Ultrastructural observations of amorphous bone mineral in avian bone.

Mineral from medullary bone of three avian species was examined with the electron microscope in order to clarify the ultrastructure of amorphous bone mineral (ABM) in a mineralized tissue. Powders from freeze-dried bone revealed bone mineral with morphology and behavior identical to synthetic amorphous calcium phosphate (ACP). These powders exhibited spherically shaped particles 180--400 A in diameter with uniform electron density when viewed at low beam intensity. Thin sections of embedded freeze-dried bone also revealed spherically shaped particles 100--350 A in diameter with electron beam sensitivity characteristic of ACP. Regions of bone mineral with irregular outline and morphology were observed which closely resemble the discoidal form of synthetic ACP. More electron-dense spherical particles (150 A in diameter) could be seen budding from these regions. Some of these buds exhibited electronlucent centers characteristic of AMB. The inorganic nature of these features of bone mineral was confirmed using ultramicroincineration. Preliminary exploration of a freeze-substitution technique showed spherical bone mineral particles which were similar in morphology to those observed in freeze-dried samples. A limited degree of preservation of cellular material was observed using this freeze-substitution technique.


Glycerolipid biosynthesis in isolated rat intestinal epithelial cells.

Intestinal epithelial cells were prepared from fasted rats by dispersion with collagenase (EC 3.4.24.3). The structural and metabolic integrity of the cells was verified by electron microscopy, a high percentage of Trypan Blue exclusion, a low degree of release of lactate dehydrogenase (EC 1.1.1.27) in the medium, and by the retention of sensitivity to agents known to modify metabolic and transport activity in everted sacs of intestinal mucosa. The isolated intestinal epithelial cells were used to study glycerolipid biosynthesis from glucose, glycerol, 2-monoacylglycerol, and free fatty acids. The cells actively incorporated the labeled precursors into glycerolipids without specific cofactor requirements. Addition of fatty acids stimulated the incorporation of both glucose and glycerol into triacylglycerols and glycerophospholipids, the greatest effect being observed with palmitate. The stimulation of monoacylglycerol acylation appeared to depend on both the nature of the monoacylglycerol and fatty acid supplied. Stereospecific analyses of the diacylglycerols formed from 2-monoacylglycerols and free fatty acids showed that 1,2-diacyl-sn-glycerols (62-70%) were the major and that 2,3-diacyl-sn-glycerols (30-38%) the minor intermediates in triacylglycerol biosynthesis. The data indicate that isolated intestinal epithelial cells exhibit a total capacity of glycerolipid synthesis and a stereochemical course of reaction which is comparable to that observed for triacylglycerol formation in everted sacs of intestinal mucosa, but much less specific than that seen in microsomal preparations of intestinal mucosa.


The transfer of glucose to steroids by chimpanzee liver microsomes.

Microsomal preparations from chimpanzee liver can transfer glucose from UDPglucose to the 17alpha-hydroxyl group of 17alpha-estradiol and of 17alpha-estradiol-3-glucuronide. A phenolic glucoside of estrone, but not of either 17alpha- or 17beta-estradiol is also formed. No formation of glucosides of  p-nitrophenol, or of diethylstilbestrol was demonstrated. The specificity of glucosyl transfer in the chimpanzee is not identical to that in either the human, the rabbit, or the sheep.


Inhibition of cardiolipin synthesis by end-products and other complex lipids in membrane preparations of Micrococcus lysodeikticus.

Using membrane preparations of Micrococcus lysodeikticus, the end-products of cardiolipin synthesis, cardiolipin and glycerol, were shown to inhibit cardiolipin synthetase at several concentrations. Other phospholipids tested for inhibitory effects, phosphatidyl-ethanolamine, phosphatidylinositol, and phosphatidic acid were also shown to inhibit cardiolipin synthesis. Phosphatidic acid was considerably more inhibitory than cardiolipin, phosphatidylethanolamine was similar to cardiolipin, and phosphatidylinositol less inhibitory at the same concentrations. A non-phosphate-containing glycolipid was also inhibitory. In contrast, glycerophosphate had no effect on cardiolipin synthesis.


The role of membrane proteins and phospholipids in the interaction of ribosomes with endoplasmic reticulum membranes.

Hydrolysis of the membrane proteins and phospholipid headgroups of rat liver rough endoplasmic reticulum membranes showed that the ribosomal binding sites involve membrane proteins susceptible to low concentrations of trypsin, chymotrypsin, and papain. Three membrane proteins having molecular weights of 120 000, 93 000 and 36 000 are found to be altered by trypsin and chymotrypsin treatment. Also the polar headgroup of phosphatidylinositol appears to play a role in the binding process.


Liver cell plasma membrane lipids and the origin of biliary phospholipid.

The liver cell plasma membranes of fed male Wistar rats were separated into a fraction rich in bile canaliculi and the remainder of the plasma membrane. Electron-microscopically, the bile canalicular fraction consisted almost exclusively of intact bile canaliculi with thier contiguous membranes. The remaining plasma membrane fraction consisted primarily of vesicles and sheets of membranes essentially free from the bile canaliculi. The bile canalicular membrane fraction contained relatively more total lipid, cholesterol, and phospholipid, and relatively less protein. Although the phospholipid composition of the two fractions was the same, the specific activity of the bile canalicular membrane phosholipids, up to 12 h following in vivo administration of [2-3H]glycerol, was always significantly greater than that of the remaining plasma membranes, and showed a biphasic response not found in the latter. The specific activity of the phosphatidylcholine, phosphatidylethanolamine and lysophosphatidylcholine of the bile canalicular membranes rose to a peak within 40 min after administration of the label, fell sharply and then rose to a second peak after 120 min. The specific activity of the sphingomyelin and phosphatidylserine plus phosphatidylinositol of the bile canalicular membranes and of all the phospholipids of the remaining plasma membranes diphasic pattern but increased steadily to reach a maximum at 120 min. The specific activity of biliary phosphatidylcholine followed a pattern identical to that of the phosphatidylcholine, phosphatidylethanolamine and lysophosphatidylcholine of the bile canalicular membrane fraction. These results show that the average rate of turnover of phospholipid in the bile canalicular membranes is considerably greater than that in the remaining plasma membrane and other cell membrane fractions; they indicate that the phospholipid of the bile canalicular membranes exists in two or more pools, turning over a different rates; and they support the concept that biliary phospholipid is derived from the bile canalicular membrane. The results also suggest that bile canalicular phospholipid may be derived from two different sources, in contrast to the remainong plasma membrane.


A comparison of lactate dehydrogenase from an ectothermic and an endothermic animal.

Adaptation to environmental temperature is examined in beef heart, beef muscle, and flounder muscle lactate dehydrogenases (EC 1.1.1.27). Low temperature adaptation in the ectothermic (flounder) enzyme is indicated by a reduced enthalpy of activation for kcat (enzyme turnover number, s-1) and increased catalytic efficiency. Also, the reaction rate at low substrate concentrations has a maximum at a lower temperature than in the endothermic enzymes. This is a result of altered bonding in the enzyme-substrate complexes. Adaptation to higher temperatures in the endothermic (beef) enzymes is  suggested by a decreased sensitivity to heat denaturation, especially in the presence of substrates. A direct correlation is found between the degree of bonding in the enzyme-substrate complexes and the decrease in rate of heat denaturation caused by addition of substrates.


Trisomy in Aedes aegypti.

A trisomic (2n=6+1) pupa of the yellow fever mosquito Aedes aegypti has been found. The trisomy involved chromosome 3 which is intermediate in size between 1 and 2. The extra chromosome formed a univalent or a trivalent during meiosis.


Limitations of horizontal partial laryngectomy.

The author considers successively: 1--The theoretical justifications for horizontal partial laryngectomy (anatomical considerations, indications as to the modalities of extension of supraglottic tumors, results concerning the cancer itself, functional results). 2--The indications of and contraindications for this type of surgery, with special reference to two types of cancers: median supraglottic carcinomas and antero-lateral epilarynx carcinomas. 3--Some specific problems, namely that of large supraglottic laryngectomy, of combined nodal surgery, and of preoperative radiotherapy. The limits of horizontal partial laryngectomy are specified on the basis of two criteria: its value as far as cancer is concerned (five-year survival rate), the quality of functional results.


Roles and limitations of conservation surgical therapy for laryngeal cancer.

This paper deals with the limitations of conservation surgical therapy for cancer in various sites. The detailed techniques are not included in this paper, but rather its pathophysiological approach to partial surgery. The technique has been reported many times previously.


Limitations of supraglottic laryngectomy and conservative neck dissection.

Limitations of supraglottic laryngectomy may arise from either the primary location, or secondary spread of the tumor in the vestibule. When a growth reaches the epilarynx a modified supraglottic laryngectomy or a more radical operation must be envisaged. Other limitations include poor bronchopulmonary condition, age of the patient, and previous radiotherapy. All these conditions may jeopardize healing and/or, even more important, functional rehabilitation. New hands to this surgery should know its limitations and should be warned against its risks. On the other hand no limitations exist to conservation neck dissection, except fixed nodes or lymph node metastases following radiation or earlier surgery. The conservative technique is as radical as the traditional technique, but respects essential structures and thus widens instead of limiting the indications for elective bilateral neck dissection.


Role and limitations of vertical hemilaryngectomy.

Two types of vertical hemilaryngectomy are discussed. The frontolateral operation is indicated for infiltrating lesions of the cord, anterior to the vocal process, which approach or involve the anterior commissure. It should not be used for carcinomas which extend posteriorly beyond the tip of the vocal process, into the ventricle or below the inferior surface of the cord. The "extended" frontolateral operation is useful for infiltrative cordal lesions showing (1) posterior extension to or beyond the tip of the vocal process, but not into the posterior commissure, (2) beginning impairment of motility without actual fixation of the cord, (3) limited and superficial extension to the ventricle or margin of the ventricular band, or (4) subglottic extension amounting to no more than a few millimeters. The lesions are removed en bloc, along with a large portion of the thyroid cartilage and usually the entire arytenoid. Accurate delineation of the gross extent of the lesion is essential if these techniques are to be successfully applied. Failures are likely if attempts are made to over-extend the indications.


Limitations of conservation surgery in carcinoma involving the arytenoid.

The various types of conservation surgery for laryngeal cancer can be systematized in one entity and be classified into four types of "laryngoplasty." Laryngeal cancer involving the arytenoid is thought to be amenable to laryngoplasty type 3. The indication and the procedures of hemilaryngectomy B, subtotal laryngectomy, and subtotal laryngopharyngectomy are described. The therapeutic results of these operations were better than that of total laryngectomy. Tumors involving both sides of the arytenoid portion rule out conservation surgery. They should be treated by the operations of laryngoplasty type 4, which means total laryngectomy and laryngeal reconstruction.


