Relative efficacy of alcohol and propranolol in action tremor.

Thirty-nine patients with a variety of diseases, including essential tremor, Parkinson's Disease, olivopontocerebellar degeneration, ataxia telangiectasia, and cervical cord injury with action tremor, were evaluated for the effect of one ounce of absolute alcohol ingestion. Tremor significantly subsided in 61.9% of E.T.; 46.6% of P.D.; one patient with A.T.; and one patient with C6 lesion. The tremor became worse in one patient with O.P.C.D. Twenty of these patients were treated with propranolol, an average dose of 92 mgm. per day, and re-evaluated three to six months later. All those who improved on alcohol improved on propranolol and the one whose tremor accentuated with alcohol had a similar response to propranolol. It is concluded that the tremorilytic effect of alcohol is neither specific for, nor limited to, essential tremor and is of no value in differentiating various neurological disorders which manifest as action tremor. It is recommended that one ounce of absolute alcohol by mouth be used as an office procedure to predict the response of patients' tremor to propranolol.


Respiratory inhibitory apraxia.

Ten patients have been described showing inability to stop breathing on command, spontaneous respiration and voluntary respiratory stimulation being unaffected. This abnormality not previously described in the literature, we feel should be named respiratory inhibitory apraxia (R.I.A.). The anatomical organization of respiration is briefly reviewed. R.I.A. is often associated with other forms of apraxia or motor impersistence. It is thought that the urinary and bowel incontinence present in some of these cases might also represent a form of inhibitory apraxia. Information is presented which supports the view that respiratory inhibitory apraxia is due to a minor hemisphere lesion, usually deepseated. Our one autopsied case showed a lesion in the descending motor pathways in the internal capsule, in middle cerebral artery branch territory, disconnecting the voluntary respiratory inhibitory center in the cortex in anterior and middle cerebral cortical branch territories.


Simultaneous recordings of visual cortex and superior colliculus field potentials in the rabbit.

The field potentials recorded simultaneously at various depth of the rabbit's visual cortex and superior colliculus were analysed following light ON and light OFF. The collicular ON and OFF potentials exhibited three slow components superimposed by fast rhythmic oscillations. Only the first slow component reversed its polarity with penetration from surface negative to positive in depth. The cortical ON and OFF responses similarly contained three slow waves which all reversed their polarity with electrode penetration: from surface positive to negative in deeper layers. The most striking difference between ON and OFF cortical responses is the absence of fast rhythmic oscillations in the cortical ON response.


Skin punch biopsies and lymphocytes in the diagnosis of lipidoses.

Skin punch biopsies and buffy coats of white blood cells were examined electron microscopically in patients suffering from a variety of storage diseases. No specific abnormalities could be detected in Gaucher's disease and adreno-leucodystrophy. While characteristic deposits were found in cutaneous nerves in globoid and metachromatic leucodystrophy, this method was deemed inferior to sural nerve biopsy. In gangliosidoses, on the other hand, pathognomonic membranous cytoplasmic bodies were common in axons of cutaneous nerves, and in generalized gangliosidoses marked vacuolation of many other cells was prominent. Specific deposits were found in various cells in skin punch biopsies and in lymphocytes of children suffering from ceroid lipofuscinoses, and in lymphocytes of their parents. This constitutes the easiest diagnostic laboratory procedure in such cases.


Effect of fundoplication on the lower esophageal sphincter.

The lower esophageal sphincter of 26 patients has been studied by infusion manometry before and after a modified Nissen fundoplication. Of these patients, 24 have remained asymptomatic (mean follow-up, 21 months). The mean time for a positive acid infusion test increased from 267 seconds before, to 617 seconds after operation. The common cavity test was positive in 22 of the 26 patients before operation but positive in only the 2 symptomatic patients after operation. Mean lower esophageal sphincter pressure (cm H20) increased from 12.6 +/- 2.0 to 16.5 +/- to 1.5 (x +/- SEM). However, in only 15 of the 26 patients was an increase in lower esophageal sphincter pressure demonstrated and in 7 a decrease in pressure was noted. One patient's symptoms recurred at 3 months in spite of an increase in lower esophageal sphincter resting pressure of 8 cm H20. The other symptomatic patient demonstrated a rise in lower esophageal sphincter pressure at 3 months of 8 cm H20, though this decreased to preoperative levels at 26 months, the time of symptom recurrence. Intraluminal pressure of the human cadaver esophagus after removal from the body was studied by means of an infusion manometric technique. Narrowing the esophageal lumen with extrinsic weight of as little as 50 g resulted in a zone of elevated pressure. The amplitude of the elevated pressure zone was directly related to the infusion rate. Elevations in lower esophageal pressure following fundoplication are probably secondary to extrinsic compression and are not necessary for a good clinical result.


Thoracicoabdominal trauma: a plan for initial management.

Trauma to multiple organs damages the body but it also disrupts the orderly process of history taking, physical examination, laboratory work, calm deliberation and treatment. The need for a rehearsed resuscitation protocol is important, as is a systematic scheme that emphasizes the skilful use of eight specific cannulas and the treatment of priority problems first.


Fulminating nonclostridial gas-forming infection: a case of necrotizing fasciitis.

Perianal infection in a 40-year-old man resulted in extensive necrotizing fasciitis of the retroperitoneal space and septic shock. Despite the fact that radiography revealed linear streaking in the belly of the psoas muscle due to gas formation, the nature of the infection was necrotizing fasciitis and not myonecrosis. This contradicts Brightmore's contention that such a finding always indicates clostridial myonecrosis. Despite the absence of Clostridium welchii, necrotizing fascitis is none the less extremely serious, usually occurring in the limbs or abdominal wall superficial to muscle layers. The case reported is unusual in that infection affected the fascia deep to abdominal muscles in the retroperitoneal space, where surgical exposure is difficult. An appropriate surgical approach afforded adequate treatment.


Duodenal obstruction related to benign biliary tract disease.

Duodenal obstruction in six patients was related to biliary tract disease and previous abdominal surgery. Causes of obstruction were adhesions to the inflamed gallbladder, adhesions to the gallbladder bed after cholecystectomy, intramural hematoma after transduodenal exploration of the common bile duct and severe pancreatitis after common bile duct exploration. Measures that may possibly prevent duodenal obstruction include early recognition and treatment of cholelithiasis, positioning of omentum between duodenum and gallbladder bed after cholecystectomy, avoidance of transduodenal exploration where possible and careful duodenal closure if necessary, avoidance of forceful dilation during common bile duct exploration, and a decrease in the number of negative explorations by increased use of cystic duct cholangiography


Spontaneous resolution of a pancreatic pseudocyst by perforation into the colon.

Spontaneous perforation of a pancreatic pseudocyst into the colon, diagnosed radiologically, resulted in spontaneous cure. Disappearance of a previously palpable abdominal mass suggested perforation of the cyst into the stomach or colon. Surgical treatment was not required. Such an occurrence has not been reported previously; it opens to doubt dicta regarding treatment of this rare complication of pancreatic pseudocyst by immediate colostomy.


Malignant degeneration of a chondromyxoid fibroma in a child.

A 10-year-old boy was treated for a chondromyxoid fibroma of the left femur by curettage, and 2 months later complete healing was demonstrated radiographically. Three years after operation, radiographs revealed replacement of the ilium, ischium and upper femur, and destruction of the left hip by a soft-tissue and bony mass; a left hemipelvectomy was performed for what was shown to be a chondrosarcoma. Follow-up 10 years after the initial operation and 7 years after the hemipelvectomy has confirmed uneventful recovery in this rare case of malignant degeneration of a proven chondromyxoid fibroma of bone.


Prophylactic role of cephaloridine in surgical wounds.

The incidence of postoperative wound infection was studied in relation to 146 surgical wounds in patients who received cephaloridine as prophylactic antibacterial therapy, and in relation to 120 wounds in another group of patients who did not receive cephaloridine. In the first group, 5 of 146 (3.4%) of wounds became infected; in the second, 25 of 120 (20.8%) of wounds became infected. The difference between the two groups is significant. Use of cephaloridine in this series of patients therefore decreased the incidence of postoperative wound infection.


Depression after myocardial infarction.

The Minnesota Multiphasic Personality Inventory was completed by 101 patients 16 to 18 months after a proved myocardial infarction. The data suggested a bimodal distribution of patients. One class of patients had a relatively "normal" personality score apart from a tendency to hypomania. The second class had severe depression, with associated hysteria, hypochondriasis and psychasthenia. The severely depressed patients were older, with a greater tendency to hypertension and angina, and a tendency to smaller gains in aerobic power despite an equal intensity of endurance training. The distinction between "normal" and "depressed" postinfarction patients seems of some clinical importance, for the two classes of patients require opposite supportive techniques--restraint and encouragement, respectively.


A practical classification of untoward drug effects.

All drug effects can be explained as results of complex interactions between the drug, the patient and his condition, and additional extrinsic factors. On the basis of these three "determinants", a practical classification of untoward drug effects (UDE) is suggested. UDE lists using this classification would fulfill the physician's informational needs better than the material with which he is presently provided.


Acute hemiplegia of childhood.

Acute hemiplegia of obscure cause occurred in 28 children: 13 had had prolonged seizures and a high temperature (considered to have been the direct cause of the brain damage); 5 had had brief seizures, a lower temperature and a depressed level of consciousness; and 10 had a nonfebrile onset of hemiplegia and were found to have vascular abnormalities.  Most of the first group were retarded and epileptic at long-term follow-up, as were about half of the second group, whereas children in the third group were of normal intelligence and epilepsy was uncommon  among them.  Hemiplegia persisted at follow-up in most of the children in each group, the proportion being at least in the third group; if cerebral angiography had demonstrated carotid stenosis or occlusion there was usually poor recovery from the hemiplegia.  Bilateral changes on plain skull films or pneumoencephalograms were associated with mental retardation.  Failure to control prolonged seizures accompanied by a high temperature predisposes to brain damage; therefore, early and vigorous management is essential.


Prognostic significance of endometrial extension from primary carcinoma of the uterinecervix.

This is a retrospective analysis of 302 patients with histologically confirmed primary carcinoma of the cervix on whom a dilatation and curettage was performed during the initial workup. The prognostic significance of endometrial extension of the tumor was investigated. Four different types of specimen were identified: 1) endometrial extension of cervical carcinoma; 2) cervical carcinoma in the currettings only; 3) a mixture of normal endometrium and and epidermoid carcinoma in the curettings; 4) no cervical carcinoma noted (negative D&C). The patients were staged according to the FIGO classification. Patients with Stage I carcinoma and positive D&Cs showed significantly lower 3-5 year survival rates (50-60 percent), as opposed to those with D&C.


Treatment of primary adenocarcinoma of the cervix.

