Fine structure of mule testes: light and electron microscopy study. Testes of 3 mules, 3 stallions, and 3 donkeys were studied by light and electron microscopic techniques. Mule testes were firm, with an average weight less than that of stallion or donkey testes. As expected, mule seminiferous tubules had few cell elements. There were 2 cell populations within the seminiferous tubules: (a) Only basal spermatogonia and Sertoli's cells in 70% of the tubules and, (b) the cells in (a) plus leptotene and pachytene primary spermatocytes in 30% of the tubules. Degenerative changes occurred at the pachytene stage and began with the appearance of nuclear vesicles and chromatin condensation. The last stages disclosed pyknotic nuclei and cytoplasmic vesicles. The interstitia were filled by connective tissue elements and few Leydig's cells. Interstitial cells had well-developed, smooth endoplasmic reticula and mitochondria, indicating a normal steroidogenic activity. Epididymides in the mule had morphologic features similar to those of the stallion or donkey, except for the presence of intertubular fibrosis. Degenerative changes in primary spermatocytes indicate metabolic alterations which could be explained by the blockage of meiosis.


Critical evaluation of oxfendazole as a swine anthelmintic. Sixty feeder pigs with mixed naturally acquired nematode infections were medicated with oxfendazole, a new benzimidazolecarbamate compound, at dose levels of 0, 3, 9, and 18 mg/kg of body weight. The dosage of 3.0 mg/kg administered once in the feed proved as efficacious as larger dosages and the same dosage administered 2 or 3 times in the removal of adult Ascaris suum, Oesophagostomum dentatum, and Metastrongylus pudendotectus. Trichuris suis adults were less susceptible. Anthelmintic efficacy was based upon comparative nematode egg counts, appearance of eggs, and recovered nematode burdens at necropsy. Signs of toxicosis were not seen in pigs given any one of the medicated feed schedules.


A 280-day tolerance-efficacy study with mibolerone for suppression of estrus in the cat. Mibolerone, a synthetic anabolic steroid, prevented estrus in domesticated cats when orally given a daily dose of 50 microng over a 180-day period. Doses of 20 microng daily and 50 microng given once a week failed to prevent estrus. Treatment with the 50-microng dose each day for 6 months had no apparent effects on subsequent estrus, mating, queening, or litter size. Kittens born to queens which had been treated did not have obvious developmental defects. Systemic metabolic changes produced by treatment were detected only in thyroid function, as revealed in dose- and time-related changes in serum cholesterol concentrations, thyroid gland weights, and thyroid histology. Clinical evidence of thyroid dysfunction was not apparent during the 6 months of treatment. Clinical and microscopic evidence of slight masculinization was apparent in cats after 3 months of treatment with 20 or 50 microng per day. Masculinizing changes consisted of thickening of the cervical dermis and clitoral hypertrophy. Behavioral changes were not observed. The apparent mechanism of action of mibolerone in the cat is the suppression of the release of pituitary luteinizing hormone.


Cardiovascular response to fentanyl-droperidol and atropine in the dog. A combination of fentanyl-droperidol was administered intravenously alone or with atropine sulfate (2 doses--0.04 or 0.02 mg/kg of body weight) to determine if stable neuroleptanalgesia could be produced in the dog. Cardiovascular responses were recorded at 5, 15, and 30 minutes. Fentanyl-droperidol given alone caused a significant increase of peripheral resistance and mean arterial pressure at 5 minutes and then a decrease of these values over a postinjection period of 30 minutes. Left ventricular dP/dt increased significantly at postinjection minutes 15 and 30. In dogs given atropine concurrently with fentanyl-droperidol, there was significant increase in heart rate and decrease in stroke volume. Also, there were significant initial increases in diastolic and mean arterial blood pressures, ventricular contractility, and coronary blood flow. The dose of 0.02 mg of atropine/kg seemed optimal for intravenous administration with fentanyl-droperidol in the dog; when the atropine dose was 0.04 mg/kg, large inotropic and chronotropic effects were produced.


Immune response of medicated turkeys vaccinated with live Pasteurella multocida. In 3 experiments, turkeys were vaccinated with the Clemson University strain of Pasteurella multocids (CU) via the drinking water for 1 or 3 days while they were fed 3 different rations. Ration 0X consisted of non-medicated turkey starter, ration 1X had 0.00625% sulfadimethoxine and 0.00375% ormetoprim added, and ration 2X had 0.0125% sulfadimethoxine and 0.0075% ormetoprim. Immune response was determined by challenge exposure with pathogenic P multocida, either serotype 3 or serotype 4, administered for 3 days via the drinking water. In experiment 1, the 1X concentration of medication reduced protective immunity by 30%. In experiment 3, the reduction was 37% for the 1X concentration and 45% for the 2X concentration. The increased concentration of medication was more efficacious in preventing infection in vaccinated and nonvaccinated turkeys.


Effects of Fusobacterium necrophorum leukotoxin on rabbit peritoneal macrophages in vitro. A method to demonstrate leukotoxic activity of Fusobacterium necrophorum in vitro is described. Continuous dialysis sac culture system, using reduced liquid medium was used to grow F necrophorum for 7-day periods. The continuous culture dialysis filtrate contained leukotoxic substance(s) which appeared to be less than 10,000 in molecular weight, heat resistant, and stable at 4 C for at least 10 days. Leukotoxic activity was demonstrated in vitro by determining the percentage of macrophages taking up trypan blue dye after these were exposed to a 1:2 solution of continuous culture dialysis filtrate and Eagle's minimal essential medium at 1, 2, 4, and 6 hours. Approximately 90% of these macrophages were destroyed during the 6-hour incubation period.


Normal laboratory values in Beagle dogs of twelve to eighteen months of age. Analyses of several body fluids are reported on Beagle dogs and compared with previous studies. All of the spinal fluid data and most of the urine data have not been previously reported in Beagles. Values of creatine phosphokinase, urine 5-hydroxyindoleacetic acid, and urine vanillymandelic acid are reported in the present study.


Tonography in the normal and glaucomatous beagle. A standard procedure for tonography was developed in the dog. Acepromazine, ketamine, xylazine, droperidol-fentamyl, and combinations of these drugs were evaluated for effects on blood pressure, intraocular pressure, and restraint of the dog for 4 minutes of Schiotz tonography of each eye. In dogs given a cepromazine (0.5 mg/kg intravenously) and then ketamine (10.0 mg/kg intramuscularly), the mean coefficient of aqueous humor out-flow in 24 normal Beagles was 0.24 mul/mm of Hg/minute (SD +/- 0.07). Mean coefficient of aqueous humor outflow in 35 glaucomatous Beagles, between 4 months to more than 37 months old, was 0.09 mul/mm of Hg/minute (SD +/- 0.04). Coefficient of aqueous humor out-flow was less in the older glaucomatous Beagle.


Clinical investigation of doxorubicin, daunomycin, and 6-thioguanine in normal cats. Cats frequently develop a variety of spontaneous hemolymphatic noeplasias. Long-term remissions in the nonlymphatic leukemic cat are difficult to obtain; therefore, more successful chemotherapeutic agents are sought for disease control. The purpose in the present study is to describe the clinical investigation of 3 antineoplastic drugs given to 23 normal cats. The cats tolerated doxorubican given at the dose level of 30 mg/m2 of body surface area once every 3 weeks; daunomycin, 15 to 30 mg/m2 once every 3 weeks; and 6-thioguanine, 25 mg/m2 for 5 days every 20 to 30 days.


Acute copper poisoning of cultured marine teleosts. Copper was accidentally introduced into seawater entering a circular outdoor tank used to culture 610 pinfish (Lagodon rhomboides) and other marine teleosts. Fish were exposed for 23 hours to copper concentrations between 0.2 parts per million (ppm) and 7.2 ppm with an average concentration of 3.3 ppm. The pinfish were lethergic by 6 hours after copper was introduced into the tank. Deaths began within 10 hours with a total of 575 (94%) deaths. Deaths stopped 11 hours after copper concentrations decreased below 0.2 ppm and signs of distress stopped in surviving pinfish by approximately 6 hours after the last death. Serum urea nitrogen, alkaline phosphatase, sodium (Na+), and potassium (K+) concentrations were increased and represent those constitutents which differ from mean control values by more than 2 standard deviations. Electrolyte imbalance, apparently resulting from impaired osmoregulation and hemolysis, was a serious terminal condition in copper-poisoned pinfish.


Pathologic manifestations of experimentally and naturally acquired lungworm infections in opossums. Experimentally induced and naturally acquired lungworm infections (Didelphostrongylus and Capillaria) were studied in opossums, Didelphis marsupialis. Each of 4 helminth-free opossums were experimentally infected with 250 3rd-stage Didelphostrongylus hayesi larvae. At 2 and 8.5 months after infection, 2 animals were necropsied. Hypertrophy of pulmonary smooth muscle characterized D hayesi infections, with lesions pronounced in the vicinity of helminths. There was mucous bronchitis characterized by peribronchial gland hyperplasia, lymphoid nodule formation, and exudation of mucus and eosinophils into bronchioles. Lung from 4 opossums harboring natural infections of D hayesi or Capillaria or both was examined. Concomitant infections produced multifocal, indurated lesions often resulting in generalized consolidation. Microscopic examination revealed pulmonic damage consisting of hypertrophy of smooth muscle, extensive peribronchiolar adenomatoid hyperplasia of alveolar epithelium, and diffuse areas of granulomatous interstitial pneumonia. Capillaria provoked well-defined granulomas, and frequently only eggs and cuticular debris were contained within the lesions. Loss of pouch young by 1 naturally infected feamle was associated with extensive verminous pneumonia.


"Spring rise" in fecal nematode egg counts in sheep in Maine. A "spring rise" in fecal nematode egg counts occurred in Maine sheep housed under conventional conditions during 2 years. Nematodes causing this increase were Haemonchus contortus in one year and Ostertagia sp in the other. The increase in egg counts was greater in sheep that lambed than in unbred sheep. Most of the eggs in the spring rise were probably produced by worms that passed the winter in developmental arrest in the sheep. Some of the female worms probably came from contaminated bedding and developed in the sheep without arrest in growth. Acquisition from contaminated bedding, therefore, should be considered as an additional factor in the epizootiologic character of the condition in Maine.


Use of 59Fe for sheep erythrocyte kinetic studies. Erythrocyte (RBC) survival time as determined by in vivo 59Fe-labeled RBC in 6 adult sheep was 111.7 (SEM +/- 8.4) days. The plasma clearance (T/2) of 59Fe was 148 (+/- 17.3) minutes and the maximum RBC uptake of 59Fe was 52.4% (+/- 3.6%). Plasma iron turnover rate was 0.356 (+/- 0.016) mg/kg/24 hours, and RBC iron turnover rate was 0.186 (+/- 0.016) mg/kg/24 hours. Blood volume measurement was 53.5 (+/- 3.9) ml/kg of body weight.


