human lymphocytes. The same antisera produced no tumor cell lysis in a complement-dependent cytotoxicity test. The specificity of the reaction was demonstrated by the inhibition of antibody-dependent lymphocyte cytotoxicity after the addition of increasing amounts of purified CEA to the antiserum and by the fact that only tumor cell lines expressing CEA on their surface were lysed. Antibody-dependent lymphocyte cytotoxicity was also observed against two colon carcinoma cell lines that expressed Blood Group A antigen, using a human serum containing anti-Blood Group A antibodies of the immunoglobulin G class. This reaction was specifically inhibited by absorption with Blood Group A red cells, whereas the anti-CEA-dependent cytotoxicity was not inhibited by absorption with red cells of different blood groups.


Different extracellular calcium requirements for proliferation of nonneoplastic, preneoplastic, and neoplastic mouse cells. The DNA-synthetic and proliferative activities of freshly isolated, nontumorigenic C3H mouse skin cells (first passage) were lowest when the extracellular free (or ionic) calcium level was reduced to between 0.05 and 0.1 mM, whereas the extracellular free calcium level in cultures of repeatedly passaged, preneoplastic C3H/10T1/2 and MCA-C3H/10T1/2 type I mouse fetal fibroblasts had to be reduced to 0.01 mM or less before the DNA-synthetic and proliferative activities were minimal. This inhibition of DNA synthesis and cell multiplication by calcium deprivation was rapidly reversed by returning the extracellular calcium level to its normal value. In contrast, the neoplastic fibrosarcoma-forming, MCA-C3H/10T1/2 type III mouse fetal fibroblasts could synthesize DNA and could multiply indefinitely even in the presence of an extremely low concentration of extracellular free calcium. Thus, the extracellular calcium requirement for DNA synthesis and proliferation appears to reflect the tumorigenic potential of the cell.


Enhancement of X-ray-induced transformation in C3H/10T1/2 cells by interferon. Interferon may enhance malignant transformation induced by X-rays in a C3H mouse embryo-derived cell line. The inhibitory effect of interferon on cell division during the proliferative phase of  the expression of the transformational damage may be of importance in the understanding of this finding.


Test for carcinogenicity of organic contaminants of United States drinking waters by pulmonary tumor response in strain A mice. The production of lung adenomas in strain A following multiple i.p. injections of selected organic water contaminants was investigated. Of the 16 contaminants tested, only bromoform produced a pulmonary adenoma response that was significantly greater than the pulmonary adenoma response of vehicle-treated control mice.


Marked differences in the tumor-initiating activity of optically pure (+)- and (-)-trans-7,8-dihydroxy-7,8-dihydrobenzo(a)pyrene on mouse skin. The ability of optically pure (+)- and (-)-trans-7,8-dihydroxy-7,8-dihydrobenzo(a)pyrene to initiate skin tumors in mice was determined with a two-stage tumorigenesis system. A single application of 50 to 200 nmoles of (+)- or (-)-trans-7,8-dihydroxy-7,8-dihydrobenzo(a)pyrene to the backs of CD-1 mice followed by twice-weekly applications of 12-O-tetradecanoyl-phorbol-13-acetate revealed that the (-)-enantiomer was 5- to 10-fold more potent than was the (+)-enantiomer as a tumor initiator at the three dosage levels tested. When the tumor-initiating activities of the (+)0 and (-)-enantiomers of trans-7,8-dihydroxy-7,8-dihydrobenzo(a)pyrene were compared to the activity of benzo(a)pyrene at an equimolar dose, the (-)-enantiomer was more active while the (+)-enantiomer was considerably less active. This is the first report of differences in the carcinogenic activity between optical enantiomers.


The emotional problems of patients with bladder cancer. In recent years the quality of survival has become a more dominant factor in our treatment and management of patients with cancer. As a result, it has become necessary to think in terms of a multidisciplinary approach to total rehabilitation. A part of this rehabilitation is of necessity determined by the emotional responses that patients exhibit to their disease and to its treatment. A useful model that has been applied to patients with other diseases and to patients with other types of cancer seems equally applicable to patients with bladder cancer. In my work with cancer patients I became aware that, after the initial fear of death was dealt with, the patients all responded to their dilemma with stereotyped and predictable responses (emotional patterns of behavior). It was apparent that these responses were determined by the degree to which the patient's life style was altered by the disease and/or treatment.