Between 1956 and 1971, a total of 74 cases of adenocarcinoma of the cervix was treatedin the Division of Radiation Therapy of the University of Maryland Hospital.  Radical radiation therapy was followed by routine surgery early in the study;after 1967, surgery was used only for radiation failure.  The likelihood of local control and 5-year survival was not improved by the routine addition of surgery to radical radiation, although the incidence of serious complications was markedly elevated.  The results of treatment of adenocarcinoma of the cervix by radiation therapy alone are not significantly different from those achieved with squamous cell carcinoma.  Surgery should be used as a salvage procedure in case of failure, rather than on a routine basis.


Evaluation of University of Florida split-course technique for various head and neck squamous cell carcinomas.

A retrospective evaluation was made of 260 patients with squamous carcinoma of various head and neck sites in order to determine the relative value of continuous vs. split-course irradiation techniques.  There was no obvious difference in local control or survival in early-stage cases.  The continuous technique resulted in a better local control and survival rate in moderately advanced and advanced stages.  Complications were less severe in the split-course cases.


Cellular radiosensitivity of a rat brain tumor.

The response of a model brain tumor system in the rat to single doses of x irradiation has been studied.  Solid tumors were exposed in situ, removed, dissociated into single cells, and grown in tissue culture.  The fraciton of surviving clonogenic cells wasdetermined as a function of x-ray dose level for tumors irradiated in anesthetized, air-breathing rats, and for tumors irradiated in rats killed 5 minutes prior to irradiation bynitrogen gas asphyxiation.  The parameters of the survival curve for tumors irradiatedin air-breathing rats were: DO= 295 rads; DQ= 350 rads; and n= 3.0.  Similarly, thesurvival parameters for tumors irradiated in nitrogen gas asphyxiated rats were: DO=625 rads; DQ=675 rads; and n= 3.0. The survival curve of these tumor cells when assayed in in vitro tissue culture only has parameters of: DO= 225 rads; DQ= 350 rads; andn= 4.7.  Data form the in vivo to in vivo tumor cell survival curves suggest that there is no significant hypoxic fraction in this brain tumor.


The role of radiation therapy in the treatment of astrocytomas.

One hundred forty-seven patients with astrocytoma were treated between 1942 and 1967. There were 25 postoperative deaths.  The 14 patients in whom the tumor was thought tohave been completely removed were not irradiated and all survived 5 years or longer.  Seventy-one of the 108 patients with imcompletely excised lesions received radiation therapy.  The 5-year survival rate for those with imcomplete resection alone was 19%, compared to 46% when irradiation was given.  Based on observations up to 20 years, after incomplete removal postoperative irradiation significally prolonged useful life and may have lead to permanent control in some.  There was no evidence of radiation damage.  Most of these tumors were fibrillary astrocytomas, and the results apply particularly to this histologic type.  Only 1 of 11 patients with gemistocytic astrocytoma survivied 5 years. The survival rate for Grade i tumors was appreciably greater than for Grade iilesions; in both grades, it was improved by irradiation.


Radiation tolerance of the spinal cord.

A series of nine cases of radiation myelopathy seen at the University of California, San Francisco (UCSF) is reviewed, and their treatment data converted into nominal single doses (NSD) and equivalent single doses (ED).  The 1% incidence level of myelopathy in the thoracic cord is 1015 rets (ED), and the 50% incidence level is 1476rets (ED).  Caution should be used when utilizing a rapid fractionation schedule; it appears that 2000 rads in 5 fractions and 3000 rads in 10 fractions is a safe regimen for the thoracic spinal cord.


Improved survival of patients with intracranial ependymomas by irradiation: doseselection and field extension.

Intracranial ependymomas, treated with postoperative irradiation, have a 50% 5-year survival.  Although this rate is good, the issue remains whether the quantity and qualityof survival can be improved by the proper selection of treatment factors, i.e. field size and dose-time relationship.  Factors influencing survival and the effect of doseand field selection are considered in 28 intracranial ependymomas.  Nineteen spinal primaries are also presented for comparison.  Only 10% of the patients treated with dosesbelow 1350 rets are alive, vs. 56% of the patients treated with higher doses.  Only21% of the patients treated with partial brain irradiation survived, vs. 57% of those treated with whole brain irradiation.  Although an apparent 5-year survival difference was found for the low-grade (63%) vs. high grade (13%) tumors, high-dose whole brain irradiation had a significant effect only on the survival of thehigh-grade tumors.  Local recurrences accounted for the majority of theraputic failures.  Ten consecutive patients were treated with elctive craniospinal irradiation and exhibited a 3-year survival of 70%, vs. 35% in those treated conventionally to the brain only.  Elective craniospinal irradiation, advocated for the high-grade, particularly infratentorial, ependymomas, has currently yieldedhigher survival, lower recurrences, good tolerance, no complications, and excellent quality of survival.  Further documentation of this treatment approach is suggested.


The influence of dose fractionation on acute and late reactions in patients with postoperative rasiotherapy for carcinoma of the breast.

The clinical courses of 418 patients with breast carcinoma who received postradical-mastectomy radiation therapy to the internal mammary and supraclavicular are were revied in order to determine the normal tissue tolerance with various time-dose fractionation radiation regimens.  Four different dose fractionation schedules wereemplyed during the period 1958-1968 at Memorial Hospital: 4900 rads/10f/32d; 4410 rads/9f/30d; 5000 rads/13f/18d; and 4600 rads/20f/28d.  There was no difference in the acute reaction or local recurrence rate of the tumor in the irradiated area among the four fractionation schemes, but the late effects were quite different.  Patients who received 4900 rads in 10 fractions in 32 days had a 33% incidence of subcutaneous fibrosis.A similiar dose in 13 fractions in 21/2-3 weeks resulted in about 17% subcutaneous fibrosis.  No significance of these clinical findings is discussed in relation to the various formulae currently being used to express time-dose equivalence.


Preoperative irradiation for carcinoma of the rectum and rectosigmoid colon: reportof a National Veterans Administration randomized study.

In 1964 the Veterans Administration Surgical Adjuvant Group (VASAG) initiated a large-scale, controlled, randomized protocol to study the role of low-dose preoperative irradiation (2000-2500 rads/10fractions/12 days) in patients with operable adenocarcinoma of the sigmoid colon and rectum.  This report analyzes the data in 700 patients, all at 5-year risk.  There appears to be a definate benefit to irradiated patients who undergo abdominoperineal resections, when compared with the controls.  This advantage is reflected in improvement of 5-year survival, and reduction in lymph node invasion, local recurrence, and distant metastases.  A second protocol has been initiated in 30 VA hospitals employing a higher dose (3150 rads) to extended portals (toL2) to male patients who require abdominoperineal resections.


The effect of radial radiotherapy on delayed hypersensitivity and the inflammatory response.

Delayed hypersensitivity to DNCB and the inflammatory response to croton oil wereevaluated in 144 and 121 patients respectively, prior ro and 3 to 6 months following curative radiotherapy.  Eighty-one patients had in vitri lymphocyte transformation by PHA;59 (41%) were nonreactors to DNCB and 27 (22%) to croton oil; 29 of 59 (49%) initiallyreactive became anergic.  Similiar improvement of the inflammatory response was obtained.  Patients who became DNCB-reactive following radiotherapy had the same favorable prognosis as those who were initially reactive.  Radiotherapy did not adversely affect either delayed hypersensitivity or the inflammatory response.  There was a 50% decrease in PHA stimulation and lymphocyte count after treatment.  No correlation was found between DNCB reactivity and lymphocyte transfermation prior to or following radiotherapy. The evaluation of the effect of radiotherapy on cell-mediated immunity depends on the tests used.


The role of splenic irradiation in chronic lymphocytic leukemia.

Fourteen patients with chronic lymphocytic leukemia (CCL) who received a total of 23 courses of splenic irradiation (SI)for various combinations of painful splenomegaly, progressive leukocytosis, lymphocytosis, thrombocytopenia, and anemia are reviewed.  Splenic midplane doses ranged from 200-1750 rads (3-14 days), while the most typicalregimen was 300-450 rads in two to three fractions of 150 rads given over 3-8 days.  Response to SI was rated according to a scoring system which evaluated the splenic and hematologic response, as well as the response of disease-related symptons.  According to this scoring system, most patients demonstrated a significant relief of painful splenomegaly, along with improvement of hemogram and bone marrow parameters.  While those patients judged as failures to prior chemotherapy or total body irradiation showed improvement following SI, those who had received minimal therapy prior to SI showed a superior response.  SI, in those showing a satisfactory response, was repeated successfully in several patients (up to six times in one instance).  The onset and duration of response to SI, dose-reponse data, survival, clinical and hemotologic responses, and the possible mechanism of action of SI are discussed with reference to the available literature.


Primary carcinoma of the ureter: a prognostic study.

Fourty-one patients with primary invasive carcinomas of the ureter were seen at Memorial Hospital from 1947 to 1972.  Overall survuval patterns were similiar in 19 patients with and 22 patients without prior or concomitant urothelial cancers elsewhere in the urinary tract, with 5-year survival rates, as estimated by the product-limit methos, of41% for both groups.  Prognosis was determined primarily by anatomical stage of ureteral cancer.  In 11 Stage A (submucosal) patients, 7 Stage B (muscular), 12 Stage C (periureteric fat), and 9Stage D (extraureteral), the similarly estimated 5-year survival rates were 91%, 43%, 23%, and nil, respectively.  None of Stage A cases had metastases for periods ranging from 5 to 11 years after surgery alone.  Seventy-eight percent of patients with more advanced stages died within 3 years of treatment, withmetastases mainly in pelvic and para-aortic lymph nodes.


Measures to minimize small intestine injury in the irradiated pelvis.

Small intestine injury causes long-term suffering and high mortality.  Five of 187of our patients had developed serious small intestine injury.  Four patients had corrective surgery.  Three patients died.  All were women.  Subsequently, all patients who received definitive pelvic irradiation had small intestine roentgenograms to determine itslocation and mobility. Female patients, thin patients, and elderly patients had larger amounts of small intestine in the whole pelvis, a deeper cul de sac, and a greater incidence of relatively immobile small intestine.  Patients with relativelyimmobile small intestine in the treatment field may be predisposed to injury.  There wasno relationship of the incidence of relatively immobile small intestine to prior pelvicsurgery. We used the findings from the small intestine roentgenograms to modify indiviually the radiotherapy regimen so as to minimize the risk for small intestine injury.  Patients were placed in the prone position to displace the small intestineout of the treatment fields used for booster dose irradiation.  The treatment field wasmodified to exclude the small intestine.  The total tumor dose delivered was determinedby expectations for curve vs. complications.  To date, none of the patients in this study group has developed small intestine injury.  Cadaver studies showed the feasibility of elective shortening of the pelvic cul de sac.  The small intestine can be displacedaway from the bladder, prostate, or cervix.


The risk of tumor induction in man following medical irradiation for malignant neoplasm.