Operative trauma to the spleen.

Operative trauma to the spleen. A patient is discussed in whom splenic rupture followed an elective uncomplicated cholecystectomy by two weeks. There was no known direct operative or postoperative trauma to the spleen and the spleen was microscopically normal with no evidence of organizing hematoma. Possible mechanisms of operative splenic injury to the spleen are reviewed and the importance of recognition of this entity by surgeons is emphasized.


Complications of repeated operations to control esophageal reflux (esophagogastrocutaneous and esophagogastropericardial fistulas). Iatrogenic injury to the distal esophagus and gastric cardia with subsequent fistula formation may occur at times following repeated direct operative attempts to control reflux esophagitis. At re-operation when this problem occurs, the indirect operative procedure of Roux-en-Y diversion is a safe, effective and preferable method to control persistent reflux esophagitis.


Atherosclerosis obliterans in healthy young adults. A healthy young man developed unilateral claudication due to an isolated lesion of his external iliac artery, At operation, this was found to be obliterated by atherosclerotic change of long standing. No evidence of systemic disease was noted. A vascular reconstruction has allowed the patient to return to an active life style.


Gastric rupture as a result of blunt trauma. Six patients with gastric rupture secondary to blunt abdominal trauma are presented in detail. The salient features of each case are briefly discussed, along with a further analysis of the review by Yajko and colleagues. The aggressive operative treatment of these patients is emphasized along with the vigorous attention that must be given to postoperative intra-abdominal sepsis caused by the initial contamination leading to morbidity and mortality.


Complications following operation in the previously irradiated abdominopelvic cavity. Twenty-three patients underwent 32 operations in a previously irradiated abdominopelvic cavity. The overall postoperative morbidity rate was 62% and the mortality rate was 6.2%. The data from this series do not support small bowel bypass as the only acceptable method of management of surgical radiation enteritis. Wide resection with primary anastomosis reduces the long-term morbidity of the "blind-loop syndrome", perforation and fistulization of the isolated loop, and does not have an increased mortality.


Arterial scan versus radiographic angiography in detection of shallow arterial ulcers. A comparison of 99m-technetium albumin aggregated arterial scan and radiographic angiography for detection of shallow intimal carotid artery ulcerations was made in a series of 12 anesthetized dogs, having a total of 16 acute arterial ulcerations. Radiographic angiography showed positive findings related to presence of stenosis or mural thrombosis in 12 instances. Direct visualization of ulceration was only exceptionally encountered. Arterial scan detected 14 of 16 intimal ulcers. The radionuclide method was reliable even in absence of stenosis or when only minimal mural thrombosis was present. Moreover, autopsy scan of the isolated arterial segments detected all 16 intimal lesions. These results indicate that the arterial scan was a more reliable method for detection of shallow arterial ulcers in this experimental model than radiographic angiography, especially when arterial lumen stenosis or mural thrombosis were minimal or absent.


Hyperplasia of Brunner's glands of the duodenum. Hyperplasia of Brunner's glands is a rare phenomenon. It may cause obstructive symptoms, anemia or the patient may be totally asymptomatic. The diagnosis can be confirmed with endoscopic examination and upper gastrointestinal series. The surgical treatment or hyperplasia of the Brunner's glands should be conservative since the lesion is not premalignant. If complications do occur, local excision is the treatment of choice. Two patients are reported who had Brunner gland hyperplasia as an incidental finding at exploration for pancreatic pseudocyst, and a brief review of the literature is made.


A simple method of determining the boundary between the antrum and corpus of the stomach. Part 2. Electric vagal stimulation in dogs. Our previous work demonstrated that electric vagal stimulation in conjunction with Congo red as a dye-indicator is the faster and more reliable method for delineating the corpus-antral boundary in rats. Based on this finding, our study was extended to a larger animal model, the dog; this study proved the electrical vagal stimulation along with Congo red is a reliable method for adequate demonstration of this corpus-antral boundary. No untoward effect on the cardiac function was observed during stimulation of subdiaphragmatic portion of vagal trunks.


The laboratory diagnosis of the carrier state for classic hemophilia. Estimation of the titer of procoagulant antihemophilic factor (AHF) and the concentration of AHF-like antigens, as detected by heterologous antiserum, provides a method for diagnosis of the carrier state of classic hemophilia. Among 87 obligate carriers, 82, or 94%, could be identified as carriers by logarithmic discriminant analysis because the titer of procoagulant AHF was disproportionately lower than the concentration of AHF-like antigens relative to normal women. Women who were the mothers of individuals with isolated cases of hemophilia appeared to be carriers in at least 85% of cases, suggesting that the frequency of cases due to fresh mutations is low. The validity of the techniques used is supported by the frequency with which the carrier state was recognized in daughters of hemophiliacs who themselves had no sons. Corrected for the known error of the method, 48% of 75 such daughters appeared to be carriers, not significantly different from their anticipated chance of 50%.


Hypocomplementemia in chronic idiopathic urticaria. A discrete evoking factor or presumed pathophysiologic mechanism is not recognized in the majority of patients with chronic urticaria or angioedema. Two cases are reported in which chronic urticaria was the main manifestation of an immune cutaneous vasculitis associated with hypocomplementemia attributable to classic and alternative mechanisms of complement activation. Among 72 consecutive patients evaluated for chronic urticaria, 10 additional patients with idiopathic urticaria were found to have hypocomplementemia. Of these, two had evidence of classic and alternative mechanisms of complement activation, five had evidence of only classic pathway activation, and three evidence of predominately or exclusively alternative pathway activation. Circulating immune complexes were found in the majority of patients with classic pathway activation. Hypocomplementemia may provide clues to pathophysiologic mechanisms operative in some patients with chronic urticaria.


Treatment of invasive aspergillosis: relation of early diagnosis and treatment to response. Aspergillus infections in patients with cancer are difficult to diagnose, and such diagnoses are frequently made at necropsy. Earlier therapy has been proposed to provide better response. We reviewed 17 consecutive patients with documented aspergillosis to determine the impact of earlier diagnosis and prompt treatment with amphotericin B. Sixteen had hematologic malignancies, and all had marked granulocytopenia. Six were diagnosed and treated within 96 h of the appearance of infiltrates. Three of these six had complete resolution of all signs and symptoms of aspergillus infection. The other three had a partial response to therapy despite continued granulocytopenia. All 11 patients in whom antifungal therapy was either delayed (six) or not given (five) for at least 2 weeks after the infiltrate was present diet with progressive aspergillosis aggressive diagnostic methods to establish the diagnosis of aspergillosis are warranted so that antifungal therapy can be started early, which may then be successful in resolving these potentially fatal infections.


Pericarditis and perimyocarditis associated with active Mycoplasma pneumoniae infection. In 13 patients an association existed from 1970-73 between Mycoplasma pneumoniae infection and acute pericarditis (in eight) or perimyocarditis (in five). In 12 patients the association was moderately probable, with a fourfold rise in complement-fixing antibody titers between acute and convalescent phase sera being noted. In the last patient, a lesser-order association was found using only convalsecent phase serum. The presence of influenza, herpes simplex, Coxsackie B, or adenovirus was excluded by serologic testing. Acute illness was variable, with four patients developing heart failure. Long-term evaluation (mean, 47 months) found eight patients asymptomatic and three symptomatic. Two patients died. Residual effects of the Mycoplasma infection seemed at least partially responsible in one compromised patient and in one who died. Mycoplasma infection should be considered in the presence of acute cardiovascular decompensation, especially when preceded by upper respiratory infection, and added to the possible causes of idiopathic cardiomyopathy.


Osmolality changes during hemodialysis. Natural history, clinical correlations, and influence of dialysate glucose and intravenous mannitol. We studied the influence of both a high-glucose-concentration dialysate (717 mg/dl) and intravenous mannitol (1g/kg) on the serum osmolality changes in stable patients on chronic dialysis. During regular dialysis, serum osmolality fell 10 mosmol/kg H2O. This fall was reduced to 5.2 mosmol/kg H2O when the high-glucose-concentration dialysate was used, and to 4.3 mosmol/kg H2O when intravenous mannitol was used. When the two methods were combined, the serum osmolality fall was reduced to 1.7 mosmol/kg H2O. The clinical signs of disequilibrium declined (from 67% to 10%) in parallel with the decline in serum osmolality changes. This fall was independent of the ultrafiltration rate. A high-glucose-concentration dialysate and intravenous mannitol can each reduce the osmolality changes that occur during hemodialysis, but when used alone, intravenous mannitol is more effective of the two. The reduction of osmolality changes also leads to reduction of the mild clinical signs usually associated with disequilibrium.


Percutaneous transhepatic cholangiography with the "skinny" needle. A rapid, simple, and accurate method in the diagnosis of cholestasis. One hundred thirty-one cases of cholestasis were evaluated by "skinny" needle percutaneous transhepatic cholangiography. The biliary tree was seen in 129 patients (98.5%). When the biliary tree was dilated, a success rate of 100% was obtained. In the 45 patients with nondilated ducts, visualization was achieved in 95.6%. There was no mortality, early operation, or transfusion requirement associated with the procedure. This form of cholangiography is a simple, reliable, and extremely high-yield technique for evaluating patients with cholestasis.


Intravascular hemolysis, thrombocytopenia, leukopenia, and circulating immune complexes after jejunal-ileal bypass surgery. Two years after jejunal-ileal bypass surgery for obesity, a 25-year-old man developed intravascular hemolysis, thrombocytopenia, and neutropenia. The patient's erythrocytes were coated with complement components (C4/C3) and his serum induced complement-dependent immune lysis of chromium-51-labeled platelets. Serum [125I]-C1q binding activity (a measure of the presence of immune complexes) was increased, and serum C4 and C3 hemolytic titers were depressed. Immune complex-mediated complement activation apparently accounted for the blood cell destruction in this patient.


Furosemide-induced reduction in ionized calcium in hypoparathyroid patients. Six hypoparathyroid patients were treated with oral furosemide for 4 days. All six had a significant decrease in serum ionized calcium level from the control period (4.05 +/- 0.31 mg/dl) to the furosemide period (3.46 +/- 0.26 mg/dl, P less than 0.005). In five patients, the decrease in serum ionized calcium concentration occurred in association with an increase in urinary calcium excretion. Although serum immunoreactive parathyroid hormone level increased in five patients, it was insufficient to return serum ionized calcium to control levels. Changes in serum phosphorus, magnesium, or pH could not account for the observed decrease in serum ionized calcium concentration. It is necessary to monitor serum ionized calcium level in hypoparathyroid patients during furosemide therapy.