Limitations of surgery in the treatment of head and neck cancer.

The limitations of head and neck surgery are discussed. The overall survival rate has not improved to any great extent in the past 30 years. A plea is made for careful selection of patients who are to undergo massive debilitating surgery.


Combination of extranasal and intranasal approach in surgery of the nasal pyramid and nasal septum (decortication).

Decortication of the nose is a combination of the extranasal and intranasal approach to the surgery of the nasal pyramid and nasal septum. It consists of temporary removal (decortication) of the skin from the infrastructure of the tip and dorsum of the nose through a columellar and slot incision. This method makes it possible to perform very precisely a wide spectrum of surgical interventions under visual inspection and thus guarantees the basic surgical postulate of good surveillance over the operative field. We have used this method in 500 cases and evaluated the esthetic and functional results of the operation in all of these patients. This modified technique meets all the requirements of functional rhinosurgery.


Performance of hearing aids and wearer satisfaction--a two year study.

A group of 66 patients with newly acquired hearing aids were seen at intervals over a two year period. The hearing aids' performance was measured at each visit. Many aids were not performing according to manufacturer's specifications but their performance as a group did not deteriorate over two years. Aids required on average nearly two repairs in the two year period, eyeglass aids being slightly more fragile than other types. Most of the patients were satisfied with their aids and their degree of satisfaction was found to correlate with their aided speech discrimination score.


Biological activity of phthalated endotoxin.

Glycolipid (GL) was extracted from a heptoseless mutant of Salmonella minnesota by a mixture of phenol, chloroform, and petroleum ether. The GL was subjected to treatment with either acetic anhydride or phthalic anhydride; a portion of the GL was untreated. Both of the chemically treated preparations as well as the parent GL were examined for biological activity in the following systems: mouse lethality assays, rabbit pyrogenicity assays, and rabbit skin assays. The results of these studies indicated that both treated preparations were less toxic in mice than the parent GL. Compared with saline-treated controls, rabbits pretreated with either of the modified preparations exhibited a reduced pyrogenic response to a subsequent challenge dose of the homologous material but no reduction when challenged with the parent GL. Pretreatment with the unaltered GL rendered rabbits tolerant to the homologous material and to some degree to the modified preparations. Rabbits immunized witn any of the three Gl preparations exhibited dermal toxicity responses comparable with those in untreated animals. Based on these findings, it was concluded that treating GL with either phthalic anhydride or acetic anhydride results in a product which is less toxic in mice and less pyrogenic in rabbits than the parent GL, but which also exhibits a loss of ability to render rabbits tolerant to challenge with untreated GL.


Scanning and transmission electron microscopy of three strains of Bifidobacterium.

Scanning and transmission electron microscopy was applied for a morphological study of three strains of Bifidobacterium grown on solid or liquid media. The pronounced pleomorphism of the cultures previously observed by light microscopy was confirmed. A possible sequence of the morphological events during transformation from one to another pleomorphic form is proposed for B.  bifidum and B. longum. Ultrastructural differences such as the formation of extensive mesosomal complexes in B. longum and characteristic plasmalemma particles only observed in the B. bifidum mutant are described and discussed.


[Numerical analysis of telluric nonfermenting Gram-negative bacteria].

A numerical analysis was carried out from a set of 165 telluric Gram-negative bacterial strains. The results allowed to join up 130 of them divided into eight phenons. Two of these phenons represent on their own 70% of the classified strains. The first of these phenons (52 strains) can be assimilated to the genus Pseudomonas in the fluorescent group; the second one (41 strains) offers some analogies with the Acinetobacter. A representative strain type of the latter phenon was retained for later taxonomic comparisons.


Heterotrophic nutrition of the marine pennate diatom Navicula pavillardi Hustedt.

Navicula pavillardi Hustedt, a marine, littoral, pennate diatom, can grow in the dark on glutamate or on the complex organic supplements tryptone or yeast extract. Growth on glutamate in the dark took place without an initial lag phase, whereas growth on tryptone began only after a 2-day lag phase that could be abolished by the simultaneous presence of glucose. Lactate inhibited growth in the dark on glutamate, but not photoautotrophic growth. Relatively low concentrations of glutamine inhibited photoautotrophic growth. The observed doubling time for heterotrophic growth on glutamate or tryptone was about 70 h, compared with a doubling time of 24 h under optimal photoautotrophic conditions. Glucose did not decrease the doubling time in the dark on tryptone. The assimilation efficiency for glutamate was 41%. The estimated necessary uptake rate for glutamate to account for the observed heterotrophic doubling time on glutamate was close to those measured with isotope techniques. The kinetic parameters for glutamate uptake, which followed Michelis-Menten kinetics, were Ks = 0.018 mM, and Vmax = 7.0 X 10(-10) mumol per cell per minute. Although several amino acids served as sole nitrogen sources for photoautotrophic growth and were demonstrated by the use of isotope techniques to enter the cells, they could not be used as substrates for growth in the dark. Glucose was not taken up to a significant extent except by cells grown in the presence of tryptone. Lactate was taken up only by dark-grown cells. Results of preliminary studies on the metabolic fate of several uniformly labeled amino acids are presented.


Valine, malic, and pyruvic dehydrogenase tests in the differentiation of Bacteroides.

Fifteen isolates of Bacteroides were tested for their ability to dehydrogenate a variety of amino acids and organic acids. A simple and rapid method was developed for detecting dehydrogenase activity using 2,3,5-triphenyl tetrazolium chloride as a hydrogen acceptor and indicator. The results indicate that the tests for valine, malic, and pyruvic acid dehydrogenases have value in differentiating organisms in this genus.


The use of irradiated vaccine in immunization against experimental murine toxoplasmosis.

Trophozoites from the peritoneal cavities of mice infected with the RH strain Toxoplasma gondii were given irradiation in doses of 5, 10, 15, and 20 kiloroentgens (kr). CD-1 strain mice that received intraperitoneal inoculation of trophozoites irradiated with 5 kr all died of toxoplasmosis, but the mice that received trophozoites irradiated with the higher doses all survived. The survirors that were examined were found to be free of toxoplasmic cysts. Single doses of these irradiated vaccines provided good protection to subsequent virulent challenge. This protection was 100% in the first 3 weeks after the immunization. Survivors of the first challenge were also solidly protected against a subsequent rechallenge.


Adaptation to cycloheximide: in vitro studies with filamentous fungi.

Adaptation of numerous strains of filamentous fungi to cycloheximide was noted upon first transfer during serial exposure to the antibiotic. The extent of this adaptation varied in part with the species and strain of the microorganism. Exposure on subsequent transfers caused no change in the growth of any of the strains used. These results suggest that nonzoopathogens may be differentiated from zoopathogens on the basis of adaptation to cycloheximide and that such adaptation might also be found in other eucaryotes.


Nitrification on a coral reef.

We report that the algal pavement just behind the reef crest at Enewetak Atoll produces nitrate at measurable rates. In situ and in vitro incubations with N-Serve indicate that the autotrophic pathway involving two separate organisms is effective in this oxidation of ammonia to nitrate. Significant nitrification is indicated throughout the reef environment; Nitrobacter agilis has specifically been identified as at least one of the organisms responsible for the terminal oxidation of nitrite to nitrate.


Relationships in Canada between mortality from tumours of the gastrointestinal tract and marital status.

The mortality from tumours of the gastrointestinal tract in the Canadian population in 1970-72 was 16% higher in single than in married men (on the basis of age-adjusted rates), 25% higher in widowed men and 28% higher in divorced men. All these differences were unlikely to be due to chance. The rates were 4% higher for single women, 14% higher for widows and 22% higher for divorced women, compared with the married. The differences for single and divorced women were not significant. Substantial excess mortality was found in the unmarried for tumours of the mouth, pharynx and esophagus, and rectum; for tumours of the stomach and colon the excess was small or nonexistent. This variation between sites suggests that systematic errors in the census data used as denominators are not responsible for the high mortality for the unmarried from certain tumours. The effect is found in conditions for which treatment can have made little difference (e.g., a 75% excess mortality for tumours of the esophagus in single men compared with married) and in conditions for which differences in the use of medical facilities may have been important (e.g., a 44% excess mortality for tumours of the rectum in widowers).


Prevalence of hepatitis B antibodies in hospital personnel.

Of 426 hospital staff tested for hepatitis B surface antibody (anti-HBS) by the radioimmunoassay method 57 (13.4%) had positive results. Laboratory staff had the highest prevalence, followed by nurses, and both values were significantly higher than that of administrative staff. Clerical staff working in laboratories were at the same risk for hepatitis B as general-duty nurses. Significantly more staff with anti-HBS had a history of hepatitis (19.3%) compared with staff without anti-HBS (79%), and significantly more staff with a history of hepatitis had anti-HBS (25.6%) compared with staff without such a history (12.0%). History of blood transfusions was not related to prevalence of anti-HBS. The risk for hepatitis B is greater in hospital staff who are in direct contact with patients or handle patients' blood and other specimens. However, contact with patients is less important than contact with patients' blood and other specimens.


Evaluation of late results of surgical treatment of congenital heart disease.

Operative mortality from surgical treatment of congenital heart disease has been steadily reduced over the past 20 years. During the same period it has become clear that a proportion of survivors have residual clinical problems. Some of these are due to imperfect repair, a number being due to factors beyond present surgical control, and some are the consequence of associated cardiovascular defects. The chief problems can be elucidated by an analysis of the results of surgical treatment of six cardiac malformations: simple pulmonary valve stenosis, coarctation of the aorta, secundum atrial septal defect, isolated ventricular septal defect, tetralogy of Fallot and transposition of the great arteries. Recognition of the sequelae of preoperative hemodynamic strain and apparently minor associated malformations is important, and it is possible to anticipate such factors. Long-term follow-up of patients after operation is particularly important.


The choice of estrogen preparations in the treatment of prostatic cancer.

A total of 154 patients with carcinoma of the prostate received estrogen therapy with diethylstillbestrol (DES), chlorotrianisene or ethinyl estradiol. During a mean follow-up period of 26 months the incidence of complications -- thromboembolic episodes, fluid retention and gynecomastia -- was recorded. Although the incidence of cardiovascular complications was significantly higher in the DES group, the differences in mortality between the groups were not significant. The differences in incidence of fluid retention and gynecomastia also lacked significance. All three compounds produced adrenal cortical hyperplasia as indicated by the increased serum cortisol values.


