Bactericidal activity of macromomycin, a antitumor antibiotic. Susceptibility testing by the broth dilution method showed that all the gram-positive but only some of the gram-negative bacteria tested were susceptible to the antitumor antibiotic, macromomycin (MCR; NSC 170105). The minimal inhibitory concentration for the susceptible organisms was less than 3 mug/ml. The action of MCR was bactericidal; however, at very high concentrations (50 mug/ml and above) some organisms occasionally escaped death. None of the escaped organisms was resistant to MCR. In combination with other commonly used antibiotics, MCR displayed partial synergy for Pseudomonas aeruginosa (from a minimal inhibitory concentration of >100 to 12.5 mug/ml with 100 mug of chloramphenicol per ml) and for Bacillus pumilus and Staphylococcus aureus (from 1.6 to 0.4 mug/ml and below) with polymyxin B. As with mammalian cells, (125)I-labeled MCR was irreversibly bound to both gram-positive and -negative bacteria. Treatment with trypsin of the (125)I-labeled MCR-exposed cells did not release the bound MCR or reverse its lethal effect. When in solution in a protective buffer at 4 degrees C, MCR was stable for up to 45 days; at 37 degrees C, however, 25% of its bactericidal activity was lost in 72 h. Loss of activity was enhanced 16-fold in the presence of both heated and unheated pooled human sera. Urine had no effect on the activity of MCR.


Five-hour minimal inhibitory concentration test of four antimicrobial agents for Pseudomonas aeruginosa. A rapid 5-h minimal inhibitory concentration procedure and a minimal bactericidal concentration procedure are described for testing Pseudomonas aeruginosa against gentamicin, tobramycin, amikacin, and carbenicillin. The rapid test results were demonstrated to correlate with the conventional tube dilution procedure.


Rapid penicillinase paper strip test for detection of beta-lactamase-producing Haemophilus influenzae and Neisseria gonorrhoeae. A modified 1-min iodometric paper strip test for penicillinase activity was developed for detection of beta-lactamase-producing isolates of Haemophilus influenzae and Neisseria gonorrhoeae. The test is simple to perform and uses reagent-impregnated strips that may be stored for 1 year or more prior to use.


Synergy between ticarcillin and tobramycin against Pseudomonas aeruginosa and Enterobacteriaceae in vitro and in vivo. The antibacterial activities of ticarcillin, carbenicillin, tobramycin, and gentamicin and of combinations of these antibiotics were measured against Pseudomonas aeruginosa and other gram-negative bacilli in vitro and in experimental mouse infections. Synergistic effects were produced by the penicillin/aminoglycoside combinations in growth inhibition tests and in bactericidal tests against many of the bacteria tested. Combinations of ticarcillin + tobramycin were more active in vitro than carbenicillin + gentamicin against P. aeruginosa but were no more active than the latter against other gram-negative bacilli. Ticarcillin + tobramycin and carbenicillin + gentamicin also demonstrated synergistic activities against P. aeruginosa, Escherichia coli, and Enterobacter cloacae in experimental mouse infection models. Thus, the penicillin/aminoglycoside combinations produced greater protective effects than the individual antibiotics against lethal intraperitoneal infections and also were more effective in reducing kidney counts of viable bacteria and kidney abscess formation in experimental pyelonephritis infections. As was the case in vitro, ticarcillin + tobramycin was more effective than carbenicillin + gentamicin against the experimental P. aeruginosa infections. The results of these in vitro and in vivo studies suggest that combined therapy with ticarcillin and tobramycin may be warranted in the treatment of serious infections due to P. aeruginosa and Enterobacteriaceae.


New species of Aspergillus producing sterigmatocystin. A number of species belonging to the genus Aspergillus were evaluated for their toxicity to ducklings and the ability to produce sterigmatocystin. Three new species capable of producing sterigmatocystin were found, namely, Aspergillus aurantio-brunneus, Aspergillus quadrilineatus, and Aspergillus ustus. All three were toxic to ducklings. The production of sterigmatocystin by Aspergillus rugulosus was confirmed, and the toxicity of Aspergillus stellatus and Aspergillus multicolor is described.


Diluent sensitivity in thermally stressed cells of pseudomonas fluorescens. Thermally injured cells of Pseudomonas fluorescens were unable to produce colonies on Trypticase soy agar (TSA) after dilution with 0.1% peptone. Nutritional exigency could not be used as the criterion for this injury, since varying the composition of the plating medium had little effect on the number of colonies that developed. The injured cells had no requirement for compounds known to leak out during the heat treatment in order to recover. The cells did not exhibit injury if dilution preceded heat treatment on the plating medium, demonstrating that the heat treatment sensitized the cells to the trauma of dilution. Substitution of 0.1% peptone with growth medium as the diluent largely offset the previously observed drop in TSA count. Little difference in survival was observed when monosodium glutamate or the balance of the defined medium was used as the diluent. The diluent effect was ionic rather than osmotic. The presence of cations was important in maintaining the integrity of the injured cell, and divalent cations enhanced this protective effect. The role of these cations at the level of the cell envelope is discussed.


Study of inhibition of outgrowth in Bacillus cereus T by ethyl picolinate. The effects of ethyl picolinate on germination, outgrowth, and sporulation of Bacillus cereus T were studied in a synthetic medium containing glucose. Ethyl picolinate specifically inhibited at two stages, outgrowth and sporulation. The initiation of germination and cell division was not affected. The inhibition of outgrowth by ethyl picolinate could be reversed by enrichment of inoculum with aspartic acid, asparagine, lysine, phenylalanine, and tyrosine among the amino acids and by oxalacetate. Nicotinic acid and nicotinamide also possessed this ability. Ethyl picolinate failed to block outgrowth when added to cultures incubated for a short time after inoculation. Enrichment of the medium with lysine plus zinc sulfate stimulated sporulation in the presence of ethyl picolinate to a significance degree.


Microbiological transformations of delta6a10a-tetrahydrocannabinol. A screening program was conducted to find microorganisms that catalyze transformation reactions with cannabinoids. Three hundred fifty-eight cultures, consisting of 97 bacteria, 175 actinomycetes, and 86 molds, were incubated in media containing 0.5 mg of Delta(6a,10a)-tetrahydrocannabinol (Delta(6a,10a)-THC) per ml. After 120 h of cultivation, ethyl acetate extracts of the cultures were examined by thin-layer chromatography (TLC) for transformation products. About 18% of the cultures modified Delta(6a,10a)-THC. The ability to modify the substrate did not predominate among any particular group of microorganisms. After purification, the products from three cultures were analyzed by high-resolution mass spectrometry, 100-mHz proton magnetic resonance spectrometry, ultraviolet spectrometry, and infrared spectrometry. These spectral data indicated that a Mycobacterium sp. oxidized Delta(6a,10a)-THC to cannabinol and a diastereomeric pair of 6a-hydroxy-Delta(10,10a)-THC isomers; a Streptomyces sp. and a Bacillus sp. oxidized Delta(6a,10a)-THC to 7-keto-Delta(6a,10a)-THC and 4'-hydroxy-Delta(6a,10a)-THC, respectively. The occurrence of these products and the presence of others that have not yet been isolated or identified indicate that microbial transformation may be a useful tool for the preparation of new cannabinoids that have desirable pharmacological properties.


Evaluation of a new presumptive medium for group D streptococci. A new medium designated as D streptococcus-enterococcus broth was formulated and evaluated for the enrichment and isolation of strains of serological group D streptococci. This medium was made by modifying Todd-Hewitt broth. Most-probable-number multiple-tube and membrane filter techniques were employed to estimate the numbers of enterococci in known cultures, wastewater, and other samples. Preliminary most-probable-number counts with this medium were as much as 3 logs higher than those counts obtained from four other media with which it was compared. The methodology for using this medium to estimate the numbers of group D streptococci in water is discussed.


Effect of combined heat and radiation on microbial destruction. A series of experiments at several levels of relative humidity and radiation dose rates was carried out using spores of Bacillus subtilis var. niger to evaluate the effect of heat alone, radiation alone, and a combination of heat and radiation. Combined heat and radiation treatment of microorganisms yields a destruction rate greater than the additive rates of the independence agents. The synergistic mechanism shows a proportional dependency on radiation dose rate an Arrhenius dependency on temperature, and a dependency on relative humidity. Maximum synergism occurs under conditions where heat and radiation individually destroy microorganisms at approximately equal rates. Larger synergistic advantage is possible at low relative humidities rather than at high relative humidities.


Methyl bromide as a microbicidal fumigant for tree nuts. Methyl bromide (MeBr) has broad microbicidal activity, but its use as a disinfectant for food is limited by the resulting bromide residues. Increasing the MeBr concentration, exposure temperature, or exposure period of a treatment tended to increase both the microbicidal efficacy of MeBr and the bromide residues. Its sporicidal activity was less at high than at low relative humidity within the range of 20 to 99%. Both the efficacy and the resulting residues of a MeBr treatment varied inversely with the load of product in a fumigation chamber due to sorption of the fumigant. Fumigation tests with almond kernels inoculated with Escherichia coli or Salmonella typhimurium indicated that MeBr can be used to disinfect whole nut kernels without resulting in excessive bromide residues, although the MeBr level necessary is higher than that normally used for insect control.


Serial propagation of Pneumocystis carinii in cell line cultures. Pneumocystis carinii was propagated on three cell lines routinely cultured in many laboratories; the method is practical and convenient. Organisms produced were found to be reactive to Pneumocystis antisera. Studies of antigenic relationships, life cycles, and diagnostic methods will be made easier by these cultures.


Penetration of bacteria into meat.

Penetration of bacteria into meat. Bacteria are confined to the surface of meat during the logarithmic phase of growth. When proteolytic bacteria approach their maximum cell density, extracellular proteases secreted by the bacteria apparently break down the connective tissue between muscle fibers, allowing the bacteria to penetrate the meat. Non-proteolytic bacteria do not penetrate meat, even when grown in association with proteolytic species.


Pemphigus antibody action on skin explants: kinetics of acantholytic changes and stability of antigens in tissue cultures of normal monkey skin explants. The pathogenic effect of the intercellular antibodies of pemphigus was studied by using organ culture of monkey skin. Skin explants that were grown on sera with intercellular antibody titers of 320 or greater fixed these antibodies within one day as demonstrated by direct immunofluorescence for IgG. None of the control sera gave such staining patterns. Following the binding of intercellular antibodies, characteristic histologic changes appeared, notably separation of individual epidermal cells and acantholysis. These histologic changes became more marked in two to five days. During this period, the bound antibodies and the intercellular antigens decreased and disappeared. These temporal relationships of immunofluorescence and histologic findings suggest that pemphigus antibodies play a role in the induction of acantholysis.


