Approach for an equation of state for adsorbed protein surfaces. A surface equation of state for polymer interfaces is obtained by equating the chemical potentials of the solvent in the bulk and surface phases. This equation of state contains the fraction of the surface covered with polymer and the surface activity coefficient as parameters. These parameters may be obtained by measuring the thermodynamic modulus of elasticity. Moreover, if the amount of protein in the interface is known the degree of folding in the surface may be evaluated. The present theory was applied to adsorbed beta-lactoglobulin in surfaces. The results are in agreement with qualitative equations and results from other sources.


Hydrodynamic structure of bovine serum albumin determined by transient electric birefringence. Birefringence relaxation studies on bovine serum albumin (BSA) reveal transient decay described by a double exponential process. The values of the relaxation times lead to estimation of the size of the equivalent ellipsoid of revolution for BSA. Previous measurements of transient birefringence for BSA have shown a single relaxation process, since the apparatus used in obtaining those data was not fast enough to detect the faster relaxation process.


[Frequency and gradient dispersion of viscosity of dna solutions]. Gradual and frequential dispersion of the viscosity of DNA solution in cattle splin was studied. Zimm--Crothers rotational viscosimeter was used for determining gradient relationship of viscosity eta (g). To determine frequential dispersion of viscosity a magnetelecttric vibrational viscosimeter with the working range of 16-100 Hz was produced. It was found that gradient relationship of specific viscosity coincides within the measuring error with the frequential relationship of actual component of specific viscosity at dynamic measurements. The influence of the alternating force with the frequency f is equivalent to the effect of stationary applied force producing the gradient g=k with f, where k=3,2 plus or minus 0,4.


[Letter: Mechanism of K+-dependent swelling of Ehrlich ascites carcinoma cells]. Limiting values of cell volume measured under the conditions of K+-dependent swelling of EAC cells decrease when the medium is hypertonized, the extent of the decrease depending more on the type of the agent applied than on the concentration gradient. According to the action efficiency the agents studied can be placed in the following sequence: Ca2+, succrose Na+,K+. It is concluded that Ca ions strengthen the cell membrane, while K+ ions produce a contrary effect.


[Application of the periodic action method to studying the kinetics of carbon monoxide binding by myoglobin]. To investigate the enzymatic and chemical reactions in a regime of forced oscillations an apparatus has been constructed which permits to observe amplitude-frequency and phase-frequency characteristics of objects in the range of frequencies 10-2-10-6 c/s. This apparatus offers a means of recording stationary and transition kinetics of reactions after the jump-perturbation. Reaction kinetics of CO-binding by whale myoglobin has been studied in the regime of forced oscillations. A phase-frequency characteristics of the system and frequency dependence of averaged (for many periods) myoglobin concentration is measured. Each of the two relationships permits to measure the relaxation times. This possibility has been theoretically predicted earlier by the author.


Primary carcinoma of the gallbladder.

Primary carcinoma of the gallbladder. The clinical and pathological features of 100 consecutive cases of primary carcinoma of the gallbladder, treated by a single surgical unit, are described. Females out numbered males by 2 to 1. The maximum incidence was seen in the fifth decade. Pain and the presence of a mass were the two most common clinical features. Gallstones coexisted in 45 per cent of cases. Ancillary investigations were of little value in preoperative diagnosis. At laparotomy biopsy alone was possible in a large number of cases owing to the advanced disease process. Two patients were alive 5 years after surgery. Prophylactic cholecystectomy for all diseased gallbladders, whether symptomatic or not, is advocated.


[Frequently recurring cyclic megakaryocytopenic-thrombopenic purpura]. An unusual case of acquired frequently remitting megakaryocytopenic-thrombocytopenic purpura in a 54 years old man is reported. The patient could be followed up anamnestically for 11 years. Regularly megakaryocytopenic-thrombocytopenic periods were followed by complete remissions. Platelet survival was normal in thrombocytopenic and in remission phases. No immune mechanism could be detected. The exceptional position of this disorder within the inhomogenous group of intermittent thrombocytopenic purpuras is discussed, and the common criteria with three other published cases are pointed out.


[Fluorimetric study of phospholipase A-induced efflux of Ca2+ from liposomes]. Artificial vesicles, liposomes, were prepared from the total fraction of phospholipids of rat liver mitochondria. Electron microscopy showed that the structure of liposomes depended on cation composition of the medium in which they were formed. Fluorescence of chlorotetracycline increased in the suspension of liposomes loaded with Ca+2 due to the formation of Ca+2-chlorotetracycline-phospholipid membrane complex. Incubation of liposome suspension with phospholipase A in the presence of EDTA resulted in a decrease of chlorotetracycline fluorescence indicating a break of the integrity of liposome membranes and Ca+2 efflux.


Hemangiosarcoma of spleen with spontaneous rupture. A 76-year-old man with spontaneous rupture of a hemangiosarcoma of the spleen, together with a review of 49 previously reported patients with splenic hemangiosarcoma are presented. Twenty-three were women and 27 were men. All but 3 were adults. Fiften were 50-59 years of age. The symptomatology was nonspecific. Three patients developed microangiopathic hemolytic anemia. Spontaneous rupture occurred in 17 cases (34%). Metastases occurred in 42 cases; they were primarily hematogenous and most often in the liver. Lymph node involvement, however, was present in 13 cases. The prognosis of untreated splenic hemangiosarcoma is poor. Three of 25 patients survived a year or more after the initial onset of symptoms. Splenectomy before rupture is advisable, since 5 of 19 patients survived at least 1-5 years. It is suggested that hemangiosarcoma be considered in patients with: 1) splenomegaly without evidence of malignant lymphoma or leukemia, 2) splenomegaly with microangiopathic hemolytic anemia, and 3) unexplained intraperitoneal hemorrhage.