Quality-of-care assessment by process and outcome scoring. Use of weighted algorithmic assessment criteria for evaluation of emergency room care of women with symptoms of urinary tract infection. Weighted process criteria based on chart review using an algorithmic method, and weighted outcome criteria based on a telephone interview with the patient, were used to assess the quality of emergency room care of women with symptoms of acute urinary tract infection. Outcomes at a municipal hospital were significantly better than those at an affiliated voluntary hospital. Process and outcome scores were positively correlated at both hospitals; the data suggest, but do not themselves adequately test, the hypothesis that the correlation is curvilinear and that there is a threshold process score below which a poor outcome is extremely likely. Our findings suggest that weighted algorithmic assessment criteria are a useful method for quality assessment and are consistent with the testable hypothesis that the greatest improvements in outcome may result from raising the quality of care from a poor to an adequate level rather than from an adequate to an optimal level.


Diagnosis and treatment of physical disease in the mentally ill. The inability of patients to provide a history and to cooperate in physical or technological examinations makes the diagnostic and therapeutic management of physical disease in psychotic and especially in elderly psychotic patients very difficult. There needs to be a disproportionate dependence upon laboratory and radiologic information in diagnosis of physical disease in such patients. Observations by personnel on the psychiatric wards are essential to identification of probable physical illness. An experience with 1800 patients on a medical service now approved for Medicare and Medicaid in a state psychiatric hospital clearly shows the impossibility of applying the norms of care set for a general hospital in such a setting. Additionally, the clinician is faced with ethical considerations of which actions are justifiable and which are not in the search for a diagnosis. The Professional Standards Review Organization, judicial verdicts requiring care for the mentally ill equal to that of other citizens, and the hazards of professional liability do not permit these considerations to be taken lightly. An organization that has responsibility for the mentally ill might well address itself to finding answers to these questions.


The book of job and the modern view of depression. The Biblical Book of Job is a wisdom book. Wisdom, in this context, refers to the intellectual discipline taught by the sages of ancient Isreael to provide professionals with a realistic approach to the problems of life. Chapter 30 of the Book of Job is a key-index of ideas that, if followed through the book, discloses a modern scientifically accurate description of a depression that, at times, was life-threatening. There are practical clinical clues to distinguish between normal mourning and depression, as well as aids to the differential diagnosis of somatic symptoms that may arise from depression. A timeless model of the scope and limitations of the professional relation between patient and comforter is also presented. Part of the wisdom of the Book of Job is to use depression as an example of a life-threatening illness to provide an unexcelled standard of clinical observation and medical intervention.


Analysis of serum for gentamicin by radioimmunoassay. A radioimmunoassay method for the determination of combined gentamicin isomers in serum has been adapted and tested. The procedure uses tritiated gentamicin, rabbit antiserum against a gentamicin-human albumin conjugate, and dextran-charcoal separation. The day-to-day coefficient of variation is 6 percent, and frozen samples are stable for at least one month. There was no cross reactivity of the antiserum with any of several tested antibiotics or with any of some tested commonly used non-antibiotic hospital pharmacy preparations. The results from the procedure correlated well with those of an enzymatic radioacetylation technique and, except for a significant incidence of "outliers", with a microbiological assay. As expected, patient serum values, both maximum and minimum, showed no correlation with dose size.


Nickel concentrations in human parotid saliva. Measurements of nickel were performed by electrothermal atomic absorption spectrometry upon specimens of parotid saliva from 38 healthy adults. The mean concentration of nickel in saliva was 1.9 +/- 1.0 microng Ni per liter (range = 0.8 to 4.5). There was no significant correlation between the concentrations of salivary nickel and protein. No significant differences were observed between the mean concentrations of nickel in saliva samples from (1) men vs women; (2) health center employees vs naval recruits; (3) caries-free subjects vs caries-prone subjects; or (4) non-smokers vs cigarette smokers.


Adenosine diphosphate-induced platelet aggregation in the states of hypercoagulability. One hundred and seven patients with congestive heart failure, myocardial infarction and arteriosclerotic heart disease were studied by adenosine diphosphate-induced platelet aggregation, fibrinogen levels and ethanol gelation test. Both increases and decreases in platelet aggregation were observed. A significantly high percentage of patients showed a decreased platelet aggregation which was especially marked in the more acute as opposed to the less acute phase. In addition, most patients exhibited a marked shift from abnormal to normal platelet aggregation or vise-versa within a short time period. This pattern of platelet aggregation suggests an active role of platelets in the states of hypercoagulability. The hypercoagulability of these patients was further substantiated by a high percentage of positive ethanol gelation tests and high fibrinogen levels.


An ultrastructural analysis of renal arterioles in benign and malignant essential hypertensions. Renal arterioles from five patients with benign essential hypertension (BH) and four patients with malignant essential hypertension (MH) were studied using electron microscopy (EM). This study suggests a concept of different pathogenetic mechanism for renal vascular (arteriolar) lesions in BH and MH (i.e., benign and malignant nephrosclerosis, respectively) and does not support the idea of progression of benign nephrosclerosis to malignant nephrosclerosis. The EM technique has proven to be important in demonstrating specific changes characteristic of severe (or malignant) hypertension in the early stage in patients without clinical manifestation of MH, thus predicting a possibility of progressive histological change and clinical deterioration. This technique has clarified the light microscopy appearance of hyaline which continues to be an intriguing feature in the study of renal pathology.


Analytical evaluation of a folate radioassay: application to serum and erythrocyte measurements. Many commercial folate radioassay kits have proven to be unsatisfactory in the past. It has been found that the Schwarz/Mann Iodine-125 folate radioassay kit permits a rapid measurement of folic acid requiring only a single incubation at pH 9.3. At this pH, the relative affinity of pteroylglutamic acid (FA) and N-5-methyltetrahydrofolic acid (MTFA) for the milk binder is the same. Additionally, the radioassay was compared to the L. casei microbiological assay. Regression analysis of 42 patients' sera, folate concentration 0.9 to 14 mug per liter, gave a correlation coefficient of 0.963. At folate concentration greater than 14 mug per liter, results with the radioassay in many instances were lower than those with the microbiological assay. In some cases the discrepancy was as much as 50 percent. However, the recovery of pure FA, MTFA and mixtures of the two at concentrations of 20 to 40 mug per liter was satisfactory by radioassay. This would indicate that the microbiological assay is inaccurate at higher folate concentrations. It has been shown that the Schwarz/Mann folate kit is accurate, precise, unaffected by icterus and lipemia. It is, therefore, suitable for both serum and red blood cell folate assay. A discrepancy has been observed between serum and red cell folate concentration; consequently, a simultaneously assay of both folates is suggested.


Classification of enzymes and current status of enzyme nomenclature and units. Recommendations for the nomenclature and coding of enzymes as presented by the International Union of Pure and Applied Chemistry and the International Union of Biochemistry are summarized and discussed. Units for reporting catalytic concentration of enzymes are briefly reviewed and abbreviations that have been proposed for enzyme names are also described.


[Study of growth from 0 to 18 months. Are infants followed in maternal and child welfare clinics a valuable reference for this study]. 1,017 healthy french children have been surveyed in the P.M.I. centres of the 14th ward of Paris. Birth weight in function of duration of pregnancy has been studied. Standard measures of growth from two weeks to 18 months regarding weight, length and head circumference of boys and girls have been established. The biometric data of the children have been compared with some biological and social factors of their parents.


[Familial pseudohypoaldosteronism (apropos of 5 cases)]. The first clinical and scientifically confirmed description of familial pseudohypoaldosteronism is reported in 5 patients, one of whom is an adult. The rarity of the disorder may be over-estimated. The 5 patients described had anextreme liking for salt which they had consumed in large amounts and tolerated without any side effects, regardless of the quantity and the period of time involved. The results of sodium balance and othr studies carried out several months are reported. Several investigations are normal (effect of spironolactone, DOCA) supporting the evidence to data against a decreased sensitivity of the distal renal tubule to aldosterone and presenting new evidence in favour of impaired proximal tubular resbsorption of sodium; persistent hypercalcuria occurred and histological changes were maximal in the proximal tubule. The 5 patients described despite apparent clinical well-being all had biological hypoaldosteronism, and some of them a chronic salt losing state without detectable renal impairment. These findings suggest that the long-term evolution of the disease has to be reconsidered.


[Diffuse hamartomatous tumor of the cerebellum and brain stem in children]. A child presented in the first year of life with a slowly evolving cerebellar syndrome which resulted in death at the age of 4 1/2 years. A peculiar tumour involving the cerebellum and brain stem and destroying the cerebellar anatomy is described. This was considered to be a form of gangliomatous hamartoma.


[Maternofetal deficiency hyponatremia caused by a selt-free diet during a multiple pregnancy]. Quadruplets born at 36 weeks gestation were severely hyponatraemic. The mother had been given a sodium deficient diet for the majority of pregnancy and she herself was also hyponatraemic with hyperaldosteronism. This patient illustrates the problems of sodium restriction during pregnancy and the need for its supervision.


[Pharmacologic bases of use of benzodiazepines in peréinatal medicine]. The pharmacokinetics of benzodiazepines have been studied because these drugs are often given to pregnant mothers and occasionally to neonates. The rate of absorbition is faster in the neonate than in the mother by all routes of administration but the drugs accumulate, the degree being related to gestational age, the more premature the greater the accumulation. There is preferential storage in some organs including the heart. Diazepam and chlordiazepoxide are metabolised to active compounds but oxazepam, nitrazepam and flurazepam are inactivated. Individual susceptibilities are important. These drugs readily pass across the placenta but the teratogenic risk is uncertain. They are also excreted in the breast milk in quantities which justify their withdrawal during breast feeding. On the basis of the findings therapeutic protocol is proposed.


[Bournevilles tuberous sclerosis. clinical and genetic study of 59 cases in children]. The authors have studied 59 cases of Bourneville's tuberous sclerosis, including 44 sporadic cases from healthy parents and 9 familial patients. They stress the very high frequency of spasms in the infants of less than one year (80 %), the frequency and the early appearance of retinal phakomata and areas of depigmentation and the interest in the early detection of intracranial calcification. They discuss the specificity of the individual manifestations: cutaneous, retinal and systemic and they consider the problem of genetic counselling.