A review of second malignant tumors (S.M.T.s.) following therapeutic irradiation formalignant neoplasm in man is presented.  Twenty-one new cases are added to the literature, 3 of adenocarcinoma of the endometrium following treatment for squamous cell carcinoma of the cervix, 3 of adenocarcinoma of the large bowel following treatment for squamous cell carcinoma of the cervix, 9 S.M.T.s in irradiated oropharyngeal or oral mucosa, and 6 single cases of S.M.T.s following radiotherapy of cancers in other sites. It is suggested that there is relatively low incidence of S.M.T.s among patients formalignancy, but the risk remains to be established by an appropriate study of controland test populations.


The effect of age on the long-term response of bone marrow to local fractionated irradiation.

The in-field long-term progressive response of bone marrow to localized fractionatedradiation to the thoracic spine was observed in weanling and adult rats.  Total dose levels were 1800,3600 and 5400 rads given in daily fractions of 180 rads for 900 rads perweek, to simulate the clinical setting.  Animals were sacrificed at 4 1/2, 6, 8, and 10months after irradiation, and bone marrow from within the treated area was ezamined.  The juvenile marrow repopulated to higher levels of cellularity and after higher doses of radiation than the adult.  The degree of marrow depletion, time of repopulation, and ultimate level of repopulation after cessation of irradiation were all dependent on the total dose.  Some of the suggested clinical implications include re-evaluation of the lower doses given to children.


Erythrocytosis and renal cell carcinoma with pulmonary metastases: case report with 18-year followup and brief discussion of literature.

A 42-year-old man was admitted to the Cancer Division of the American MedicalCenter in February, 1956.  He had been treated for erythrocytosis beginning 6 months previously, with reduction of his hemoglobin to relatively normal values.  On admission, a huge tumor of the left kidney was palable.  Roentgenograms of the  chest demonstrated small bilateral pulmonary metastases.  A left nephrectomy was per formed, followed by wedge resection of a total of 16 metastatic nodules, 12 of which showed metasticrenal carcinoma.  It is now 18 years since the last thoracotomy and the patient is well, leading a normal life, and free of any demonstrable malignant disease and any evidenceof erythrocytosis.


Epitheloid sarcoma masquerading as Peyronie's disease.

A 23-year-old man presented witha firm plague-like penile mass with pain and deviationon erection, suggesting Peyronie's disease.  Biopsy was erroneously interpreted asconformatory.  Irradiation was the one mode of therapy which gave temporary subjective relief.  Extension of the mass and focal gangrene led to rebiopsy 6 years later.  Epithelioid sarcoma was present microscopically, and was also found on re-examination of theoriginal biopsy.  Partial penectomy was performed, but pulmonary metastasis had alreadyoccurred.  He was treated with chemotherapy and radiotherapy and is now alive with disease 2 years later.  The diagnosis of possible penile sarcoma, although rare, must be considered in patients with symptons suggesting Peyronie's disease, especially if the lesion continues to enlarge.


Estimation of tumor cell kill from Gompertz growth curves.

The Gompertz function has been found to mathematically describe several types of biologic growth, including the growth of transplantable animal tumors. If it is assumed that treatment (chemotherapy, surgery, x-irradiation) of tumors causes cell death in a relatively short interval and that the surviving cells begin to repopulate immediately, producing tumor growth according to the Gompertz function before treatment, then the displacement in time between the two growth curves may be quantitatively related to tumor cell kill. This theory is developed and compared to bioassay values. It is also shown that tumor cell kill may be determined graphically without the necessity of computer-generated estimates of the Gompertz parameters.


"Platinum-pyrimidine blues" and related complexes: a new class of potent antitumor agents.

The diaquo species of cis-dichlorodiammineplatinum (II) reacts with pyrimidines and substituted pyrimidines to form very soluble, deep blue complexes. These are believed to consist of mainly monomeric species containing one pyrimidine moiety liganded to each platinum. The structures of the complexes are largely uncertain at this time. We describe the methods of synthesis and characterization of approximately 70 such complexes, with a suggested classification scheme, incluse complexes show superior activity against the ascites Sarcoma 180 tumor in Swiss mice when compared to cis-dichlorodiammineplatinum (II). Initial screening test results and dose schedule-dependency results indicate we can achieve 100% cures in this tumor-host system. Activity is also shown against the Rauscher leukemia, Ehrlich ascites, and ADJ/PC6A tumors. Microscopic histopathologic studies of resulting kidney lesions show that the "platinum-uracil blue" complex causes only minor focal damage to the proximal convoluted tubules at a therapeutic dose level, rather than the generalized degenerative changes so prominent and dose limiting noted with cis-dichlorodiammineplatinum (II).


Antitumor activity of tetraacetylglucosamine mustard.

1,3,4,6-Tetra-O-acetyl-2-(di-2-chloroethyl)amino-2-deoxy-D-glucopyranose is active against L1210 leukemia, giving over 100% increased life-span at optimal dose. Against P388 leukemia, it gives 200% increased life-span with long-term survivors. The compound is most active when given i.p., but shows some activity when given s.c. than p.o., and is more potent (therapeutic and toxic effect) than mechlorethamine on both a molar and a mg basis. Of importance, the schedule dependency for the administration of 1,3,4,6-tetra-O-acetyl-2-(di-2-chloroethyl)amino-2-deoxy-D-glucopyranose in L1210 leukemia differs from most alkylating agents in that it is best given by multiple daily injections rather than as a single large injection on Day 1. This characteristic can be attributed to the amino-glucose moiety.


Inhibition and enhancement of Friend leukemia virus by pyran copolymer.

Inhibition or enhancement of Friend leukemia virus disease could be produced by treatment of mice with the immunopotentiator, pyran copolymer. The result depended on the route of inoculation of the drug. Prophylactic administration of the drug i.p. retarded splenomegaly, reduced splenic foci, and increased survival time of mice infected with Friend leukemia virus. Conversely, when the same dose and regimen of pyran was administered i.v., splenomegaly was enhanced, splenic foci were increased, and survival time was decreased. Histopathological examination of the spleens of mice revealed that i.p. pyran administration caused a marked increase in the splenic marginal zone with some increase in erythropoiesis in the red pulp, while i.v. pyran administration did not markedly change the splenic marginal zone but caused an early and sustained increase in erythropoiesis in the red pulp.


Concanavalin A (NSC-143504): its action on experimental tumor cells and possible use in cancer chemotherapy.

Concanavalin A administered to mice kills KHT fibrosarcoma tumor cells and, less markedly, AKR lymphoma cells at doses where it has no effect on numbers of normal hematopoietic colony-forming cells. Concanavalin A in vitro agglutinates suspensions of cells prepared from murine and human tumors but does not agglutinate suspensions prepared from most normal mouse tissues. In spite of its selective action against tumor cells at the cellular level, the possibilities for using concanavalin A therapeutically are limited by its unusual toxicity, a generalized vasculitis that is seen in the mouse, monkey, and pig at approximately the same dose level.


Effects of high-molecular levan on the growth and spread of lymphoma in AKR mice.

Levan was shown to inhibit lymphoma development in AKR mice. Growth of tumor was inhibited at the site of injection and in metastases. Levan caused a decrease in the incidence of tumors, and it reduced pleomorphism, mitoses, and invasiveness of tumors in comparison to nonlevanized mice. Evidence of tumor cell destruction was observed in levanized mice. The effect of levan on tumor development was dose dependent.


Development of resistance to rubidazone (NSC-164011) in Ehrlich ascites tumor in vivo.

Resistance to a new daunorubicin derivative, rubidazone, was developed in a strain of Ehrlich ascites tumor in vivo by treatment with the drug during 11 weekly passages of the tumor. In this tumor reciprocal cross resistance was found between rubidazone, daunorubicin, adriamycin, vincristine, and vinblastine. There was no change in sensitivity of the tumor to methotrexate, 6-mercaptopurine, BCNU, cytosine arabinoside, or actinomycin D. The resistance was stable for at least 71 weeks when treatment was continued and for at least 18 weeks when treatment was discontinued. Rubidazone given intraperitoneally was significantly less toxic than daunorubicin and adriamycin; unlike the findings with daunorubicin and adriamycin, late toxic death (after 30 days) often occurred. In the wild-type tumor the growth inhibition with rubidazone was less than with daunorubicin and adriamycin on an equal weight basis. The LD10 of rubidazone inhibited the growth to nearly the same extent as did the LD10 of daunorubicin and adriamycin.


Problems in the clinical use of the microcytotoxicity assay for measuring cell-mediated immunity to tumor cells.

Lymphocytes from control donors and from cancer patients have been tested for antitumor, cell-mediated immunity against various melanoma and breast cancer target cell cultures with a microcytotoxicity assay. Control lymphocytes inhibited growth of target cells with high frequency, particularly with cell line, as opposed to short-term, cultures. Inhibition was not found for all target cells tested at a given time with a single preparation of lymphocytes. Sequential studies over a 2-year period with lymphocytes from the same control donors showed fluctuations of inhibition against the same target cells. With serial passage, the target cells also changed in their susceptibility to destruction by control lymphocytes. Lymphocytes from melanoma patients were more inhibitory than control lymphocytes for one melanoma target cell culture but not for two other melanoma cultures. Significant inhibition by lymphocytes from melanoma patients was not seen against two cultures derived from breast cancer patients. Lymphocytes from breast cancer patients were more inhibitory than control lymphocytes for 4 of 5 breast cancer cultures and they did not inhibit two melanoma cultures. Significantly specific inhibition was seen with short-term, but not cell line, breast cultures. The over-all data show specificity for target cells of the appropriate histological type. However, the high and inconstant reactivity of control lymphocytes in this assay suggests that nonspecific inhibition of tumor target cells by patient lymphocytes is found in many experiments. It is concluded that the microcytotoxicity assay is not suitable for clinical studies, since sequential data obtained in individual patients are difficult to interpret.


Intestinal tumors in mice treated with a single injection of N-nitroso-N-butylurea.

N-Nitroso-N-butylurea was injected once at dosage levels of 150 or 75 mg/kg into 3- or 6-week-old male C57BL/L mice. Intestinal tumors occurred in 100% of the mice that survived more than 15 weeks after injection with a high dose of N-nitroso-N-butylurea at 6 weeks of age, and in 35 to 70% of the mice in other treatment groups. These intestinal tumors were seen primarily at the junction of the pylorus and duodenum and in the anterior portion of the small intestine, with a few in the cecum, colon, and rectum. The tumors at the junction were not very invasive tumors and frequently appeared as polypoid growths. Tumors not at this location were adenocarcinomas that invaded all the layers of the gut wall but which did not metastasize. Colorectal tumors were adenomas and adenocarcinomas. N-Nitroso-N-butylurea also induced tumors of the stomach, hematopoietic system, lung, and liver.