Promotion of experimental breast carcinoma in rats treated with extract of liver from rats with portacaval shunt. Liver homogenates or extracts of liver homogenates from rats in which portacaval shunt had been performed were found to have a significant growth-promoting effect on 7,12 dimethyl (a) benzanthracene (DMBA)-produced breast carcinoma in rats.  Tumor potentiation was manifested by increased incidence of animals developing tumors, increased number of tumors, and increased tumor size, when compared with animals receiving injections of shunted or control liver. These observations suggest the existence of a tumor-stimulating factor in liver from which portal blood has been completely and chronically diverted by portacaval shunt. The demonstration of tumor growth stimulating factor(s) present in shunted liver, together with previously reported observations of the modification of the growth of various types of tumors in animals with a portacaval shunt, suggests that the liver is capable of playing an important role in tumor-host interactions. The portacaval shunt appears to be useful as a technique in elucidating ways that liver function may influence tumor growth.


Complications and total care of a child with acute leukemia. The complications which occur in a child with acute leukemia depend on the stage of the disease and the therapeutic regiman. Most children will present with some manifestation of marrow failure. An occasional child will have marked leukocytosis and disturbance of organ function due to massive leukemic infiltrates. Metabolic disturbances such as hyperuricemia and hyperphosphatemia-hypocalcemia may develop, expecially after therapy is initiated. The myelosuppression and immunosuppression due to drug toxicity may result in opportunistic infections. Other toxicities which can occur with a chemotherapeutic regimen are numerois and varied, and the physician must be cognizant of them in order to minimize damage. Therapy to the central nervous system, either for subclinical or clinical disease, has been associated with a variety of symptoms ranging from meningismus to paraplegia and death. To prevent the development of these complications, and to manage them effectively if they occur, the physician must be knowlegeable about their etiology, clinical and laboratory manifestations, and treatment.


Trends in surgery for childhood rhabdomyosarcoma. Total excision remains the aim of surgical management for rhabdomyosarcomas in children, but effective adjunctive treatment for the majority of these tumors permits preservation of function and appearance in situations where formerly these were sacrificed. Control of residual tumor in microscopic quantity can now be achieved by chemotherapy in the majority of cases. The coordination of surgery, irradiation, and chemotherapy has reduced the need for ultra-radical extirpative procedures. The following conservative trends have evolved: 1) Amputations are rarely indicated for tumors of the extremities. 2) Primary irradiation for orbital tumors may obviate exenteration and save the eye. 3) Multilating excisions for head and neck tumors can often be avoided. 4) Prophylactic regional lymph node dissections are usually not indicated. 5) Primary tumors of the bladder or prostate can usually be managed by anterior exenteration.


Diagnosis and surgical treatment of childhood brain tumors. As the most frequent solid tumor occurring in childhood, brain tumors constitute an important segment of pediatric oncology. Neurologic manifestations may be deceptively mild and easily overlooked or misinterpreted, particularly in the very young, because of the remarkable resiliency of the immature central nervous system and the skull's ability to expand throughout the pre-adolescent years. The majority of childhood tumors produce increased intracranial pressure, usually the consequence of obstructive hydrocephalus. Specific neurologic deficits correspond to the tumor's location. The posterior fossa harbors two-thirds of childhood tumors, and each of the four common tumors in this location produces a characteristic syndrome. Supratentorial tumors occupy the cerebral hemisphere, the suprasellar area, and the pineal gland. Diagnostic studies have reached a state of great sophistication and precise anatomical localization. Surgery, either alone or with adjuvant radiotherapy, cures no more than one-third of all tumors; for the remainder, it has a diagnostic and palliative role. The introduction of operative microsurgery has advanced the art, particularly in the surgical treatment of craniopharyngiomas and pinealomas, but any significant improvement in the treatment of brain tumors as a group seems unlikely to be achieved by surgery alone.


Examination of some factors affecting sensitivity and reproducibility in radioimmunoassay of thyrotropin. Conditions for measuring human thyrotropin by radioimmunoassay have been investigated, to improve the sensitivity and reproducibility of the assay. The Chloramine-T method of iodination was used, the reaction time being 20 s. Doubling the iodination reaction volume from 55 to 95 mul did not affect the immunoreactivity. Purification of labeled hormone by use of anion-exchange resin followed by silica (Quso G-32) instead of Sephadex gel-filtration or anion exchange alone yielded a product that was the least (less than 4%) contaminated with Na125 I. Human serum albumin (2.5 g/liter) in phosphate-buffered saline, instead of bovine serum, should be used as diluent for unknowns; within-assay variance was 3% for the former, 62% for the latter. The assay worked equally well for both pregnant and nonpregnant patients, with use of 50 to 100 mul of serum per determination. A five-week-old labeled hormone yielded higher values than did two-week-old material. Use of sequential saturation techniques (tracer added on day 3) resulted in a greater than 50% drop in B/B-0 ratio in the standard curve between 0.39 and 50 microunits/ml. Somatotropin, choriomammotropin, and procine insulin did not cross react in this system. Human follitropin and lutropin did cross react, and this cross reaction could not be prevented with as much as 40 international units of human choriogonadotropin per tube. With a total reaction volume of 0.4 ml before addition of second antibody, 0.2 ml of 10-fold diluted second antibody yielded a standard curve with lower nonspecific binding and higher maximum precipitation than one constructed by using only 20 mul of second antibody. In 29 euthyroid patients the mean thyrotropin value was 5.7 microunits/ml (range 2.8-11); nine hypothyroid patients had a mean of 112 microunits/ml (range 38-267); and 13 hyperthyroid subjects showed suppressed thyrotropin with a mean of 3.1 microunits/ml (range 2.2-4.5). Following these suggestions, one can expect a more highly purified immunoreactive tracer and a more sensitive assay than is obtained with the procedure from the Pituitary Agency of the National Institutes of Health.