Biliary-bronchial fistula demonstrated by endoscopic retrograde cholangiography.

Endoscopic retrograde cholangiography is valuable in the evaluation of biliary tract disorders. A 50-year-old Italian woman developed biloptysis 1 year after cholecystectomy because of intrabiliary rupture of a hydatid cyst with secondary infection, which resulted in intrathoracic rupture and communication with the bronchial tree. Endoscopic retrograde cholangiography showed the cause and pathway of the fistulous tract by outlining the biliary tree, abscess cavity and communication with the right upper lobe bronchus. This technique appears to be well suited to the investigation of patients with biliary-bronchial fistula.


External biliary fistula.

External biliary fistulas, once common, are now rare: before the present report of 4 cases only 27 cases have been reported in the English literature since 1900. Review of the records of four patients with external biliary fistula confirmed its occurrence in patients over 50 years of age and the variable site for operning of the fistulous tract. Cholecystectomy provided successful treatment in three of the four patients but the fourth was too ill to undergo an operation; in general, definitive treatment is cholecystectomy, together with excision of the fistulous tract if this takes a direct path through the abdominal wall from the gallbladder, or curettage if the course is devious.


Lithium intoxication: a report of two cases.

Two cases of toxic reaction to lithium carbonate are reported. The first patient displayed symptoms resembling those of organic brain syndrome which was not associated with a high serum lithium level. The influence of diuretic therapy in combination with lithium is felt to enhance the risk of intoxication. The second case depicts acute CNS toxicity with known fatal potential. Conservative lithium administration is recommended for acute manic psychosis and reference is made to the beneficial effects of urea and aminophylline on renal lithium elimination in the active treatment of lithium poisoning.


[Perception of the father's attitudes and educational practices by delinquent and normal adolescents].

This is a study of the perception of fathers' child-rearing attitudes and practices as experienced by the child during his socialization process. Forty-one delinquent adolescents and a similar number of normal adolescents evaluated ten variables of child-rearing by means of a rating scale and projective techniques. Findings strongly support the hypotheses that a significant difference exists between the two groups and that a far greater number of delinquents perceive their fathers as ineffective, negative and pathogenic.


Medicines and mental illness in old age.

When treating an apparent psychiatric disturbance in an elderly patient it is wise to review all his medications carefully, and then to prescribe sparingly only those drugs with which one is completely familiar. Psychiatric side effects of drugs commonly used with elderly patients are reviewed.


Growth hormone treatment in hypopituitary dwarfs: longitudinal psychological effects.

This is one of a series of studies on the psychological effects of medically induced growth in a group of hypopituitary dwarfs treated with Human Growth Hormone (HGH). Before treatment these dwarfs were found to be psychologically immature and hypoactive, without any manifestation of aggressive drives, and had an underlying low self-esteem when compared with the normal population. When compared with a matched group with constitutional growth delay the hypopituitary patients were found to use denial, whereas the control group used well-organized compensatory mechanisms (9). Immaturity was found in both groups. When the hypopituitary dwarfs reached adolescence they lacked the core attributes of masculinity or femininity and manifested some ambiguity or even inversion in their choice of gender role (10). This report deals with their psychological evolution in relation to physical growth.


Amnesia following severe head injury.

This report concerns some aspects of the problem of establishing valid and reliable criteria for post-traumatic amnesia, and of relating this to other cognitive functions in the post-injury period. Of specific interest are the variability of the duration of retrograde and anterograde amnesia on attempts at repeated assessment, and the relationship between the time of return of full orientation and the end of the period of anterograde amnesia. A prospective study was undertaken of sixty consecutive patients who had suffered severe head injuries and had been admitted to the neurosurgical service of the Winnipeg General Hospital. The object of this study was to obtain information regarding organic, psychological and social indices of the future clinical course.


Neuropsychological deficits in 'primary' and 'secondary' non-medical drug users.

An extensive neuropsychological test battery, specifically designed to investigate brain-behaviour relationships, was administered to two groups of multiple drug users. The 'primary' group consisted of subjects referred fundamentally on account of drug abuse problems. The 'secondary' group were referred principally for neurological reasons, but during history taking they reported drug use. Fifty patients, in all, were examined. Results showed that people who have received psychiatric care for drug abuse and who had started using drugs at an earlier age ('primary' group) had fewer adaptive resources and limited psychometric and academic achievement skills. This effect cannont be attributed to personality differences. Both groups had noticeably greater difficulties on motor tests, fine manipulative skills, and Halstead's Impairment Index when compared with a normal control group matched for age and sex.


The Iowa 500: field work in a 35-year follow-up of depression, mania, and schizophrenia.

This paper reports a follow-up and family study being carried out at the University of Iowa College of Medicine on primary affective disorders and schizophrenia in patients hospitalized thirty-five years ago. The purpose of this project is to obtain objective data to shed light on our understanding of these two major functional psychoses -- their diagnostic validity, clinical features, course and outcome, heterogeneity, life histories, related illnesses and characteristics of familial association. This research started with 525 patients selected from inpatients consecutively admitted to the Iowa Psychopathic Hospital between 1934 and 1944, and with a stratified random sample of 160 surgical patients admitted to the University General Hospital during the same period. A specially designed structured interview form -- the Iowa Structured Psychiatric Interview (ISPI) -- is used to follow up all living index patients, and as many of their living first-degree relatives as possible. At this time, over one-third of the estimated total study population of three thousand patients and relatives have been personally interviewed, and the preliminary findings are reported here.


Fetal mesenchymal hamartoma of kidney. A tumor of secondary mesenchyme.

The ultrastructure of a fetal mesenchymal hamartoma of the kidney shows undifferentiated mesenchymal cells, whose morphological characteristics conform to those of secondary mesenchyme. In contrast to primary mesenchyme or mesoblast, which may form epithelial structures, secondary mesenchyme is incapable of doing so. While Wilms' tumor may be considered a tumor of mesoblast, the fetal mesenchymal hamartoma is believed to originate from secondary mesenchyme or some of its more mature derivatives.


Unclassified benign tumor of the rib. Osteochondroblastoma.

A previously unclassified benign tumor of the rib is reported. The tumor was found in a 21-year-old black woman who presented herself with chest pain. After its radiologic discovery the exophytic tumor was resected. Histologically, it was composed of two seemingly independent components. The more central portion of the tumor was principally an osteoblastoma, while the more peripheral and greater mass of the neoplasm was chondroblastic. The name "osteochondroblastoma" is proposed for this lesion.


A possible primary cutaneous carcinoid.

A case is described of carcinoid tumor in the skin in which all efforts failed to discover a primary tumor in the internal organs. The possibility that this is a primary cutaneous carcinoid is discussed.


Erythroplakia of the oral cavity.

Erythroplakia of the oral cavity is a specific disease entity which must be differentiated from other specific or nonspecific inflammatory oral lesions, although this can only be done in most cases by biopsy. A series of 58 cases of oral erythroplakia has been retrieved from 65,354 consecutively accessioned biopsy-surgical specimens. The disease was found to have no apparent sex predilection (31 males and 27 females) and was most frequently seen during the 6th and 7th decades. The most common site of occurrence in females was the mandibular alveolar mucosa-mandibular gingiva-mandibular sulcus, whereas this was the least common site in males. The floor of the mouth was the most common site in males, followed by the retromolar area in both males and females. The histologic findings emphasized the serious nature of the disease, since 91% of the specimens were either invasive carcinoma, carcinoma in situ, or severe epithelial dysplasia.


Eccrine sweat gland tumor of clear cell origin involving the eyelids.

A 47-year-old patient with an unusual tumor involving the right upper and lower eyelids has been followed for almost 6 years. The tumor has remained localized to the eyelids and has recurred locally following attempts at complete or partial excision. The morphological features of the tumor as seen by ordinary light microscopic methods were puzzling, and resulted in a variety of pathologic diagnoses. Light microscopic examination of plastic-embedded semithin sections, and electron microscopic examination indicate that this is a hitherto undescribed eccrine sweat gland tumor of clear cell origin. Its infiltrative growth pattern and tendency to local recurrence suggests that it may be a low-grade malignant neoplasm.


Ganglioneuroblastoma associated with malignant mesenchymoma.

A case of ganglioneuroblastoma associated with malignant mesenchymoma is reported. The tumor originated from the retroperitoneum and was comprised not only of neuroblastoma and ganglioneuroma, but also of typical rhabdomyosarcoma, liposarcoma, undifferentiated mesenchymoma, as well as cartilaginous tissue. No distinct border existed between thses different components. The histogenesis of the tumor is discussed. We suggest that the tumor derived from a remnant of neural crest (ectomesenchyme).


Glycogen-rich adenocarcinoma of minor salivary glands. A light and electron microscopic study.

A malignant glycogen-rich adenocarcinoma of palatal salivary glands is reported. Histopathology revealed nonencapsulated nests and cords of polyhedral cells showing voluminous clear cytoplasms and cellular pleomorphisms, separated by fine vascular septae. Small and large ducts were infrequently seen showing apparent transition of large ducts into clear cells. The tumor cells were PAS- and Best-carmine positive, diastase soluble, and mucicarmine and Alcian-blue negative. Ultrastructurally, the tumor cells were arranged in solid nests and cords of electron-lucent cells surrounding single or multiple lumina, and surrounded by basement lamina. Occasional fusiform electron-dense cell processes were present beneath the basement lamina. The electron-lucent cells were joined by junctional complexes, contained intracytoplasmic canals, and were filled with massive accumulations of beta glycogen particles. The electron-dense processes contained interlacing whorls of fine filaments and exhibited peripheral focal densities. The findings suggest that this glycogen-rich malignant tumor is of epithelial origin most probably of ductal cells.


Proportional mortality among alcoholics.

To test the hypothesis that there is a positive association between chronic alcoholism and carcinoma of the pancreas, the mortality experience of 1382 chronic alcoholics was studied. Analysis was limited to a comparison of observed and expected proportional mortality of different causes of death in the 894 whites who were known to have died. For carcinoma of the pancreas, 3 deaths were observed and 5.2 were expected. The observed/expected ratios for other causes of death, including other sites of cancer, were in accordance with prior studies.


Pericardial effusion secondary to mucoepidermoid carcinoma of the parotid gland. A report of an unusual case.

This paper reports a unique case of malignant pericardial effusion secondary to metastatic mucoepidermoid carcinoma in a patient who previously had mucoepidermoid carcinoma of the parotid gland and subsequently was found to have a hypernephroma of the kidney. Its distinctive features warranted the diagnosis of mucoepidermoid carcinoma as the cause of the effusion, and excluded the more aggressive hypernephroma. Although pericardectomy was ultimately necessary, there was no evidence of viable malignancy in fluid specimens obtained after an initial course of radiation therapy or in the resected pericardium. This suggests that conservative treatment with local radiation therapy might be successful in future cases of pericardial effusion in this malignancy.