Changes of the immunogenic properties of a radiation-induced mouse lymphoma following treatment with antitumor drugs.

Eight sublines of the radiation-induced lymphoma S-1033 of C57BL/10 (hereafter called B10) origin were established by exposing the cells in vivo to eight antineoplastic agents for a number of transplant generations. The parental and drug-treated sublines were tested for immunogenic properties, i.e., the ability to elicit allograft reactions in the host of origin and in congenic-resistant mice differing for the S-D or K-I-S regions of the H-2 complex. Lymphoma S-1033 and all drug-treated sublines except one were found to be essentially nonimmunogenic for B10 mice. The S-DIC subline, when exposed for 8 to 12 transplant generations to dimethyltriazenoimidazolecarboxamide, became immunogenic for syngeneic B10 mice, as judged from prolongation of survival time. Large i.v. inocula (10(7) cells) of S-1033 and of the drug-treated sublines, with the possible exception of the cyclophosphamide-treated and dimethyltriazenoimideazolecarboxamide-treated lymphomas, were more effectively rejected by K-I-S- than by S-D-incompatible mice. Dilution escape (i.e., tumor rejection after challenge with large inocula, and lethal tumor growth after injection of small inocula of lymphoma cells in allogeneic recipients) occurred in K-I-S-incompatible mice that were inoculated with S-1033 and three drug-treated (5-fluorouracil, cyclophosphamide, and pyrazocarboxamideamino) sublines. No dilution escape occurred with dimethyltriazenoimidazolecarboxamide or bischloroethylnitrosourea sublines. These data favor the hypothesis that various types of immunogenic changes of neoplastic cells may occur in tumor-bearing hosts following treatment with antineoplastic agents in vivo.


Irradiation-induced adduct formation of RNA with carcinogenic arylamine derivatives.

Radiolysis of N2O-saturated solutions of transfer RNA (tRNA) and the arylacethydroxamic acids, N-hydroxy-N-2-acetylaminofluorene and N-hydroxy-N-4-acetylaminobiphenyl; their corresponding acetamides, 2-acetylaminofluorene and 4-acetylaminofluorene; or the O-glucuronide of N-hydroxy-N-2-acetylaminofluorene resulted in adduct formation of the nucleic acid with these carcinogenic arylamine derivatives. The yield of adducts on irradiation of the arylacethdroxamic acids with tRNA was greater than that for their corresponding acetamides or the O-glucuronide. The fluorenylacethydroxamic acid and acetamide were also more reactive than the biphenyl analogs. Adduct formation resulting from radiolysis of tRNA and the arylacethydroxamic acids or the O-glucuronide proceeded with retention of both the aromatic nucleus and the N-acetyl group. The yields of adducts were much greater for irradiated mixtures than for irradiation of either component alone followed by mixing. Evaluation of the data shows that initial modification of the tRNA or the carcinogen can lead to adduct formation. In the case of primary radical attack of the nucleic acid, it has been shown that short-lived reactive RNA intermediates are responsible for a major fraction of the observed yield of adducts in the irradiated mixtures. Comparative studies showed that irradiation under conditions that favor reaction of oxidizing radicals enhanced formation of the adducts. Oxygen was shown to protect RNA from irradiation-induced binding of the arylacethydroxamine acids due to competition of O2 with the carcinogen for the reactive RNA intermediates.


Role of cellular RNA polymerases in virus-induced leukemogenesis.

Friend murine leukemia virus induces splenic enlargement and an increase in RNA polymerase activity of spleen nuclei. Actinomycin D, administered at 60 mug/kg body weight/day prevents the development os splenomegaly and the elevation of polymerase activity following infection, but it has only a slight effect on the production of virus in spleen tissue. Thus, the alteration of RNA synthesis is not a result of virus proliferation, but instead may be a manifestation of leukemic erythropoiesis. Normal erythropoiesis, stimulated by erythropoietin administration, produces a similar but transient increase in RNA polymerase activity in spleen nuclei. Erythropoietin administered before, but not after, Friend virus infection results in an enhancement of RNA polymerase activity, as measured 9 days after inoculation. This effect is most simply explained by assuming that there is a common target cell pool for both erythropoietin and Friend virus, and that this pool becomes refractory to the influence of the hormone as a result of the leukemic process.


The metabolism of benzo(alpha)pyrene in isolated rat liver cells.

Isolated rat liver cells catalyze the metabolism of benzo(alpha)pyrene (BP) with the resulting formation of phenols, dihydrodiols, and conjugates. The rate of the primary oxidative step in the process was similar to that catalyzed by isolated rat liver microsomes in the presence of a reduced nicotinamide adenine dinucleotide phosphate-generating system and responded similarly to various inhibitors, including 2-diethylaminoethyl-2,2-diphenylvalerate, metyrapone, alpha-naphthoflavone, and hexobarbital. The level of cytoplasmic, reduced nicotinamide adenine dinucleotide phosphate was not rate limiting in liver cells isolated from either fed or fasted animals. The conjugates and dihydrodiols formed were readily excreted, whereas low concentrations of phenols accumulated intracellularly. The pattern of metabolites of BP was the same in isolated rat liver cells and in the isolated perfused rat liver. 3-Methylcholanthrene treatment of the rats caused a marked increase in cellular BP metabolism as well as in cytochrome P-450 concentration. The induced hemoprotein revealed characteristics similar to those previously established with isolated liver microsomes, i.e., increase in high-spin form, enhanced affinity for BP as revealed by a lower Michaelis constant, and sensitivity to the inhibitory action of alpha-naphthoflavone. After 3-methylcholanthrene treatment, phenols and dehydrodiols constituted a larger percentage of the total metabolites, indicating a more pronounced stimulation of the oxidative than of the conjugative step of BP metabolism by induction, and the dihydrodiols now tended to accumulate intracellularly.


Glycoprotein metabolism in inflammatory and neoplastic diseases of the human colon.

Carbohydrate compositions of the membrane and cytoplasmic fractions of human normal and cancerous colonic mucosa were compared in patients with blood groups O and B. The total sugar content in both fractions was reduced in the cancer tissues to about one-third of that in the normal colonic mucosa. The sugars that are associated with mucinous glycoproteins such as fucose and N-acetylgalactosamine were reduced significantly, while sugars that are primarily associated with "serum-type" glycoproteins were relatively unchanged or reduced to a lesser extent. The activities of glycoprotein:glycosyltransferases were variable, some showing so significant change, others beinb significnatly reduced in cancerous tissues. A polypeptidyl:N-acetylgalactosaminyltransferase (an enzyme that catalyzes the transfer of the first sugar to hydroxyamino acids of the protein core of mucinous glycoproteins), a sialyltransferase (involved in the addition of sialic acid to mucinous glycoproteins), and a galactoxyltransferase (thought to be responsible for blood group B antigenicity) were reduced in the cancerous colonic tissue. In contrast, the activities of these glycosyltransferases were unchanged in the colonic mucosa of patients with granulomatosis or ulcerative colitis. Glycosidase activities in the normal, cancerous, and inflammatory tissues were the same. These results suggest that in colonic cancer tissues the synthesis of one type of oligosaccharide chain may be greatly affected, while another family of oligosaccharides may remain relatively unaffected.


Potentiation of hamster tumors by normal cells or charcoal.

Admixed spleen cells from normal animals or from animals given injections of Syrian hamster type C virus significantly potentiated the growth of the transplanted D9 lymphoma of random-bred hamsters. Potentiation was measured by an increase in incidence of tumors, a shortened latent period, and a decreased 50% tumor-producing dose of tumor cells. Intermediate doses of spleen cells (10 to 100 spleen cells per tumor cell) produced the greatest potentiation. Preincubation of admixed spleen and tumor cell suspensions in vitro was unnecessary. Immunization to isoantigens was not responsible for potentiation, since growth of a transplantable carcinoma of inbred hamsters was also facilitated by normal spleen cells. In addition, normal kidney or liver cells increased the incidence of tumors transplanted by a small number of tumor cells. Potentiation did not occur when spleen cells were injected at a site remote from the tumor cells. Since the potentiating cells might act eigher as a physical barrier to host response, or by blocking normal macrophage function, we injected charcoal with tumor cells. Simultaneous treatment with charcoal facilitated the growth of the lymphoma but not that of the carcinoma. Treatment with some doses of charcoal was also effective at distant sites. Although potentiation of tumor growth by cells or charcoal may operate through different mechanisms, these phenomena should be explored in regard to outgrowth of primary tumors, tumor immunity, or enhancement of tumor growth.


Megalocytosis and other abnormalities expressed during proliferation in regenerating liver of rats treated with methylazoxymethanol acetate prior to partial hepatectomy.

Rats were partially hepactectomized at various intervals after receiving a single injection of the carcinogen, methylazoxymethanol acetate. In treated rats, there was a delay and a reduction in the peak response of hepatic DNA synthesis. At 48 hr after partial hepatectomy, following completion of the first mitoses, almost all hepatocytes were enlarged (megalocytes) and many contained enlarged nuclei. Although at 7 days after the hepatectomy many megalocytes could still be found in centrilobular zones, the majority of the hepatocytes were of normal size or smaller. Abnormal anaphase and telophase figures containing chromosomal bridges and acentric fragments were found during the period of regeneration. In addition, discrete nests of small cells with increased cytoplasmic basophilia were evident. Megalocytes also appeared when rats were partially hepatectomized as late as 26 weeks after injection of the carcinogen. These results show that a single dose of methylazoxymethanol acetate can affect almost all hepatocytes and that latent effects persist for long periods of time.


Increased carcinogenicity of 2,6-dimethylnitrosomopholine compared with nitrosomopholine in rats.

Nitrosomorpholine was given to rats in drinking water at the rate of 4 mg/week for 30 weeks. Tumors of the liver were induced in 53% of treated animals and were of both hepatocellular and Kupffer cell origin. One-half of the treated animals were alive 75 weeks after the beginning of treatment, but only 2 survived to 104 weeks. 2,6-Dimethylnitrosomopholine was given to rats at the same molar concentration in drinking water for 30 weeks (5 mg/week). All of these animals died with tumors within 34 weeks after the beginning of treatment; these tumors were mainly in the esophagus and nasal turbinates, and only 1 liver tumor was seen. This greatly changed tumorigenicity of the dimethyl compound might be due to steric factors or to enhanced reactivity at the positions alpha to the nitroso function induced by the methyl groups at the beta position.


Characterization of a chimpanzee anti-human melanoma antiserum.