Solid phase radioimmunossay for serum ferritin. A solid-phase "direct" radio immunoassay which is highly sensitive (detecting less than 0.5 ng/ml) and reproducible is described for the assay of ferritin in human serum.  The method makes use of specific anti-ferritin antibody prepared using human liver ferritin;  It has the advantage that tubes coated with specific antiserum may be stored for later use and lends itself readily to an automated washing procedure.  The concentration of ferritin present in the serum of normal males in the population was 56,9 ng/ml (geometric mean) with a 95 percent confidence range of 18 to 180 ng/ml.  In normal females the concentration was 34.0 ng/ml (geometric mean) with a 95 percent condidence range of 10 to 143 ng/ml.


Measurement of deep body temperature from the urine. 1. The relation between urine and rectal temperatures has been studied under a variety of conditions by using the Uritemp bottle, which measures deep body temperature from the urine. 2. This device gives a reliable measurement of deep body temperature and has advantages over both rectal and oral techniques for many clinical applications. 3. The development of a new and more reliable device, the Uritemp meter, is described. Urine is voided through a specially designed and disposable unit, equipped with either a standard mercury-in-glass thermometer or any one of a range of commercially available electronic thermometers. Provision is made to collect a urine sample simultaneously if required.


Changes in the renin-angiotensin-aldosterone system and in sodium and potassium balance during development of renal hypertension in rats. 1. The relationships between the renin-angiotensin-aldosterone system, sodium and potassium balance and systolic blood pressure were studied during development of moderate (160-180 mmHg; clip i.d. 0.25 mm) and severe (200-230 mmHg; clip i.d. 0.20 mm) renal hypertension in rats with an undisturbed contralateral kidney. 2. In severely hypertensive rats renin activity in the peripheral plasma increased from day 9, by which time the systolic blood pressure was elevated to 160-180 mmHg. The rate of total corticosteroid and aldosterone production in vitro increased from day 14 and plasma renin substrate concentration increased from day 24. In moderately hypertensive rats, none of these changes occurred. 3. During the first 10 days after the application of 0.25 and 0.20 mm clips, sodium and potassium retention/g gain in body weight were higher than in sham-operated controls. During the next 10 days, the positive balance stabilized in animals with a 0.25 mm clip whereas, in animals with a 0.20 mm clip, sodium and potassium balance returned to the level of the sham-operated controls through increased renal losses. Despite these changes the systolic pressure rose further in animals with a 0.20 mm clip. 4. The initial sodium retention could be a factor in the early rise of blood pressure and could account for the delay in the rise of peripheral plasma renin activity. The subsequent loss of the retained sodium and potassium during the development of severe hypertension could have facilitated the rise in peripheral plasma renin activity, but did not initiate this rise.


Collagen in the lung: development of a technology applicable to human lung disease. Techniques have been developed to quantitate in vitro the types, rates and fates of collagen synthesized by animal and human lung. Studies are continuing in our laboratory to apply these techniques to investigate important lung disorders, including: (1) classification of the fibrotic lung disorders by the types of collagen synthesized; (2) development of an in vitro drug evaluation system to determine, in a lung biopsy, which drugs are useful in reducing the fibrotic process for a patient with interstitial disease; and (3) investigations of the influence of serum proteins on lung connective tissue synthesis and proteolysis.


Avian parathyroid glands in organ culture: secretion of parathyroid hormone and calcitonin. Parathyroid (PT) glands from 20-day-old embryonic chicks cultured in a chemically defined medium secreted a stimulator of in vitro bone resorption. This stimulator was presumed to be parathyroid hormone (PTH) because: 1) the in vitro dose response curve was parallel to that obtained with bovine PTH; 2) the activity was eluted on Sephadex G-1-- chromatography at a position similar to that for PTH; and 3) the material produced hypercalcemia in vivo in chicks. The amount of PTH-activity secreted was inversely proportional to the calcium concentration of the medium over the range of 0.75-2.25 mM. The chick PT glands also secreted an inhibitor of PTH-stimulated bone resorption in vitro. This inhibitor was presumed to be calcitonin (CT) because: 1) the in vitro dose-response curve was parallel to that obtained with synthetic salmon CT; 2) the activity was eluted on Sephadex G-50 chromatography at a position similar to that for salmon CT; and 3) the material produced hypocalcemia in vivo in rats. In contrast to what would be expected for CT secretion, the CT-activity was secreted by the PT glands in response to a low, not high calcium concentration. The data suggest that the secretion of avian PTH is similar to that of the mammalian hormone, and that the ultimobranchialectomized chick with an intact parathyroid gland may not be deficient in CT.