Human essential fatty acid deficiency: treatment by topical application of linoleic acid. An essential fatty acid (EFA) deficiency developed in a 19-year-old man who was being maintained on a long-term regimen of fat-free, intravenous hyperalimentation fluids. The EFA deficiency was reversed after 21 days by daily, topical application of linoleic acid to the patient's skin. The ratio of eicosatrienoic acid (20:3, n-9) to eicosatetraenoic acid (20:4, n-6) decreased to normal levels in the skin and serum with clinical improvement of the EFA deficiency syndrome. The cutaneous manifestations (scalp dermatitis, alopecia, and depigmentation of hair) were reversed with continued, topical application of safflower oil, which contains 60% to 70% linoleic acid.


Pseudomonal balanitis.

Pseudomonal balanitis. Significant ecologic changes in the cutaneous flora during treatment may present a challenge both diagnostically and therapeutically. The development of an erosive balanitis due to Pseudomonas aeruginosa presented an example of such a microbiologic shift in the case reported. The eruption developed during treatment with topical antibacterial, antifungal, and corticosteroid agents. The sudden exacerbation of any balanitis while under treatment should alert the physician to the possibility of superinfection.


Aetiology of acute gastroenteritis in infancy and early childhood in southern India. The aetiology of acute gastroenteritis was studied in 50 infants and young children. Bacterial pathogens were isolated in 33, enteropathogenic E. coli (EPEC), Salmonella, and Shigella being the commonest isolates. Rotaviruses were detected in the stools of 13 of the cases. All children with gastroenteritis in whom rotavirus was detected were seen during the months July to December. In 30 children who served as controls, EPEC were isolated in 6, but rotavirus was detected in none. It is concluded that infection with rotaviruses is a significant cause of morbidity in children with gastroenteritis in southern India.


Localization of gold in synovial membrane of rheumatoid arthritis treated with sodium aurothiomalate. Studies by electron microscope and electron probe x-ray microanalysis. The localization of gold in the synovial membrane of rheumatoid arthritis patients treated with sodium aurothiomalate was examined and quantitative analysis of epon-embedded sections was carried out with a wavelength dispersive x-ray microanalyser. Gold was only detected in the lysosomes of synovial lining type A cells and subsynovial mononuclear cells in the form of filamentous deposits and highly electron-dense granules, the latter being few in number. The concentration of gold within the lysosomes containing the characteristic deposits and granules was equivalent to that in epon-embedded standard specimens of freeze-dried albumin in which 2-0-87-4 mg/ml of gold was included. In addition, sulphur was detected in the lysosomes containing the filamentous deposits, but the S/Au x-ray signal ratio was not equal to that detected in sodium aurothiomalate. The significance of the coexistence of gold with sulphur in lysosomes is discussed.


Inhibition by gold of human lymphocyte stimulation. An in vitro study. Transformation of lymphocytes stimulated by mitogen, purified protein derivative, and in the mixed lymphocyte culture was significantly inhibited by gold sodium thiomate and gold sodium thiosulphate at concentrations of 5 X 10(-4) to 5 X 10(-5) mol/1.


Pancreaticoduodenal arterial aneurysms.

Pancreaticoduodenal arterial aneurysms. Experience with four aneurysms of the pancreaticoduodenal artery is reviewed and compared to the reported experience of 19 other cases. In view of the common presentation of such lesions as intra-abdominal hemorrhage preceded by non-specific abdominal pain and other digestive symptoms, it is suggested that angiography perfomed preoperatively or intraoperatively allows definitive diagnosis and leads to specific therapy.


Intravenous amino acids as the sole nutritional substrate. Utilization and metabolism in fasting normal human subjects. The fasting normal human volunteer subject provides an ideal experimental setting for the initial investigation of foodstuffs whose use is proposed for the acutely ill surgical patient. In the normal human subject many variables can be controlled; the achievement of an ideal body fuel economy is quite simple; if a favorable utilization of injected foodstuffs cannot be achieved in this setting, it is unlikely, and remains to be proven, that utilization will be satisfactory under the challenges of acute surgical trauma. In this experimental model, employing four normal human volunteer subjects, nutrition has been provided by the intravenous infusion of isotonic amino acids (FreAmine(R) II) at a 3.4% concentration. No other source of calories or nutrients was provided. In this setting, utilization was very poor; the subjects were in negative nitrogen balance throughout. The nitrogen excretion was significantly greater than the total of infused nitrogen. The changes in protein, fat and carbohydrate intermediates, as well as the alteration in hormone concentrations, suggest the following endocrine governance of fuel economy in this setting: a sharp rise in glucagon with maintenance of insulin concentration; rapid gluconeogenesis at the expense of both injected and endogenous amino acids; a progressive ketosis without any associated improvement in protein economy; fat oxidation to meet caloric need. The changes in plasma amino acid concentrations are of outstanding interest. They demonstrate changes appropriate to the infusion gradient with the exception of three amino acids whose concentrations did not respond to high infusate levels (serine, lysine, and alanine); likewise, by the fact that methionine rose remarkably though present in only low concentrations in the infusion. These data, taken with other information reported in the literature, as well as continuing studies in these laboratories, strongly suggest that the utilization of infused amino acids for protein synthesis is favored by the provision of an additional caloric source such as glucose.


Cardiac depression in bacteremia.

Cardiac depression in bacteremia. Hemodynamic and respiratory effects of a 5-hr IV infusion of Ps. aeruginosa at a dose of 10(8) organisms per ml per minute were studied in 6 dogs. Four dogs served as controls. Gramnegative bacteremia, with 70,000 +/- 1,800 organisms per ml of blood, caused a 50% reduction of cardiac output at three hrs. Peripheral vascular resistance increased significantly, but mean heart rate fell below control levels. Decline in mean systemic blood pressure from 150 +/- 5 mm Hg to 88 +/- 6 mm Hg was accompanied by a significant increase in pulmonary arterial wedge pressure with normal right atrial and pulmonary arterial pressures. Pulmonary vascular resistance also remained unchanged. With progression of the low output state and development of hypothermia, arteriovenous oxygen difference (A-V DO(2)) fell significantly. Despite a decline in functional residual capacity, venoarterial admixture diminished in the face of reduced pulmonary capillary perfusion, normal arterial Po(2) values, decline in body temperature and finally very narrow A-V DO(2). Histologically, ventricular myocardium revealed severe interstitial edema. It is concluded that myocardial dysfunction may occur early during gramnegative bacteremia, and formation of myocardial edema appears to be a significant contributing factor in myocardial failure.


Effect of antrectomy on gastric hypersecretion induced by distal small bowel resection. Seven male Rhesus monkeys, with demonstrated gastric hypersecretory response to 50% distal small bowel resection, were studied. This increase in gastric acid output had persisted for more than 6 months. Precise, anatomical antrectomy without vagotomy was performed and intestinal continuity restored by gastroduodenostomy. Complete abolition of basal acid secretion and of the secretory response to histamine occurred. These effects could not be reversed by continuous (5 hour) infusions of pentagastrin at 0.2 microgram/kg hr-1 or 2 microgram/kg hr-1. The dose response curve to gastrin pentapeptide was altered; maximum secretion was greatly reduced and occurred at 10 microgram/kg hr-1. Neither basal serum gastrin or basal serum GIP was significantly reduced by antrectomy. These findings indicate that in this animal model the antrum is essential in the genesis of the hypersecretory state. If these findings are relevant to man precise antrectomy may be the procedure of choice for symptomatic acid hypersecretion after small bowel resection or disease.


Sequential hemodynamic studies in patients having aortic valve replacement with the Ionescu-Shiley pericardial xenograft. Sequential hemodynamic studies were performed in 13 patients having aortic valve replacement with glutaraldehyde-stabilized pericardial xenografts. The investigation done preoperatively was repeated twice in all patients at mean intervals of 9.9 and 42.2 months following valve replacement. Annulus diameters of the valves were 19, 23, and 25 mm. Between the preoperative and the first postoperative investigation there was a significant increase in cardiac index (p less than 0.05) and a significant reduction in both pulmonary wedge and left ventricular end-diastolic pressures (p less than 0.01). At the first postoperative study the mean ejection systolic gradient across the three sizes of aortic pericardial xenograft was 7, 15.7, and 8.4 mm Hg at rest and 10, 19.6, and 13.7 mm Hg on exercise, respectively. The corresponding surface areas of the three valve sizes were 1.1, 1.5, and 1.62 cm2 at rest and 1.4, 2.1, and 2.03 cm2 on exercise. There was no significant change in any of these measurements between the first and the second postoperative study. The results have shown considerable postoperative hemodynamic improvement, insignificant transvalvular pressure gradients, and maintenance of the functional integrity of the pericardial xenograft up to 56 months following valve insertion.


Insidious rifampin-associated renal failure with light-chain proteinuria. A patient who was receiving rifampin treatment for tuberculosis developed heterogenous light-chain proteinuria and insidious renal failure after a period of fluid restriction. The renal damage was characterized pathologically by an interstitial nephritis with invasive tubular casts and an associated renal vein thrombosis. The possible role of the light-chain proteinuria in the pathogenesis of the renal failure is discussed.


Pulmonary and rhinocerebral mucormycosis. Successful outcome with amphotericin B and griseofulvin therapy. An ominous prognosis is associated with combined pulmonary and rhinocerebral mucormycosis (phycomycosis). We report the case of a diabetic patient with ketoacidosis who had extensive pulmonary and rhinocerebral mucormycosis that responded satisfactorily to amphotericin B and griseofulvin therapy. The affected lung is completely atelectatic and has remained so for 12 months without evidence of necrosis or abscess formation.


Deoxyribonucleic acid base composition of Simonsiellaceae. The molar percentages of guanine plus cytosine in the DNA of 51 strains of Simonsiellaceae were determined by buoyant density ultracentrifugation of cell lysates in CsCl. The DNA base ratios ranged from 41-55 mole-% guanine plus cytosine. These values fall within the range known for the Order Cytophagales, the non-fruiting gliding bacteria, and are outside the range of the Order Myxobacterales, the fruiting myxobacteria. Among the strains of the genus Simonsiella, four distinct group can be delineated on the basis of source of origin (sheep, dog, cat, human) and GC content. The neotype of Alysiella filiformis has a GC content of 45.4 mole-%.