An antiserum to human melanoma has been produced in a chimpanzee by hyperimmunization with melanoma cells from a single donor. After absorption with the peripheral blood lymphocytes of the tumor donor, this antiserum is specifically cytotoxic to melanoma cells from 14 tissue culture cell lines and to cells from 8 fetal fibroblast cell lines. Peripheral blood lymphocytes-absorbed antiserum is negative when tested against a large panel of normal peripheral blood lymphocytes and fibroblasts and against eight non-melanoma neoplastic tissue culture cell lines. Subsequent absorption with melanoma cells from any of seven sources removes all antimelonoma and all antifetal reactivity of the antiserum. Similar absorption with fetal cells removes all antifetal activity but does not completely remove the antimelanoma activity. The immunoglobulin G fraction of the antiserum is shown to possess the cytotoxic activity.


N-hydroxy-2-acetylaminofluorene inhibition of rat live RNA polymerases.

Administration of N-hydroxy-2-acetylaminofluorene (N-OH-AAF) to rats inhibits liver nuclear RNA synthesis. This effect is reflected in an in vitro inhibition of RNA synthesis by isolated whole nuclei. The decreased RNA synthesis can be accounted for entirely by an inhibition of the RNA polymerase activities quantitatively solubilized and partially purified from these nuclei. Both nucleolar and nucleoplasmic polymerases are affected. A similar inhibition of the polymerases was demonstrated in intact nuclei by inactivating the endogenous template with actinomycin D and assaying the polymerases with an added exogenous template, poly(deoxyadenylate-deoxythymidylate). Chromatin was prepared from similar nuclear preparations by two methods, differing in the extent to which they remove endogenous polymerase activity. Each chromatin preparation was transcribed with added Escherichia coli or partially purified rat liver nucleoplasmic RNA polymerase respectively. With either polymerase and either chromatin preparation, no inhibition of the template activity of chromatin isolated from N-OH-AAF-treated animals could be detected. It is concluded that N-OH-AAF is a potent inhibitor of rat liver nuclear RNA synthesis and that the mechanism of this inhibition is inactivation of the RNA polymerases. At the same time, N-OH-AAF leaves the chromatin template, at least quantitatively, intact for the synthesis of RNA. The implications of such an effect of N-OH-AAF on RNA synthesis are discussed.


Sequential irreversible, actinomycin D-sensitive, and cycloheximide-sensitive steps prior to cortisol inhibition of uridine utilization by P1798 tumor lymphocytes.

Events preceeding the cortisol inhibition of uridine utilization by corticoid-sensitive P1798 lymphocytes have been investigated. When tumor cells were incubated with 1 muM cortisol for 15 min and then washed free of steroid and reincubated in the absence of hormone, the expected decrease of uridine uptake failed to appear 1.5 hr later. In contrast, the removal of cortisol after 30 or 60 min did not prevent subsequent development of the steroid effect. Addition of actinomycin D with cortisol, or 15 min after hormone treatment was started, blocked steroid action. However, when actinomycin D was added 30 or 60 min after the initial exposure to cortisol, hormone-induced depression of uridine uptake was no longer prevented. To study the role of protein synthesis, cycloheximide was added to the tumor cell suspensions at various times after cortisol treatment was started. Cortisol suppression of uridine utilization was blocked when cycloheximide was added with the hormone or 30 min after the start of hormone treatment. Cycloheximide added together with cortisol and washed out with the steroid after 30 min did not prevent subsequent appearance of decreased nucleoside uptake. Hydroxyurea, an inhibitor of DNA synthesis, did not prevent cortisol action, even when present throughout a 2 hr exposure to the steroid. Hormone removal or actinomycin D addition after 1.5 to 2 hr (when uridine uptake was already inhibited about 25%) did not prevent intensification of the steroid effect during a subsequent 1.5- to 2-hr incubation period, while addition of cycloheximide at this time completely prevented its progression. These results suggest aht: (a) cortisol inhibition of uridine uptake by P1798 lymphocytes involves an early irreversible step and appears to require continuing RNA but not protein synthesis during the first 15 to 30 min of hormone action; (b) protein synthesis but not RNA synthesis is required between 30 and 60 min; and (c) continuing protein synthesis but not RNA synthesis or hormone presence is necessary for the preestablished cortisol effect to progress.


The identification of phorbolol myristate acetate as a new metabolite of phorbol myristate acetate in mouse skin.

Aspects of the metabolism of phorbol myristate acetate (PMA) in mouse skin were investigated. Phorbolol myristate acetate (PHMA), a potential metabolite of PMA in mouse skin, was prepared from PMA by NaBH4 reduction of the C-5 carbonyl group of PMA to a secondary alcohol. The structure of PHMA was assigned on the basis of spectral and chemical evidence. PHMA had an inflammatory effect in mouse skin equal to or slightly less than PMA, on the basis of the dermal infiltration of lymphocytes and neutrophils and interfollicular epidermal hyperplasia. Application of PHMA and PMA to mouse skin resulted in equal increases in numbers of interfollicular epidermal basal layer mitotic cells after 28 hr. PHMA was found to be nearly as effective as PMA in increasing the incorporation of [3H]thymidine into interfollicular epidermal basal layer cells after 28 hr. When tritiated PMA was applied to mouse skin, PHMA was identified as a metabolite. PHMA was present in about 1.7% of the amount of PMA. When tritiated PHMA was applied to mouse skin, PMA was not detected as a metabolite in the mouse skin extracts.


Mode of action of 9-beta-D-arabinofuranosyladenine on the synthesis of DNA, RNA, and protein in vivo and in vitro.

The influence of 9-beta-D-arabinofuranosyladenine (ara-A) and its 5'-triphosphate derivative on programmed synthesis was tested with an intact cell system as well as with isolated enzyme systems. The effect of ara-A was tested in mouse lymphoma cells (L5178Y). The compound reduces cell proliferation in low concentration by cytostasis; under high ara-A concentration of radioactive precursors into DNA, RNA, and protein showed that ara-A selectively inhibits DNA synthesis. Formation of a polysome complex is not affected by ara-A. [3H]ara-A is incorporated into DNA in an intact cell system; 1 molecule of ara-A is incorporated per 8000 molecules of deoxyadenosine. Most of the ara-A molecules appeared to be in internucleotide linkages. Incorporation of ara-A into RNA could not be detected. 9-BETA-D-Arabinofuranosyladenine 5'-triphosphate (ara-ATP) does not reduce the incorporation rate of the following enzymes, isolated from quail oviducts: DNA-dependent RNA polymerases I and II, polyadenylic acid polymerase, and poly(adenosine diphosphate ribose) polymerase. The compound was found to inhibit DNA synthesis catalyzed by DNA polymerases isolated from quail oviducts and from oncogenic RNA viruses (Rous sarcoma viruses). All the enzymes tested were inhibited by ara-ATP in a competitive way with respect to deoxyadenosine 5'-triphosphate. The highest affinity of ara-ATP, i.e., the highest inhibitory potency of the drug, was found in the assays with the eukaryotic low-molecular DNA-dependent DNA polymerase. The influence on the eukaryotic high-molecular DNA-dependent Dna polymerase was a litte less. Compared to the eukaryotic DNA polymerases, the viral enzymes (RNA-directed DNA polymerase and DNA-directed DNA polymerase) are affected to a smaller extent by ara-ATP. No effects of ara-A and ara-ATP are observed in a protein-synthesizing, cell-free system isolated from L5178Y cells.


Granulocytes as effectors in cell-mediated cytotoxicity of adherent target cells.

In the microassay for cell-mediated immunity, detachment of adherent target cells from the wells occurs to an even greater extent when tested with granulocytes than when tested with lymphocytes. Intact cells are not necessary since the sonic extract from granulocytes causes the same effect. The reaction by granulocytes appears to be mediated by enzymes, and is inhibited by the presence in the wells of hyaluronic acid. Moreover, it is limited to the detachment of adherent target cells, since neither intact nor sonically disrupted granulocytes exhibit cytotoxic activity in the chromium release assay. The detachment of target cells is also inhibited by heparin which may be used to specifically mullify the effect of granulocytes contaminating lymphocyte suspensions.


Dental abnormalities in rats after a single large dose of cyclophosphamide.

Delayed drug-related mortality in rats treated with a single high dose (75 mg/kg) of cyclophosphamide complicated experiments using this drug treatment. We observed that this delayed mortality was due to dental abnormalities including broken teeth, absent teeth, extra long teeth, and/or supernumerary teeth. These dental abnormalities developed about 140 days after treatment and, if left untreated, interfered with eating. Eventually, the untreated rats starved. Clipping their long teeth and feeding the rats powdered chow eliminated the deaths. Researchers should be aware that high doses of cyclophosphamide may result in dental abnormalities several months after the treatment.


A new approach for induction of pancreatic neoplasms.

Weekly s.c. injections of equitoxic doses of 2-hydroxy-propyl-n-propylnitrosamine, 2-oxopropyl-n-propylnitrosamine, and methyl-n-propylnitrosamine, assumed metabolites of di-n-propylnitrosamine by beta oxidation, induced low incidences of pancreatic duct adenomas in Syrian golden hamsters. Di-n-propylnitrosamine did not. Application of 2,2'-dihydroxydi-n-propylnitrosamine, another postulated intermediate of di-n-propylnitrosamine, led to development of various types of pancreatic duct adenomas and ductal carcinomas in high percentages of hamsters. In addition, a few acinar-cell carcinomas were found. The morphology of these neoplasms, their latencies, and their distribution in the different segments of the pancreas are described.


Pancreatic adenocarcinoma in inbred guinea pigs induced by n-methyl-N-nitrosourea.

Long-term studies of the carcinogenicity of N-methyl-N-nitrosourea (MNU) in inbred guinea pigs were undertaken to develop an animal model of pancreatic adenocarcinoma. MNU, freshly, dissolved in 0.015 M sodium citrate buffer, pH 6.0, was administered by gavage once weekly at a dose of 10 mg/kg body weight to inbred strain 13 guinea pigs. By 27 weeks, 54% of guinea pigs given MNU weekly died, mostly due to severe atrophy and fatty metamorphosis of the exocrine acinar cells of the pancreas. Of 34 guinea pigs that survived more than 27 weeks of MNU administration, 10 (29%) developed pancreatic adenocarcinoma between 28 and 44 weeks. Other tumors included adenocarcinoma of the stomach (two animals) and of colon (one animal), lymphoma of mesenteric nodes (three animals), and a hepatoma (one animal). Atypical changes involving acinar cells were observed in certain pancreatic lobules and included ductular or pseudoductular transformation, acinar ectasia, increased mitotic activity, and periacinar fibrosis. These changes are suggestive of acinar cell dedifferentiation, and their role, if any, in the histogenesis of pancreatic adenocarcinoma remains to be elucidated. These studies suggest the feasibility of using inbred guinea pigs for developing a satisfactory model of pancreatic adenocarcinoma. Additional studies are necessary to minimize the high mortality rate during the first 6 months and to enhance the incidence of pancreatic adenocarcinoma.


Preliminary observations on the mitogenic effect of cyclopropenoid fatty acids on rat pancreas.