Effects of ovarian steroids on plasma LH in normal and persistent estrous adult female rats. The ability of ovarian steroids to influence the release of LH was examined after ovariectomy in regularly cycling adult female rats and two types of anovulatory persistent estrous rats: rats treated with 10 mug of testosterone propionate (TP) shortly after birth, and normal rats housed under constant illumination. Two months after ovariectomy the negative feedback effects of a single injection of 20 mug estradiol benzoate (EB) on plasma LH were measured in these three preparations. Ovariectomized (ovex) normal and TP-treated but not ovex constant light-exposed rats had reduced LH values 6 hr after EB treatment. However, LH levels were reduced by the next day in rats in constant light. Positive feedback was measured in similar EB-primed rats following a second injection of EB or progesterone (P). In ovex normal rats LH values remained low in the morning but increased in the afternoon following the second steroid injection. Ovex EB-primed androgenized rats failed to demonstrate this positive feedback. Similarly, in ovex EB-primed constant light-exposed rats, LH values did not increase after a second EB injection. However, high LH values were observed in the afternoon 6 hr after P administration and again 40 and 50 hr later. These results indicate that the lack of spontaneous ovulation in androgenized rats may be due to the absence of positive feedback to ovarian steroids. Constant light-exposed ovex rats do not respond to the positive feedback action of estrogen and show a different response to P than ovex normal rats, in that a regular diurnal peak in plasma LH is not observed.


Similarity of plasma LH release in androgenized and normal rats following electrochemical stimulation of the basal forebrain. Because of conflicting data in the literature, the ovulatory response to electrochemical stimulation (ECS) of the medial septal-preoptic complex was measured in pentobarbital-treated normal proestrous and androgenized female rets. Increasing the stimulus strength increases the incidence of ovulation in normal but not in androgenized rats. However, since the ovaries of androgenized rats are relatively insensitive to LH, whereas pituitary responsiveness to LH-releasing factor is normal, the response to ECS was further evaluated in terms of plasma LH. Immunoassayable plasma LH was significantly elevated 30 min after stimulation. Peak LH levels occurred 90 min after stimulation and were returning toward baseline after 120 min. There was no difference in mean plasma LH values in normal and androgenized rats at any time. Stimulation of the diagonal band of Broca, medial septum or medial preoptic area elicited similar plasma LH responses in both groups. These results indicate that the responsiveness of the medial septal-preoptic complex to ECS is not deleteriously affected by neonatal androgen treatment.


A renin-like activity in pineal gland and hypophysis. The presence and distribution of renin-like activity in hypophysis, pineal gland and some neural tissues were studies in both normal and salt-loaded rats. The Boucher micromethod was used to detect enzymatic activity. In normal rats both the pituitary and pineal glands contained significantly higher values of renin-like activity than did the other tissues examined. In salt-loaded animals there was a significant decrease of the renin-like activity of the glandular tissue while in the other brain areas the activity increased. The results are discussed in terms of the possible physiological role of the central renin-angiotensin system in the regulation of fluid and electrolyte balance.


Plasma TSH levels, by radioimmunoassay, during the estrous cycle of the rat. Between 1000 and 1200 hr on any given morning, female rats displaying regular 4-day estrous cycles were bled by decapitation. Plasma TSH concentrations were determined by homologous radioimmunoassay (RIA), in terms of NIAMDD-Rat-TSH-I-1 standards. Mean plasma TSH concentrations plus or minus SE were 1.2 plus or minus 0.1, 1.5 plus or minus 0.2, 2.3 plus or minus 0.6, and 1.1 plus or minus 0.1 ng/ml for rats killed at proestrus, estrus, diestrus-1, and diestrus-2, respectively. Statistical analysis revealed no significant differences among the TSH levels, except for the elevation at diestrus-1, which is of borderline significance at the 0.05 probability level. Our findings, based on RIA, are in contrast to previous reports, based on bioassays, of an elevation of plasma TSH at estrus.


Failure of injected growth hormone to increase plasma glucose and insulin concentration in the puppy. The resistance of puppies to the diabetogenic effect of administered growth hormone is well known. The possibility that this involves an adaptive increase in insulin secretion was explored by determining the effect of injected growth hormone on plasma insulin concentration at various age periods. Bovine growth hormone (1 mg/kg per day for 4 days) administered to 5-month-old puppies had no effect on plasma glucose or insulin levels nor on the effectiveness of injected insulin to lower plasma glucose levels. At 8 and 12 months of age the growth hormone regimen increased plasma glucose and insulin levels and produced a resistance to the hypoglycemic effect of injected insulin. It is evident that the difference between the puppy and adult in response to growth hormone is not due to excessive secretion of insulin by the puppy.