Malignant mesenchymoma. Case report with electron microscopic study.

A malignant tumor from the thigh of a 65-year-old man was examined by light and electron microscopy. The tumor was diagnosed by light microscopy as malignant mesenchymoma with prevalence of leiomyosarcomatous or liposarcomatous components, but electron microscopic study revealed that the basic cell was similar to chondroblastic or osteoblastic cells. However, a precise ultrastructural classification of the tumor cells was not possible, since the majority of the cells had mixed features. The histogenesis, differentiation, and relation to prognosis are discussed.


Geographic pathology of occult thyroid carcinomas.

Thyroid glands obtained from patients in southeastern Canada, northeastern Japan, southern Poland, western Colombia, and from Japanese living in Hawaii were serially step-sectioned and examined microscopically using identical techniques and diagnostic criteria. The prevalence of occult papillary thyroid carcinoma was significantly higher in Japan (28.4%) and in Hawaiian Japanese (24.2%) when compared with Canada (6%), Poland (9.1%), and Colombia (5.6%). The carcinomas were all papillary except for a single follicular lesion from Colombia. There was no significant sex prevalence. Most of the patients were between 40 and 79 years of age, but there was no particular predominant decade. Only the Colombian series had a large number of younger patients, and they showed a slightly lower prevalence of occult carcinomas before age 40. Most papillary thyroid carcinomas grow slowly and probably remain occult for the life of the patient.


A case of hyperparathyroidism associated with radiation to the head and neck area.

Hyperparathyroidism due to a hyperfunctioning parathyroid adenoma developed in a 57-year-old woman exposed to radiation for facial hirsutism, in whom other tumors of the head and neck had occurred. The details of history and the implications of causal relationship are presented.


Adenocarcinoma of the nose and paranasal sinuses in woodworkers in the state of Victoria, Australia.

The case index of the Cancer Institute of Victoria (Australia) contained 19 cases of adenocarcinoma of the nose and paranasal sinuses. Eighteen of the cases were in men and 1 in a women. Routine questioning of these patients revealed an occupation involving woodworking in 7 cases, whereas among 80 cases of other malignant tumors of the nose and sinuses there were only 4 who had been woodworkers. Among the patients with adenocarcinoma of the nose and sinuses, there was a significantly higher proportion of woodworkers than in the general population. The findings are consistent with European reports associating nasal adenocarcinoma with wood dust, but whereas the workers at risk in Europe are mainly in the furniture industry, some of the workers affected in Victoria have been sawmillers or carpenters. The specific salivary patterns of tumors of mucous glands are not associated with woodworking.


Kaposi's sarcoma of the brain. A case report with necropsy findings.

A case of Kaposi's sarcoma involving the brain is described, this being the first of its kind in Uganda. The patient was a 40-year-old woman who presented with a 2-year history of skin tumors on the left ankle that proved resistant to chemotherapy. She therefore underwent an above-knee amputation. Six months after amputation, she developed signs of cerebral involvement, and 6 weeks later she died. Necropsy showed large tumor deposits in bone and lungs as well as in the brain. Histologically the cutaneous tumors consisted of the monomorphic variety, whereas the cerebral and other deposits were anaplastic. In the three cases of cerebral Kaposi's sarcoma hitherto reported from other countries, only minute foci of tumor were found in the brain.


Death from Hodgkin's disease after 18 years' complete remission.

Survival studies on series of patients with Hodgkin's disease have indicated that localized disease and certain histologic types are associated with a favorable prognosis. The permanency of cure in such instances has seldom been proved by long-term followup and autopsy; there is evidence that the disease may remain dormant for considerable periods, with the attendant risk of recurrence. A case is reported wherein a patient was relapse-free for 18 years prior to fatal recurrence, which was confirmed by autopsy.


Prognosis in thyroid carcinoma.

All cases of thyroid cancer reported to the Finnish Cancer Registry in 1958-62 were re-examined histologically; 227 cases of thyroid carcinoma were found. The correlation between survival rates and different histologic and clinical features was studied. The survival rates for papillary carcinoma were significantly higher than those for follicular carcinoma, and the latter significantly higher than those for anaplastic carcinoma. The survival rates for medullary carcinoma were similar to those for papillary carcinoma, but there were only 10 medullary tumors. Within papillary carcinoma, vascular invasion correlated with low survival rates. In follicular carcinoma, the encapsulated type, and in anaplastic carcinoma the diffuse small cell type had higher survival rates than other tumors. The extent of the primary tumor and the presence of distant metastases correlated with survival, but the presence of regional metastases did not. The presence of severe compression symptoms correlated with low survival rates. Young patients and females had higher survival rates than old patients and males. In making a postoperative estimation of prognosis it is best to take the histologic type, the extent of the primary tumor, and the presence of distant metastases into consideration. A tentative preoperative prognosis can be bases on the age and sex of the patient and the presence of distant metastases and severe compression symptoms.


Augmentation mammoplasty, irradiation, and breast cancer. A case report.

A case report is presented of mammary adenocarcinoma in association with a Silastic prosthesis and earlier irradiation exposure. Causative relationships of these two variables are discussed. Recommendations for management of the breast cancer patient with a mammary prosthesis are suggested.


Prognosis in Black children with neuroblastoma.

Thirty white and 15 black children with neuroblastoma were compared with respect to survival. Patient age and tumor stage at diagnosis were similar in the two groups, as was duration of symptoms prior to diagnosis. There was no significant difference between the white and black children in regard to median duration of survival or percentage of long-term survivors. The results of this study indicate that race is not a factor in the prognosis of this tumor.


Metastatic thymoma with myasthenia gravis and pure red cell aplasia.

Metastatic malignant thymoma developed in a 64-year-old man after a 9-year history of myasthenia gravis. Following good response to irradiation of the mediastinal mass and osseous metastasis, pure red cell aplasia developed.


Nonevaluable patients in clinical cancer research.

Analysis of controlled and uncontrolled published papers in clinical cancer therapy has revealed differences in patient data handling, which may reflect both the inherent biases of patient selection for uncontrolled studies and the primary purpose of such studies. A distinction between drug-data oriented research and patient oriented research was advanced to explain these results. If comparable results are to be developed, we urge controlled trials at all phases of treatment development.


Cancer mortality among Mormons.

Preliminary results show that the 1970-72 cancer mortality rate among California Mormon adults is about one-half to three-fourths that of the general California population for most cancer sites, including many sites with an unclear etiology. Furthermore, the cancer death rate in the predominately Mormon state of Utah is about two-thirds to three-fourths of the United States rate, and the lowest in the entire country. Mormons are a large, health-conscious religious group whose Church doctrine forbids the use of tobacco, alcohol, coffee, and tea, and recommends a nutritious diet. Initial indications are that Mormons as a whole smoke and drink about half as much as the general population, and that active Mormons abstain almost completely from tobacco and alcohol. However, they appear to be fairly similar to the general white population in other respects, such as socioeconomic status and urbanization. The significance of these findings is discussed.


A clinical staging system for multiple myeloma. Correlation of measured myeloma cell mass with presenting clinical features, response to treatment, and survival.

The presenting clinical features of 71 patients with multiple myeloma were correlated with myeloma cell mass (myeloma cells X 10(12)/m2 of body surface area) determined from measurements of monoclonal immunoglobulin (M-component) synthesis and metabolism. Bivariate correlation and multivariate regression analyses showed that myeloma cell mass could be accurately predicted from A) extent of bone lesions, B) hemoglobin level, C) serum calcium level, and D) M-component levels in serum and urine. Analyses of response to chemotherapy and survival indicated significant correlation with measured myeloma cell burden. The results were synthesized to produce a very reliable and useful clinical staging system with three tumor cell mass levels (Table 7). For clinical research purposes, multivariate regression equations were developed to predict optimally the exact myeloma cell mass. Thus, initial staging can be quantitatively related to followup using tumor cell mass changes calculated from changes in M-component production. Use of the clinical staging system sould provide better initial assessment and followup of individual patients, and should lead to improved study design and analysis in large clinical trials of therapy for multiple myeloma.


The role of cytosine arabinoside maintenance in acute nonlymphoblastic leukemia.

A series of 30 unselected patients with acute nonlymphoblastic leukemia (ANLL) was treated with combination chemotherapy, including three courses of cytosine arabinoside (Ara-C) by 5-day continuous i.v. infusion, vincristine i.v. weekly, and prednisone daily to complete remission. Ara-C was administered alone as a 5-day continuous i.v. infusion monthly for maintenance. Ten (33%) achieved a complete remission (CR). The remaining 30 (67%), including temporary partial remissions, hematologic improvements, inadequate trials, and early deaths, were all considered failures. The CR rate was 57% in those 17 cases receiving an adequate trial. After After 5 1/2 years' followup, the overall median survival, including cases failing to achieve CR, was 3.1 months. For those having adequate trials the median survival was 16.6 months, and for those achieving a CR, 36.6 months. Two patients are still alive, one at 55.2 months on maintenance therapy, and the other at 62.8 months, currently unmaintained.


Evaluation of methotrexate in the treatment of bronchogenic carcinoma.

One hundred ninety-five patients with disseminated bronchogenic carcinoma were treated with 10 different dosage schedules of methotrexate. Patient response to methotrexate therapy varied with the type of treatment, but overall, 18% of the patients treated responded to a measurable degree. Survival was not increased in any of the treatment categories when compared to a placebo group; however, the survival of patients that responded to therapy was greater than the survival of patients that did not respond to therapy. Methotrexate will find its greatest potential use in combination with other drugs proven to be effective in treating bronchogenic carcinoma.


Rupture of major vessels after radiation.

Rupture of irradiated large vessels is an uncommon complication which tends to occur in carotid, aorta, and femoral arteries, in decreasing order of incidence. It particularly affects men subjected to surgery and radiotherapy for epidermoid carcinomas in oropharynx, esophagus, or genitalia. Contrary to some opinion, radiation is not the most significant cause; this "spontaneous" arterial rupture can occur without radiation. The perforation is not associated with tumor invasion of arterial wall, as has been claimed. Surgical complications, especially necrosis of skin flaps, infection, and fistulas are most important etiologically. Only 2 of the 11 instances that we describe appear to have been caused mainly by radiation. The perforation is usually fatal, but several patients, including 3 of our series (2 carotid, 1 femoral), have been saved by hospital personnel aware of this complication. To prevent it, prolonged exposure or infection of arteries, whether irradiated or not, should be avoided.