Male Sprague-Dawley rats were fed a diet containing 500 ppm of cyclopropenoid fatty acids (CPFA). After 2 weeks, mitotic activity and [3H]thymidine incorporation into DNA were significantly increased in pancreatic acinar cells. Continued feeding of the diet for 4 and 8 weeks led to decreasing mitotic activity which was still significantly increased over that of control animals. Focal necrosis of acinar cells was histologically apparent after 8 weeks of CPFA ingestion. Ultrastructural evidence of focal cytoplasmic injury was detected in acinar epithelial cells as early as 2 weeks after feeding of the CPFA diet was begun. A difference in dose response to CPFA appears to exist between Sprague-Dawley and Fischer F344 rats in that the latter, a dose of CPFA (5 mg/100 g body weight), did not evoke a mitogenic response.


Survival of functional pancreatic acinar tissue in circumfusion organ culture enhanced by chemically defined medium with hydrocortisone.

Pancreatic explants from perinatal or 1-week-old rat circumfusion organ cultured with an insulin-free variant of Trowell's Medium T8 survive functionally, as judged from tissue amylase content, for about 3 days. When hydrocortisone 21-sodium succinate, 1.0 mg/liter, is added to the chemically defined medium, high levels of anylase persist for longer periods. Explants from 7-day-old animals, circumfusion cultured with hydrocortisone-supplemented medium, maintain tissue amylase levels equal to or greater than those of uncultured control pancreas for at least 5 days of culture, and over this period they release amylase into culture medium at a stable rate. Methods for maintaining functional pancreatic acinar tissue in culture provide a new biological model for in vitro analysis of the early defects of potential chemical carcinogens on this target organ.


The pancreatic epithelial cell in vitro: a possible model system for studies in carcinogenesis.

A technique is described for the isolation and maintenance in culture of pancreatic epithelial cells from human embryos or neonatal guinea pigs. Dissociated cells from human material were essentially uncharacterized. Those obtained from guinea pig pancreas were subjected to centrifugal fractionation, and the resulting populations consisted of up to 95% exocrine cells. Suspensions of dissociated cells were incubated with gyration for 16 to 18 hr, and cell aggregated that formed were placed in stationary culture. Two-dimensional colonial aggregates developed on the plastic substratum. Cells comprising such colonies formed junctional complexes and could be maintained for 20 to 40 days in vitro. This system may provide a useful model for studies on pancreatic exocrine cell physiology and carcinogenesis.


Spontaneous release of DNA by human blood lymphocytes as shown in an in vitro system.

Human blood lymphocytes released DNA in vitro in the absence of any stimulation. Once purified from the complex appearing in the supernatant, this DNA exhibited typical characteristics as shown by its UV absorption curve, its deoxyribose coloration, and its sensitivity to DNase. Elution patterns on hydroxyapatite columns indicated that the excreted DNA is double stranded. The released DNA was smaller than the cellular DNA; its molecular weight ranged from 3.5 x 10(5) to 3.7 x 10(6) daltons. The DNA appearing in the supernatant does not seem to be due to dead or dying cells since: (a) the same amount of DNA was found in the medium whether the incubation lasted 2 hr or as long as 16 hr; (b) cell death rate had no effect on the amount of extracellular DNA; (c) when the lymphocytes were centrifuged and placed in a new medium several times in a row, a similar amount of extracellular DNA was isolated from each of the successive supernatants, whereas if, after centrifugation, the lymphocytes were put back in their original medium, no increase in the amount of extracellular DNA was observed, suggesting an active regulatory mechanism independent of a mechanical effect; (d) it took more than 1 hr for extracellular DNA to reach its maximum concentration, a fact that also argues against a mechanical effect; (e) the specific activity of the released DNA was different from that of the cellular DNA, depending on the time of labeling; and (f) the cells that had excreted DNA kept their functional integrity, as shown by their fully maintained capacity to increase DNA synthesis after stimulation. The extracellular DNA hybridized specifically with cellular DNA. The hybridization curves indicate that the DNA excreted is highly complex, and they suggest that it is composed of part of the newly synthesized DNA. The higher specific activity of the released DNA, compared with that of the cellular DNA after a long labeling period, can be explained only by a preferential release of the newly synthesized DNA.


1-beta-D-arabinofuranosylcytosine-induced malignant transformation of hamster and rat cells in culture.

The potent antileukemic chemotherapeutic drug, 1-beta-D-arabinofuranosylcytosine (ara-C), was observed to transform malignantly secondary hamster fetal cells as well as established rat cells in vitro. Results indicated that: (a) hamster cells altered by treatment with ara-C are morphologically indistinguishable from cells transformed with benzo(a)pyrene; (b) the ara-C-induced transformation can be observed under conditions of little or no inhibition of DNA synthesis and little or no cytotoxicity; (c) the transformation can occur with only a 6-hr exposure to ara-C; and (d) the transformation of hamster cells seems to require cellular DNA synthesis for cells in S phase are much more sensitive to ara-C-induced transformation than are G1-arrested cells. Representative transformed colonies of hamster and rat cells produced rapidly growing fibrosarcomas in inoculated newborn hamsters and rats, respectively. Control cells remained nontumorigenic.


Inhibition and recovery of DNA synthesis in host tissues and sensitive and resistant B16 melanoma after 1-(2-chloroethyl)-3-(trans-4-methylcyclohexyl)-1-nitrosourea, a predictor of therapeutic efficacy.

Single doses of 1-(2-chloroethyl)-3-(trans-4-methylcyclohexyl)-1-nitrosourea caused transient suppression of [3H]thymidine incorporation into DNA in bone marrow and gastrointestinal mucosa and more prolonged inhibition of such incorporation in B16 melanoma. A single dose of 1-(2-chloroethyl) -3- (trans-4-methylcyclohexyl) -1-nitro-sourea, 16 mg/kg, doubled the mean life-span after treatment of C57BL times DBA/2F1 male mice bearing 12-day-old B16 melanomas. Subsequent doses timed to minimize toxicity and maximize antitumor effect, however, produced no further prolongation of survival, and studies with B16 melanoma previously expsed to 1-(2-chloroethyl)-3-(trans-4-methylcyclohexyl)-1-nitrosourea demonstrated that the suppression of [3H]thymidine incorporation into DNA was no longer prolonged beyond that seen with normal host tissues. The loss of clinical efficacy was accompanied by a loss of differential suppression of [3H]thymidine incorporation into DNA between the tumor and host tissues.


Tumor induction relationships in development of transplantable cancers of the colon in mice for chemotherapy assays, with a note on carcinogen structure.

In an effort to establish an animal colon tumor model suitable for biological and chemotherapy studies, 82 colon tumors were induced and transplanted in four different inbred strains of mice. Four colon tumors survived the first transplant and are now in serial passage. All are suitable for chemotherapy trials. Two tumors are highly metastatic, and at least one of these is known to be suitable for surgery-chemotherapy adjuvant studies. The effective colon carcinogens contained a (see article) molecular similarity.


Physiological disposition of 2,2'-anhydro-1-beta-D-arabinofuranosyl-5-fluorocytosine in humans.

[2-14C]-2,2'-Anhydro-1-beta-D-arabinofuranosyl-5-fluorocytosine ([14C]AAFC) was given to 7 patients i.v. and to 3 patients p.o. at doses of 2 or 20 mg/kg. After i.v. administration of [14C]AAFC, 20 mg/kg, maximum plasma levels of up to 42.5 mug [14C]AAFC equivalents per ml of plasma occurred. A rapid exponential decrease of the radioactivity resulted in an initial half-line of 0.5 to 1.5 hr for the first part and a half-line of 8 to 24 hr for the second part of the curve. Most of the radioactivity was unchanged starting material. In plasma, 1-beta-D-arabinofuranosyl-5-fluorocytosine was found for only a short time and at low levels after i.v. injection. Its deamination product, 1-beta-D-arabinofuranosyl-5-fluorouracil (AFU), too, showed up in minor quantities. A small amount of 2,2'-anhydro-1-beta-D-arabinofuranosyl-5-fluorouracil was also detected. Administration p.o. of the 14C-labeled drug (2 mg/kg) resulted in the slow appearance of radioactivity in plasma. It peaked at 6 to 18 hr and slowly disappeared with a half-life of 12 to 18 hr. In a fasting patient, [14C]AAFC, 20 mg/kg, administered p.o. resulted in its rapid absorption into the bloodstream and in elevated levels in plasma for 48 hr. The unchanged drug and AFU were the predominant substances identified in plasms. Radioactivity after i.v. injection was found primarily in urine; only small amounts were recovered in expired air (to 2.4%) and traces were found in feces. The predominant urinary excretions product was the unchanged drug (average, 79%). The rest was AFU (average, 12.4%), 1-beta-D-arabinofuranosyl-5-fluorocytosine (average, 3.9%), and 2,2'-anhydro-1-beta-D-arabinofuranosyl-5-fluorouracil (average, 1.9%). After p.o. administration of the labeled drug, the calculated absorption was 32%. Urine contained about 50% unchanged drug and 40% AFU; the remainder was composed of 1-beta-D-arabinofuranosyl-5-fluorocytosine and the deaminated anhydro compound, 2,2'-anhydro-1-beta-D-arabinofuranosyl-5-fluorouracil. The rate of excretion in the urine was slow. In general after both i.v. and p.o. administration, spinal fluid contained considerably less radioactivity than plasma at shor intervals after the administration. At longer intervals after i.v. and p.o. administration, spinal fluid contained comparable or even considerably higher levels of radioactivity when compared to the levels in plasma. Autopsies performed 6 to 25 days after i.v. or p.o. drug administration revealed that radioactivity remained in kidney, spleen, small intestine, liver, and lung.


Tumor size dependency in the radiation response of the Lewis lung  carcinoma.

Tumor cell survival characteristics were assessed following 60Co gamma-irradiation of the Lewis lung carcinoma as 500-cu mm s.c. tumors or as 0.5-cu mm (1 mm in diameter) pulmonary metastases. Cells in the small pulmonary tumors were markedly more radiosensitive (D0 = 106 rads; hypoxic fraction less than 0.005) than were those in large s.c. tumors (final D0, 315 rads; hypoxic fraction, 0.36). When pulmonary metastases were excised and irradiated intact under well-oxygenated conditions in vitro, the hypoxic fraction rose to 0.30. This implies that, under normal in situ conditions, these nodules contain a microvascular system that achieves adequate oxygen supply to the great majority of tumor cells. Thus, the tumor cells within these small metastatic implants were more sensitive to irradiation, largely due to better oxygenation, and may be more sensitive to chemotherapy, due to better drug availability.


Biochemical and morphological changes in hepatic nuclear membranes produced by N-hydroxy-2-acetylaminofluorene.