Synergism between Trichuris suis and the microbial flora of the large intestine causing dysentery in pigs. The role of the microbial flora of the large intestine in experimental Trichuris suis infection was studied by comparing the clinical syndrome in conventionally reared (CR) pigs, specific pathogen-free pigs, and gnotobiotic pigs. Thedisease in CR pigs was characterized by a severe mucohemorrhagic enteritis; in contrast, a mild catarrhal enteritis was observed in specific pathogen-free and gnotobiotic pigs. Spirochaetes and vibrio-like organisms were observed only in CR pigs and increased during the clinical phase of the disease. The clinical syndrome was not transmitted by oral administration of intestinal or fecal material from infected CR pigs to CR pigs free of T. suis. Smaller numbers of T. suis produced diarrhea in CR pigs and significantly reduced the growth rates of infected animals; clinical signs and the reduction in growth rate was prevented by incorporating an antibacterial substance (dimetridazole) in the food. Although clinical trichuriasis closely resembles swin dysentery, the two syndromes seem to be distinct. The present results suggest that a microbial component acts synergistically with T. suis to produce the severe clinical syndrome in CR pigs, but identification of the microbial component and the mechanism by which clinical signs are produced await further studies of the bacterial flora of the large intestine of pigs.


Prostatic effects of a nonsteroidal antiandrogen. The possible mechanisms for the antiprostatic effects of a nonsteroidal antiandrogen, SCH 13521 (4'-nitro-3'-trifluoromethylisobutyranilide), were investigated in rats and dogs. The influence of administered SCH 13521 on the deposition of the radioactivity associated with labeled testosterone, dihydrotestosterone, and estriol (E-3) in the prostate and other tissues of the dog, and rat was determined in short term experiments. SCH 13521 definitely interfered with the localization of the radioactivity of these steroids in the prostate and indicated a competitive situation between SCH 13521 and the steroids. Even though in vitro binding data were generally in accord with in vivo results, he descrepancies regarding E-3 and the more intense effects of SCH 13521 observed in vivo, as compared to those in vitro, lead us to suggest that a metabolite of the compound may also play an important competitive role in vivo. Of particular interest was the competition between SCH 13521 and estrogens in vitro (estradiol-17-beta) and in vivo (E-3). Sch 13521 greatly decreased the volume of prostatic secretion whereas id minor effects on prostatic 5-alpha-reductase and arginase activities. The latter is surprising, since both enzymes are very highly androgen-dependent. Thus, even though the mechanisms of action of SCH 13521 on the prostate may involve competition with androgens at the cellular level, we think that its competition with some estrogens points to a more complicated action than observed with other antiandrogens.


Studies on vesicourethral reflexes. I. Urethral sphincteric responses to detrusor stretch. For the purpose of studies on vesicourethral reflexes, a model was established wherein the vesicosphincteric unit is divided into two separate compartments. In one group, the compartments were formed by single mucosal separation but muscular continuity was maintained. In the other, complete interruption of muscular continuity was performed by full bladder wall thickness separation. To maintain the proper nerve and blood supply to the upper compartment, the separation was made below the ureteral orifices and urinary diversion was accomplished through ureterosigmoidostomy. Postoperative cystographic studies demonstrated the complete separation. Detailed pre- and postoperative physiologic studies were done. Spontaneous contraction of the upper compartment excited contraction of the detrusor component of the lower compartment where muscular continuity was maintained. Contraction of the upper compartment induced reflex relaxation of the sphincteric element, regardless of whether the muscular continuity was maintained or interrupted. Flaxedil and Regitine both led to a drop in resting urethral pressure and abolished the drop in urethral pressure in response to contraction of the upper detrusor compartment. Although muscular contraction might be a contributory factor, it is clear that a nerve-mediated spinal reflex exists that induces drop in urethral resistance in response to detrusor contraction.


New azasteroidal antifungal antibiotics from Geotrichum flavo-brunneum. II. Isolation and characterization. A novel group of antibiotics, comprising microbiologically-active structurally-related factors A25822A, B, D, H, L, M and N, produced by culturing Geotrichum flavo-brunneum NRRL 3862 under submerged aerobic fermentation conditions was isolated by extraction. The individual factors were separated and purified by chromatography and crystallization. The major factor, A25822B, a 15-aza-24-methylene-D-homocholestadiene is a white crystalline compound, C25H45NO. The antibiotics are highly active against fungi and marginally active against bacteria.


Induction of sugar transport in chick embryo fibroblasts by hexose starvation. Evidence for transcriptional regulation of transport. Incubation of chick embryo fibroblasts in glucose-free medium resulted in a dramatic increase in the rate of 2-deoxy-D-glucose transport. The greatest increase in rate occurred during the first 20 hours of incubation in glucose-free medium and was blocked by actinomycin D, dordycepin, or cycloheximide. The conditions of 2-deoxy-D-glucose concentration and time of incubation with the sugar were determined where transport rather than phosphorylation was rate-limiting in sugar uptake. These studies demonstrated that the transport of 2-deoxy-D-glucose was rate-limiting for only 1 or 2 min when the concentration of sugar in the medium was near the Km for transport, i.e. 2mM. No difference was found in the level of hexokinase activity in homogenates prepared from cells incubated glucose-free medium or standard medium when either 2-deoxy-D-[14C]glucose or D-glucose was used as substrate. A kinetic analysis of the initial rates of 2-deoxy-D-glucose transport by Lineweaver-Burk plots showed that the Vmax for sugar transport increased from 18 to 95 nmol per mg of protein per min when fibroblasts were incubated in glucose-free medium for 40 hours. The Km remained constant at 2 mM. Analysis of the initial rates of 3-omicron-methyl-D-glucose transport by Lineweaver-Burk plots further substantiated that the increase in sugar transport was due to an increase in the Vmax for transport with the Km remaining constant. The activation energy for the transport reaction calculated from an Arrhenius plot was 17.4 Cal per mol for cells cultured in the standard medium and 17.2 Cal per mol for cells cultured in the glucose-free medium. These results are consistent with the interpretation that the Vmax increase observed in hexose-starved cells is due to an increase in the number of transport sites.