Hyperkalemia complicating splenic irradiation of chronic lymphocytic leukemia.

Hyperkalemia is an infrequent complication of the therapy of malignant disease. In previously reported cases, hyperkalemia following the institution of chemotherapy has been associated with acute renal failure. In this report, we present a patient with chronic lymphocytic leukemia who developed hyperkalemia following splenic irradiation. Necrosis of tumor cells, either as direct or an abscopal effect appears to be implicated as a cause of hyperkalemia. It seems appropriate to monitor potassium levels when therapy of a responsive tumor is instituted.


The effects of therapeutic radiation on colonic mucosa.

Low to moderate doses of therapeutic irradiation are capable of producing bizarre cytologic changes in the mucosal cells of colorectal crypts. This may be associated with eosinophilia of the lamina propria and with eosinophilic crypt abscesses. The bizarre cells generally line intact crypts and do not exhibit invasive behavior. The change subsides within 1-2 months at the dose levels studied (approximately 2000 rads).


Radiation-adriamycin interactions: preliminary clinical observations.

This communication reports four patients treated with the anthracycline adriamycin and irradiation either concurrently or sequentially, who demonstrated apparent augumentation of radiation reactions. Two of these patients demonstrated the so-called "recall" phenomenon.


Correlation of delayed hypersensitivity responses with chemotherapeutic results in advanced Hodgkin's disease.

Delayed skin test responses to several or all of a battery of six antigens were evaluated in 64 patients with disseminated Hodgkin's disease before, during, and after multiple-agent chemotherapy. Before therapy, 53% of patients had one or more positive skin tests, as compared to 55% during intensive chemotherapy, 79% during maintenance therapy, and 100% after discontinuation of all treatment. Pretreatment skin tests were of no value in predicting clinical response to chemotherapy. Response rates, duration of response, and survival were similar among anergic patients and patients with positive skin tests before treatment. There were too few patients who remained anergic after intensive induction chemotherapy to permit a correlation of immunologic reactivity with course. We conclude that skin test responses to recall antigens, before or during aggressive treatment, are more indicative of Hodgkin's disease activity (and also, of the immunosuppressive effects of treatment routines) than of prognosis.


Lymphocyte response to autologous tumor antigen(s) and phytohemagglutinin in ovarian cancer patients.

Peripheral blood lymphocytes from 11 patients with ovarian carcinoma were incubated several times with autologous tumor extract as well as phytohemagglutinin (PHA) at different concentrations. Lymphocytes from 10 normal, healthy, age-matched females were also studied similarly. Tritiated thymidine incorporation into DNA of transformed lymphocytes was measured. No significant response was obtained with lymphocytes from ovarian cancer patients or normal individuals in the presence of tumor extracts. Of 6 tumor extracts tested, 5 did not have any cytotoxic effect on autologous or on homologous normal healthy lymphocytes. Hence, with this test system employed, no evidence for a cell-mediated immunity to autologous tumor in ovarian cancer patients was observed. PHA, on the other hand, induced vigorous blastogenic response in several ovarian cancer patient's lymphocytes, as well as in all normal healthy lymphocytes, indicating no inherent defect in the T-lymphocyte system of ovarian cancer patients. Only 2 of the patients, when tested for delayed hypersensitivity reaction against a battery of recall antigens and keyhole limpet hematocyanin, were completely anergic. None of the patients responded when skin tested with their own tumor extracts.


Immunologic aspects of human thyroid cancer. Humoral and cell-mediated immunity, and a trial of immunotherapy.

Immunologic studies were performed on 16 patients with thyroid cancer. Circulating leukocyte counts increased, parallel to development of the terminal stage of disease, but total lymphocytes decreased. Serum immunoglobulin and complement were high, even though almost all patients showed negative antithyroid antibodies. Delayed skin hypersensitivity to bacterial and viral antigens and lymphocyte responsivity to PHA were not impaired at the initial stage of disease, but were impaired in terminal illness. Cell-mediated immunity (CMI) to tumor antigens(s) was measured using the assays of lymphotoxin, migration inhibition factor, and peripheral leukocyte migration inhibition. A few patients showed significant response to tumor antigen, but not to homogenates of Graves' thyroid gland. Active immunotherapy was applied to three patients. Two patients, who were in the terminal stage of illness, could not develop generalized CMI; immunization did not alter the patients' rapid downhill course. One patient developed in vitro evidence of CMI against cancer tissue antigens, associated with decrease in tumor size. Four months after immunization, CMI was impaired in autologous plasma culture, but not in cultures in allogenic normal plasma.


Thymosin-induced increase in E-rosette-forming capacity of lymphocytes in patients with malignant neoplasms.

Thymosin, a thymic hormone, restores decreased cellular immunity under various experimental conditions. The purpose of this study was to evaluate the effect of thymosin on the E-rosette-forming capacity in cancer patients. Peripheral blood lymphocytes obtained from eight patients with various malignant neoplasms were examined. One of these patients received thymosin intramuscularly; his E-rosette formation was examined serially. Eight normal adults served as controls. Patients with advanced stages of malignant tumors showed decreased E-rosette-forming capacity, which increased significantly under the influence of thymosin. Patients with less-advanced tumors, as well as normal adults, demonstrated normal E-rosette formation without further effects by thymosin. The in vivo administration of thymosin to one patient was followed by a marked increase of the E-rosette-forming capacity. These results suggest that the administration of thymosin, both in vivo and in vitro, significantly increases the circulating T-lymphocyte levels and/or functions in patients with advanced malignant neoplasms.


Clear cell sarcoma with melanin pigment.

A patient with a clear cell sarcoma of tendons and aponeuroses arising in the left sacral area is reported. The tumor grossly contained two black foci that had pigment with the staining characteristics of melanin on light microscopy. Electron microscopy of these areas showed the pigment to be within melanosomes. This is the second report of a neoplasm diagnosed as clear cell sarcoma in which melanin was demonstrated; the possibility that the tumor represents a soft tissue variant of malignant melanoma is discussed. It is suggested that clear cell sarcoma is a heterogeneous entity, and that a number of different soft tissue neoplasms may present with a clear cell pattern, making diagnosis and classification difficult.


A classification of transplantable tumors in Nb rats controlled by estrogen from dormancy to autonomy.

Transplantable tumor lines were previously established from a variety of estrogen-induced tumors in Nb rats, including tumors of the adrenal cervix, salivary gland, and pancreas, a lymphoma, and a liposarcoma. Spontaneous tumors, however, were found to arise in untreated females and showed the same characteristics. Tumor growth was dependent upon or influenced by estrogen when assessed in estrogenized and unconditioned hosts. Intermitten estrogenization was effective, but tumor growth took place more slowly. The type of response observed led to a new classification of five types of hormone-responsive tumors including tumors inhibited by estrogen. Estrogen-dependent tumor cells might remain dormant indefinitely and not grow in unconditioned animals until stimulated to grow by estrogen. The growth rate of hormone-dependent adrenal carcinomas was related to the amount of estrogen. Tumor growth started more rapidly in the presence of low estrogen dose levels in old rats used as hosts than it did in young rats. Breast carcinomas required the largest amount of estrogen for growth, whereas ovarian thecomas would grow in normal females but not in males. The growth rate in conditioned hosts of most transplanted tumors (some have maintained hormone dependency over 10 years) increased over successive generations. Progression, however, towards a more autonomous state after repeated transplantations was remarkably slow, and a sudden change to autonomy was rarely noted. In contrast, transplants of 9,12-dimethylbenz(a)anthracene-induced mammary carcinomas progressed rapidly to autonomy. Fould's concept of progression (2, 3) has been discussed but the described classification of tumors under hormone influence apparently allows a more detailed analysis of definition of different types of progression.


Tumor-blocking and -inhibitory serum factors in the clinical course of canine venereal tumor.

An in vitro method for growing colonies of canine transplantable venereal tumor cells in a semisolid agar medium is described. Using autologous or pooled homologous normal dog sera as a feeder layer, 49.3 +/- 3.5 and 47.5 +/- 4.5 tumor colonies were obtained, respectively, when 2 X 10(4) tumor cells were plated. With this assay system, assessment of colony counts provided an accurate and rapid technique for monitoring serum factors in tumor-bearing dogs that inhibited colony formation, or blocked the inhibition of colony formation. In 52 dogs given tumor transplants, a direct correlation was demonstrated between serum activity tested in vitro and the in vivo growth characteristics of the tumor. Tumor cells preincubated with serum from dogs with active tumor growth consistently showed normal colony growth when cultured in agar containing colony-inhibitory sera (blocking effect). In vivo regression was characterized by loss of serum blocking and the development of serum colony-inhibitory activity in culture. In metastatic disease, only blocking activity could be identified, while persistent local invasive disease was characterized by low levels of both blocking and inhibitory serum activity. The sensitivity of this technique coupled with its in vivo predictive capabilities provides a model for monitoring serological responses to a naturally occurring neoplasm in a large, randomly bred animal.


Conditions modifying development of tumors in mice at various sites by benzo(a)pyrene.

The modifying roles of age, sex, and strain of mice on the incidence, multiplicity, and spectrum of tumors induced by benzo(a)pyrene have been investigated. The first-generation (F1) hybrids of C57BL/6J X C3HeB/FeJ and C3HEB/Fej X A/J mice of both sexes were given single i.p. injections (75 or 150 mug/g) of benzo(a)pyrene at 1, 15, or 42 days of age. Experimental animals were allowed to live their life-spans, while animals in control groups were killed at 52, 90, 142, or 170 weeks of age. Animals treated with benzo(a)pyrene died, in general, by the 100th week of age due to development of liver, lung, stomach and lymphoreticular tumors. Few of the control animals died during that same observational period. The age of mice at the time of exposure to the carcinogen modified development of tumors at all the sites. The sex of animals influenced the development of liver and lymphoreticular tumors. The C3HeB/FeJ X A/J F1 hybrids developed lung tumors more readily than did the C57BL/6J X C3HeB/FeJ F1 mice, which had significantly more liver tumors and neoplasms of the lymphoreticular system than the former strain. No strain difference was observed in regard to tumors at other sites. Higher doses of benzo(a)pyrene were more effective in inducing lung, liver, and stomach tumors. In addition, 5 cases of pancreatic ductal adenoma and adenocarcinoma were observed in carcinogen-treated mice.


Comparative toxicity of N-hydroxy-2-acetylaminofluorene in several strains of rats.