Exercise performance in 6-to-11-year-old boys with Duchenne muscular dystrophy. To determine effects of severe muscular dystrophy on the performance of dynamic exercise, cardiorespiratory responses to incremental work on a bicycle ergometer and isokinetic limb strength measurements were compared for 13 dystrophic boys and 13 normal, untrained boys. The dystrophic boys (D) were matched to the normal boys (N) on the basis of age (8.4 yr), height (125 cm) and weight (25.7 kg). At rest, the dystrophic group had higher heart rates (HR) (D = 102; D = 31; N = 39 ml), with no difference in oxygen uptake (VO2), calculated cardiac output (Q), pulmonary ventilation (VE), or respiratory exchange ratio (R). During submaximal work, VO2, SV, Q and VE were lower in D. During maximal work, D had lower peak values for work rate (D = 400; N = 600 kg/min), endurance (D = 41; N = 60 ml), Q (D = 5.2; N = 11.0 liters/min), VE (D = 8.2; N = 36.9 liters/min), and R (D = 0.84; N = 0.99). Arm and leg strengths (four flexion and four extension motions) were lower in D, but muscle girths were not necessarily smaller. The findings indicate exercise performance in D was below normal and limited by low cardiorespiratory capacities, diminished leg strength, and perhaps reduced peripheral oxygen utilization. Duchenne muscular dystrophy, even in its early stages, apparently affects the work capacity of cardiac and pulmonary muscles as well as limb muscles.


An organizational typology of treatment facilities for the physically handicapped. The lack of a viable organizational typology is cited as a factor in the current absence of comparative research among physical rehabilitation facilities. A short discussion of approaches to typology construction is given which concludes that theoretically grounded typologies are most likely to produce useful insights. Subsequently, a fourfold typology of treatment facilities-maintenance, processing, changing and rehabilitation-is generated from a theory of variations in patient handling.


Patient education by videotape after myocardial infarction: an empirical evaluation. Patients recovering from myocardial infarction (MI) or other heart diseases at St. Francis Hospital, Hartford, Ct, were educated by videotape or by staff lectures on alternating weeks. Both programs included the following: risk factors for MI, medications, diet, MI symptoms and life style changes. Patients were interviewed before and after the educational program. The MI patients under the age of 60 scored equally well on an informational test irrespective of the type of education program experienced. Older MI patients were significantly more likely to complete the educational program when it was given by videotape; those discontinuing attendance at lectures were less psychologically motivated to participate but were not necessarily more ill. Overall, higher education was the single most significant predictor of superior scores following patient education. Implications for the coronary care ward of the success of videotape in educating MI patients are discussed.


Paraplegia following trauma in a patient with kyphosis. The reported incidence of paraplegia following acute spinal trauma in patients with kyphosis is very low. One such case is presented here and discussed. Although deterioration of spinal cord function in patients with kyphosis who have not sustained trauma is treated by surgical decompression and stabilization of the spine, a more conservative approach may be beneficial in those patients with a history of trauma. Thus, the treatment appears to follow that now recommended for acute spinal injuries in patients with a premorbidly normal spine.


Isolation of an influenza virus, similar to A/Port Chalmers/1/73 (H3N2) from a common murre at Sakhalin Island in U.S.S.R. (Strain A/common murre/Sakhalin/1/74). An influenza A virus isolated from the cloaca of a common murre was characterized antigenically as H3N2 and was most closely related to the A/Port Chalmers/1/73 strain. Serological studies of sera collected from common murres in the area of virus isolation showed that 21 per cent of the birds had antibodies to Hong Kong influenza virus.


Temperature-sensitive mutants of measles virus produced from persistently infected HeLa cells. A persistent infection with the Edmonston strain of measles virus was established in HeLa cells in the absence of measles virus antibody (HeLaPI cells). By hemadsorption or immunofluoresnce virtually 100 per cent of the cells possessed measles virus components. HeLaPI cells produced no interferon and were not resistant to superinfection with Newcastle disease virus. HeLaPI cells contained both smooth (15--18 nm) and rought (20--35 nm) nucleocapsids as detected by electron microscopy. The virus produced from the HeLaPI cells (MVPI) varied in titer between 1.5 X 10(2) and 5.5 X10(4) PFU/ml, had a smaller plque size and was more heat resistant than wild-type measles virus. MVPI was also found to be temperature-sensitive. The temperature-sensitivity of MVPI was determined by the efficiency of plaquing at 33 degrees and 39 degrees C in Vero cell monolayers. When HeLaPI cells were incubated at 33 degrees C, there was a 50-fold increase in virus production as well as a slight increase in the percentage of cells forming infectious centers compared to HeLaPI cells grown at 37 degrees C. MVPI readily established a persistent infection in HeLa cells which also rleased temperature-sensitive virus.


Failure of poxvirus replication in the presence of an inhibitor of nucleolar RNA synthesis. In vaccinia virus infected cells the appearance of a late enzyme RNA polymerase was prevented by MPB, an inhibitor of nucleolar RNA synthesis, although inductions of the early enzymes thymidine kinase and DNA polymerase were not affected. It is inferred the nucleoli may be involved in the replication of vaccinia virus.


Vaccinia virus cytotoxin.

Vaccinia virus cytotoxin. Extracts of vaccinia-infected HeLa cells were rendered free from infectious virus by centrifugation followed by membrane filtration and were shown to be toxic to uninfected HeLa cells in the presence of hypertonic MgSO4, used as a macromolecular uptake inducer, under conditions which did not kill control cells. Extracts from uninfected cells were nontoxic. This biological test was adapted to a semi-quantitative assay which was used to monitor the purification of the cytotoxic factor by DEAE-cellulose and Sephadex G-100 chromatography. The cytotoxic factor was purified 100-fold, shown to be of molecular weight 30 -- 100,000 daltons, acidic and completely inactivated by soluble trypsin but not by ribonuclease under conditions believed to degrade both single- and double-stranded RNA species. It was demonstrated to be virus specific by approrpiate immunosorbent chromatography. Extracts were also prepared from vaccinia-infected HEp-2, RK and W-K cells respectively. A virus-specific factor, toxic to uninfected HeLa cells, with similar chromatographic properties to that isolated from infected HeLa cells, was isolated from these three additional cell lines. The concept of virus induced cytotoxins, substances which exert their toxic effect in the host cells in which they are made, is discussed.


[Local immune response in mouse experimental airborne influenza: immunoglobulin concentrations, antibody levels of immunoglobulin classes and anamnestic response in bronchial washings (author's transl)]. Sera and bronchial washings from normal mice and from mice previously infected with influenza virus were analyzed for their concentration of four immunoglobulin (Ig) classes by the method of single radial immunodiffusion and for their content of specific antibody of these Ig classes by the immunofluorescent method. IgA were not detected in bronchial washings from normal mice. The IgA/IgG ratio was not higher than 0.33 in infected mice. The IgA level increased briefly in infected bronchial washings, but the levels of IgG2 and particularly of IgG1 exhibited a longer and higher increase. The maximal increase of these three Ig classes occurred by the seventh day and might have derived from transsudated serum. IgM was not identified in normal bronchial washings, but immunofluorescence detected IgM in infected washings, which also showed specific antibody in each of the four Ig classes. IgG2 and IgM contained antibody in greatest, IgA in lower concentration. Our results indicate that the most part of bronchial antibody was produced locally, since the ratio of Ig concentration to antibody titre of each Ig class was consistently higher for bronchial washings than for sera. An anamnestic secretory antibody response could not be demonstrated in bronchial washings.


Interactions of tocopherols and ubiquinones with monolayers of phospholipids. 1. The penetration of alpha-tocopherol and seven of its derivatives, and five compounds in the ubiquinone series, having differing chain lengths, into monolayers at the air/water interface of 11 different synthetic phospholipids and cholesterol was investigated; the properties of mixed monolayers of the tocopherols and of ubiquinones with phospholipids were also studied. 2. Penetration of alpha-tocopherol into diarachidonylglycerylphosphorycholine was approximately constant for molar ratios of tocopherol/phospholipid ranging from 0.4:1.0 to 2.0:1.0. 3. Tocopherols with shorter or longer side chains than alpha-tocopherol had a lesser ability to penetrate monolayers of phospholipid molecules with 16 or more carbon atoms in their acyl chains. 4. All the tocopherols penetrated more readily as unsaturation in the phospholipids was increased, and their penetration into mixed monolayers of phospholipids was greatly facilitated by the presence of relatively small quantities of unsaturated phospholipid molecules. 5. There was relatively little interaction between the tocopherols and cholesterol, or between the ubiquinones and phospholipids. 6. The possible significance of the observed interactions between alpha-tocopherol and polyunsaturated phospholipids is discussed in relation to the biochemical actions of alpha-tocopherol in vivo. 7. It is suggested that fluidity of the lipid bilayer in membranes containing polyunsaturated phospholipids may allow alpha-tocopherol to interact in a dynamic manner with a number of phospholipid molecules.


Action of human lysosomal elastase on the oxidized B chain of insulin. The specificity of action of the lysosomal elastase of human neutrophil leucocytes on the oxidized B chain of insulin is similar to that of pig pancreatic elastase, but is more directed towards valine than alanine as the residue contributing the carboxyl group of the cleaved bond. The most susceptible bonds are Val-12-Glu-13 and Val-18-Cys(O3H)-19. Other bonds hydrolysed are Ala-14-Leu-15, Ser-9-His-10 and Cys, (O3H3)-7-Gly-8. Tables listing amino acid composition, N-terminal residue, and yields of isolated peptides have been deposited as Supplementary Publication SUP 50 075 (8 pages) at the British Library Lending Division, Boston Spa, Wetherby, West Yorkshire LS23 7BQ, U.K., from whom copies can be obtained on the terms indicated in Biochem. J. (1977) 161, 1.


Ionic-strength-dependent changes in the structure of the major protein of the human erythrocyte membrane. The effect of ionic strength on the proteolysis by trypsin of the major membrane-penetrating protein (polypeptide 3) in the erythrocyte membrane was studied. Both the intracellular and extracellular regions of the protein are susceptible to trypsin proteolysis under hypo-osmotic conditions, whereas under iso-osmotic conditions the extracellular region of the protein is resistant to trypsin, and the intracellular region yields only two cleavage products with trypsin. Studies of the fragments obtained from polypeptide 3 by trypsin digestion under iso-osmotic conditions of 'ghosts' radioiodinated with lactoperoxidase confirmed our earlier conclusions that the polypeptide chain of polypeptide 3 traverses the membrane twice. Ionic-strength-dependent changes were also observed in the incorporation of iodine by lactoperoxidase into the individual extracellular tyrosine sites of the protein. These results show that polypeptide 3 undergoes ionic-strength-dependent changes in structure.