The effect of N-hydroxy-2-acetylaminofluorene on the ultrastructure and synthesis of hepatic neclear membranes was evaluated in partially hepatectomized rats. The incorporation of L-[4,5-3H]leucine into two nuclear membrane fractions increased within 2 hr after hepatic resection and reached a peak at 20 hr. After partial hepatectomy, the decay of radioactivity in nuclear membrane proteins labeled with L-[4,5-3H]leucine revealed similar half-lives for the two membrane fractions when compared to those obtained from sham-operated animals. The protein concentration of the nuclear membrane fraction of higher density decreased sharply within 2 hr after partial hepatectomy and remained low throughout a 20-hr postoperative period. Polyacrylamide gel electrophoresis of both nuclear membrane fractions showed a similar composition. Nine proteins were resolved, varying from 21,000 to 190,000 daltons. The two major protein bands were in the range of 50,000 and 70,000 daltons, respectively. Treatment of partially hepatectomized animals with N-hydroxy-2-acetylaminofluorene showed marked dilation of the nuclear envelope and rough endoplasmic reticulum in situ upon electron microscopic examination. Vacuolization and evagination of the perinuclear membranes were also noticeable in isolated nuclei obtained from carcinogen-treated rats. Inhibition by N-hydroxy-2-acetylaminofluorene of the incorporation of L-[4,5-3H]leucine into the nuclear membranes was dose-dependent and remained depressed throughout a 60-min labeling period. These results suggest that the inhibitory effects on RNA and protein synthesis previously shown to be produced by this arylhydroxylamine hepatocarcinogen may lead to disruption of the morphology and synthesis of the nuclear envelope.


Amphotericin B-induced sensitivity to actinomycin D in drug-resistant Hela cells.

HeLa cells, which were selected for resistance to actinomycin D on the basis of decreaded penetration of the antibiotic into the cells, were treated with nontoxic concentrations of the polyene antibiotic amphotericin B. In the presence of amphotericin B, the cells became sensitive to the effects of actinomycin D, as demonstrated by loss of cell viability, typical morphological changes, and effects on the pattern of RNA synthesis. Furthermore, we were able to demonstrate that amphotericin B increased the amount of (3H)actinomycin D incorporated into HeLa cells as determined by both radioactive counts and radioautography. Amphotericin B might be useful in overcoming the resistance is based on decreased entry of the agents into cells.


Carcinogenic activity of di- and trifunctional alpha-chloro ethers and of 1,4-dichlorobutene-2 in ICR/HA swiss mice.

Four bifunctional and one trifunctional alpha-chloro ethers were tested for carcinogenicity. These compounds were bis-1,2-(chloromethoxy)ethane (Compound I), bis-1,4-(chloromethoxy)butane (Compound II), bis-1,6-(chloromethoxy) hexane (Compound III), bis-1,4-(chloromethoxy)-p-xylene (Compound IV), and tris-1,2,3-(chloromethoxy)propane (Compound V). trans-1,4-Dichlorobutene-2 (Compound VI) was tested along with the five alpha-chloro ethers. All six compounds were tested in female ICR/Ha Swiss mice for 502 to 569 days, depending on survival, by skin application or s.c. and i.p. injection. There were 30 or 50 mice/group. The i.p. and s.c. injections were given once weekly at 0.1 or 0.3 mg of compound dissolved in 0.05 ml tricaprylin for Compounds I to V and 0.05 mg/0.05 ml tricaprylin for Compound VI for the duration of the tests. The skin applications, three times weekly, were at doses of 0.3 or 1.0 mg/0.1 ml cyclohexane for the alpha-chloro ethers and 1.0 mg/0.1 ml acetone for Compound VI. Vehicle and no treatment controls were carried out together with the test compounds. Significance values (p) were calculated for all the compounds tested. Three compounds, I, IV and V, gave notable tumor incidences by all three routes of administration. Compounds II, III, and VI were either inactive by one or more routes of administration or gave low tumor yields.


Photochemotherapy of glioma cells by visible light and hematoporphyrin.

Malignant tumors take up and retain hematoporphyrin to a much greater extent than do normal tissues. Porphyrins are photodynamic agents that sensitize cells so that they are damaged by exposure to light. Treatment with hematoporphyrin followed by irradiation with light can destroy glioma cells in culture in less than 8 min and gliomas growing s.c. in rats in about 40 min. Photochemotherapy may become useful in the management of malignant tumors that are resistant to current methods of treatment.


Expression by human neuroblastoma cells of an antigen recognized by naturally occurring mouse anti-brain autoantibody.

A number of human tumor cell lines of both neural and nonneural origin have been assayed for the expression of an interspecies brain antigen (mouse brain antigen 2), detected by naturally occurring antibodies in normal mouse sera. These experiments indicate that mouse brain antigen 2 is present on four human neuroblastoma cell lines but not on other human tumor cell lines tested, including four glial cell lines and a retinoblastoma. These findings demonstrate the value of naturally occurring antibodies in normal mouse sera for the detection and serological classification of human tumor antigens and indicate that considerable caution should be used in attempts to distinguish tumor-specific and tissue-specific antigen expression on human neuroblastoma cells.


Circular dichroism studies on alpha- and beta-ketosides of 5-acetamido-3,5-dideoxy-D-glycero-D-galacto- nonulopyranosonic acid (N-acetylneuraminic acid) and of some of its derivatives.

The circular dichroism spectra of a number of N-acetylneuraminic acid derivatives in aqueous solution were studied.  For all compounds, the Cotton effects were found to be in the spectral range of the acetamido and carboxyl chromophores.  The c.d. curves of the methy, ethyl, and allyl alpha-D-ketosides are characterized by a broad, positive band centered at lambda similar to 195 nm with a slight skew towards the higher wavelengths and weak bands between lambda 225 and 255 nm, whereas the methyl beta-D-ketoside and the corresponding methyl ester show only an intense positive band with a broad shoulder in the same spectral range.  5-Acetamido-3,5-dideoxy-D-glycero-beta-D-galacto-nonulopyranose, its methyl beta-D-ketoside, and 5-acetamido-3,5-dideoxy-D-glycero-D-galacto-nonulopyranosonamide containing only the acetamido chromophore showed one single positive Cotton effect centered at lambda similar to 192 nm.  The c.d. spectrum of 5-acetamido-3,5-dideoxy-D-glycero-D-galacto-nonulopyranosonic acid confirms the beta-D configuration of the free acid in aqueous solution, whereas the shape of the c.d. curve of O-(N-acetyl-alpha-D-neuraminopyranosyl)-(2yields3)-O-beta-D-galactopyranosyl-(1 yields 4)-D-glucopyranose resembles that of the methyl, ethyl, and allyl alpha-D-ketosides 2-4.


Deoxyfluoroketohexoses: 4-deoxy-4-fluoro-D-sorbose and -tagatose and 5-deoxy-5-fluoro-L-sorbose.

4-Deoxy-4-fluoro-alpha-D-sorbose (6) was prepared in crystalline form by the action of potassium hydrogen fluoride on 3,4-anhydro-1,2-O-isopropylidene-beta-D-psicopyranose (3) followed by deacetonation.  Under identical conditions, 3,4-anhydro-1,2-O-isopropylidene-beta-D-tagatopyranose (7) underwent epoxide migration to give 4,5-anhydro-1,2-O-isopropylidene-beta-D-fructopyranose (12), which after deacetonation yielded 4-deoxy-4-fluoro-D-tagatose (15) and 5-deoxy-5-fluoro-alpha-L-sorbopyranose (16), the latter as the crystalline, free sugar.  The action of glycol-cleavage reagents on the isopropylidene acetals of the deoxyfluoro sugars was consistent with the assigned structures.  The structures were established by 13-C n.m.r. studies of the free deoxyfluoro sugars 6 and 16 and of the isopropylidene acetal 13, and by 1-H n.m.r. studies on the acetylated isopropylidene acetals 5 diacetate, 13 diacetate, and 14 diacetate.  5-Deoxy-5-fluoro-L-sorbose (16) was biologically active, producing in mice effects characteristic of deoxyfluorotrioses and of fluoroacetate.  4-Deoxy-4-fluoro-D-tagatose (15) and 4-deoxy-4-fluoro-D-sorbose (6) produced no apparent effects in mice up to a dose of 500mg/kg.  The implications of these findings with respect to transport, phosphorylation, and the action of aldolase on ketohexoses are discussed.


The ischemic leg: a clue to dissecting aneurysm.

The acute ischemic leg may be an important clue to the diagnosis of a dissecting aneurysm. Seventeen of 65 patients with dissecting aneurysms presented with this picture. The ischemic leg was often the dominant feature and obscured the correct diagnosis. The possibility of aortic dissection should be considered in any patient with an acutely ischemic leg, especially if there is any accompanying chest, neck, or back discomfort. The presence of an ischemic limb is not necessarily an indication for surgical repair of the dissection itself. Of seven patients treated medically, five survived with limbs intact, and in all five the pulses returned within one month.


Coronary artery disease in young women.

Although unusual, coronary artery disease does occur in young women. It may be present to a severe degree between ages 20 and 30, but a typical history of angina pectoris by no means assures the presence of significant coronary artery disease. Proof that oral contraceptives predispose to coronary artery disease is lacking. It seems wise not to recommend them to young women with other known risk factors. Significant risk factors in a group of 1000 women under age 50 studied by cinecoronary arteriography for the evaluation of chest pain included cigarette smoking, hypertension, hypercholesteremia, and glucose intolerance. Combinations of factors increased the risk. Electrocardiographic abnormalities in themselves did not seem to increase the risk of coronary artery disease, but did seem to enhance it in combination with other factors. Electrocardiographic evidence of transmural myocardial infarction without significant coronary artery disease was more common in young women than in young men. Cinecoronary arteriography may possibly be performed after resolution of more severe lesions related to lysis of emboli or thrombi. Special conditions may temporarily increase myocardial oxygen requirements. Angia-like chest pain has been described in patients without significant coronary artery disease. Many have normal electrocardiograms and no known risk factors. Spasm has been mentioned among many possible causes, but is very difficult to tell whether or not underlying atherosclerotic lesions may be present. Whatever the cause, the prognosis for patients with angina-like chest pain and normal coronary arteriography seems excellent; early death is a rarity and improvement is common.


Myocardial revascularization in patients aged 65 and older.

Experiences with 265 consecutive patients 65 years of age and older who underwent myocardial revascularization surgery have been presented. With proper patient selection, expeditious surgery, and good postoperative management, good surgical results can be obtained that are comparable to any good risk group of patients. This study suggests that age alone is not a contraindication to aggressive surgical treatment of coronary artery disease.


Cell kinetics methods for clinical use: assessment of autoradiography and 125I-iodo-deoxyuridine with mouse Ehrlich tumour.