Male Spraque-Dawley rats were 6 or 7 times more susceptible than females to the acute toxic effects of a single i.p. injection of N-hydroxy-2-acetylaminofluorene. The N-hydroxy compound were equally toxic in male and female Fischer rats and about twice as toxic to male as to female Wistar rats. A negative correlation between the 50% lethal dose of N-hydroxy-2-acetylaminofluorene and hepatic N-hydroxy-2-acetylaminofluorene sulfotransferase activity was found. These data substantiate earlier indications that the level of the liver sulfotransferase is an important factor in determining the degree of toxicity of N-hydroxy-2-acetylaminofluorene. It is suggested that the reported sex difference in the hepatocarcinogenicity of N-hydroxy-2-acetylaminofluorene might be peculiar to the Sprague-Dawley rat.


The reduction of N-hydroxy-4-acetylaminobiphenyl by the intestinal microflora of the rat.

The role of the intestinal flora in the conversion of N-hydroxy-4-acetyl-aminobiphenyl (N-OH-AABP) to 4-acetylaminobiphenyl has been examined. This reaction, which reverses the metabolic activation of the parent carcinogen, can be demonstrated in cultures of some bacteria indigenous to the intestinal microflora. These include cultures of Clostridium sp., Clostridium perfringens, Peptostreptococcus productus I, and Bacteroides fragilis ss. thetaiotaomicron and ss. vulgatus. In contrast, cultures of Lactobacillus plantarum and Escherichia coli show little or no capacity for this reaction. The reduction of N-OH-AABP is also carried out by homogenates of liver, kidney, and brain. On a weight basis, the cecal flora is considerably more active in reducing N-OH-AABP than are homogenates of tissues of the gastrointestinal tract. The cecal flora also has a greater activity for reducing N-OH-AABP than the stomach flora, an observation which may relate to the induction of tumors in the forestomach but not in the cecum of rats fed this compound. The products of the metabolism of N-OH-AABP have been compared in germ-free and conventional animals. Glucuronide conjugates of N-OH-AABP are found in the cecal contents and feces only of the germ-free rats, while 4-acetylaminobiphenyl is found in the feces only of conventional rats. These results suggest that the flora, by hydrolyzing glucuronides and reducing N-OH-AABP, may influence the level of metabolities of 4-acetylaminobiphenyl which are critical for carcinogenesis.


Carbamoylation of amino acid, peptides, and proteins by nitrosoureas.

Incubation at approximately physiological conditions of amino acids, peptides, and proteins with 1-(2-chloroethyl)-3-cyclohexyl-1-nitrosourea or cyclohexyl isocyanate resulted in carbamoylation of the alpha-amino groups of amino acids, the terminal amino groups of peptides and proteins and the epsilon-amino groups of lysine moieties. Carbamoylation of the alpha-amino groups and the terminal amino groups occurred as readily as, or more readily than, the carbamoylation of the epsilon-amino groups. Carbamoylation of the amino groups of amino acids or peptides by 1,3-bis(2-chloroethyl)-1-nitrosourea or 1-(2-chloroethyl)-3-cyclohexyl-1-nitrosourea altered the electrophoretic mobility of those compounds. Cyclization of (2-cloroethylcarbamoyl)-amino groups to form (2-oxazolin-2-yl)amino groups occurred at room temperature, and the resulting oxazolinyl compounds migrated electrophoretically similarly to the parent compounds. Since such cyclization did not occur with cyclohexylcarbamoylamino groups, treatment of amino acids, peptides, or proteins with 1,3-bis(2-chloroethyl)-1-nitrosourea might result in less permanent alteration of the respective charges on the resulting products than would treatment with 1-(2-chloroethyl)-3-cyclohexyl-1-nitrosourea or other nitrosoureas lacking a 2-chloroethyl group on N-3. The relevance of these differences in charge to differences in physiological effects is not presently known. Although the present study does not establish a definite relationship between carbamoylation of any specific protein and the physiological effects of nitrosourea, it does reinforce and expand the existing evidence that carbamoylation of proteins is a proteins is a process that must be considered in efforts to explain the physiological effects of these agents, and it points to terminal amino groups of proteins as possible primary sites of carbamoylation.


Tissue-specific synthesis of methotrexate polyglutamates in the rat.

Purified tritium-labeled methotrexate was administered to rats and Sephadex G-15 gel chromatography was used to study the formation of poly-gamma-glutamyl metabolites of methotrexate in different tissues. These metabolities were recognized as tritium-labeled antifolate fractions and were identified by their response to serum pterolypolyglutamate hydrolases and by cochromatography with authentic standards. After doses equivalent to 15 to 20 mg for man, rat liver and kidney were found to contain both 4-amino 10-methylpterolyglutamyl-gamma-glutamic acid and 4-amino-10-methylpterolyglutamyl-gamma-glutamyl-gamma-glutamic acid. With one-third of the dose, only 4-amino 10-methylpteroylglutamyl-gamma-glutamic acid was found. Synthesis of methotrexate polyglutamates in liver and kidney was limited to the interval immediately following methotrexate administration and appeared to occur by sequential addition of single glutamyl residues to so-called "free" intracellular methotrexate. No synthesis of methotrexate polyglutamates was demonstrated in small intestine or thymus. After formation, 4-amino-10-methylpteroyglutamyl-gamma-glutamic acid disappeared from liver and kidney with a half-time of 6.5 days. The effect of these metabolites of methotrexate of cell metabolism is unknown.


Surface properties of phorbol esters and their interaction with lipid monolayers and bilayers.

The potent tumor-promoting agent 12-O-tetradecanoylphorbol-13-acetate (TPA) is surface active and was found to occupy a limiting area of 62 sq A/molecule in monolayers at the air-water interface. The interfacial tension of aqueous TPA solutions is decreased by increasing the bulk-phase TPA concentrations up to 2 x 10(-6) M,beyond which no further decreases were observed. This concentration is in agreement with the apparent solubility limit previously obtained. The apparent aqueous solubility limit of the more hydrophobic phorbol-didecanoate is 5 x 10(-8) M. Interaction of TPA with egg phosphatidylcholine monolayers at the air-water interface was shown by an increase in the surface pressure of the monolayer from 22 dynes/cm, initial film pressure, to 34 dynes/cm 90 min after introduction of TPA into the aqueous subphase. It was shown by gel filtration chromatography that a similar phorbol derivative, tritiated phorbol-didecanoate, binds to phospholipid vesicles. Differential scanning calorimetry also indicated that the addition of either TPA or an inactive stereoisomer, 4-alpha-phorbol-didecanoate, to phospholipid bilayers results in a marked reduction of the enthalphy of the minor transition of dipalmitoylphosphatidylcholine liposomes. Several fluorescence polarization probes for membrane fluidity indicate that TPA does not affect this membrane parameter. Further, the presence of TPA induces no measurable change in the cation permeability of phospholipid vesicles, the conductance of planar bilayer membranes, or the electrophoretic mobility of negatively charged liposomes. The lack of a specific effect with bilayers alone, combined with the documented physiological effects at low TPA concentrations, point to the possibility of a specific membrane component as the receptor for TPA at the plasma membrane.


Schedule-dependent synergistic cytotoxicity of arabinofuranosylcytosine with adriamycin or 3,6-bis(5-chloro-2-piperidinyl)-2,5-piperazinedione in cultured cells.

The purpose of this study was to demonstrate that in vitro cell culture systems could serve a useful purpose in providing some guidelines in formulating drug schedules for combination cancer chemotherapy in the clinic. Using monolayer cultures of Chinese hamster ovary cells as the test system, we screened the combinations of 1-beta-D-arabinofuranosylcytosine (ara-C) + adriamycin, ara-C + 3,6 - BIS(5 - CHloro - 2 - piperidinyl) - 2,5 - piperazinedione, melphalan + 5-fluorouracil, and melphalan + ara-C as to the effect of drug sequence on the plating efficiency of these cells. Schedule-dependent therapeutic synergism was observed only when ara-C treatment was followed by either adriamycin or 3,6-bis(5-chloro-2-piperidinyl)2,5-piperazinedione but not vice versa. No synergistic effects were observed in the combination of melphalan with 5-fluorouracil or ara-C. The basis for this synergism appeared to be that ara-C was primarily acting as a synchronizing agent to set up the cells in S phase so that subsequent treatment with an S-phase-specific drug, such as adriamycin or 3,6-bis(5-chloro-2-piperidinyl)-2,5-piperazinedione, would produce the maximum cell kill. On the other hand, the lack of synergism in the combinations of melphalan with 5-fluorouracil or ara-C was due to the lack of S-phase specificity for melphalan. On the basis of these data, we postulate that schedule-dependent synergism could be expected if the first agent renders the cells more sensitive (for example, by selectively blocking cells in one of the phases of the cell cycle) to the action of the second drug.


The nature of adriamycin-induced cytotoxicity in Chinese hamster cells as revealed by premature chromosome condensation.

The short- and long-term effects of adriamycin treatment on the cell cycle kinetics and chromosome structure of Chinese hamster ovary cells were investigated. Adriamycin treatment, either pulse or continuous, did not delay the progression of G1 cells into S phase but did prolong the duration of S and G2 phases. This effect was dose dependent, and the prolongation of G2 period was greater than that of S. The frequency of chromosomal aberrations induced by adriamycin was studied in the mitotic as well as in the prematurely condensed chromosomes (PCC) of the treated cells. The types of aberrations observed were chromosome stickiness, chromatid exchanges, breaks, and gaps. The frequency of aberrations continued to increase as cells progressed to mitosis even after the removal of drug from the medium. At prolonged intervals (24 to 72 hr) after a brief or extended treatment, the labeling indices of the treated populations decreased, indicating an eventual block in the cell cycle progression. Visualization of chromosomes of these blocked interphase cells by the PCC method revealed that normal G2 PCC were totally absent. We observed that the PCC of these blocked cells were abnormal in the following respects: (a) some of the G2 PCC were extensively damaged; (b) many of the S PCC failed to incorporate [3H]thymidine; (c) a significant fraction of these PCC were poorly condensed and hence it was difficult to determine whether they were in G1 or G2. These results indicate that there is a good correlation between damage to the genome and the inhibition of cell cycle progression after adriamycin treatment.


Cholera toxin effects on cell growth accompanied by selective alterations in metabolite uptake and modification of cell surface proteins.