The structure of the major protein of the human erythrocyte membrane. Characterization of the intact protein and major fragments. Polypeptide 3, the major membrane-penetrating protein of the human erythrocyte membrane, was characterized, together with two major fragments derived by specific proteolysis of the native protein in the membrane. One fragment (fragment 3f) was obtained from thermolysin cleavage in the extracellular region of the protein, and the other (fragment T1) was derived from tryptic cleavage in the intracellular region of the protein. The results of N- and C-terminal group analysis suggest that fragment 3f contains the N-terminal region of polypeptide 3 and fragment T1 contains the C-terminal part of the molecule. The carbohydrate contents of the polypeptides suggest that carbohydrates are present in three regions of the molecule, much of this carbohydrate being present in the C-terminal part of the molecule. This region of the protein also contains the receptors for concanavalin and the lectins from Phaseolus vulgaris and Ricinis communis, and our results suggest that there is heterogeneity in the carbohydrate chains present in the C-terminal region of polypeptide 3. These data are related to the folding of polypeptide 3 in the erythrocyte membrane.


Purification and immunochemical characterization of monoamine oxidase from rat and human liver. 1. Monoamine oxidase from rat and human liver was purified to homogeneity by the criterion of polyacrylamide-gel electrophoresis in the presence of sodium dodecyl sulphate. 2. The enzyme activity was extracted from mitochondrial preparations by Triton X-100. The enzyme was purified by (NH4)2SO4 fractionation followed by chromatography on DEAE-cellulose, Sepharose 6B, spheroidal hydroxyapatite, and finally chromatography on diazo-coupled tyramine-Sepharose. 3. Distinct differences occur in the chromatographic behaviour of the two enzymes on both DEAE-cellulose and spheroidal hydroxyapatite. 4. It is unlikely that the purification of the enzymes on tyramine-Sepharose is due to affinity chromatography and reasons for this are discussed. 5. The purified enzymes did not oxidize-5-hydroxytryptamine and the relative activities of the enzymes with benzylamine were increased approx. 1.25-fold compared with the enzyme activities of mitochondrial preparations. 6. Immunotitration of enzyme activity in extracts of mitochondrial preparations from rat liver was carried out with 5-hydroxytryptamine, tyramine and benzylamine. The enzyme activities were completely immunoprecipitated by the same volume of antiserum. Similar results were obtained with the antiserum to the enzyme from human liver.


Action of human cathepsin G on the oxidized B chain of insulin. The specificity of cathepsin G, a serine neutral proteinase from human neutrophil leucotyes, was determine dby its action on the insulin B chain. The most susceptible bonds were Phe-24-Phe-25, Leu-15-Tyr-16 and Tyr-16-Leu-17. Other bonds hydrolysed were Leu-6-Cys(O3H)-7, Leu-11-Val-12, Leu-17-Val-18 and Phe-25-Tyr-26. These results suggest that the specificity of cathespin G is closer to that of pig chymotrypsin C than ox Chymotrypsin A. Tables listing amino acid composition, N-terminal residue, and yields of isolated peptides have been deposited as Supplementary Publication SUP 50 075 (8 pages) at the British Library Lending Division, Boston Spa, Wetherby, West Yorkshire LS23 7B2, U.K., from whom copies can be obtained on the terms indicated in Biochem. J. (1977) 161,1.


Identity of rat kidney histidine-pyruvate aminotransferase with glutamine-oxo acid aminotransferase. Histidine-pyruvate aminotransferase and glutamine-phenylpyruvate aminotransferase were purified from rat kidney by the same procedure. The ratio of the two activities remained constant during purification and was unchanged by a variety of treatments of the purified enzyme. Glutamine was found to act as a competitive inhibitor of histidine-pyruvate aminotransferase. These results suggest that rat kidney histidine-pyruvate aminotransferase is identical with glutamine-oxo acid aminotransferase. Identity of the two enzymes in other tissues of rats is discussed.


Partial amino acid sequence of rat pre-prolactin. Rat pituitary mRNA was used to direct the cell-free synthesis of pre-prolactin labelled with [4,5-3H]leucine and either [35S] methioninc or [35S] cystine. Sequence analysis of the labelled protein indicates that pre-prolactin has 29 amino acid residues joined to the N-terminus of the prolactin sequence. Leucine residues were found at positions 13, 14, 15, 16, 21 and 22, methionine residues at positions 1, 17 and 18, and a cysteine residue at position 24 of the precursor sequence, and this partial sequence shows considerable similarity with other precursors that have been sequenced.


The binding of copper ions to copper-free bovine superoxide dismutase. Copper distribution in protein samples recombined with less than stoicheiometric copper ion/protein ratios. Samples of superoxide dismutase containing less than stoicheiometric amounts of Cu2+ were obtained by either partial re-addition of Cu2+ to the Cu2+-free protein or partial removal of Cu2+ by controlled CN-treatment. In these samples the distribution of the metal between the two identical sites on the two subunits was studied by quantitative gel electrophoresis and found to be statistical only in the process of copper removal by CN-. In the other case the distribution fits a model of co-operative interaction between the two sites, where the sites are equivalent for the binding of the first Cu2+ ion, but the occupation of the first site lowers the activation energy of the binding of the second Cu2+ ion. This indicates that binding of Cu2+ ion at its site on one subunit brings about conformational changes that facilitate Cu2+ binding on the other subunit. These results may relate to possible intersubunit interactions during the catalytic activity.


The binding of copper ions to copper-free bovine superoxide dismutase. Properties of the protein recombined with increasing amounts of copper ions. 1. E.p.r. (electron-paramagnetic-resonance), proton-relaxation and u.v.-absorption parameters, and enzyme activity of samples of Cu2+-free bovine superoxide dismutase recombined with different amounts of Cu2+ up to the stoicheiometric [Cu2+]/protein] ratio were investigated after attainment of equilibrium in the recovery process. 2. The e.p.r. spectra were identical with the spectrum of the native protein at all [Cu2+]/[protein] ratios. The relaxation rate of the water protons (T1) and the u.v. absorption increase as linear functions of the added Cu2+. 3. On the other hand, in recombination experiments in the range pH 7.6-10.5 the enzyme activity shows a non-linear increase as the [Cu2+]/[protein] ratio rises. The experimental curves can be interpreted in terms of the model of co-operative binding of Cu2+ to the two sites proposed on the basis of the electrophoretic analyses of the samples, and show that the specific activity of the molecules containing only one Cu2+ ion is twice as high as that of the molecules with two Cu2+ ions. 4. These results support the hypothesis of an anti-co-operative interaction between the two sites during the activity, which allows only one Cu2+ ion to function in catalysis.


Preparation and characterization of armadillo submandibular glycoproteins. The nine-banded armadillo (Dasypus novemcinctus mexicanus Peters) was chosen for this study so that a comparison could be made of the salivary mucus glycoproteins of an ancient mammalian species with those derived from previously studied, more highly evolved species. Two mucus glycoproteins, armadillo submandibular glycoprotein A and armadillo submandibular glycoprotein B, were prepared from the armadillo submandibular gland by a modification of the method of Tettamanti & Pigman (1968) (Arch. Biochem. Biophys. 124, 41-50). The composition of glycoprotein A is the simplest one among the known mucus glycoproteins. Six amino acids constitute 98.5 mol/100mol of the protein of glycoprotein A and 82 mol/100 mol of that of glycoprotein B. These are serine and threonine (which make up 40-50% of the molar amino acid composition), glutamic acid, glycine alanine and valine. Proline is absent from glycoprotein A and comprises only 2.3% of glycoprotein B. For both glycoproteins, the protein content, as determined by the method of Lowry, Rosebrough, Farr & Randall (1951) (J. Biol. Chem 193, 265-275), with bovine serum albumin as standard, was nearly 60% higher than when determined by the sum of the amino acids. The ratios of total mol of amino acid/total mol of carbohydrate are 1:0.63 for glycoprotein A and 1:0.68 for glycoprotein B, N-Acetylneuraminic acid and N-acetylgalactosamine, in a molar ratio of about 0.35:1.00, are the principal carbohydrates present in both glycoproteins. Neutral sugars seem to be absent from glycoprotein A, but galactose and fucose are present in glycoprotein B. The carbohydrate side chains in glycoprotein A are composed of about two-thirds monosaccharide and one-third disaccharide residues, whereas those of glycoprotein B are more complex. For both glycoproteins, essentially all of the N-acetylgalactosamine was attached O-glycosidically to the hydroxyamino acid residues of the protein core. The linkage of N-acetylneuraminic acid glycoprotein A was extremely sensitive to dilute acid and neuraminidase. Glycoprotein B has chemical properties similar to those of glycoprotein A. However, whereas glycoprotein A was susceptible to both Clostridium perfringens and Vibrio cholerae neuraminidases, only the latter enzyme had an effect on glycoprotein B at pH 4.75. Both glycoproteins were homogeneous by cellulose acetate electrophoresis and ultracentrifugal analyses. The apparent mol.wts. of glycoprotein A and glycoprotein B were 7.8 X 10(4) and 3.1 X 10(4) respectively.


Folding domains and intramolecular ionic interactions of lysine residues in glyceraldehyde 3-phosphate dehydrogenase. 1. Treatment with methyl acetimidate was used to probe the topography of several tetrameric glyceraldehyde 3-phosphate dehydrogenases, in particular the holoenzymes from rabbit muscle and Bacillus stearothermophilus. During the course of the reaction with the rabbit muscle enzyme, the number of amino groups fell rapidly from the starting value of 27 per subunit to a value of approx. five per subunit. This number could be lowered further to values between one and two per subunit by a second treatment with methyl acetimidate. The enzyme remained tetrameric throughout and retained 50% of its initial catalytic activity at the end of the experiment. 2. Use of methyl [1-14C]acetimidate and small-scale methods of protein chemistry showed that only one amino group per subunit, that of lysine-306, was completely unavailable for reaction with imido ester in the native enzyme. This results is consistent with the structure of the highly homologous glyceraldehyde 3-phosphate dehydrogenase of lobster muscle deduced from X-ray-crystallographic analysis, since lysine-306 can be seen to form an intrachain ion-pair with aspartic acid-241 in the hydrophobic environment of a subunit-subunit interface. 3. Several other amino groups in the rabbit muscle enzyme that reacted only slowly with the reagent were also identified chemically. These were found to be located entirely in the C-terminal half of the polypeptides chain, which comprises a folding domain associated with catalytic activity and subunit contact in the three-dimensional structure. Slow reaction of these 'surface' amino groups with methyl acetimidate is attributed to intramolecular ionic interactions of the amino groups with neighbouring side-chain carboxyl groups, a conclusion that is compatible with the reported three-dimensional structure and with the dependence of the reaction of ionic stength. 4. Very similar results were obtained with the enzymes from B. stearothermophilus and from ox muscle and ox liver, supporting the view that the ion-pair involving lysine-306 and aspartic acid-241 will be a common structural feature in glyceraldehyde-3-phosphate dehydrogenases. The B. stearothermophilus enzyme was fully active after modification. 5. No differences could be detected between the enzymes from ox muscle and ox liver, in accord with other evidence that points to the identify of these enzymes. 6. The pattern of slowly reacting amino groups in the enzyme from B. stearothermophilus, although similar to that of the mammalian enzymes, indicated one or two additional intramolecular ionic interactions of lysine residues that might contribute to the thermal stability of this enzyme.