Temperature range variants of Bacillus megaterium. Facultatively and obligately thermophilic variants were isolated from 3 out of 12 tested mesophilic Bacillus megaterium strains. The variants occurred at a frequency of 10(-8)-10(-9). The ability to grow at elevated temperatures was cured by means of treatment with acridine orange. Stable  revertants were isolated from facultatively and obligately thermophilic variants. An unknown type of megacin was produced by the facultative thermophiles. This megacin attacked mesophilic and obligately thermophilic strains. The thermophiles displayed a few divergent taxonomic characteristics but a close relationship between the strains was indicated by the megacin spectrum and sensitivity to phage. Arrhenius plots revealed that the strains could be considered as temperature range variants and that the temperature characteristic increased with growth at a higher temperature range. The case for a plasmid involvement in the phenomenon is discussed.


Pharmacotherapy for organic brain syndrome in late life. Evaluation of an ergot derivative vs placebo. Evaluation of treatment modalities, including pharmacotherapy, for organic brain syndrome (OBS) has been difficult because of sampling and methodological problems, and comparisons of research studies are all but impossible. In this study, an ergot derivative, a combination of dihydroergocornine mesylate, dihydroergocristine mesylate, and dihydroergokryptine mesylate (Hydergine) was compared with placebo, using a double-blind technique in a sample of nursing home residents with evidence of OBS. An 18-category symptom rating scale was used for periodic assessment over a six-month interval. Comparisons of the two groups of subjects disclosed that the Hydergine-treated group showed statistically significantly more improvement in most of the variables measured, especially during the last three months of treatment. Furthermore, sophisticated analysis revealed that positive changes in cognitive function cannot be accounted for as a mere reflection, or "halo" effect, associated with improved mood and general sense of well-being.


[Experiments on rhesus-monkeys (Macaca mulatta) with the carcinogens methyl- and ethylnitrosourea (author's transl)]. Application of methyl- and/or ethylnitrosourea during intrauterine or postnatal life did not induce tumours in Macaca mulatta. Most of the animals survived 5 to 10 years after the beginning of the experiments and are in good health at present. Complete postmortem examinations were performed in all monkeys who died.


The influence of Listeria monocytogenes cells on the primary immunologic response in irradiated mice. The influence of killed Listeria monocytogenes cells on the primary immunologic response in mice irradiated with 300 or 500 R was studied. The immunologic response of the mice to sheep red blood cells used as antigen was assessed at the cellular level (by counting PFC) and humoral level. Injection of killed Listeria monocytogenes cells before irradiation of the mice diminished the immunosuppressive effect of roentgen radiation. Injection of the cells after irradiation accelerated regeneration of immunologic reactivity in the irradiated mice.


Problem oriented medical record: a predetermined problem list for spinal cord injury. The Problem Oriented Medical Record is ideally suited to the documentation and management of patients with spinal cord injury, with their involvement of multiple organ systems and need for long term follow-up. The classic format described by Weed has been modified by use of a predetermined problem list, subdividing spinal cord injury into eight problems. We find this method to be of value in organizing the records and managing patients, from the acute phase to the follow-up phase, and additionally in organizing the records of long term patients when seen in follow-up. The problems listed are: 1--neurologic injury; 2--spinal skeletal injury; 3--other injuries; 4--general medical conditions; 5--urologic; 6--gastrointestinal; 7--sexual function; 8--rehabilitation. As complications or other problems arise, they are added to this list. Presented are two case histories, one of a long term patient and the other of a patient with new acute spinal cord injury to illustrate the application of this method.


Nutritional therapy based on positive caloric balance in burn patients. Oxygen consumption and caloric expenditure was 1 1/2 to 2 times normal in 15 major burn patients from the time of burn to the time of surface coverage. This hypermetabolic state was quite consistent hour-to-hour and day-to-day, and correlated best with the extent of full-thickness burn. Nutritional management and caloric intake can be intelligently planned using simple spirometry and indirect calorimetry to measure caloric requirements. Weight gain and prompt healing can be achieved by positive caloric balance. Based on daily metabolic studies, a positive caloric balance feeding protocol has been used in the treatment of 556 patients. This regimen, in combination with many other factors in physiologic and surface care, has resulted in high survival rates, short hospitalization, and rapid rehabilitation.


Pharmacological prophylaxis in the kindling model of epilepsy. Review of antiepileptic drug assessment to date by means of the kindling model of epilepsy suggests that it fills a gap that is evident in standard screening methods such as maximum electroshock or pentylenetetrazol screening tests. However, in order to obtain a comprehensive profile of antiepileptic drugs regarding prophylaxis of developing seizures and treatment of well-established seizures through the kindling preparation, it is desirable to have the following: (1) standardization of kindling techniques, (2) examination of drug effects on developing as opposed to developed seizures, (3) use of a variety of animal species, involving different functional brain sites, and (4) monitoring of plasma levels of the drug administered. It is envisaged that judicious use of the kindling preparation might also enable us to gain some insight into the mechanisms by which drugs produce their prophylactic or therapeutic effect.


Optic disc edema in raised intracranial pressure. I. Evolution and resolution. Progressively growing intracranial space-taking lesions were simulated in 32 rhesus monkeys by balloons introduced into the subarachnoid space of the temporal region. Optic disc edema (ODE) first appeared at the lower pole, then the upper pole, then the nasal part, and last the temporal part of the disc; severity of edema generally followed sult, most severe at the lower pole (P less than .005). Fluorescein fundus angiography showed that swelling of the optic disc preceded the vascular changes associated with ODE. Raised intracranial pressure for 24 hours, or less, could cause ODE. The atrophic part of the optic disc did not develop ODE. The studies indicate that swelling of the optic disc is the first sign of raised intracranial pressure and is due to swelling of the nerve fibers in the optic disc; the various associated vascular changes are secondary.


Optic disc edema in raised intracranial pressure. II. Early detection with fluorescein fundus angiography and stereoscopic color photography. Optic disc edema (ODE) due to chronic intracranial hypertension was produced experimentally in rhesus monkeys. Serial studies of fundus changes at frequent intervals, by routine ophthalmoscopy, steroscopic color photography, and fluorescein angiography, revealed that swelling of the optic disc was the first sign of ODE. Other early signs were striation of nerve fibers on the optic disc margins and peripapillary retina, blurring of the disc margins, hyperemia of the disc and capillary dilation, hemorrhages, and other retinal vascular changes; these usually appeared in that sequence. The classically described signs of early ODE were almost always absent. A normal fluorescein fundus angiogram during the incipient stage did not rule out ODE. Stereoscopic color fundus photography was the most sensitive means of detecting early ODE. Fluorescein angiography did not show changes till edema was of a mild to moderate degree; routine ophthalmoscopy was the least reliable method.


Atropine inhibition of echothiophate cataractogenesis in monkeys. One cynomolgus and three vervet monkeys were treated topicall twice daily in one eye with 63 microgram of echothiophate iodide and 750 microgram of atropine sulfate. The opposite eyes were treated with echothiophate alone. The eyes treated with echothiophate alone all developed posterior and/or anterior subcapsular cataracts within 26 to 66 days. The echothiophate plus atropine-treated cynomolgus eye developed a cataracts of much delayed onset and lesser severity compared to its echothiophate-only-treated fellow. None of the echothiophate plus atropine-treated vervet eyes developed cataracts during the 121 (two eyes) or 231 (one eye) days of treatment. After 121 days of echothiophate plus atropine treatment, two vervet eyes were switched to echothiophate alone. Both eyes developed unequivocal posterior and anterior subcapsular lens opacities, one within 51 and one within 71 days after the switch. We conclude that atropine inhibits echothiophate cataracto-genesis in vervet and cynomolgus monkeys.


[Trigemino-facial reflex as a diagnostic aid in Recklinghausen's neurofibromatosis]. The blink reflex evoked by unilateral electrical stimulation was studied in 7 patients with Neurofibromatosis and 50 normal persons. Studing data of the early R1 reflex, there was no difference between normal persons and patients with von Recklinhausen disease. A slightly, but significantly increased reflex latency of the direct R2 was recognized in the majority of the patients with Neurofibromatosis.


Sexuality and contraceptive acceptability. A woman's sexuality may influence the particular choice and acceptance of a contraceptive method which, in turn, may also affect her sexual response. The type of contraceptive recommended should be determined following both physical and psychological assessment of the woman. A method that is not psychologically acceptance is likely to result in discontinuation. Positive and negative effects of the oral contraceptive pill are considered. Pharmacologically induced symptoms are difficult to specify. There appears to be a much greater incidence of psychological problems.


The care of the diabetic child in general practice. One child in a thousand under the age of 15 years has clinical diabetes. Such children require insulin once or twice daily. Their diabetes is usually best controlled with a diet that provides adequate nutrition for growth and a carbohydrate allowance that balances the hypoglycaemic effect of their insulin. With good initial training in the care of their child, intelligence and emotional stability, families usually cope well with diabetes and allow him to lead a normal life during childhood.


Fluorescein angiography and light microscopy studies of retinas irradiated by oxygen nuclei. Fluorescein angiography and light microscopy techniques were utilized in the rhesus monkey to evaluate retinal damage following oxygen ion irradiation. Particle flux ranged from 1.3 x 10(7) particles/cm2 to 1.5 x 10(8) particles/cm2. Earliest evidence of damage was detected with fluorescein angiograms 24 h after exposures to 7.7 x 10(7) particles/cm2. No evidence of the hypothesized microlesion was found by either technique.


Relative biological effectiveness (RBE) of heavy ions (O+8) for producing retinal lesions. The relative biological effectiveness (RBE) for oxygen ions (250 MeV/nucleon) for producing an ischemic retinal lesion in the rhesus monkey has been shown to be greater than 10. The RBE concept and the use of the standard methodology is discussed with respect to other published reports.


Purine nucleoside phosphorylase polymorphism in sheep erythrocytes. A polymorphism of purine nucleoside phosphorylase is described in sheep erythrocytes. Two isozymes were distinguished electrophoretically, one with high activity (NP-1) and one with low activity (NP-2). Breeding data suggest that the two isozymes are the product of two codominant alleles, NP1 and NP2. The Km's for inosine did not differ between NP-1 and NP-2; however, NP-2 had a lower pH optimum and was relatively unstable when incubated at 48 C.


Codominant autosomal inheritance of polymorphic red cell acid phosphates of lemurs and some properties of the enzymes. Red cell acid phosphatase phenotypes of 207 captive animals of the genera Lemur, Hapalemur, and Prophithecus were determined by starch gel electrophoresis and phosphatase-specific staining. In Lemur fulvus, three phenotypes, designated A, B, and AB, were observed. In each of the species L. catta, L. macaco, L. mongoz, and L. variegatus, a single phenotype was observed, In Hapalemur griseus, three phenotypes were found: A,B, and AB. In Propithecus verreauxi, a single phenotype was found. Examination of breeding records in conjunction with the results of the electrophoretic analyses supports the conclusion that the erythrocytic acid phosphatases in this group of nonhuman primates are the products of at least two codominant autosomal alleles. There is a wide range of specific activities of the acid phosphatases as determined by colorimetric assays. The values range from 60.6 micronmoles of p-nitrophenol released per gram of hemoglobin per 30 min in Lemur catta to 429.1 micronmoles in Propithecus verreauxi. The enzymes of L. fulvus and P. vereauxi were purified approximately 400-fold, and Michaelis-Menten constants were determined on the purified preparations. For L. fulvus phenotype A, Km = 0.8 mM; for L. fulvus phenotype B, Km = 0.8 mM; and for P. verreauxi, Km = 0.6 mM; the substrate in each case was p-nitrophenylphosphate.