Exposure of Chinese hamster ovary cells to cholera toxin at or below mug levels causes a marked morphological changes and increased adhesion and orientation of the cells. Such changes are paralleled by alterations in surface proteins as indicated by the cholera toxin-mediated modifications detectable by lactoperoxidase-catalyzed radioiodination of outer proteins. Mild tryptic treatment of cells prelabeled with [3H]glucosamine revealed a different kinetics of release of external glycoproteins in cells exposed to the toxin. An alteration in a specific glycoprotein species in cholera toxin-treated cells became evident by polyacrylamide gel electrophoresis followed by fluorography of 3H-labeled cellular glycoproteins. The effects of cholera toxin on surface proteins and growth of the cells occurred in the absence of a modification in amino acid uptake or incorporation of precursors into protein. However, thymidine uptake and glucosamine incorporation were inversely affected to toxin treatment. Some of the effects of the toxin appeared to be antagonized by colchicine.


9-beta-D-arabinofuranosyladenine 5'-phosphate metabolism and excretion in humans.

9-beta-D-Arabinofuranosyladenine (ara-A) was converted chemically to the 9-beta-D-arabinofuranosyladenine 5'-phosphate (ara-A-5'-P) and administered i.v. to four cancer patients in seven experiments. Urinary excretion and plasma levels of radioactivity were monitored for 24 hr in each case. Radioactivity present as unchanged ara-A-5'-P, ara-A, and the deamination product of ara-A, 9-beta-D-arabinofuranosylhypoxanthine, was determined. Excretion was, as in earlier studies with ara-A, given i.v., largely as 6-beta-D-arabinofuranosylhypoxanthine. However, in contrast to the 88 to 97% excretion of ara-A and products in 24 hr when ara-A was given by i.v. push, excretion was 41.47 to 79.1% in 24 hr when ara-A-5'-P was given. With the exception of one experiment at a low dose, where plasma ara-A levels were significant for 6 hr, the plasma levels of ara-A were sustained at significant levels for 24 hr after a single dose of ara-A-5'-P. The doses of ara-A-5'-P given were well tolerated by the four patients. Indications are that this derivative provides important advantages (solubility and sustained blood levels) over ara-A.


The release of high-molecular-weight alkaline phosphatase and leucine aminopeptidase into the media of cultured human cells.

Using exclusion from Sepharose 4B as our criterion, we have found a high-molecular-weight form of alkaline phosphatase and of leucine aminopeptidase which are released into the culture media by the FL amnion cell line. A low-molecular-weight form of leucine aminopeptidase is also found to contribute to the total levels of this enzyme in the media. The levels of these enzymes increased during the growth cycle of the culture, paralleling the increase in cell density, suggesting that the two events may be related. This phenomen in culture suggests a possible explanation for the appearance of similar enzyme forms in patient serum and fluids originating from diseased tissue.


Enhancement of spontaneous C3H/HeJ mammary tumorigenesis by long-term polyadenylic-polyuridylic acid therapy.

Female C3H/HeJ mice received weekly s.c. injections of 2 mg polyadenylic-polyuridylic acid. Therapy was initiated at either 2 or 9 months of age. In both cases, poly-adenylic-polyuridylic acid-treated animals developed the spontaneous mammary carcinoma associated with this strain more rapidly. Because the opposite result was formerly observed for AKR spontaneous leukemia, the data indicate the polyadenylic-polyuridylic acid has no generalized antineoplastic effect upon spontaneous tumors genetically associated with specific murine strains.


The relationship among transport, intracellular binding, and inhibition of RNA synthesis by actinomycin D in Ehrlich ascites tumor cells in vitro.

Uptake of actinomycin D (AD) was characterized in Ehrlich ascites tumor cells in vitro. The time course of uptake consists of an initial component that represents a unidirectional flux of AD into the cells, following which the net uptake rate falls to achieve a velocity that is constant over a 45-min interval of observation. The velocity of influx exceeds that of the late uptake phase by a factor of 2. Influx is 1st order over an AD concentration range of 0.02 to 14 muM. Influx is highly temperature sensitive (Q10 = 4.5); this is related only in part to the high temperature dependence (Q10 =2) of the oil-adqueous partition coefficient for this agent. Within 4 min after exposure of cells to AD, a component of intracellular AD appears which exceeds the concentration of extracellular AD and rapidly leaves the cells (t 1/2 congruent to 3 min) when the cells are resuspended into an AD-free medium. The level of this component of intracellular drug is unaffected by metabolic poisons, and the apparent chemical gradient is attributed to loose binding. While transport of AD does not appear to be energy-dependent, the data do not clafity whether translocation across the cell membrane is diffusional or carrier mediated. In addition to a loosely bound intracellular component, analysis of the unidirectional net efflux of AD indicates at least 3 other exit components; one with a t 1/2 of 35 min, a very tightly bound component, and another representing drug that is free within the intracellular water, The bulk of AD taken up is tightly bound to intracellular constituents, and the rate of accumulation of this cellular component accounts almost entirely for the net rate of uptake of AD during the late uptake phase. Because influx exceeds the rate of tight binding within the cell by a factor of only 2, it is unlikely that osmotically active AD achieves thermodynamic equilibrium across the cell membrane over the interval of these experiments. Rather, prior to saturation of binding sites within the cell, the level of free AD achieved is probably below the extracellular concentration. Since the rate of binding should be influenced by the level of free intracellular AD, factors that increase influx and bring the intracellular AD level closer to equilibrium should also increase the rate of intracellular binding. Tween 80 augments both influx and AD binding within the cell, an effect that may be related to augmented transport alone, but an additional stimulatory effect on the binding process is also possible. Association of AD with only a small number of the very-high-affinty intracellular binding sites, in the absence of free or loosely bound AD, results in approximately 35% inhibition of [2-14C]uridine incorporation into RNA within 50 sec after exposure of cells to 1 muM AD. Within 10 min. 75% inhibition is achieved indicating that association of AD with only a small fraction of the total high-affinity sites is sufficient to suppress RNA synthesis.


Bulk transfer of fluid in the interstitial compartment of mammary tumors.

Venous blood leaving a solid tumor showed higher erythrocyte concentration than did aortic blood. Net fluid loss of efferent blood as calculated from hematocrit differences was 2.7 to 6.7% of flow volume, 4.5 to 10.2% of perfusing plasma volume, or 0.14 to 0.22 ml fluid per hr per g in 2 to 5 g transplanted MTW9 and Walker 256 mammary carcinomas, and primary N-nitrosomethylurea- and 7,12-dimethylbenz(alpha)anthracene-induced mammary carcinomas of rats. Net fluid loss was directly related to blood flow but inversely related to tumor size. Increased hydrostatic pressure in tumor interstitial space was a consistent finding. Micropore chambers embedded in transplanted tumors drained 4 to 5 times more interstitial fluid than did identical chambers in the s.c. tissue. It is concluded that: (a) convective currents are present within the interstitial spaces of tumors; (b) the magnitude of fluid transfer can be measured by the difference in hemoconcentration between afferent and efferent tumor blood; and (c) the volume of this fluid transfer is not altered by hormone-induced tumor regression. The increased hydrostatic pressure of tumor interstitial spaces is interpreted as being due to absence of an anatomically well-developed lymphatic network. The bulk transfer of fluid within interstitial spaces is comparable to lymphatic drainage and should be considered in assessing drug concentration and distribution in solid tumors.


Fatty acid synthetase from a mouse mammary adenocarcinoma.

Fatty acid synthetase was isolated from transplantable mammary adenocarcinomas carried by C3H mice. Comparison of physicochemical and immunological properties of this enzyme with those of the fatty acid synthetase isolated from normal glands of lactating C3H mice indicates that the enzymes are similar in all respects. The products of the purified fatty acid synthetase from normal and neoplastic tissues were the same, mainly long-chain fatty acids. The normal gland was found to contain 30 times more enzymes per g tissue than did the neoplasm; this may account in part for the relatively low rat of fatty acid synthetase found previously in this neoplasm.


Selective destruction of cultured tumor cells with uncontrolled nuclear division by cytochalasin B and cytosine arabinoside.

Cultures of normal human and hamster and malignant human and hamster cells respond to cytochalasin B (CB) differently. The neoplastic cells become highly multinucleated with continuous nuclear division, while cytoplasmic division is prevented. These cells exhibit uncontrolled nuclear division. The normal cells show control of nuclear division, since CB treatment results in only binucleation, although cytoplasmic division is prevented. The CB-treated normal cells also show reduced incorporation of [3H]thymidine. When rapidly growing normal or neoplastic cells of either species are treated with cytosine arabinoside (ara-C), all cells are killed within 10 to 20 days. If the normal cells are treated with ara-C in the presence of CB, the cells survive for at least 35 to 40 days, suggesting that CB can protect normal cells from the destructive effects of ara-C. However, if malignant cells are treated with ara-C in the presence of CB, all cells are destroyed in 25 days. Although CB affords some protection to tumor cells, it is relatively small and appears only to effect a delay in ultimate cell death. Treatment of any of the cell lines with CB alone results in some loss of viability, but the drug is reversible although less so for the malignant cells, In additional experiments, are-C plus CB-treated normal and neoplastic cells were reversed from the drugs and propagated in normal medium at various periods after the initiation of ara-C treatment. ara-C plus CB-treated normal cells could be freely reversed at any time after the start of ara-C treatment up to at least 35 days. After reversal, cell growth resumed, and the total number of cells returned to that present prior to drug treatment. The tumor cells could not be freely reversed from ara-C plus CB, even when the ara-C was present for only 5 days. In this case the cells seemed to be destroyed at the same rate as in cultures where both drugs were kept in the medium. These results suggest that CB can protect normal but not malignant cells from the toxic effects of ara-C and offer a method to specifically destroy tumor cells.


The effect of dead cells on the activity of actinomycin D against mouse sarcoma 180 ascites.

It has been demonstrated that cells killed by heat or irradiation have four times greater affinity for actinomycin D (AMD) than do viable tumor cells. By using a double labeling technique, we were able to show that, with increasing amounts of AMD bound in cells, the incorporation of RNA precursors is proportionally decreased. However, in the presence of nonviable cells or of native DNA, the AMD-induced inhibition of [3H]uridine incorporation is markedly reduced. This reduction does not occur if DNase is added to the system. The accumulation of dead cells in the tumor vicinity during the natural course of tumor growth or therapy must be taken into consideration in planning therapeutic regimens. We suggest that, in combined chemo- and radiotherapy, increased effectiveness of AMD may be obtained by its use prior to irradiation, thereby assuring its direct access to the tumor cells. The addition of DNase could eliminate or greatly diminish the dead cell competition for the drug.


Decrease of epidermal histidase activity by tumor-promoting phorbol esters.