A long-term study of reversible and progressive urinary bladder cancer lesions in rats fed N-[4-(5-nitro-2-furyl)-2-thiazolyl]formamide. Hyperplasia and, ultimately, neoplasia of bladder epithelium were produced by feeding 0.2% N-[4-(5-nitro-2-furyl)-2-thiazolyl]formamide (FANFT) to weaning male Fischer rats. Hyperplasia induced by FANFT feeding for 2, 4, or 6 weeks, followed by feeding control diet until the end of the 84-week experiment, was reversible, and at the end of the experiment bladder epithelium in these animals was normal by light and scanning electron microscopy. Hyperplasia produced by 8 or more weeks of FANFT feeding was irreversible and by 84 weeks had resulted in bladder tumors in all animals fed FANFT for 12 or more weeks and in 4 of 5 and 6 of 7 animals fed FANFT for 8 and 10 weeks, respectively. The epithelial changes after 6 and 8 weeks of FANFT were similar by light microscopy but different by scanning electron microscopy. Pleomorphic microvilli seen with scanning electron microscopy are the hallmark of irreversible, and possibly progressive, epithelial proliferative change.


Development of tissue culture procedures for predicting the individual risk of recurrence in bladder cancer. We are using three correlated approaches in tissue culture to develop procedures for distinguishing between histologically similar tumors and to develop distinctions that we hope can be correlated with a favorable outcome or with recurrence of more serious disease. Our procedures involve study of the growth of resected tumor tissue in a three-dimensional matrix of collagen-coated cellulose sponge. Using bladder cancer cell lines we are also studying the patterns of cytotypic zonation that appear in response to prolonged exposure to continuous gradients of oxygen tension and of temperature. Finally, we are using vitamin A and modifiers of cyclic adenosine 3':5'-monophosphate as molecular probes to alter the morphological expression of tumors in matrix and in gradient cultures. We have studied over 80 specimens of clinical cancer in matrix culture. Tumors of similar histopathology grow with distinctly different architecture in the matrix of collagen-coated sponge. We must now determine whether these patterns in vitro can be correlated with the course of individual patients.


Cell surface changes associated with malignant transformation of bladder epithelium in vitro. Three properties of cell surface membranes of normal bladder epithelium and of two malignant urothelial lines transformed in vitro by Dr. Y. Hashimoto and Dr. H. S. Kitagawa were analyzed by means of morphological and chemical tools. Under scan electron microscopy, normal bladder epithelium displayed an appearance devoid of the pleomorphic, abundantly distributed, microvillous structures seen on the neoplastic cells. The molecular weight profiles of proteins dispersed from purified plasma membrane fractions demonstrated quantitative differences in the content of three molecular-weight classes between the native and the transformed cells. More striking differences were seen upon two-dimensional analysis of proteins solubilized from these two cell types, using 3 M KCl. These findings suggest that further investigations of the chemical moieties appearing on the cell surface early after transformation may enhance our understanding of proteins amenable for chemical and/or immunological attack to achieve control of the progression of bladder neoplasia.


Biochemically detectable tumor markers in urine of bladder cancer patients. Potential biochemical markers excreted in the urine of bladder cancer patients have been considered, with the conclusion that none alone has yet proven to be useful as a screening procedure for the detection of urothelial cancer. Quantitative fluctuations in urinary levels of several of these markers in combination, such as pseudouridine, beta-amino-isobutyric acid, and fibrinogen degradation products, appear to be valuable in the assessment of the treatment of bladder cancer patients and in helping to predict recurrences in these patients.


Immunological responsiveness in patients with bladder cancer. Delayed cutaneous hypersensitivity tests, especially skin tests with dinitrochlorobenzene, are impaired increasingly as the amount of tumor increases. Recall antigens are less sensitive indicators of disease. Therapy, especially radiotherapy, also depresses cell-mediated immunity. Removal of tumor, however, allows these tests to return to normal. Dinitrochlorobenzene skin testing can contribute significantly to prognostic evaluation. An important facet of the tumor-host relationship is measured, and this reflects factors that are independent of tumor staging. Combination of tumor staging and dinitrochlorobenzene-delayed hypersensitivity testing can provide a strong indication of the clinical course, especially for the year following initial treatment of invasive or metastatic transitional cell carcinoma of the bladder.