The prepupal salivary glands of Drosophila melanogaster. Evidence is presented in support of the concept that the larval salivary gland of Drosophila melanogaster  continues to function as an important secretory organ throughout prepupal stages and after pupation. Just after puparium formation, and at other later periods, the glands appear to be in the process of disintegration, but each time they recover until after pupation. Nuclear blebbing occurs through the time of survival of the glands, but is shown not to involve transport of RNA out of the nucleus. Transport in and out of the nucleus is clearly rapid and in a steady state as compared to the massive and intermittent export of cytoplasmic substance into the lumen of the gland.


Transcriptional changes in pupal hypoderm in Drosophia melanogaster. Transcriptional changes with development of the imaginal thoracic hypoderm of Drosophila melanogaster were evaluated by studies on pulse labeling and total composition with respect to protein. Evidence presented indicates that certain glycoproteins produced by prepupal salivary glands are important for bristle development long after the salivary gland is histolyzed. Striking changes in protein synthesis with development can be correlated with major hypodermal functions such as bristle formation, chitin deposition, and a moltlike process which occurs around 52 hr after puparium formation.


Enzymatic variation at seven loci in nine natural populations of Drosophila melanogaster. Allozyme polymorphisms at seven loci have been studied in nine natural populations of Drosophila melanogaster from the Saône and Rhône valleys sampled in 1973 and 1974. A great deal of polymorphism was observed; an individual was on the average heterozygous at 20.2% of its loci. The populations were genetically very homogeneous throughout the region sampled. The number of ovariolae per female varied from one group of populations to another depending on their geographical separation. Yet the number of ovariolae remained constant from one year to the next. The results show that migration alone cannot explain the homogeneity of the allozyme frequencies. It seems reasonable to conclude that selection plays a major role in maintaining the homogeneity of populations living in proximal biotopes.


The immunological characterization of several human ribonucleases by using primary binding tests. RNAases (ribonucleases), purified from four human tissues, as well as bovine pancreatic RNAase (RNAase A), were studied by immunodiffusion methods and by two different primary binding tests. The enzymes fell into two groups immunologically, those purified from plasma and pancreas in one and those from spleen and liver in the other. No antigenic cross-reaction between the two groups was detected by any of the immunoassays used. There was a slight antigenic cross-reaction between the human and bovine pancreatic RNAases. The liver and spleen RNAases were immunologically identical by all criteria used, whereas a small but consistent antigenic difference between the human plasma and human pancreas enzymes was detected. The significance of this difference between the human plasma and pancreas RNAases is discussed in relation to similarities and differences in their properties.


High-sensitivity sequence analysis of peptides and proteins by 4-NN-dimethylaminoazobenzene 4'-isothiocyanate. A manual high-sensitivity sequencing method is described, in which 4-NN-dimethylaminoazobenzene 4'-isothiocyanate is used for the stepwise degradation of amino acid residues from the peptides. The 4-NN-dimethylaminoazobenzene 4'-thiazolinones of amino acids that were released, after conversion into their thiohydantoin derivatives, were identified by t.l.c. on polyamide sheets. This new method is simple and sensitive, and requires only 2-10nmol of peptides or proteins for extended sequence analysis. The method was tested on the sequence analysis of a hexapeptide (Leu-Trp-Met-Arg-Phe-Ala), bradykinin, glucagon and native lysozyme. Results show that the proposed procedure is a sensitive method for the sequence determination of short peptides as well as for the partial sequence determination of intact proteins.


Separation and properties of two arylamidases from rat cardiac-muscle extracts. Two main arylamidase activities were separated from a particle-free supernatant of rat heart by chromatography on DEAE-Sephadex. Although both enzymes hydrolysed L-leucine 4-nitroanilide, only peak-II enzyme hydrolysed L-lysine 4-nitroanilide. A third minor peak (Ia) contained an enzyme that was active mainly on the L-lysine 4-nitroanilide. The mol.wts. of the enzymes in peaks I and II were approx. 257000 and 105000 respectively. The pH optimum was approx. pH7.0 for peak-I enzyme and 7.0-8.0 for peak-II enzyme. Both enzymes were inhibited by addition of puromycin, p-hydroxymercuribenzoate, o-phenanthroline and bivalent metal ions. Addition of dithiothreitol resulted in stimulation of both activities. Dialysis against o-phenanthroline resulted in inhibition of peak-I and -II enzymes, but after dialysis against EDTA only peak-II enzyme was inhibited.


Effects of adenine nucleotides and phosphate on adenosine triphosphate sulphurylase from Anabaena cylindrica. Production of adenosine 5'-[35S]sulphatophosphate by a partially purified ATP sulphurylase from Anabaena cylindrica was inhibited by AMP, ADP and P1. Decreases in enzyme activity in the presence of these inhibitors were reversed by increasing the concentrations of ATP. The adenine nucleotides inhibited the enzyme competitively with respect to ATP. In the presence of P1, ATP showed a positive co-operative effect on enzyme activity. The inhibition by P1 was enhanced by increasing concentrations of MG2+. The effects of the adenine nucleotides and the interaction of P1 and Mg2+ on ATP sulphurylase activity are discussed in relation to the regulation of sulphate assimilation via the energy metabolism of the alga.


Effect of buformin on the regulation of glycogen metabolism in the skeletal muscle of normal rats. In view of the experience that after short- or long-term oral administration of biguanides to rats no glycogenolysis in the muscle tissue occurs or even an increase of the muscle glycogen content and a high rate of incorporation of radioglucose into the muscle glycogen, the glycogen synthetase activity of skeletal muscle in a concentrated muscle homogenate and the effect of isoproterenol on glycogenolysis were investigated in normal rats after short-term oral administration of 1-butylbiguanide (buformin). The basal synthetase activity and the ATP inhibition of the enzyme were not affected by buformin, but the cyclic AMP mediated inactivation was significantly inhibited after buformin pretreatment. This inhibition was reversed by running the assay in the presence of Mg2+. Buformin also inhibited the isoproterenol induced glycogenolysis of the skeletal muscle tissue. From our results we suppose that biguanides may influence the regulation of glycogen metabolism by inhibiting the inactivating synthetase kinase and the activating phosphorylase kinase. It is possible that divalent cations like Mg2+ as an activator of kinase reactions are concerned.


Comparative effects of aspirin and diflunisal on prostaglandin synthetase from human platelets and sheep seminal vesicles. 1. We have compared the effects of diflunisal on prostaglandin (PG) synthetase from human platelets and sheep seminal vesicles (SSV) by measuring malonaldehyde, PG products, and rabbit aorta contraction. 2. Aspirin inhibits PG synthetase by covalently acetylating the enzyme. Inhibition is dependent on time and aspirin concentration. Aspirin is 37-fold more potent in inhibiting enzyme in human platelets compared with enzyme from SSV. 3. Diflunisal inhibits PG synthetase from intact human platelets and SSV equally (50% inhibition at 3-5 micronM). The drug does not covalently modify the enzyme, and inhibition is not time dependent. Diflunisal probably acts at a site similar to aspirin since the drug will inhibit acetylation of the enzyme by aspirin. 4. The results suggest that diflunisal is relatively less inhibitory to platelet function than is aspirin.


Zinc is associated with the beta subunit of DNA-dependent RNA polymerase of Bacillus subtilis. The Bacillus subtilis DNA-dependent RNA polymerase holoenzyme and core enzyme each contain approximately two atoms of zinc per molecule. When the dissociated subunits of the enzyme are passed through a blue dextran-Sepharose affinity column, only the beta subunit binds to the column. The total zinc content of the enzyme is tightly bound to the beta subunit. Dialysis studies suggest that the two zinc ions differ in the strength of their association with the beta subunit. The presence of zinc in beta is consistent with several other lines of evidence which indicate that this subunit is dirrectly involved in phosphodiester bond formation. The blue dextran-Sepharose column procedure should be useful in future studies of the dissociation and reassociation of the enzyme since the method is rapid and provides excellent recovery of the beta subunit as well as the alpha and beta' subunits of the RNA polymerase.


Cyanide-resistant electron transport in sporulating Bacillus megaterium KM. The NADH oxidase activity of stage V mother-cell membranes, isolated from sporulating Bacillus megaterium KM, shows a greater inhibition by cyanide and displays this response at lower concentrations of cyanide than the stage V forespore inner membrane. Comparison of the effects of various respiratory inhibitors reveals that the difference in cyanide sensitivity between these membranes is located on the oxidase side of the 2-heptyl-4-hydroxyquinoline N-oxide-sensitive step. Both membranes contain cytochromes a+a3, b-562, b-555, c and d, with three potential oxidases: cytochromes a+a3, o and d. Cyanide difference spectra suggest that cytochromes b-562 and d may be the components involved in the cyanide-resistant electron transport pathway. Membrane ascorbate-N,N,N',N'-tetramethylphenylenediamine and ascorbate 2,6-dichlorophenolindophenol oxidase activities are highly sensitive to cyanide. Evidence is presented for terminal branching of the respiratory chain with branches differing in cyanide sensitivity. The cyanide sensitivity of the NADH oxidase of membranes prepared from various stages of sporulation is compared. Morphogenesis of the mother-cell plasma membrane to a cyanide-sensitive form during stages II and III of sporulation is postulated.


Effects of temperature and bovine serum albumin on lysis of erythrocytes induced by dilauroylglycerophosphocholine and didecanoylglycerophosphocholine. The effects of the incubation temperature and bovine serum albumin on hemolysis induced by short-chain phosphatidylcholine were examined. The rate of hemolysis of human, monkey, rabbit, and rat erythrocytes by dilauroylglycerophosphocholine showed biphasic temperature-dependence: hemolysis was rapid at 5-10 degrees C and above 40 degrees C, but slow at around 25 degrees C. In contrast, the rate of lysis of cow, calf, sheep, pig, cat, and dog erythrocytes did not show biphasic temperature-dependence, but increased progressively with increase in the incubation temperature. Bovine serum albumin increased the hemolysis of human erythrocytes induced by dilauroylglycerophosphocholine or didecanoylglycerophosphocholine: it shortened the lag time of lysis and reduced the amount of phosphatidylcholine required for lysis. A shift-down of the incubation temperature from 40 to below 10 degrees C also shortened the lag time of lysis of human erythrocytes induced by dilauroylglycerophosphocholine and reduced the amount of phosphatidylcholine required for lysis.