Intramolecular ionic interactions of lysine residues and a possible folding domain in fructose diphosphate aldolase. 1. Treatment with methyl acetimidate was used to probe the topography of the tetrameric fructose 1,6-diphosphate aldolase from ox liver. A single treatment with imido ester in the presence or absence of 20mM-fructose 1,6-diphosphate caused the number of amino groups in the enzyme to fall to approx. 30% of the starting number (assumed to be 30 per subunit). The catalytic activity of the aldolase modified in the presence of fructose 1,6-diphosphate was unaffected, whereas that of the enzyme modified in the absence of substrate fell by about 20%. 2. Use of methyl [1-14C]acetimidate and small-scale methods of protein chemistry showed that the amino group of lysine-27 (the numbering is that of the highly homologous rabbit muscle enzyme) is essentially unavailable for amidination in the native enzyme and is therefore predicted to be buried in a hydrophobic environment, probably in the form of an ion-pair with a negatively charged side-chain carboxyl group. All the other lysine residues that reacted poorly with methyl acetimidate in the native enzyme (a total of 7) were found to be within the primary structure bounded by lysine-107 and lysine-227. An important member of this group of lysine residues displaying aberrant reactivity is lysine-227, which is known to form an imine with the substrate as part of the catalytic mechanism of the enzyme. 3. The results of the amidination experiments can be correlated in an interesting way with previous studies of thiol-group modification in the aldolases. Taken together, and arguing in part by analogy with the results of identical experiments with glyceraldehyde 3-phosphate dehydrogenases where the three-dimensional structure is known [Lambert & Perham (1977) Biochem. 4. 161. 49-62], they suggest that the region of primary structure from residues 107-227 may form the whole or part of a three-dimensional structural feature, perhaps a folding domain. A three-dimensional structure deduced from X-ray-crystallographic analysis will be needed to interpret these findings more closely. 4. The amino groups of lysine residues are commonly thought to reside at the 'surface' of protein structures. The patterns of specific lysine residues in glyceraldehyde 3-phosphate dehydrogenases and in aldolases that have been found to react poorly with methyl acetimidate in the native enzymes can be attributed to intramolecular ionic interactions deep in hydrophobic pockets and at the protein 'surface'. Such ionic interactions may contribute significantly to the stability of a given protein.


The effect of temperature on the individual stages of the hydrolysis of non-specific-p-nitrophenol esters by alpha-chymotrypsin. Precise studies were performed on the effect of temperature on the rate and equilibrium parameters characterizing the individual stages of the alpha-chymotrypsin-catalysed hydrolysis of non-specific p-nitrophenol esters at pH 7.40 and 8.50. At both pH values the results indicate that a sharp kinetic anomaly is observed in Arrhenius plots of these parameters for the binding and acylation stages of the process, but not for the deacylation stage. Detailed comparison with other kinetic studies was made, and a comparison with thermal transitions observed in alpha-chymotrypsin by using physical techniques was attempted. A detailed discussion of possible causes of the anomalies is given.


Macromolecular distribution near the limits of density-gradient columns. Some applications to the separation and fractionation of glycoproteins. 1. Expressions are derived for the distribution at density-gradient equilibrium of macromolecules whose densities are (a) close to the values characterizing the solution limits or (b) outside the span of the gradient. 2. Density-distribution predicted by the expressions agree with those obtained by rigorous methods. 3. The distribution equations are applied to hypothetical mixtures of proteins and glycoproteins in commonly used density-gradient media to simulate separation and fractionation conditions. 4. It is shown that CsBr, although less efficient than CsCl for fractionation, is nevertheless adequate for most purposes; in analytical experiments it may often have advantages over CsCl. Limitations on the use of LiBr are explored. 5. An expression is derived which allows the variance of the partial specific volume of the macromolecular component to be determined from the variance of the buoyant density. It is shown that the relative resolving powers of different salts is expressed by their values of the quantity (formula: see text). 6. The equations are applied to a well-characterized glycoprotein preparation at equilibrium in CsCl and in Cs2SO4:it is shown that the much wider distribution in CsCl than in Cs2SO4 is explicable in terms of the variance in buoyant density and the solvation properties of the salts. 7. Limitations of the expressions arise when dispersity in density is represented by a low apparent molecular weight; realistic simulations can then only be obtained when the component is fully banded.


Affinity-chromatographic isolation and some properties of troponin C from different muscle types. 1. The formation of a complex between troponin I and troponin C that is stable in 6M-urea and dependent on Ca2+ was demonstrated in extracts of vertebrate striated and smooth muscles. 2. A method using troponin I coupled to Sepharose is described for the rapid isolation of troponin C from striated and smooth muscles of vertebrates. 3. Troponin C of rabbit cardiac muscle differs significantly in amino acid composition from troponin C of skeletal muscle. The primary structures of troponin C of red and white skeletal muscle are very similar. 4. The troponin C-like protein isolated from rabbit uterus muscle has a slightly different amino acid composition, but possess many similar properties to the forms of troponin C isolated from other muscle types. 5. The electrophoretic mobilities of the I-troponin C complexes formed from components isolated from different muscle types are determined by the troponin I component.


Calculation of steady-state rate equations and the fluxes between substrates and products in enzyme reactions. 1. Two methods are described for deriving the steady-state velocity of an enzyme reaction from a consideration of fluxes between enzyme intermediates. The equivalent-reaction technique, in which enzyme intermediates are systematically eliminated and replaced by equivalent reactions, appears the most generally useful. The methods are applicable to all enzyme mechanisms, including three-substrate and random Bi Bi Ping Pong mechanisms. Solutions are obtained in algebraic form and these are presented for the common random Bi Bi mechanisms. The steady-state quantities of the enzyme intermediates may also be calculated. Additional steps may be introduced into enzyme mechanisms for which the steady-state velocity equation is already known. 2. The calculation of fluxes between substrates and products in three-substrate and random Bi Bi Ping Pong mechanisms is described. 3. It is concluded that the new methods may offer advantages in ease of calculation and in the analysis of the effects of individual steps on the overall reaction. The methods are used to show that an ordered addition of two substrates to an enzyme which is activated by another ligand will not necessarily give hyperbolic steady-state-velocity kinetics or the flux ratios characteristic of an ordered addition, if the dissociation of the ligand from the enzyme is random.


A simple enzymic method for the synthesis of adenosine 5'-[alpha-32P]triphosphate on a preparative scale. A simple, rapid and inexpensive method is described for the enzymic synthesis of [alpha-32P]ATP from [32P]Pi on a preparative scale with an overall yield of 53%. The final product contained all of the detectable radioactivity (less than 99.9%) in the alpha position and has been shown to behave identically with commerically availabe [alpha-32P]ATP during the synthesis of 3':5'-cyclic AMP in the reaction catalysed by adenylate cyclase.


Differences in catalytic properties between cerebral cytoplasmic and mitochondrial hexokinases. 1. Clear kinetic differences between cytoplasmic and mitochondrial forms of type-I cerebral hexokinase were demonstrated from experiments performed under identical conditions on three (cytoplasmic, bound mitochondrial and solubilized mitochondrial) preparations of the enzyme. 2. Whereas the Michaelis constant for glucose (KmGlc) was consistent, that for MgATP2- (KmATP) was lower in the cytoplasmic than in the two mitochondrial preparations. The substrate dissociation constants (KsGlc and KsATP) were both higher in the cytoplasmic than in the mitochondrial preparations. A further difference in the substrate kinetic patterns was that KmATP=KmATP for the cytoplasmic enzyme, in contrast with the mitochondrial enzyme, where KmATP was clearly not equal to KsATP [Bachelard et al. (1971) Biochem. J. 123, 707-715]. 3. Dead-end inhibition produced by N-acetyl-glucosamine and by AMP also exhibited different quantitative kinetic patterns for the two enzyme sources. Both inhibitions gave Ki values similar or equal to those of Ki' for the cytoplasmic activity, whereas Ki was clearly not equal to Ki' for the mitochondrial activity. 4. All of these studies demonstrated the similarity of the two mitochondrial activities (particulate and solubilized), which were both clearly different from the cytoplasmic activity. 5. The analysis gives a practical example of our previous theoretical treatment on the derivation of true inhibition constants. 6. The results are discussed in terms of the function of cerebral hexokinases.


Characteristics of hepatic serine-pyruvate aminotransferase in different mammalian species. 1. Serine-pyruvate aminotransferase was purified from mouse, rat, dog and cat liver. Each enzyme preparation was homogeneous as judged by polyacrylamide-disc-gel electrophoresis in the presence of sodium dodecyl sulphate. However, isoelectric focusing resulted in the detection of two or more active forms from enzyme preparations from dog, cat and mouse. A single active form was obtained with the rat enzyme. All four enzyme preparations had similar pH optima and molecular weights. 2. Both mouse and rat preparations catalysed transamination between a number of L-amino acids (serine, leucine, asparagine, methionine, glutamine, ornithine, histidine, phenylalanine or tyrosine) and pyruvate. Effective amino acceptors were pyruvate, phenylpyruvate and glyoxylate with serine as amino donor. The reverse transamination activity, with hydroxypyruvate and alanine as subtrates, was lower than with serine and pyruvate for both species. Serine-pyruvate aminotransferase activities were inhibited by isonicotinic acid hydrazide. 3. In contrast, both dog and cat enzyme preparations were highly specific for serine as amino donor with pyruvate, and utilized pyruvate and glyoxylate as effective amino acceptors. A little activity was detected with phenylpyruvate. The reverse activity was higher than with serine and pyruvate for both species. Serine-pyruvate amino-transferase activities were not inhibited by isonicotinic acid hydrazide.