Surveillance, initial assessment, and subsequent progress of patients with superficial bladder cancer in a prospective longitudinal study. National Bladder Cancer Collaborative Group A (NBCCGA). Through a bladder cancer surveillance protocol, a sizable population of patients suitable for characterizing the dynamics and patterns of recurrent superficial transitional cell carcinoma has been identified. One hundred thirty-three patients have been followed after their first occurrence of transitional cell carcinoma (Stage O or A).  Only patients whose tumors were considered to be completely removed by transurethral resection and who were not treated by any other means were included. Although a full year of follow-up is not yet complete, recurrent bladder carcinoma has been noted in 44 (33%). Most of these recurrences were identified at the first follow-up cystoscopy and the majority were present at the site of the initial lesion. Tumor size, grade, and stage did not appear to influence recurrence, whereas multiplicity did. The relevance of these findings to various theories about the mechanism of recurrence is discussed.


Histopathological evaluation of the inhibition of rat bladder carcinogenesis by 13-cis-retinoic acid. An objective system for histopathological and statistical evaluation of rat bladder lesions induced by the carcinogen, N-methyl-N-nitrosourea, is described. This system has been used to measure the inhibitory effects of 13-cis-retinoic acid on the development of bladder cancer in female Wistar/Lewis rats. 13-cis-Retinoic acid caused significant inhibition of development of both preneoplastic and neoplastic lesions in bladder epithelium.


The importance of synergy between weak carcinogens in the induction of bladder cancer in experimental animals and humans. It is now well established that the interaction of multiple environmental factors may increase the incidence of some human cancers more than exposure to a single carcinogen. With an in vivo experimental rat model, we have demonstrated a synergistic effect in bladder carcinogenesis between a subcarcinogenic dose of the strong bladder carcinogen, N-methyl-N-nitrosourea, and saccharin- or cyclamate-containing diets. If these artificial sweeteners are capable of interacting with other environmental bladder carcinogens, their potential for increasing the incidence of human bladder cancer is greater than many more potent chemical carcinogens, because of their wide distribution as food additives to all sections of the population. Retrospective epidemiology shows no evidence of such risk from saccharin at current levels of consumption. No comparable studies are available for cyclamate, which was consumed in greater quantities but for relatively few years. It is emphasized that it is possible for interaction between multiple factors to contribute to the incidence of human bladder cancer as it does in other human organs and in other animal species.


Characteristics of human prostatic cell cultures. In preliminary experiments, the respective requirements of prostate-derived epithelial cells were compared with those of human lung fibroblast line (WI38). Optimal levels for cysteine and glutamine were similar but not identical. The response of these two cell types to the macromolecular fraction of serum (dialyzed) was reciprocal. These results suggest that serum contains a dialyzable factor or factors necessary for the growth of prostatic epithelial cells.


Separation and characterization of epithelial cells from prostates and prostatic carcinomas: a review. Our investigation of normal, hyperplastic, and neoplastic prostatic tissue during the past 2 1/2 years has produced several findings which have been published or accepted for publication. (a) Cells from hamster prostates with intense histochemically demonstrable acid phosphatase activity (HDAP) after fixation with formaldehyde which we believe to be epithelial cells can be obtained in 97.2% +/- 0.8% purity by velocity sedimentation in a previously described isokinetic density gradient; (b) similarly, cells with HDAP, many of which contain lipofuscin granules, can be obtained as 81.0% +/- 12.2% of nucleated cells from hyperplastic human prostates and as 86.4% +/- 9.4% of nucleated cells from human prostatic carcinomas; (c) more cells were obtained from human hyperplastic prostates and prostates with prostatic carcinoma per gram of tissue with the aid of Pronase than were obtained with trypsin, collagenase, or mechanical methods; (d) more cells per gram of tissue were obtained from surgically removed prostates than from prostates obtained at even very rapid autopsies, and a much larger proportion of the cells from surgically removed prostates were viable as assessed both by dye exclusion and by plating efficiency; (e) none of several substrates and inhibitors which we tested were highly specific for acid phosphatase from purified prostatic epithelial cells compared with several other kinds of purified human cells; and (f) purified hamster prostatic epithelial cells incorporate large amounts of tritiated thymidine in 72-hour cultures.