Selective compartmentation of the hydration products of carbon dioxide in liposomes, and its role in regulating water movement. Using liposomes, some possible consequences of auxin-stimulated proton secretion in plant tissues are modelled. Liposomes, made specifically permeable to protons and potassium ions, but containing at least one impermeant, swell at a rate which is proportional to the availability and hydration rate of carbon dioxide; the gas functions as a source of proton and dehydrated anion and this enables a net transfer of KHCO3 to take place.


Self-regulation of membrane fluidity. The effect of saturated normal and methoxy fatty acid supplementation on Tetrahymena membrane physical properties and lipid composition. Tetrahymena cells elongated and desaturated massive supplements of palmitic or lauric acid at nearly twice the rates employed by unfed cells, thereby maintaining constant the physical properties of their membrane lipids. However, when a mixture of the 9- and 10-monomethoxy derivatives of stearic acid was administered, these compounds were incorporated without further metabolism. The marked fluidizing effect of the phospholipid-bound methoxy-fatty acids elicited an immediate reduction in fatty acid desaturase activity, the pattern of change being very similar to that induced by supplements of polyunsaturated fatty acids. The modulation of fatty acid desaturase activity by methoxy-acids clearly seems to be governed by membrane fluidity rather than by some form of end product inhibition of the type which might have been postulated to explain the similar effect caused by polyunsaturated fatty acids.


Kinetics of bidirectional active sodium fluxes in the toad bladder. The kinetics of isotopic Na+ flows was studied in urinary bladders of toads from the Dominican Republic. Initial studies of the potential dependence of passive serosal to mucosal 22Na+ efflux demonstrated the absence of isotope interaction and/or other coupling with passive Na+ flow. The electrical current I and mucosal to serosal 22Na+ influx were then measured with transmembrane potential clamped at deltapsi=0, 25, 50, 75 or 100 mV. Subsequent elimination of active Na+ transport with mucosal amiloride permitted calculation of the rates of active Na+ transport JaNa and active and passive influx leads to JaNa and leads to JpNa. The results indicate that for Dominican toad bladders mounted in chambers only Na+ contributes significantly to transepithelial active ion transport; hence JaNa=Ja. Ja was abolished at deltapsi=E=96.3+/-1.9 (S.E.) mV. As deltapsi approached E, active efflux comes from Ja became demonstrable. At deltapsi=100 mV, comes from Ja exceeded leads to Ja, so that Ja was negative. Experimental values of leads to Ja agreed well with theoretical values predicted by a thermodynamic formulation: leads to Jaexp=0.985 leads to Jatheor (r=0.993). The dependence of leads to Ja on deltapsi is curvilinear.


Occurrence and function of membrane teichoic acids. Membrane teichoic acids, sometimes described as lipoteichoic acids, are important but not major components of nearly all Gram-positive bacteria. They appear on the outer surface of the cytoplasmic membrane and possess antigenic properties. Several functions have been ascribed to these glycerol phosphate polymers, including the binding of divalent cations required for optimal activity of membrane-bound enzymes, and the control of certain lytic enzymes. A substance that is identical or closely similar to membrane teichoic acid, lipoteichoic acid carrier, plays an important part in the  biosynthesis of wall teichoic acid; it accepts polyol phosphate residues from CDP-glycerol or CDP-ribitol to form a polyol phosphate chain which is then transferred after the incorporation of a tri(glycerol phosphate) linkage unit, to the growing glycan chain of peptidoglycan.


The structure of Tetrahymena pyriformis mitochondrial DNA. I. Strain differences and occurrence of inverted repetitions. We have analysed the structure of the mtDNAs of six amicronucleate Tetrahymena pyriformis strains, belonging to at least four phenosets, as defined by Borden et al. (Borden, D., Whitt, G.S. and Nanney, D.L. (1973) J. Protozool. 20, 693--700). 2. The mtDNAs of all strains are linear, but they differ in size, in their fragmentation by endonuclease EcoRI and in overall sequence; less than 20% sequence homology was found by DNA-DNA hybridization in all combinations tested, except for the mtDNAs from strains T and ST which are indistinguishable. 3. In spite of these marked sequence differences the mtDNAs of all strains share two structural peculiarities: ragged (gnawed) duplex ends and a duplication-inversion, which varies in length between 0.3 and 1.2 micrometer, depending on the strain. In four strains the duplication-inversion is terminal, allowing formation of single-stranded DNA circles with a duplex tail; in two strains it is subterminal. 4. The ragged ends and sub-terminal position of the duplication-inversion in some of the Tetrahymena mtDNAs do not fit any of the current models for the replication of linear mtDNAs.


The structure of Tetrahymena pyriformis mitochondrial DNA. II. The complex structure of strain GL mitochondrial DNA. 1. Isolated mtDNA from Tetrahymena pyriformis strain GL is a linear duplex molecule with an average molecular weight of 32.6 - 10(6) and without internal gaps or breaks. Denaturation of this DNA results in single strands with a duplex hairpin at one end. The length of this hairpin varies between 0 and 5 micrometer within one preparation. 2. Uder renaturation conditions the single strands with hairpins are able to circularize in two ways, depending on the length of the hairpin. Circularization is also observed after partial digestion with exonuclease III of native strain GL mtDNA. 3. All these data fit a model (see Fig.2) in which the DNA is heterogeneous in length at both ends. At the left end a 10-micrometer duplication-inversion is present; part of this duplication-inversion is complementary to a region at the right end of the molecule. 4. The analogy between the structural peculiarities of strain GL mtDNA and of some linear viral DNAs is stressed.


Inhibition of amino acyl tRNA synthetase activity by copper complexes of two metal binding ligands. N-Methyl isatin beta-thiosemicarbazone and 8-hydroxyquinoline. Copper complexes of N-methyl isatin beta-thiosemicarbazone, 1-formyl isoquinoline thiosemicarbazone and thiosemicarbazide inhibit amino acyl tRNA synthetase activity. Copper complexes of 8-hydroxyquinoline and 8-mercaptoquinoline also inhibit. The 1 : 1 ligand-metal complex is significantly more active than the 2 : 1 complex. The free ligand alone and copper sulfate alone have little, if any, effect. These complexes have no effect on the ATP-PPi exchange reaction and do not cause deacylation of amino acyl tRNAs. This indicates that the process inhibited by these complexes is the amino acylation reaction. This is the first report that these copper binding ligands can inhibit enzymatic processes which involve nucleic acids but which are not viral, bacterial or mammalian cell polymerases.


Purification of human granulocyte catalase in chronic myeloid leukemia. Human granulocyte catalase (hydrogen peroxide:hydrogen peroxide oxidoreductase, EC 1.11.1.6) was purified from chronic myeloid leukemia cells. The purification procedure included heat precipitation, ammonium sulphate fractionation, DEAE-Sephadex chromatography, gel chromatography on Sephadex G-200 and isoelectric focusing with an approximate yield of 30% and a 1000-fold purification. The molecular weight of the subunit obtained by sodium dodecyl sulphate electrophoresis was 65 800. So20,w was 11.6 +/- 0.24. The pH-optimum was 6.6-6.7 and the spectrum showed a major peak at 405 nm and shoulders at 500, 540 and 625 nm typical for catalase. The electrophoretic mobility was towards the anode at pH 8.6 and identical to normal granulocyte and erythrocyte catalase. These three species of catalase gave the reaction of identity on immunodiffusion and crossed immunoelectrophoresis. The content of catalase and its activity of isolated granulocytes were approximately identical in normal and chronic myeloid leukemia granulocytes while the specific activity of leukemic catalase was higher than normal. No difference in catalase content was found between mature and immature leukemic granulocytes.


Effects of lipids on the activity of ferrochelatase. Removal of lipids from submitochondrial particles or detergent-solubilized mitochondrial preparations of rat liver resulted in a 90% loss of ferrochelatase (protochemeferro-lyase, EC 4.99.1.1) activity. The addition of either a fatty acid or phospholipid restored enzyme activity; the extent of reactivation being correlated with the degree of unsaturation of the fatty acid or acyl chain and independent of the polar head group of the phospholipid, Arrhenius plots of the ferrochelatase activities of submitochondrial particles and detergent-solubilized mitochondrial preparations showed transition temperatures of 37 and 28.5 degrees C, respectively. Ferrochelatase of submitochondrial particles or detergent-solubilized preparations had an absolute requirement for Ca2+. The ferrous salt of oxalic acid, a Ca2+ chelator, was a very poor substrate for these preparations. In contrast, ferrochelatase activities of fatty acid- or lipid-supplemented acetone extracts of these preparations were not dependent on the presence of Ca2+ and ferrous oxalate served as substrate for these extracts.20


Affinity chromatography of lysyl hydroxylase on concanavalin A-agarose. Lysyl hydroxylase (peptidyllysine, 2-oxoglutarate: oxygen 5-oxidoreductase, EC 1.14.11.4) has a high affinity for columns of concanavalin A-agarose, which was markedly reduced in the presence of alpha-methyl-D-mannoside, suggesting that the enzyme is a glycoprotein. Once bound, the enzyme could not be eluted with the glycoside alone, whereas an effective elution was achieved by a combination of alpha-methyl-D-mannoside and ethylene glycol. The data thus suggest that hydrophobic interaction stabilized the complex of the enzyme with the column. This information was applied to obtain a lysyl hydroxylase purification of about 3000-fold with a recovery of more than 10% from extract of chick embryos by relatively simple steps.


Hemoglobin Izu (Macaca): beta83 (EF 7) Gly leads to Cys. A new hemoglobin variant found in the Japanese monkey (Macaca fuscata). Recently, Ishimoto, Kuwata and Shotake reported a polymerizing hemoglobin found in Japanese monkey (Macaca fuscata) (J. Anthropol. Soc. Nippon 83, 233-243 (1975)). They separated the variant hemoglobin by gel filtration and from finger print results and from the fact that beta-mercaptoethanol dissociates the polymer deduced the substitution of an amino acid(s) by cysteine in the betaT10 peptide. We have purified the variant hemoglobin by ion-exchange chromatography using carboxymethyl cellulose after the protection of the reactive thiol groups with cystamine, and purified the betaT10 peptide and demonstrated that the usual glycine at beta 83 (EF 7) is substituted in the variant hemoglobin by cysteine.