A novel method for the synthesis of nucleoside triphosphates labelled with inorganic [32P]phosphate specifically in the beta-position. A method was developed for the introduction of [32p]Pi specifically into the beta-position of ATP and GTP. The method is based on two separate reactions involving (a) phosphorolysis of poly(A) or poly(G) [Soreq, Nudel, Salomon, Revel & Littauer (1974) J. Mol Biol. 88, 233-245] in the presence of [32P]Pi and (b) conversion of the labelled diphosphate into the corresponding triphosphate by transferring the active phosphate group from 1,3-diphosphoglycerate in a coupled reaction as decribed by Glynn & Chappell [(1964) Biochem. J. 90, 147-149]. Radioactivity in the beta- and gamma-phosphate groups of the labelled triphosphate was measured by using polynucleotide kinase. No detectable radioactivity was found in the gamma-phosphate group. The suitability of this method for the synthesis of other nucleoside triphosphates specifically labelled in the beta-position is discussed.


Evaluation of benzofuroxan as a chromophoric oxidizing agent for thiol groups by using its reactions with papain, ficin, bromelain and low-molecular-weight thiols. 1. Benzofuroxan (benzofurazan 1-oxide, benzo-2-oxa-1,3-diazole N-oxide) was evaluated as a specific chromophoric oxidizing agent for thiol groups. 2. Aliphatic thiol groups both in low-molecular-weight molecules and in the enzymes papain (EC 3.4.22.2), ficin (EC 3.4.22.3) and bromelain (EC 3.4.22.4) readily reduce benzofuroxan to o-benzoquinone dixime; potential competing reactions of amino groups are negligibly slow. 3. The fate of the thiol depends on its structure: a mechanism is proposed in which the thiol and benzofuroxan form an adduct which, if steric factors permit, reacts with another molecule of thiol to form a disulphide; when the thiol is located in the active site of a thiol proteinase and steric factors preclude enzyme dinner formation, the adduct reacts instead with water or HO- to form a sulphenic acid; attack on the sulphur atom of the adduct by either a sulphur or oxygen nucleophile releases o-benzoquinone dioxine. 4. Benzofuroxan contains n o proton-binding sites with pKa values in the range 3-10 and probably none in the range 0-14; o-benzoquinone dioxine undergoes a one-proton ionization with pKa=6.75.5. o-benzoquinone dioxime absorbs strongly at wavelengths greater than 410nm, where absorption by benzofuroxan, proteins and simple thiol compounds is negligible; 416 nm is an isosbestic point (epsilon 416 = 5110 litre. mol-1-cm-1); epsilon430=3740+[1460/(1+[H+]/Ka)] where pKa=6.75. 6. The possibility of acid-base catalysis of the oxidation by active-centre histidine residues of the thiol proteinases is discussed.


A consideration of the effects of added solutes on the activity of bovine superoxide dismutase. Pulse radiolysis was used to study the effects of changing the concentration of buffer and added salts on the activity of bovine superoxide dismutase. The results are discussed with reference to other work and are interpreted mainly in terms of ionic-strength effects. The effects are best considered as empirical parameters, which are essential for comparative activity measurements. The specific activity of the enzyme was higher than found in previous pulse-radiolysis studies.


Evidence of tumorigenic activity of candidate cell substrates in vaccine production by use of antilymphocyte serum. Candidate cell substrates for vaccine production must be controlled for the absence of tumorigenic activities. This test is done in most laboratories by heterotransplantation of living cells in Syrian hamsters either subcutaneously into newborns or into the cheek-pouch tissue of older ones. The necessity of a long observation period and high rate of intercurrent mortality among newborn hamsters are the disadvantages of this method. The method described by the authors is based on the use of three-week-old mice. Suspensions of living cells (at least 10(6)) were inoculated simultaneously with an antilymphocyte serum of bovine origin. The application of the antilymphocyte serum is repeated twice. Different heteroploid cell strains were studied and their tumorigenic activities were demonstrated. In no case tumors developed after inoculation of diploid cell strains. The results showed that the use of immunodepressed mice is an efficient method for the determination of the cell culture tumorigenic activity.


OSHA response to occupational health personnel needs and resources. Because of the nonavailability of sufficient numbers of qualified industrial hygienists to assume roles as health compliance officers in the Occupational Safety and Health Administration, a three - year career development program for trainee industrial hygienists has been initiated. The present and projected personnel needs in the occupational health field in the U.S. are presented, together with a general description of the OSHA career development program. The program is a response to current needs; it will be curtailed when university graduates are available to meet Agency needs.


Evaluation of "criteria for film badge performance" ANSI N13.7-1972. To determine if ANSI N13.7-1972 is a useful method to test the accuracy of film badge processors to measure radiation doses to workers in the nuclear industry, a set of exposed dose values is compared to several sets of typical, but imaginary, reported dose values. The ANSI Standard is found to be a complex procedure with no direct test for accuracy. Because of these and other limitations of the Standard, it is concluded that ANSI N13.7-1972 is unacceptable for routine testing of film badges.


Subcellular distribution of zinc in rat prostate studied by x-ray microanalysis: I. Normal prostate. Zinc is distributed subcellularly throughout the lateral prostate of the rat in both the stromal and epithelial elements. The connective tissue appears to be a major store of zinc. Within the epithelium, the highest concentrations of the element are found in the lysosomes, nucleoli, nuclear chromatin, secretory granules and luminal secretion. Histochemical studies indicate that the metal is bound relatively tightly within the nucleoli (associated with RNA) and in the secretory products of the cytoplasm. Changes in tissue zinc concentration, observed by other workers, following changes in various external stimuli, may not necessarily be reflected by proportionate changes in epithelial concentrations. The role of zinc in the epithelium is considered to be at least two-fold: firstly, for incorporation into vital cellular mechanisms necessary for cell maintenance and, secondly, for involvement in secretory products. It is also possible that the metal participates in the physiology of the sub-epithelial stroma.


A scanning electron microscopic study of alkali-digested human and rabbit alveoli. Elastic fibers are fairly uniform in the size around all alveolar orifices and taper and branch continuously in the alveolar wall to form a spiderweb-like structure. Unusually large gaps of the elastic fiber network in the alveolar wall frequently show coiled ends or tissue debris at the margin. The latex-filled capillaries interweave and intersect with the elastic fibers and in many of these places appear to angulate or narrow acutely; these angulations may alter with lung inflation, affecting pulmonary vascular resistance. Disruption of fine elastic fibers in the alveolar wall could be related to the development of pulmonary emphysema.


Left ventricular performance in the critically ill premature infant with patent ductus arteriosus and pulmonary disease. Left cardiac dimensions and an index of left ventricular performance, the percent shortening of the internal diameter (%SID) of the left ventricle, were evaluated in premature infants who were asymptomatic, others with pulmonary disease and others with patent ductus arteriosus (PDA). In contrast to controls, left atrial and/or left ventricular end-diastolic dimensions were increased in all infants with clinical criteria of significant PDA. Postoperative dimensions decreased significantly. Percent SID values for normal premature infants (m=33.5%; SD=3.5%) and those with pulmonary disease alone did not differ significantly. In those with clinical criteria of PDA, who were subsequently found to have echocardiographic evidence of left cardiac enlargement, values for %SID were increased. As expected %SID values for individual patients represented a wider range of left ventricular function and/or afterload than for controls. Upon spontaneous or surgical closure of the PDA, %SID returned to normal. A PDA which is associated with left cardiac enlargement exhibits increased %SID, whereas decreasing %SID in the presence of increased dimensions suggests deteriorating myocardial performance. Echocardiography provides valuable insight into the cardiac status of these infants and may contribute to their medical and/or surgical management.


Detection of edema associated with myocardial ischemia by computerized tomography in isolated, arrested canine hearts. This study was undertaken to determine if computerized tomography (CT scanning) with an EMI cranial scanner could detect edema associated with myocardial ischemia in canine hearts. A localized area of decreased density in the posterior papillary muscle and surrounding myocardium was detected on serial 8 mm CT scan slices of each heart after 60 min of circumflex artery occlusion and 45 min of reflow of blood. The wet/dry weight ratios and previous electron microscope studies of the ischemic posterior papillary muscles revealed edema accumulation. After 1 hour of arterial occlusion and 12 hours of reflow (which produces extensive necrosis and a decrease in the wet/dry ratio) lesions were still discernible but were less consistently as severe. Permanent ligation of the left anterior descending coronary artery and major collateral arteries for 6 hours also resulted in a lesion of decreased density in the distribution of the occluded arteries. Thus, CT scanning can detect, and is a potential means for sequential noninvasive quantitation of myocardial edema associated with ischemia.


Single dose response curves of normal cells in situ. Analysis of data on the response of normal tissues (skin, lung, spinal cord, intestine, testis) to single and multifractionated irradiation schedules revealed the characteristics of the single dose survival curves of target cells responsible for skin, lung, and spinal cord reactions, and of stem cells of colonic, jejunal, gastric mucosae, and testis exposed in situ to relatively low doses of x- or gamma-rays. The single dose survival curves of these cells seem to be complex (unlike those of cells studied in vitro), that is, simply exponential in the initial portion up to approximately 250 rads, and downward-bending thereafter in a curve which includes one or two linear portions on a semilogarithmic scale.


Pseudoblocks and portal localization.

Pseudoblocks and portal localization. A new method of rapid simulation including design, verification and patient marking for shaped fields is described. This system utilizes a "pseudoblock" constructed from the Styrofoam components of the Cerrobend blocking system. The procedure is precise; errors in block aperture or patient positioning can readily be seen and corrected before the block is cast. In addition, the use of the pseudoblock does not significantly increase the time the patient spends in the simulator on his initial visit.


Delta check tables for the Technicon SMA 6 continuous-flow analyzer. To provide a data base for the establishment of a method to detect potentially incorrect SMA 6 test results, we analyzed 2970 pairs of SMA 6 results. For each test result, as well as for the urea nitrogen/creatinine ratio and the anion gap, we estimated to probability density function of the amount of change since the most recent test of the same type. We made separate estimates for each of three ranges of the current test result and two ranges of time-spacing between the tests. Tables of nonparametric estimates of selected points from each of the six distributions for each test result and calculated quantity were developed. These tables could be used to identify unlikely test results, either manually or with a laboratory computer.


Replication process of the parvovirus H-1. VI. Characterization of a replication terminus of H-1 replicative-form DNA. The linear duplex replicative form (RF) DNA of the parvovirus H-1 has been characterized with respect to cleavage by the bacterial restriction endonuclease of Escherichia coli, EcoRI. RF DNA has a single cleavage site 0.22 genome length from the left end of the molecule. The molecular weight of H-1 RF DNA determined by gel electrophoresis is 3.26 X 10(6). H-1 RF DNA has been found to dimerize by hydrogen-bounded linkage at the molecular left end, and in some molecules the viral strand is covalently linked to the complementary strand. Some 10% of monomeric RF DNA also has a covalent linkage between the viral and complementary strands at the left end. The EcoRI-B fragment, containing the left end of the RF molecule, appears to be a replication terminus by its labeling characteristics for both RF and progeny DNA synthesis. These findings suggest that the left end of H-1 RF DNA has some type of "turn-around" structure and that this end is not an origin for DNA synthesis.