Interaction of lanthanide ions with bovine factor X and their use in the affinity chromatography of the venom coagulant protein of Vipera russelli. The substitution of trivalent lanthanide ions for Ca(II) in the Ca(II)-DEPENDENT ACTIVATION OF BOVINE Factor X by the coagulant protein of Russell's viper venom was studied at pH 6.8. Factor X contains two high affinity metal binding sites which bind Gd(III), Sm(III), and Yb(III) with a Kd of about 4 X 10-7 M and four to six lower affinity metal binding sites which bind Gd(III), Sm(III) with a Kd of about 1.5 X 10-5M. In comparison, 1 mol of Factor X binds 2 mol of Ca(II) with a Kd of 3 X 10-4M and weakly binds many additional Ca(II) ions. No binding of Gd(III) to the venom protein was observed. Dy(III), Yb(III), Tb(III), Gd(III), Eu(III), La(III), AND Nd(III) cannot substitute for Ca(II) in the Ca(II)-dependent activation of Factor X by the venom protein at pH 6.8. Kinetic data consistent with the models of competitive inhibition of Ca(II) by Nd(III) yielded a Ki of 1 to 4 X 10-6M. The substitution of lanthanide ions for Ca(II) to promote protein complex formation of Factor X-metal-venom protein without the activation of Factor X facilitated the purification of the coagulant protein from crude venom by affinity chromatography. Using a column containing Factor X covalently bound to agarose which was equilibrated in 10 mM Nd(III), Tb(III), Gd(III), or La(III), the coagulant protein was purified 10-fold in 40% yield from crude venom and migrated as a single band on gel electrophoresis in sodium dodecyl sulfate. These data suggest that lanthanide ions complete with Ca(II) for the metal binding sites of Factor X and facilitate the formation of a nonproductive ternary complex of venom protein-Factor X-metal. Tb(III) fluorescence, with emission maxima at 490 and 545 nm, is enhanced 10,000-fold in the presence of Factor X. The study of the participation of an energy donor intrinsic to Factor X in energy transfer to Tb(III) may be useful in the characterization of the metal binding sites of Factor X.


The structure of ferrocytochrome b5 at 2.8 A resolution. Crystals of cytochrome b5 reduced by sodium dithionite are isomorphous with the oxidized form. An electron density difference map between the two forms was calculated at 2.8 A resolution. There are no changes in main chain conformation or internal side chain orientation upon reduction. However, an ion becomes attached at the entrance of the heme crevice causing displacement of a surface lysine side chain on an adjacent molecule. The ion, identified as a cation by the nature of its coordinating ligands, appears to neutralize one of the heme propionate groups which is partially buried. It is proposed that the negatively charged propionate serves to neutralize the net formal positive charge on the heme iron in the oxidized cytochrome and that the neutralization of the heme iron upon reduction then leads to binding of a cation to the propionate.


Chemical modification of amino acid residues associated with the delta-4-3-ketosteroid-dependent photoinactivation of delta-5-3-ketosteroid isomerase. The photoinactivation of the Δ (5)-3-ketosteroid isomerase of Pseudomonas testosteroni in the presence of 3-oxo-4-estren-17β-yl acetate and air is accompanied by destruction of histidine and aspartate (or asparagine). The first order rate constant of photoinactivation of the enzyme is equal, within experimental error, to the first order rate constant for the destruction of a single aspartate (or asparagine) residue and is considerably greater than the first order rate constant for the destruction of a single residue of histidine. When the photolysis is carried out under anaerobic conditions, only aspartic acid (or asparagine) is destroyed as enzyme is inactivated. Both inactivation and aspartate (or asparagine) destruction occur to a greater extent in the absence of oxygen than in its presence. The destruction of histidine, on the other hand, is found to be strictly oxygen-dependent. These results suggest that photochemical modification of a single residue of aspartate (or asparagine) is largely, if not entirely, responsible for photoinactivation of the enzyme under these conditions. When irradiated in the presence of 3-oxo-4-entren-17β-yl acetate, performic acid-oxidized bovine pancreatic ribonuclease does not suffer any detectable destruction of its aspartic or asparaginyl residues but does undergo significant destruction of its histidine residues. These observations suggest that the aspartate (or asparagine) residue modification found with isomerase is an active site-directed photochemical reaction, whereas the modification of histidine may not be.