Selective stimulation of venous prostaglandin E 9-ketoreductase by bradykinin. The effects of bradykin on prostaglandin metabolism in canine mesenteric vessels were examined. Bradykinin stimulated microsomal prostaglandin synthesis in both artery and vein; this stimulation was more pronounced when [14C] hosphatidylcholine rather than [14C] arachidonate was used as the substrate for prostaglandin synthetase. This suggested that bradykinin enhanced a membrane phospholipase. In addition, bradykinin selectively stimulated prostaglandin E 9-ketoreductase activity from veins but not arteries. This may explain the finding that bradykinin induces the release of prostaglandin E compounds from arteries but prostaglandin F compounds from veins.


Vitamin A receptors. I. Comparison of retinol binding to serum retinol-binding protein and to tissue receptors in chick retina and pigment epithelium. 1. A simple, efficient three-step method for purification of serum retinol-binding-protein is described with homogeneity obtained after chromatography on DEAE-Sephadex, CM-Sephadex and Sephadex G-100. 2. Evidence is presented indicating that retinol receptors present in the cytosol fraction of chick retina and pigment epithelium are separate and distinct from purified retinol-binding protein. Fluorescence characteristics are different in tissue cytosol and serum as assessed by sucrose density gradient analysis. Tissue retinol receptors do not interact with human serum prealbumin although the prealbumin readily complexes with purified chicken retinol-binding protein. Likewise, no binding to serum retinol-binding protein antibody could be detected by sucrose density gradient analysis, in immunoprecipitation experiments or by double immunodiffusion. It thus appears that specific retinol receptors are present in neural retina and pigment epithelium that are different from serum retinol-binding protein.


[Primary structure of tRNA Thr 1a and b from brewer's yeast]. One of the two major species of brewer's yeast tRNA threonine (tRNA Thr 1) has been purified by countercurrent distribution followed by two chromatographic steps (respectively on a Sepharose 4B and a BD-cellulose column). Complete digestion with pancreatic and T1 RNases and a partial hydrolysis with T1 RNase followed by the isolation and determination of the nucleotide sequences of the resulting fragments permitted the derivation of its primary structure. tRNA Thr 1 is in fact a mixture of two subspecies differing only by a A49-U65 base pair in 50 per cent of the molecules which is replaced by a G49-C65 pair in the other 50 per cent. These two subspecies consist of 76 nucleotide residues including 14 minor nucleotides. They show a characteristic m3C at the 3'terminal end of the anticodon loop, an anticodon I-G-U followed by t6A and C48, uncompletely modified (50 per cent) to m5C within the 5 nucleotides long extra-arm. The minor nucleotides m2G m2 2G are located at positions in which they generally occur in the tRNA structures as does m1A within the T-psi-C loop.


Sympathetic adrenergic blockade effects upon operantly conditioned blood pressure elevations in baboons. Sympathetic adrenergic nervous activity during operantly conditioned hypertension was evaluated by assessing the effects of specific alpha-(phentolamine or phenoxybenzamine) and beta- (propranolol) adrenergic blockers in baboons reinforced for increasing diastolic pressure in daily, 12-h sessions. In the first 10 min of control (no blockade) sessions, mean heart rate increased 24 bpm (21%) above the value for the 10 min immediately prior to the beginning of the sessions; systolic pressure increased 27 mm Hg (22%) and diastolic pressure increased 24 mm Hg (31%). Beta-blockade eliminated the tachycardia but did not attenuate the increased blood pressure. Alpha-blockade did not attenuate the increased blood pressure significantly either. Combined alpha- and beta-blockade did significantly attenuate the increase in diastolic pressure, but consistent, significant increases in systolic pressure (17 mm Hg, 17%) and diastolic pressure (16 mm Hg, 26%) still occurred. The results support the participation of the sympathetic adrenergic nervous system in producing operantly conditioned blood pressure changes, but the results are also consistent with the additional participation of nonadrenergic factors in operantly conditioned hypertension.


Diffusion of 133Xe through frog skins, toad bladders, and water boundary layers. We have measured the total permeability coefficients P as a function of stirring frequency omega for 133Xe through frog skins and toad bladders. The permeability coefficients for the frog skins and toad bladders proper are, respectively, Pm = (3.9 +/- 0.8) X 10(-4) cm/s and (7.4 +/- 4.2) X 10(-4) cm/s. "Unstirred" water layer thickness delta is determined concurrently, from the frequency dependence of P(omega); the result for frog skin is delta = (0.060 +/- 0.016) square root of omega(rad/s) cm. The stirring frequency range is from omega = 7.5 rad/s (72 rpm) to 55 rad/s (530 rpm). The results support the conclusions that the principal barrier to Xe diffusion in these epithelia is inter- and intracellular water, and that the diffusion is passive and rapid. The experimental method may be straightforwardly adapted to the measurement of diffusion or counterdiffusion of any gamma-radioactive soluble or partly soluble solute through any flat membrane or through a solvent. We estimate the amount of total body-absorbed radioactivity due to environmental 133Xe to be 50 fCi for an ambient concentration of 2.6 pCi/m3 of air.


The determination of the electron density profile of the human erythrocyte ghost membrane by small-angle x-ray diffraction. Diffraction patterns of stacked hemolyzed erythrocyte ghosts in the wet state were recorded. Three orders of a surprisingly high first-order periodicity of 600 A were detected. The scattering curves were evaluated by the Q-function method, including lattice distortions of the one-dimensional multilamellar system. The resulting electron density profile of the membrane in the wet state is strongly asymmetric. It consists of an asymmetrical bilayer-type part and an excess of positive relative electron density at the inner, cytoplasmic, side of the membrane. The extension of the whole membrane profile in the wet state is 100--120 A. We suggest that the innermost positive density peak mainly represents the loosely bound protein components spectrin and actin, located at the cytoplasmic side of the membrane and sometimes seen as "fuzzy" material on electron micrographs.


Denaturation of phycocyanin by urea and determination of the enthalpy of denaturation by microcalorimetry. Denaturation of the protein phycocyanin in urea solution was investigated by microcalorimetry, ultraviolet and visible spectroscopy, circular dichroism and sedimentation equilibrium. The results consistently demonstrated that in the presence of 7 M urea this protein is completely denatured. By assumings a two-state mechanism, an apparent free energy of unfolding at zero denaturant concentration, (formula: see text) was found to be 4.4 kcal/mole at pH 6.0 and 25 degrees C. By microcalorimetry the enthalpy of denaturation of phycocyanin app was found to be -230 kcal/mole at 25 degrees C. The relatively large negative enthalpy change results from protein unfolding and changes in protein solvation.


[Effect of sodium aurothiomalate in acute toxic hepatitis]. The effect of lysosomotropic agent--sodium aurothiomalate on liver damage and lysosomal changes during acute toxic hepatitis was studied. Combined administration of sodium aurothiomalete led to less extensive necrosis and to widening dystrophic areas in the hepatic parenchyma. Solubilization of the acid RNA-ase activity was decreased, and nonsedimentable activities of beta-galactosidase, cathepsin D were the same as in acute CCl4-hepatitis.


[Change in the metabolism of 7,12-dimethylbenz(a)anthracene under the influence of vitamin A]. The in vitro metabolism of 7,12-dimethylbenz(a)anthracene in its incubation with the liver microsomes, the liver and mammary gland homogenates of rats, kept on vitamin A-enriched diet, was studied. Vitamin A inhibited the formation of lipophilic metabolites and increased the generation of water-soluble metabolites. The amount of lipophilic metabolites extracted from the microsomes and the liver and mammary gland homogenates were decreased by a factor of 2.2 and 5, respectively. The amount of unmetabolized DMBA in the liver microsomes was the same in control and experimental animals.


[Interaction of 2,6-dimethyl-3,5-dicarbethoxy-1,4-dihydropyridine with enzymes of the NADP.H2-specific electron transport chain of rat liver microsomes]. 2,6-Dimethyl-3,5-dicarboethoxy-1,4-dihydropyridine (DHP) interacts with NADPH-dependent electron transfer system of rat liver microsomes: it forms a complex with the terminal oxidase-cytochrome P-450, according to type I, and inhibits clearly the activity of NADPH-cytochromes c-reductase and mitindione dimethylesterase. DHP repeatedly administered in vivo rendered no inducing influence upon the microsomal enzymes.


[Activation of respiration in liver mitochondria by catecholamines]. Catacholamines activate in vivo and in vitro the rat liver mitochondria respiration (VO2) at high piruvate, 2-oxoglutarate and succinate concentrations. The effect was equally characteristic of free oxidation (state 4), phosphorylation (state 3), and even of VO2 uncoupled by DNP. EDTA and bovine serum albumin addition failed to eliminate the effect of the oxidoreductases activation by catecholamines.


Comparative effects of glyceryl trinitrate on venous and arterial smooth muscle in vitro; relevance to antianginal activity. 1. A quantitative in vitro study has been made of the actions of glyceryl trinitrate and sodium nitrite on vascular smooth muscle (dog femoral artery and saphenous vein; rat portal vein); these have been compared with the actions of papaverine, isoprenaline, salbutamol, pentaerythritol tetranitrate and trimetazidine. 2. Glyceryl trinitrate was more active on the saphenous vein than on the femoral artery in inhibiting noradrenaline and potassium-induced tone. 3. Unlike glyceryl trinitrate, sodium nitrite and isoprenaline, papaverine and diazoxide inhibited noradrenaline-induced contractions of venous and arterial smooth muscle to the same extent. 4. The selective dilator effects of glyceryl trinitrate on venous smooth muscle may explain its action in alleviating the pain of angina pectoris. It is suggested that the use of these three vascular smooth muscle preparations (arterial, and veins with and without spontaneous myogenic activity) is a useful initial screening procedure for prospective antianginal drugs acting by venodilatation.


Traumatic spreading depression syndrome. Review of a particular type of head injury in 37 patients. From 1476 hospital admissions with head injury, 37 patients were selected on the basis of distinctive clinical features which appeared to share a single and benign aetiology. In head injuries of this type, transient neurological disorders resulted from trivial or rather mild head injuries. These disorders, which included headache, nausea and vomiting, pallor, somnolence, irritability and restlessness, stupor, hemiparesis and aphasia, appeared after a lucid interval which was usually less than two hours; then the patient either recovered, or went on to develop convulsive attacks. Recovery without convulsions was usual in children and adolescents; convulsions occurred both in infants and younger children. The symptoms were not attributable to cerebral compression but were probably due to a self-limiting cortical phenomenon. It appears that convulsive attacks occurring within a few hours of this type of head injury may not be significant as precursors of post-traumatic epilepsy. It is suggested that there is a close relation between the convulsive attacks and the non-convulsive symptoms seen in this type of injury and that both are based on a common process which has the characteristics of the spreading depression of Leão. This type of head injury should be classified as a distinct clinical entity, in which no surgical treatment is required and the prognosis is good.