Superficial Ehrlich tumours in mice were used to assess how much information on cell kinetics could be obtained from only the simplest techniques of autoradiography, and in situ monitoring of 125I-iodo-deozyuridine. These techniques were selected as being readily applicable to clinical situations. Intradermal tumours were studied from the earliest stages of rapid growth to large slow growing tumours with necrotic cores, as well as tumours undergoing regression. For comparison, intramuscular tumours were studied with systemic injections of radioactive DNA precursors. It was found that extensive information on cell production and loss rates was obtained from a single injection of tritiated thymidine followed by a single biopsy, or alternatively in vitro labelling of one minute biopsy specimen, and measurements of only the labelling index, together with a knowledge of the tumour's growth characteristics. Following a single localized injection of 125I-iodo-deoxyuridine, the loss rate of radioactivity measured in situ for a period of about 1 week provided an index of cell loss rate from the tumours.


Cell population kinetics in the rat jejunal crypt.

Cell kinetics in the jejunal crypt of the male Wistar rat were studied using autoradiographic techniques with tritiated thymidine and a stathmokinetic technique with vincristine. The migration rate measured by following the movement of the 50% peak on the labelling index distribution curve with time after injection of tritiated thymidine gave a value of 1-43 +/- 0-14 (SE) cell positions per hour, compared with a value from a cumulative birth rate of 1-78 cell positions per hour. Tht crypt column length was 32-9 +/- 0-2 cells and the column count was 22-3 +/- 0-2. This measurement gave a total crypt population of 734 cells, compared with an estimate of 650 +/- l from direct observation of squashed, microdissected crypts. In each crypt 22-5 +/- 0-5 mitoses were present, and the crypt cell production rate was 32 cells per crypt per hour; this latter value was confirmed using two independent techniques. The crypt growth fraction calculated from the durations of phases of the cell cycle and the labelling index was 0-62. A value of 0-61 was found from the labelling index distribution curve. As assessed from crypt squashes, there were 403 proliferating cells per crypt.


Temperature-sensitive hypoxanthine-guanine phosphoribosyltransferase in mutant Chinese hamster cells.

By selecting variants of Chinese hamster cells that were resistant to 6TG at 39 degrees C, but which would continue to grow in HAT medium at 33 degrees C, we have isolated cell lines with thermosensitive phenotypes for HGPRT controlled properties. THE HGPRT produced by those clones is temperature sensitive. These results indicate that the selected clones carry missense mutations in the structural gene for the enzyme.


Cell kinetics of irradiated experimental tumors: relationship between the proliferating and the nonproliferating pool.

The role of nonproliferating cells in tumor regeneration has been studied after subcurative doses of low L.E.T. irradiation. Radiation was applied in a single dose at three different levels; 0-47, 0-94 and 1-88 krad. Studies included estimation of the absolute number of cells per tumor, differential cell counts, and autoradiographic determination of kinetic variables, employing transplantable mouse mammary adenocarcinoma DBAH. Quantitative changes of morphologically defined proliferating and nonproliferating cell pools were followed at different time intervals after irradiation. Irradiation resulted in reduction of the number of cells in both pools, with apparent sparing of nonproliferating cells. The regenerative period started with a gradual increase in the number of cells in the proliferating pool, whereas the number of cells in the nonproliferating pool continued to fall in tumors irradiated with 0-94 and 1-88 krad. In the late phase of tumor regrowth, the increasing number of cells in the non proliferating pool corresponded to its replenishment by cell transition from the proliferating pool. In an effort to clarify whether cell transition from the nonproliferating to the proliferating pool may take place during the regrowth of radiation perturbed tumors, cell loss rates from both pools were estimated using experimental data. In addition to cell loses from the tumor as a whole, the 'net loss rate' of the nonproliferating pool reflects the rate of cell transition from the nonproliferating to the proliferating pool, minus the rate of transition in the opposite direction. A similar definition applies to cell loss rates from the proliferating pool. The results showed: (1) high losses in both pools, with excess losses in the proliferating during the early phase after irradiation; (2) in the early stage of regrowth after irradiation, the cell net loss rate f-or the nonproliferating pool increased, in contrast to the behavior of cell loss rate for the proliferating pool and the average cell loss rate for the tumor as a whole; (3) in the late stage of regrowth a decrease in net loss rate for the nonproliferating pool reflects the excess production of nonproliferating cells over control tumors. These results suggest that cell transition from the nonproliferating to the proliferating pool takes place at the beginning of tumor regrowth after subcurative single-dose irradiation.


Catecholamine-containing nerve terminals of the eccrine sweat glands of macaques.

The external layer of the median eminence (ELME) and the arculate nucleus of male rats were studied with the Falck-Hillarp technique and electron microscopy of aldehyde-OsO4 or KMnO4 fixed after various types of hypothalamic deafferentation experiments with the Halász knife. Special reference was paid to the monoamine systems and the results can be summarized as follows. 1. The main monoaminergic input to the ELME comes from the arcuate nucleus-periventricular area via a dorsal approach. A horizontal transection through the arcuate nucleus decreases the percentage of monoamine boutons i.e. boutons with small granular vesicles, from 31.6% in the controls to 4.4% in the lesion group, whereas only a small effect is seen after anterior (or complete) deafferentations. 2. A major input to the ELME enters the basal hypothalamus at the anterior-lateral aspects (see Réthelyi and Halász, 1970). The fibers cut after anterior deafferentations in all probability mainly come from cell bodies localized in the anterior hypothalamus or even further rostrally but some may represent NA axons ascending from the lower brain stem. 3. The degeneration course of nerve endings in the ELME both after anterior deafferentations as well as after lesions in the arcuate nucleus is rapid (within 2-3 days) and morphologically characterized by an initial aggregation of large dense cored vesicles seemingly to electron dense bodies within the boutons and probably also to a closer spacing of the small electron lucent synaptic vesicles (see Raisman, 1972). This type of degeneration seems to take place both in monoamine and non-monoamine neurons. 4. Degenerating boutons are found in the arcuate nucleus after anterior and complete deafferentations. Thus, the anterior hypothalamus may exert an "indirect" control of the pituitary gland via synapses on arcuate neurons although quantitatively the "direct" influence through the projection to the ELME is of more importance. 5. After anterior deafferentations enlarged axons containing large amounts of large dense cored vesicles and other organelles are found caudally of the cut indicating the existence of rostral projections from the medial hypothalamus.


Nerve terminals and epithelial cell variety in the human lacrimal gland.

Pieces of tissue taken from three human lacrimal glands were examined electron microscopically. Secretory cells varied according to the electron density and structure of their secretion granules. Secretory cells were arbitrarily categorized as light, medium or dark based on their granule content. Acini were composed of two or all three categories of cells together with myoepithelial cells and lymphocytes. A minority ogic, sympathetic terminals. The appearance of most interstitial and all parenchymal fibres was consistent with that of cholinergic, parasympathetic terminals. Parenchymal terminals were confined to ducts, terminal tubule areas and to serous (dark) cells. A large proportion of parenchymal terminals lay adjacent to myoepithelial cells in the ducts and terminal tubule regions but terminals observed among serous cells were rarely in contact with myoepithelial cells. A possible parasympathetic control of serous secretion, granule production and duct contraction and the autonomy of mucous cells is discussed.


Projections to the median eminence and the arcuate nucleus with special reference to monoamine systems: effects of lesions.

The external layer of the median eminence (ELME) and the arcuate nucleus of male rats were studied with the Falck-Hillarp technique and electron microscopy of aldehyde-OsO4 or KMnO4 fixed material after various types of hypothalamic deafferentation experiments with the Haláz knife. Special reference was paid to the monoamine systems and the results can be summarized as follows. 1. The main monoaminergic input to the ELME comes from the arcuate nucleus-periventricular area via a dorsal approach. A horizontal transection through the arcuate nucleus decreases the percentage of monoamine boutons i.e. boutons with small granular vesicles, from 31.6% in the controls to 4.4% in the lesion group, whereas only a small effect is seen after anterior (or complete) deafferentations. 2. A major input to the ELME enters the basal hypothalamus at the anterior-lateral aspects (see Réthelyi and Halsáz, 1970). The fibers cut after anterior deafferentations in all probability mainly come from cell bodies localized in the anterior hypothalamus or even further rostrally but some may represent Na axons ascending from the lower brain stem. 3. The degeneration course of nerve endings in the ELME both after anterior deafferentations as well as after lesions in the arcuate nucleus is rapid (within 2-3 days) and morphologically characterized by an initial aggregation of large dence cored vesicles seemingly to electron dense bodies within the boutons and probably also to a closer spacing of the small electron lucent synaptic vesicles (see Raisman, 1972). This type of degeneration seems to take place both in monoamine and non-monoamine neurons. 4. Degenerating boutons are found in the arcuate nucleus after anterior and complete deafferentations. Thus, the anterior hypothalamus may exert an "indirect" control of the pituitary gland via synapses on arcuate neurons although quantitavely the direct influence through the projection to the ELME is of more importance. 5. After anterior deafferentations enlarged axons containing large amounts of large dense cored vesicles and other organelles are found caudally of the cut indicating the existence of rostral projections from the medial hypothalamus.


Pigment movements in fish melanophores: morphological and physiological studies. V. Evidence for a microtubule-independent contractile system.

A pressure device has been used to obtain information about the forces involved in the maintenance of the aggregated state of melanophores of the angelfish, Pterophyllum scalare. Single aggregated melanophores of isolated scales were submitted to mechanical compression with forces ranging from 50-320 mup. As a function of the gradually increasing force melanophores disperse their pigment, the degree of dispersion being proportional to the intensity of the force. When microtubules are destroyed by treatment with 0.3 mM vinblastine in KCl solution, pigment dispersion in response to the external force is similar to that observed in KCl alone. After changing the medium to NaCl solution, melanin granules remain concentrated in the cell center; the force-induced melanosome dispersion, however, is significantly enhanced. Distinctly lower forces are required to produce an expansion equivalent to that observed in KCl solution. When the medium is changed to vinblastine-KCl again, the dispersion in response to the external force resembles that obtained before NaCl treatment. Removal of Ca++ and Mg++ ions by treatment with 2 mM EDTA or EGTA in Ringer's solution containing 0.1 mM adrenalin produces a remarkable enhancement of melanosome dispersion in response to increasing external force. This effect of EDTA or EGTA is completely reversible. When the medium is changed to normal Ca++-Ringer's, the force/dispersion curve resembles that obtained before EDTA or EGTA treatment. It is concluded that a state of equilibrium exists between the external force and an opposing force produced by the melanophore. The differences in the opposing force under different experimental conditions may be due to a "contractile component". This component seems to be independent of microtubules, as indicated by vinblastine experiments. It "contracts" under aggregating stimulus and "relaxes" under dispersing stimulus. From the data presented in this paper, the order of magnitude of the pressure developed by the contractile component in the completely aggregated state was calculated as between 5-7 p/cm2 in the relaxed state and about 20 p/cm2 during contraction. These values are comparable to those observed in other nonmuscular cells.