Sodium transport by the acetylcholine receptor of cultured muscle cells. Activation of the acetylcholine receptors of cultured muscle cells by carbamylcholine increases the rate of passive 22-Na+ uptake into the muscle cells up to 20-fold. The Na+ transport activity of the receptor desensitizes during exposure to carbamylcholine. The rate and extent of desensitization is reduced by lowering the assay temperature from 36 degrees to 2 degrees, allowing accurate measurements of initial rates of Na+ transport by the receptor. Activation of the receptor by carbamylcholine and acetylcholine is significantly cooperative (Hill coefficients of 1.4 to 2.0). Inhibition by D-tubocurarine is not cooperative. The carbamylcholine-induced Na+ transport activity of the receptor is inhibited 50% by 4 muM D-tubocurarine, 100 muM atropine, or 1.6 nM diiodo-alpha-bungarotoxin but is not affected by tetrodotoxin. The initial rate of Na+ transport by the receptor is temperature-independent between 2 degrees and 36 degrees. Receptor Na+ transport is saturable by Na+ at 2 degrees with an apparent Km of 150 plus and minus 20 mM. Saturation by Na+ not observed at 36 degrees at the concentrations tested. Saturation by Na+ is observed at 2 degrees both under conditions of net Na+ influx and under conditions of isotopic exchange at equilibrium. The receptor does not catalyze obligatory exchange diffusion at a detectable rate. Comparison of binding of [125-I]diiodo-alpha-bungarotoxin with rates of Na+ transport indicates a turnover number of 2 times 10-7 ions per min per receptor. These results are discussed in terms of the mechanism of Na+ transport by the receptor.


Changes in adenylate energy charge in Ehrlich ascites tumor cells deprived of serum, glucose, or amino acids. The adenylate energy charge in Ehrlich ascites tumor cells increases when cells are cultivated in serum-limiting medium and decreases when they are incubated in glucose- or amino acid-limited media. Protein synthetic rates decrease in cells deprived of serum, glucose, or amino acids. Supplementation of deprived cells with respective nutrients restores normal protein synthetic rates and adenylate energy charge values. Serum-deprived cells incubated in depleted serum media do not increase their rates of protein synthesis and their adenylate energy charge remains elevated. These results suggest that serum factors regulate protein synthetic rates by mechanisms other than those regulating the availability in cells of glucose or of amino acids. The increased rates of utilization of glucose and of amino acids following the addition of serum are probably due to increased biosynthetic requirements.


The pathway of adenylate catabolism in Azotobacter vinelandii. Evidence for adenosine monophosphate nucleosidase as the regulatory enzyme. Cell-free, dialyzed extracts from Azotobacter vinelandii rapidly dephosphorylate [U-14C]ATP to labeled ADP and AMP, which is then degraded to hypoxanthine, the end product of AMP catabolism under the experimental conditions which were used. The intermediates of the pathway from ATP to hypoxanthine have been identified by thin layer chromatography and quantitated by the 14-C content. The concentrations of intermediates present during the production of hypoxanthine are consistent with AMP nucleosidase being responsible for AMP degradation in these extracts. This result was confirmed in experiments which utilized rabbit antibody prepared against purified AMP nucleosidase. The antibody inhibited AMP nucleosidase activity in cell-free extracts but did not inhibit adenine demanase or adenosine deaminase from the same extracts. In the presence of antibody prepared against purified AMP nucleosidase, the dialyzed extracts showed a marked reduction in the production of hypoxanthine from ATP. Other enzymes which could be responsible theoretically for the conversion of AMP to hypoxanthine were not detected by standard assay procedures. These results are consistent with AMP degradation proceeding by way of AMP nucleosidase to yield adenine and ribose 5-phosphate. The adenine is then converted to hypoxanthine by adenine deaminase. Both of these enzymes were present in sufficient quantities to account for the observed rates of hypoxanthine formation. The rate of hypoxanthine formation decreases during the time course of the [U-14-C]ATP degradation experiments, even though the concentration of AMP remains high. This decrease in the rate of hypoxanthine formation as a function of time is attributed to the decreasing ATP and increasing P0-4 concentrations, since ATP is an activator of AMP nucleosidase and P0-4 is an inhibitor. These observations suggest that the in vivo activity of AMP nucleosidase could also be regulated by changes in the relative ratios of ATP:AMP:P0-4.


The amino acid sequence of a hypothalamic peptide, neurotensin. The amino acid sequence of neurotensin, a hypotensive peptide isolated from acid-acetone extracts of bovine hypothalami, has been established as less than Glu-Leu-Tyr-Glu-Asn-Lys-Pro-Arg-Arg-Pro-Tyr-Ile-Leu-Oh. (The nomenclature and symbols follow the suggestions of the IUPAC-IUB Commission on Biochemical Nomenclature (1972) J. Biol. Chem. 247, 977). This was accomplished by sequence analyses performed on the intact peptide as well as its isolated tryptic, chymotryptic, and papain-generated fragments. The results of enzymic hydrolyses were consistent with the specificities of the enzymes used and indicated that all of the amino acids are unsubstituted and in the L configuration. The absence of non-amino acid constituents was further supported by analyses of electrophoretic mobility-molecular weight relationships of neurotensin and its fragments.


Histone III. VI. Two forms of calf thymus histone III. Microheterogeneity in the amino acid sequence of pea embryo histone III at residue 96 has been established previously. It has been indicated that calf-thymus contains two forms of histone III, with 1 or 2 residues of cysteine, respectively. Evidence is presented here that these two forms are also due to microheterogeneity at residue 96 with one form containing a cysteine residue and the other a serine residue.