Acute dystonia as an idiosyncratic response to neuroleptics in baboons. Among 25 baboons, Papio papio, 2 consistently showed acute dystonic reactions, with mouthing, compulsive gnawing and limb and trunk dystonia, following the intravenous administration of neuroleptics and related drugs (haloperidol, 0-6-1-2 mg/kg; pimozide 0-5-2-5 mg/kg; chlorpromazine 5-25 mg/kg; metoclopramide 1-5-1-7 mg/kg; oxyperomide 0-25-1-0 mg/kg). The syndrome was not seen after thioridazine (3-7 mg/kg). The dystonic responses occurred within 1-2 h of drug injection and lasted for 2-24 h. They were abolished for 1-3 h within 1-2 min of the intravenous injection of acetylcholine antagonists (benztropine 0-2 mg/kg; hyoscine 0-02 mg/kg). Pre-treatment with a combination of reserpine (2 mg/kg) and alpha-methylparatyrosine (2 X 200 mg/kg) substantially reduced the dystonic response to haloperidol. A second larger dose of haloperidol (5 mg/kg), given 60-90 min after 0-5 mg/kg) initially reduced the intensity of the dystonic response, but after 29 min induced vomiting and generalized seizures in the idiosyncratic baboons. The hypothesis is advanced that the dystonic responses result from release of dopamine on to a sub-population of receptors in the striatum that are relatively insensitive to blockade by neuroleptics.


Neurotensin: central nervous system effects of a hypothalamic peptide. The central administration of neurotensin, an endogenous hypothalamic tridecapeptide, produces a marked dose-related decrease in body temperature of mice and rats at an ambient temperature of 25 degrees C. This effect is even more pronounced when mice are placed at 4 degrees C to increase the rate of decline of body temperature. Other sequelae observed after central administration of neurotensin are decreases in locomotor activity in rats and a marked dose-related enhancement in pentobarbital-induced mortality, sedation and hypothermia. This latter effect was shown to be due to a significant reduction in the metabolic degradation of the barbiturate. None of the above-mentioned effects are observed after peripheral neurotensin administration, suggesting that this peptide does not readily cross the blood-brain barrier. Neurotensin appears to be one of a growing list of neuropeptides that can affect CNS function.


Activity of neurones in the inferotemporal cortex of the alert monkey. The activity of neurones in the inferotemporal cortex of the alert rhesus monkey was recorded while the monkey was shown visual stimuli, which included both food and non-food objects for comparison with the activity of neurones in the lateral hypothalamus and substantia innominata. In the anteroventral part of the inferotemporal cortex, neurones were found with visual responses which were sustained while the animal looked at the appropriate visual stimuli. The latency of the responses was 100 msec or more. The majority (96/142 or 68%) of these neurones responded more strongly to some stimuli than to others. These units usually had different responses when objects were shown from different views, and physical factors such as shape, size, orientation, colour and texture appeared to account for the responses of some of these units. Association of visual stimuli with a food reward (glucose solution) or an aversive taste (5% saline solution) did not affect the magnitude of the responses of the neurones to the stimuli either during the learning or after the period of learning. Nor did feeding the monkey to satiety affect the responses of the neurones to their effective stimuli.


Vertical eye movement related unit activity in the rostral mesencephalic reticular formation of the alert monkey. Eye movement related unit activity was recorded in the rostral mesencephalic reticular formation (MRF) of the alert monkey. Most units (78 out of 117) were activated with a short activity burst starting before the eye movement and were otherwise silent. The activity was the same whether movements occurred spontaneously in the light or dark, or were fast phases of vestibular or optokinetic nystagmus, and could be related to parameters of a vector representing the eye movement such as amplitude, position changes along certain planes or direction of movements. Units coding position changes or direction of movement had their preferred direction always close to the vertical. Other units (18 out of 117) showed some tonic activity, which was also only related to vertical eye position. It is suggested that this region of the rostral MRF acts an an immediate supranuclear structure, mediating eye movements in the vertical plane.


Oxidant effect of acetylphenylhydrazine: a comparative study with erythrocytes of several animal species. Red cells from several animal species were treated with acetylphenylhydrazine and a comparative study of the oxidation of hemoglobin (Hb) and glutathione (GSH) made. Wide interspecies differences were observed but the oxidation of GSH paralleled that of Hb. Added glucose protected both Hb and GSH from oxidation; GSH by itself exercises a protective effect on Hb. The characteristic rates of oxidation of GSH in the different species can be observed only in the presence of oxyhemoglobin but not carboxyhemoblobin or methemoglobin. The oxidation of Hb appears to be the primary event, the oxidation of GSH being a consequence thereof.


Possible mechanism for the frequency-dependent increases in the cardiac contraction in monkey. The effects of various interventions on the frequency-dependent increases in the contractility of the papillary muscles of monkeys were investigated. Ouabain (10(-6)M) and KCl-free Krebs-Ringer solution, which are known to inhibit membrane Na+,K+-ATPase (EC 3.6.1.3), abolished the frequency-dependent increases in the contractility of the papillary muscles. Epinephrine (4.5 X 10(8)M) or quinidine (1.3 X 10(-5)M), which are known not to inhibit the membrane Na+,K-ATPase at these concentrations, did not alter the frequency-dependent increases in the contractility. These results indicate that the frequency-dependent increases in the contractility might be mediated through an inhibition of the sarcolemmal Na+,K+-ATPase.


Radioimmunodiffusion technique for determining diphtheria antitoxin. A radioimmunodiffusion technique for detecting low levels of diphtheria antitoxin was developed. Diphtheria toxoid was labelled with 125I to facilitate detection of lines of precipitation by the use of X-ray film, the lower limit of detection being 0.001 unit per millilitre of diphtheria antitoxin.


Punch sampling technique for quantitative identification of tritiated D-amphetamine. A punch sampling, tissue solubilization, and scintillation counting technique has been devised for the quantitative identification of tritiated d-amphetamine in monkey brain tissue. The method is superior to autoradiography in identifying this water soluble radiochemical, and is easier and more anatomically precise than radiochemical assay of brain homogenates. This rapid and simple technique allows one to survey the whole brain by sampling unit volumes of individual structures verified by light microscopy. The procedure consists of six steps which may be spaced as the laboratory schedule permits: obtaining tissue blocks, preparation for sectioning, punch sampling and sectioning, preparing samples for scintillation counting, histological verification of sample sites, and analysis of scintillation count data. The technique has been successfully used to demonstrate a gradient of radioactivity following experimental intraventricular injections of 3H amphetamine.


Elective postoperative radiotherapy for locally advanced colorectal cancer. A preliminary report. Preoperative radiotherapy in colorectal carcinoma invalidates surgical staging and delays performing the surgical resection. Postoperative radiotherapy does neither. From October 1972, to December 1975, 40 patients at high risk for local recurrence (B2 and C) received postoperative radiotherapy. Lesions that were located in the rectum, rectosigmoid and low sigmoid colon were given 4600 rads in four and a half weeks through an inverted T-shaped field which encompassed the pelvic and paraortic nodes. Patients with tumors located above mid-sigmoid were treated to the entire abdominal cavity by the moving strip technique. Of 19 patients with rectal and rectosigmoid lesions, 14 (74%) are alive without evidence of disease. Two had local recurrence in the treated area. Of 21 patients with lesions above the mid-sigmoid, four have failed locally, while 11 (52%) are alive without evidence of disease. One of these 40 patients died to radiation enteritis. Although the follow up period is short, the results suggest that a moderate dose of radiation may prevent local recurrence in patients with locally advanced colorectal cancer.


The significance of needle biopsy after irradiation for stage C adenocarcinoma of the prostate. The presence of malignant cells in needle biopsy specimens following irradiation for adenocarcinoma of the prostate has been used to criticize or defend this treatment. At Walter Reed Medical Center, 38 consecutive patients with Stage Cadenocarcinoma underwent definitive irradiation between August 1970 and March 1973. The median dose to the pelvis was 7000 rads in 31 fractions in 43 days (2030 ret). Post treatment examination included palpation of the prostate and transperineal biopsy of the most suspicious areas. Gradual disappearance of the palpable tumor occurred in all patients. Two men have had clinical evidence of re-growth of prostatic cancer. Thirty-three patients have had up to seven biopsies each for a total of 139, an average of four biopsies per patient. There were 49 positive and 90 negative biopsies. Positive biopsy rate correlated only with the interval after irradiation--60% at six months, 37% at one year, 30% at 18 months, and approximately 19% after two and one-half years. There was no correlation of biopsy results with pre-irradiation estrogen or orchiectomy, with time-dose-fractionation relationships, or with prognosis. These biopsies provide interesting data about the regression rate of prostatic adenocarcinoma, but they have no significance for the individual patient. They have, therefore, been eliminated from follow-up studies.


Use of body scanner in radiotherapy treatment planning. CT body scans obtained on 98 patients before, during, and after radiotherapy were evaluated for their utility in radiotherapy treatment planning and in follow-up after radiotherapy. Twenty-two patients were studied after irradiation. Four received additional treatment, and continuing or planned treatment was withheld from another four on the basis of CT data. Tumor extent was clearly delineated on CT scan in 48/76 cases (63%), suggestive in 25/76 (33%), and not seen in only three (4%). Utilizing CT data relative to all other available tests, in 75 patients, total treatment volume was altered in 34 (45%), tumor coverage was marginal or inadequate in 35 (47%), and volume of normal tissue irradiated was changed in 34 (45%). CT scan data was judged essential for treatment planning in 41, or 55%, of patients studied. Unsuspected areas of tumor involvement were seen in 32 of 75 cases (43%). Use of the CT scan as a patient contour for radiotherapy treatment planning and alternative techniques for inputting the CT data to treatment planning computers are discussed. A simple inexpensive device to accomplish this is described. Speculations are made regarding the impact of CT scanners on transverse axial tomography units and treatment simulators as well as the potential application of the technique in brachytherapy dose computation.


Preliminary results of aggressive multimodality therapy for metastatic osteosarcoma. Ten patients with metastatic osteosarcoma were treated at the Joint Center for Radiation Therapy, the Sidney Farber Cancer Institute, and the Children's Hospital Medical Center from 1973 to 1976. Patients were treated with an aggressive multimodality approach with included surgery, chemotherapy, and radiation therapy. Three of 10 patients are alive with no evidence of disease, five are alive, with disease, and two are dead of disease. The median survival is 12+ months. Local control data for radiation combined with high dose methotrexate in metastatic osteosarcoma is shown.