Stereometric craniometry.

Stereometric craniometry. A method is described whereby three-dimensional co-ordinates of points on a cranium can be recorded in terms of azimuth, elevation and radial distance from a selected point. These co-ordinates can be used to create two-dimensional representations of single crania, the differences between many crania or growth stages of individuals or series of individuals. The co-ordinates can be used in more conventional analytic ways in the same way as cartesian co-ordinates.


Reconstruction of duodenal wall defects with the use of a gastric "island" flap. Many surgical techniques have been proposed for the repair of duodenal wall defects; however, none of these has gained popularity among surgeons. This is a report of our experimentation with the construction of a gastric wall "island" flap, which consists of a segment of greater curvature supplied by the gastroepiploic vessels. In the dog model, the flap allowed patching of large duodenal wall defects without any postoperative complications.


cis-dichlorodiammineplatinum(II) chemotherapy in experimental murine myeloma MOPC 104E. Data are presented indicating marked antineoplastic activity for cis-dichlorodiammineplatinum(II) in MOPC 104E myeloma. One-eighteenth of the dose that produced 100% cures can be combined with noncurative, low doses of cyclophosphamide and 1,3-bis(2-chloroethyl)-1-nitrosourea to produce antineoplastic activity of the same degree as that produced by much higher dose regimens which regularly produce cures. Since, in the past, results of therapeutic trials in plasma cell tumors in humans have paralleled results in this animal model, clinical trials of cis-dichlorodiammineplatinum in multiple myeloma appear warranted.


Comparison of the esterase and human plasminogen activator activities of various activated forms of human plasminogen and their equimolar streptokinase complexes. A comparison was made of the esterase and activator activities of the various activated forms of human plasminogen and their streptokinase complexes with Nalpha-Cbz-L-lysine-p-nitrophenyl ester as the substrate. The steady state kinetic properties of Glu- and Lys-plasmins, and Glu- and Lys-plasminogen-streptokinase complexes were identical, while the Lys-plasmin-streptokinase complex showed a 2-fold increase in Km with the same kcat and a 3-fold increase in Ki for the competitive inhibitor leupeptin. Lys-plasminogen (zymogen with an active site) was prepared which incorporated 0.7 mol of [3H]idisopropyl phosphorofluoridate and 0.43 mol of p-nitrophenyl-p'-guanidinobenzoate/mol of protein. The Km for Lys-plasminogen was 3-fold higher than that of Lys-plasmin, and its maximum velocity 10-fold lower. The steady state kinetic parameters of a plasmin-derived light (B) chain (CmCys)3, and a derived equimolar light (B) chain-streptokinase complex (CmCys)3, isolated from human plasmin and equimolar plasmin-streptokinase, or plasminogen-streptokinase, complexes, respectively, were determined. When the light (B) chain-streptokinase complex is isolated from its parent complexes, there is a complete retention of the original parent's esterase activities, with respect to Km and kcat, and interaction with the competitive inhibitors benzamidine and leupeptin. The plasmin-derived light (B) chain does not retain its parent esterase activities. This chain has very similar kinetic properties to Lys-plasminogen except that streptokinase, in an equal molar amount, does not impart full esterase activity to the light (B) chain whereas the zymogen can be completely activated by streptokinase. The kcat of the plasmin-derived light (B) chain, and its streptokinase complex can be enhanced by 50 and 30%, respectively, in the presence of 10(-4) M leupeptin, a competitive inhibitor of plasmin, attesting to the increased structural flexibility within the active site of this enzyme species. Urokinase hydrolyzes Nalpha-Cbz-L-lysine p-nitrophenyl ester efficiently with a kcat/Km of one-third that of plasmin. The human plasminogen activator activities of various activated forms of human plasminogen and their equimolar streptokinase complexes were compared in a kinetic assay. The Lys-plasmin-streptokinase complex, and streptokinase were the least active of the activator species and were approximately equal in their activator activities. Glu- and Lys-plasminogen-streptokinase complexes had approximately 1.5 times the activity of streptokinase, whereas the equimolar light (B) chain-streptokinase complexes had approximately 2- to 3-times the activator activity of streptokinase. Since the esterase activity remained unchanged, this indicates a greater degree of specificity in the active site of the equimolar light (B) chain-streptokinase activator complex. Urokinase proved to be a poor activator species...


Antibody-mediated suppression of tumor growth. I. Suppression by murine IgG1 isolated from alloantiserum. The tumor suppressive activity of murine IgG1 antibody was studied in vivo. IgG1 was isolated from hyperimmune alloantisera against a murine lymphoma, 6C3HED, by absorption with heat-killed, formalinized Staphylococcus aureus. Cowan strain I, followed by DEAE ion exchange chromatography and Sephadex G-200 gel filtration chromatography. The isolated IgG1, which had no detectable IgM, IgA, IgG2a, IgG2b or IgG3, could suppress the growth of the tumor. In addition, DEAE and Sephadex G-200 column profiles of the in vivo tumor suppressive activity showed good correlation with the profiles of total IgG1 immunoglobulin and anti-tumor antibody assayed for IgG1 heavy chains and kappa light chains. The IgG1 tumor suppressive activity was not diminished after heating at 56 degrees C for 30 min.


The anemia of prematurity. Factors governing the erythropoietin response. We performed sequential studies in 45 premature infants (birth weights less than 1500 g) from 7 to 120 days of age to determine factors governing the erythropoietin response to a declining hemoglobin concentration. The hemoglobin level and the plasma erythropoietin showed a significant inverse correlation (r = 0.50, P less than 0.001), as did, even more strikingly, the plasma erythropoietin response and the infants' oxygen-unloading capacity (r = 0.55, P less than 0.001). In infants with "right-shifted" oxygen-hemoglobin dissociation curves (hemoglobin F less than 30 per cent) hemoglobin levels fell 2 to 3 g per deciliter lower than those in infants with "left-shifted" curves (hemoglobin F greater than 60 per cent) before comparable erythropoietin responses occurred. It appears that premature infants respond appropriately to alterations in oxygen unloading capacity and that the position of the oxygen-hemoglobin dissociation curve and not the hemoglobin concentration alone has a major role in modulated erythropoiesis.


Human hexosaminidase isozymes: chromatographic separation as an aid to heterozygote identification. The correct identification of Tay-Sachs heterozygotes requires a reliable procedure for separation and quantiation of the hexosaminidase isozymes. The most commonly employed method involves thermal inactivation of the heat labile hexosaminidase A assay of residual enzyme activity. This procedure, however, consistently yields a significantly lower absolute and relative activity of hexosaminidase A and a higher activity of the thermostable components (B and I) in comparison with the results obtained by DEAE-cellulose chromatography. DEAE-cellulose chromatographic separation of the hexosaminidase isozymes in serum following thermal inactivation reveals the presence of relative and absolute increase in the activity of the B and I components in addition to loss of the heat-labile A isozyme. Because the conversion of hexosaminidase A into thermostable forms by heating may vary according to the conditions employed, the thermal inactivation procedure may lead to ambiguity in heterozygote identification. This difficulty can be minimized by fractionation of the hexosaminidase isozymes by DEAE-cellulose chromatography followed by assay of the individual components. In addition to the Tay-Sachs carrier state, other conditions can alter the distribution of the hexosaminidase isozymes in tissues and body fluids. For example in serum of patients with juvenile diabetes mellitus there is a characteristic elevation of hexosaminidase B and less consistently, of hexosaminidase A. Since the activity of hexosaminidase A in serum of diabetics fractionated by ion exchange chromatography is at least as high as the activity in serum of healthy non-carriers, patients with diabetes can be easily differentiated from Tay-Sachs heterozygotes. Similarly, the distribution of the hexosaminidase isozymes in serum is altered during pregnancy, where there is usually a significant rise in hexosaminidase A and I (P). However, during pregnancy activities of hexosaminidase A and I in serum of obligate Tay-Sachs carriers are only 50% of the values observed in non-carriers at comparable gestational periods. Since the absolute activities of hexosaminidase A in serum of pregnant carriers obtained by ion exchange chromatography do not overlap with the activities in serum of non-carrier pregnant women at comparable gestational periods, this method has obvious advantages for identification of pregnancies where the fetus may be at risk for Tay-Sachs disease.


Topographic organization of the projections from cortical areas 17, 18 and 19 onto the thalamus, pretectum and superior colliculus in the cat. The distribution of cortical projections from areas 17, 18 and 19 to the lateral thalamus, pretectum, and superior colliculus was investigated with the autoradiographic tracing method. Cortical areas 17, 18 and 19 were demonstrated to project retinotypically and in register upon the dorsal lateral geniculate nucleus, medial interlaminar nucleus, lateral zone of the lateral posterior complex, nucleus of the optic tract and superior colliculus. Area 19 was shown to project retinotopically upon the pulvinar nucleus. Clear retinotopic organization was not demonstrable in the projections of areas 17, 18 and 19 to the reticular complex of the thalamus and ventral lateral geniculate nucleus, or in the projection of area 19 to the anterior pretectal nucleus. The cortical projections were employed to define the retinotopic organization of the nucleus of the optic tract, pulvinar nucleus, and later zone of the lateral posterior complex. The cortical projections show the vertical meridian to be represented caudally, with the lower visual field represented laterally, and the upper visual field medially, within the nucleus of the optic tract. The projections of area 19 to the pulvinar nucleus demonstrate the lower visual field to be represented rostrally and the upper field caudally in this mucleus; the vertical meridian to be represented at the lateral border and the visual field periphery to be represented at the medial border of the pulvinar nucleus. Cortical projections to the lateral zone of the lateral posterior complex demonstrate the lower visual field to be represented rostrally and the upper visual field caudally; the vertical meridian to be represented at the medial limit and the visual field periphery at the lateral border of the termination zones. On the basis of the experimental findings a new terminology is introduced for the feline lateral posterior complex. Divisions are proposed which correspond to zones with demonstrably distinct afferent input. The pulvinar nucleus is defined by the distribution of projections from area 19. Three flanking divisions are defined within the lateral posterior complex; a lateral division recipient of projections from area 17, 18 and 19, and interjacent division recipient of projections of the superficial layers of the superior colliculus, and a medial division flanking the tectorecipient zone medially.