The potent skin tumor promoter (12-O-tetradecanoyl phorbol-13-acetate (TPA) stimulates epidermal macromolecular synthesis as well as proliferation, but little is known of specific functional aberrations produced by TPA. This report presents results of a study on the effects of TPA on epidermal histidase (L-histidine ammonia lyase), an enzyme found in normal epidermis but not in dermis or in mouse squamous cell carcinomas. Histidase activity was assayed on postmitochondrial supernatants obtained from hairless mouse epidermis after removal by keratotome. Topical TPA treatment at doses active in tumor promotion (1.7 to 17.0 nmoles/application) produced dose-dependent decreases in epidermal histidase specific activity at 19 hr posttreatment. The onset of the decrease occurred at 12 hr with recovery to control level specific activity by 5 days, showing kinetics similar to those obtained for stimulation of DNA synthesis. This decrease in histidase could not be attributed to a general inhibition of soluble protein synthesis or to the appearance of an inhibitor of histidase activity. The strong promoter TPA produced a greater histidase decrease than did the moderate promoter and mitogen 12,13-didecanoyl phorbol at equimolar dose, while phorbol, a nonpromoter and nonmitogen, produced no effects on histidase. The relationship of this histidase depression to tumor promotion and not initiation is further indicated by the finding that (a) Tween 60, a structurally unrelated tumor promotor, also produced a decrease in histidase; and (b) the tumor initiator urethan and an initiating dose of 9,10-dimethybenz(a)anthracene showed no effects on histadase activity.


Isolation of cis- and trans-4-methylcyclophosphamide and antitumor evaluation in vivo.

4-Methylcyclophosphamide was synthesized and separated into cis and trans isomers by column chromatography. Isolation of these isomers permitted individual evaluation against murine leukemia L1210 in vivo and assessment of possible differences in antileukemic activity. Results indicate no appreciable difference in activity of the isomers, suggesting essentially equal facility for activation by mouse liver microsomes in vivo.


Carcinogenicity of N-nitroso-3,4-dichloro- and n-nitroso-3,4-dibromopiperidine in rats.

The carcinogenic potencies of 3,4-dichloro- and 3,4-dibromonitrosopiperdine were compared with that of nitrosopiperidine by feeding to groups of 15 male rats in drinking water. A treatment of 15 weeks with a total of 0.5 mmole of the dichloro compound led to death of all animals before 24 weeks with tumors of the tongue, pharynx, esophagus, nonglandular stomach, nasal turbinates, trachea, bronchi, and bronchioles. Treatment of 27 weeks with the dibromo compound, a total of 1.0 mmole, caused death of all the animals by 41 weeks, with the same types of tumors. One-half of the rats treated with an almost 3-fold higher daily dose of nitrosopiperidine, 3.9 mmoles total, were alive at 40 weeks, and all were not dead until 55 weeks. Most of these animals died with tumors of the tongue, pharynx, esophagus, and nonglandular stomach and with squamous cell tumors and olfactory carcinomas of the nasal cavity, but there were no tumors of the respiratory tree. Substitution of chlorine or bromine in nitrosopiperidine greatly increased the carcinogenicity of the compound.


Synthesis of p-nitrobenzyl and p-nitrophenyl 1-thioglycopyranosides.

Reaction of 2,3,4-trio-O-acetyl-alpha-L-fucopyranosyl bromide (1) with thiourea (2), followed by reductive cleavage of the product, gave 2,3,4-tri-O-acetyl-1-thio-beta-L-fucopyranose (4). Reaction of 4 with p-nitrobenzyl bromide followed by O-deacylation yielded p-nitrobenzyl 1-thio-beta-L-fucopyranoside (6). Similar reaction conditions were used for the synthesis of p-nitrobenzyl 1-thio-beta-D-fucopyranoside (11) and 1-thio-alpha-D-mannopyranoside (16). A facile preparation of O-acylated p-nitrophenyl 1-thioglycopyranosides was achieved by condensing the appropriate glycosyl halide with sodium p-nitrobenzenethioxide in N,N-dimethylformamide.


The galactan sulphate of the red alga Polysiphonia lanosa.

The structure of the galactan sulphate of P. lanosa has been established by a combination of methylation, treatment with alkali, and partial methanolysis of the alkali-treated polysaccharide to give derivatives of agarobiose. The polysaccharide belongs to the agar class, in which 3-linked derivatives of beta-D-galactose alternate with 4-linked derivatives of alpha-L-galactose in a repeating sequence. In addition to D-galactose itself, the 3-linked units include 6-O-methyl-D-galactose, D-galactose 6-sulphate, and a hitherto unreported unit, 6-O-methyl-D-galactose 4-sulphate. The 4-linked units include L-galactose 6-sulphate, 2-O-methyl-L-galactose 6-sulphate, and 3,6-anhydro-L-galactose.


Fragmentation analysis of extracellular acid polysaccharides from seven Rhizobium strains. Part I. D-glucuronic acid-containing oligosaccharides.

The extracellular, bacterial polysaccharides from seven Rhizobium strains have been submitted to partial hydrolysis with acid. Several neutral oligosaccharides, some containing pyruvic acid, were isolated together with D-glucuronic acid-containing oligosaccharides. The polysaccharide from Rh. meliloti did not contain glucuronic acid. For the other six strains, the following components were characterized: 4-O-(beta-D-glucopyranosyluronic acid)-D-glucuronic acid, 4-O-(beta-D-glucopyranosyluronic acid)-D-glucose, and O-(beta-D-glucopyranosyluronic acid)-(1leads to4)-O-(beta-D-glucopyranosyluronic acid)-(1leads to4)-D-glucose. These results indicate the presence of chains containing two beta-(1leads to4)-linked D-glucuronic acid residues, beta-linked to D-glucose at position 4.


Studies on synthesis of 1,2-cis-D-galactosides with partially benzylated intermediates.

The 1-bromides of p-nitrobenzoylated 2-O-benzyl- (1) and 2,3- (8) and 2,6-di-O-benzyl-D-galactose (14) and 2-O-benzyl-D-glucose (20) were treated under Koenigs-Knorr conditions with 6-(trifluoroacetamido)-1-hexanol. Examination of the products by p.m.r. spectroscopy, g.l.c., and mass spectrometry revealed that, whereas the major product derived from 14 was the pyranoside (19), the glycosides derived from both 1 and 8 were mainly furanosides. The corresponding glycoside from 20 was entirely the pyranoside (23).


Comparison of clinical and hemodynamic assessment of cardiac disability.

Clinical versus hemodynamic assessment of cardiac disability was compared in 187 patients with either coronary artery or valvular disease classified using the New York Heart Association classification (excluding class IV). The following variables were analyzed during rest and exercise: arteriovenous oxygen content difference (CavO2), cardiac index (CI), pulmonary artery pressure (PAP), pulmonary artery wedge pressure (PAWP), pulmonary arteriolar resistance index (PARI) and heart rate (HR). 180 comparisons of variables were made. Hemodynamic evaluation paralleled clinical evaluation only 19 times during rest and 12 during exercise. There were 5 instances of clinical and hemodynamic distinction between class I and II, 14 between class I and III and 12 between class II and III. Correlation was best in mitral stenosis. The most useful variable was CI followed by PAP and CavO2. HR was not helpful. This study emphasizes that subjective clinical disability may not correlate with physiologic derangement in certain patients.


Cardiac effects of sodium selenite.

The classical idea that selenium is toxic to the heart at levels higher than available in a balanced diet is not supported by experimental work. In mice, treatment with sodium selenite increased the LD50 of ouabain and 2,4-dinitrophenol, and increased the tolerance to nitrogen inhalation. Although sodium selenite had no effect on the dog heart with circulation intact, there was a reduction in coronary vascular resistance in the heart-lung preparation. In the isolated ventricular segment perfused with blood, the administration of sodium selenite caused a positive inotropic effect which appeared even after blockade of beta-adrenergic receptors and in segments perfused with a Krebs-bicarbonate solution that was deficient in oxygen. These results cannot be explained merely as the correction of a selenium deficiency but rather as a positive influence of sodium selenite on the heart that has been acutely stressed by oxygen lack, ouabain, or 2,4-dinitrophenol.


Myocardial contractility in primary nonobstructive myocardiopathy.

Left ventricular myocardial function was studied in 13 patients with primary nonobstructive myocardiopathy and compared to the values determined in two control groups. Peak rate of rise of ventricular pressure (max dp/dt) and rate of the proportional rise of ventricular pressure at max dp/dt [(max dp/dt)p-1] were significantly decreased and interval t-max dp/dt was significantly prolonged in primary myocardiopathy, indicating that decreased contractility is characteristic of this disease. Left ventricular pre-ejection period (PEP) was significantly prolonged and PEP/LVET (left ventricular ejection time) ratio was significantly higher in patients with primary myocardiopathy. Increased end diastolic volume (EDV) and end diastolic pressure (EDP) were not characteristic of this diease. Best correlations were found between (max dp/dt)p-1 and t-max dp/dt (r = 0.71). Good correlations between PEP/LVET and internal indices of myocardial contractility qualify this ratio as a simple and useful index of myocardial contractility in various heart diseases.


Left ventricular end-diastolic volume in advanced ischemic heart disease; comparison between healthy subjects and patients with mitral stenosis.

The left ventricular end-diastolic volume (EDV), end-systolic volume (ESV) and systolic ejection fraction (SV/EDV) were determined in 6 healthy subjects, 21 patients with ischemic heart disease and 8 patients with mitral stenosis by the left ventricular 133Xe washout technique. Cardiac and stroke indexes as well as left ventricular work and stroke work indexes do not differ in all three groups. A significantly higher EDV and ESV together with a low SV/EDV suggested impaired left ventricular function in patients with ischemic heart disease.


Studies on the hemodynamic changes induced by electrical stimulation of the vagosympathetic trunks in the dog.

The effects of electrostimulation of the cephalic endings of the vago-sympathetic trunks on the systemic and perfusion pressure of the hind limb of the dog depend upon frequency, intensity and duration of the stimulus. The electrostimulation performed with stimuli of high frequency, low intensity and moderately long duration (rectangular waves of 1.5 V, 0.7 msec, 100 Hz) consistently elicited systemic hypertension and vasoconstriction. This response was completely abolished by pretreatment with guanethidine or hexamethonium. On the other hand, electrostimulation with stimuli of low frequency, high intensity and long duration (5 V, 1 msec, 10 Hz) produced systemic hypotension and vasodilatation in the perfused hind limb. These effects were completely blocked by atropine or hexamethonium, while guanethidine was unable to induce any modification. These findings suggest that electrostimulation of vagosympathetic trunks can evoke different hemodynamic responses depending upon the characteristics of the stimulus. The occurrence of a  vasodilatation specifically blocked by atropine demonstrates that a purely cholinergic mechanism underlies such a reflex response.