Stages IIB and IIIB Hodgkin's disease. Results of combined modality treatment. Between April 1969, and December 1974, 23 IIB and 26 IIIB surgically staged patients with Hodgkin's disease were treated at the Joint Center for Radiation Therapy. Stage IIB patients received either mantle and para-aortic-splenic pedicle, or total modal irradiation (TNI) alone or with the addition of combination chemotherapy. Relapse-free survival is 83% and overall survival 88%. Eleven patients received combination chemotherapy in addition to mantle and para-aortic irradiation, and both the relapse-free and overall survival are 100%. Of the stage IIIB patients, seven received TNI alone with four relapses, and 19 were treated with TNI and MOPP with two relapses. These relapse rates are significantly different (p less than 0.05). The relapse-free and overall survival for all stage IIIB patients is 66% and 84% respectively. These data imply that irradiation alone is not adequate treatment for stage IIIB Hodgkin's disease, and that with the addition of combination chemotherapy both the disease-free and overall survival is similar to that of early stage Hodgkin's disease without systemic symptoms. The ideal management of stage IIB Hodgkin's disease is less certain; it is our plan to study the efficacy of combined modality treatment.


Hodgkin's disease stage I and II. A comparison between two different treatment policies. A retrospective analysis was performed on 145 patients with Stage I and II Hodgkin's disease treated over an 11-year period. Sixty-two patients (Group I) received a mantle field without systematic irradiation of the para-aortic lymph nodes. Eight-three patients (Group II) received radiotherapy according to the folowing policy: all Stage IB and IIB and all mixed cellularity and lymphocytic depletion types received total nodal irradiation while stage IA and IIA nodular sclerosing and lymphocytic predominance cases received irradiation to a mantle field and to the para-aortic lymph nodes. The characteristics of the two groups were roughly comparable in age range, sex, staging, histopathologic subtypes and total irradiation doses. All patients had lymphangiograms although not all underwent staging laparotomies. The staging laparotomy did not appear to have an influence within each group. The extent of irradiation did significantly affect both the incidence of further manifestation of disease as well as survival rates. The frequency of lymph node extension, organ extension and local recurrence for Group I was 24%, 14%, and 3%, while for Group II it was 4%, 6%, and 6%, respectively. The seven-year absolute survival rate for Group I was 57% while for Group II it was 93%.


Early nodal and extra-nodal non-Hodgkin's lympnomas. There were 54 Stage I and 38 Stage II patients with non-Hodgkin's lymphomas treated with curative radiotherapy between 1964 and 1975. The initial biopsy material was reclassified according to the criteria of Rappaport el at.10 The frequency of nodular vs diffuse histological pattern was 26% vs 74%. The survival and recurrence free survival were significantly superior for the nodular pattern as compared with diffuse in both Stages I and II. The sites of first recurrence were evaluated. Fourteen out of 20 Stage I and 18 out of 26 Stage II patients recurred in one of the nodal areas only as their first site of recurrence. In 50%, this was in contiguous lymph node areas only. Extra-nodal non-Hodgkin's lymphomas survival and recurrence-free survival rates at five years were identical.


Particle radiation therapy: experimental basis and clinical application. Conventional radiation therapy can eradicate cancers within tissues of their origin and regional spread with conservation of anatomic structure, thus preserving function and cosmesis. New treatment methods may improve the therapeutic ratio either by increasing the frequency of tumor control or lessening the treatment-related morbidity, or both. There are several physical and biological reasons why particle radiation therapy may increase tumor cell killing without increasing normal tissue sequelae. After preliminary basic research, clinical trials of fast neutron and proton teletherapy were started. Over 700 patients were treated with fast neutron beams in 3 U.S. research programs. These studies will be extended to include negative pi mesons and heavy particles.


Ovarian cancer: use of multiple modality programs involving surgery, radiation therapy and chemotherapy. Appreciating the past reports of effectiveness for various therapeutic modalities in ovarian cancer, the Gynecologic Oncology Group activated eight protocols. Three involved epithelial lesions with randomized multimodality trials alone or in combination. The other protocols were devoted to registration of rare tumor case reports. Conclusions are still difficult to reach due to inconsistencies in pathologic diagnoses and deficiencies in radiation therapy, chemotherapy and surgery inherent in the initial phases of group development by diverse specialist in oncology. Adjuvant therapy for early cancer seems to have no advantage. Single drug, melphalan therapy may be as effective as multi-drug, irradiation or combined drug-irradiation therapy and less toxic. For the rare tumors, preliminary results suggest therapeutic advantage for at least one triple drug program in malignant teratoma. With the lessons of the past, it is anticipated that new studies briefly described herewith may be more effectively applied.


Cytogenetic and immunoglobulin markers of human leukemic B-cell lines. This study describes the establishment of two permanent leukemic B-cell lines (BALM-1 and -2) originating from the blood cells of a patient with a B-cell type of acute lymphoblastic leukemia. The cells of BALM-1 and -2 exhibited cell surface markers compatible with B-cell origin. The identity between the leukemic cells in vivo and those of the BALM lines was established on the basis of the karyotypic picture, including marker chromosomes 14q+ and t(12;17;?) and cell surface immunoglobulins (K, delta, and mu chain determinants).


Detection and microbiologic assays of antimetabolites. The biochemical considerations applicable in the detection of new antimetabolites are very similar to those used in developing microbiologic assays for these drugs. An in vitro system for detection of antimetabolites was developed based on different sensitivities to such drugs of bacteria cultivated in complex versus completely synthetic media. A number of new antimetabolites of amino acids, vitamins, and purines or pyrimidines were isolated and microbiologic assays for these drugs were developed. As an example of the technology utilized and possibilities available, an assay for cytosine arabinoside was described. It can detect 0.5-microgram/ml concentrations of this drug even in the presence of other antitumor or antibacterial antibiotics.


Microbiologic assays of cancer chemotherapeutic agents. Among various assay methods available for the study of drug distribution kinetics in cancer chemotherapy, a method that determines the drug in its active form is the most desirable. The microbiologic assay method, with its rapid, simple, and inexpensive procedures, fulfills such a requirement. Besides the two commonly used chemotherapeutic agents, methotrexate and cytosine arabinoside, microbiologic assays are available for other chemotherapeutic agents, such as 6-mercaptopurine, 5-fluorouracil, actinomycin D, adriamycin, and daunorubicin. Assays have been developed for citrovorum factor and 5-methyltetrahydrofolate in the presence of high levels of methotrexate. With the introduction of combination chemotherapy, microbiologic assay methods using resistant mutants, sensitive to a single drug but resistant (either due to mutation or due to culture conditions) to all other drugs involved in the combination, could well be the most efficient for the study of distribution kinetics. Several such assay methods employed in our laboratory are discussed.


Dose-dependent metabolism of methotrexate in man and rhesus monkeys. Humans and rhesus monkeys receiving high-dose methotrexate (MTX) (greater than 50 mg/kg) excrete significant quantities of the metabolite, 7-hydroxy-MTX. This metabolite, though 200-fold less potent than MTX as an inhibitor of mammalian dihydrofolate reductase, is of very limited aqueous solubility, and thus may contribute to the renal toxicity of the high-dose regimen. The metabolite was not observed in previous pharmacologic studies in which conventional doses of MTX were administered (less than 10 mg/kg).


Characterization of 2-amino-2,6-dideoxy-D-glucose as a constituent of the lipopolysaccharide antigen of Pseudomonas aeruginosa immunotype 4. The title lipopolysaccharide was freed from its lipid A component by mild, acid hydrolysis, to give a polysaccharide fraction that was subsequently hydrolyzed completely to afford a mixture of neutral sugars and amino sugars. The amino sugars were separated, and identified as 2-amino-2-deoxy-D-galactose, 2-amino-2,6-dideoxy-galactose as a 2:1 mixture of the D and L enantiomers, and 2-amino-2,6-dideoxy-D-glucose. A reference sample of 2-amino-2,6-dideoxy-D-glucose was synthesized by an improved preparative route. Among the lipopolysaccharide antigens of the seven recognized immunotypes of Pseudomonas aeruginosa, 2-amino-2,6-dideoxyglucose is also characterized as a constituent of two others, types 3 and 5.


The vascularization of the neural stalk and the pars nervosa of the hypophysis in the toad, Bufo bufo (L.) (amphibia, anura). A comparative light microscopical and scanning electron microscopical study. The angioarchitecture of the neural stalk and the encephaloposthypophysial portal system of the hypophysis of the toad, Bufo bufo (L.), was studied using three different methods. The neural stalk is mainly supplied by branches of the arteria infundibularis superficialis which form a widemeshed vascular network. Dorsally this network continues into the plexus of the pars nervosa. The vascularization of the pars nervosa is made up of the encephalo-posthypophysial portal system. This portal system consists of a hypothalamic branch (=portion), a mesencephalic and a mesencephalicbulbar branch (=portion). The hypothalamic branch was found to drain the pars ventralis of the tuber cinereum as well as more dorsal regions of the diencephalon. The mesencephalic-bulbar trunk enters the hypothalamic branch. The resulting common stem of the encephalo-posthypophysial portal vein the curves around the retroinfundibular communicating artery, crosses its ventral side and runs caudally. The secondary capillary plexus of the pars nervosa is characterized by well defined capillary plexus of the pars nervosa is characterized by well defined capillary networks which are located at the periphery of the parenchyma of the pars nervosa, thus forming a rostral, dorsal and ventro-caudal net. The central region of the parenchyma of the pars nervosa is supplied only by main branches of the encephalo-postpophysial portal vein. The venous drainage of the pars nervosa is via the vena hypophysea transversa.


Subsurface cisterns in the Cynomolgus retina. Subsurface cisterns were found in retinal neurons of the Cynomolgus monkey. They were located in amacrine, bipolar and ganglion cells and were associated with the rough endoplasmic reticulum. Subsurface cisterns were not found in Müller cells. The possible functional significance of the subsurface cisterns is discussed.


Factors for efficiency of the Salmonella/microsome mutagenicity assay. Factors were studied which modify the enzymatic capacity of mouse liver microsomal mixed-function oxidase to convert vinylidene chloride (1.1-dichloroethylene) (VDC) into mutagens in the Salmonella/microsome mutagenicity test. A microsomal fraction incorporated in soft agar layer converted VDC into mutagens during 7 h at a constant rate; these were detected with S. typhimurium TA100. In absence of VDC the enzymatic activity declined gradually to nil after 14 h of incubation at 37 degrees C. The presence of EDTA greatly enhanced the microsome-mediated mutagenicity of VDC and led to prolonged enzymatic viability, but only when liver fractions from phenobarbitone (PB) pretreated mice were used. The efficiency of the plate incorporation assay for the detection of mutagens is discussed in comparison with assays in liquid suspension.