Quantitative evaluation of cell orientation in culture. A quantitative method to assess mutual orientation of cells in cultures on a substrate includes the following operations: (1) the cellular groups to be evaluated are chosen; (2) position of the long axis for each nucleus of the group is determined; (3) the axis OX is arbitrary chosen for every group and the angles alphai between the long axis of every nucleus i and the axis OX are measured. Every nucleus i corresponds to a vector of unit length ei with the angles 2alpha. D, the mean of the vectors ei for every cell group is calculated. This value of D is compared with a set of values of D computed according to a model of mutual orientation studies in a simulation experiment. In this model the group of n vectors consists of a fraction of Kn parallel vectors (o less than or equal to K less than or equal to I) and of (I minus K)n randomly oriented vectors. K corresponding to the computed D which is equal to the experimental value of D is considered as an index of orientation for the group. Contact orientation with respect to the relief of the substrate may be evaluated as a root mean square deviation sigma0 to the angles between the long axes of cell nuclei and the direction of relief. Examples of the measurements of K and sigma0 in cell cultures are given.


Linkage of the loci for glucose-6-phosphate dehydrogenase and for inosinic acid pyrophosphorylase to the X chromosome of the field-vole Microtus agrestis. It has been proposed that there are strong selective pressures which have acted during the evolution of mammals to conserve the linkage of genes on the X chromosome. If so, loci that are known to be X-linked in one mammalian species should be X-linked in others. The loci for glucose-6-phosphate dehydrogenase (E.C. 1.1.1.49) and for inosinic acid pyrophosphorylase (E.C. 2.4.2.8) are known to be X-linked in a variety of mammals. The linkage of these loci to the X chromosome of the field-vole, Microtus agrestis, is indicated by the pattern of segregation of these loci in hybrid cells derived by fusion of mouse cells with vole lymphocytes.


Effects of intraruminal and intra-abomasal additions of cod-liver oil on milk fat production in the cow. Four Holstein cows were used in a modified switchback design to show the effect on milk-fat synthesis of (1) control diet, (2) 29.23, 3.92, 6.48, 30.65, 3.44 control diet plus 255 g/day of cod-liver oil in rumen, and (3) control diet plus 225 g/day of cod-liver oil in abomasum. Mean milk yield (kg/day), fat production (kg/day), and milk-fat percentage were (1) 21.87, .79, 3.63; (2) 22.99, .70, 3.05; and (3) 23.02, .75, 3.28. Compared to the post-treatment control period, only one cow decreased in milk-fat percentage on 3 while all cows decreased in milk-fat percentage on 2. Proportions of carbons 16:0, 16:1, 18:0, 18:1, 18:2, and trans-monoene fatty acids of the milk fat were (1) 30.31, 3.24, 10.15, 30.48, 2.59, 7.91; (2) 26.37; and (3) 26.30, 3.72, 10.56, 30.10, 2.96, 9.84. Proportions of rumen volatile fatty acids were not significantly affected by treatment. Proportions of carbons 16:0, 18:0, 18:1, 18:2, and 18:3 fatty acids in the rumen ingesta were (1) 17.28, 47.92, 21.71, 10.59, 2.46; (2) 19.55, 25.90, 37.21, 11.97, 5.39; and (3) 16.90, 50.80, 19.66, 9.11, and 3.23. Fatty acid composition of blood-serum phospholipids were affected by treatment.


Vitamin E adequacy of vegetable oils.

Vitamin E adequacy of vegetable oils. Young male rats were fed diets containing 20 per cent fat in the form of soybean oil, corn oil, safflower oil, or hydrogenated shortening, and their vitamin E status was assessed for twenty-seven weeks. On the basis of growth rate, in vitro red cell hemolysis, plasma creatine phosphokinase activity, and testicular development, soybean oil, corn oil, and shortening provided adequate vitamin E. Rats fed safflower oil had slight red cell hemolysis but were normal in other respects. When the tocopherols in corn oil were reduced by half, vitamin E status still appeared normal. Tissue levels of alpha- and gamma-tocopherols were determined in all groups, and the limitations of the dietary E:PUFA ratio are discussed.


Host-derived antibody-forming cells in lethally irradiated mice. The technique of adoptively transferring an immune response by injecting donor cells into irradiated recipient mice is based on the assumption that the host is rendered immunologically inert by irradiation. We present evidence that these mice are not inert; in the presence of donor cells of various types they mount a substantial immune response against both heterologous erythrocytes and dinitrophenylated bovine serum albumin. This restoration of the host requires donor cells for its expression and can account for up to 100% of the total response. The problems associated with this finding are discussed.


Effects of human C 1 inhibitor on complement-mediated human leukocyte chemotaxis. Human C1 inhibitor (C1 INH) enhances the chemotactic responsiveness of human leukocytes to lipopolysaccharide (LPS), AgAb complex, and zymosan-activated plasma or serum when added to the compartment of the chemotaxis chamber containing the cells. It seems to affect cells directly and causes an increased number of leukocytes to respond to the chemotactic factors at early time intervals. Spontaneous motility does not appear to be affected. Results of studies employing anti-C3 and anti-C5 sera seem to indicate that the chemotactic factor derived from C5 is involved in the C1 INH induced enhancement of chemotaxis. In fact, preliminary experiments utilizing trypsinactivated C3 as a chemotactic source indicate that C1 INH causes inhibition of chemotactic response to C3a. C1 INH is the first naturally occurring plasma component reported to cause enhanced chemotactic responsiveness.


A radioimmunoassay for human antigen-antibody complexes in clinical material. Soluble antigen-antibody complexes have been detected by a radioimmunoassay with rheumatoid factor as the primary antibody to bind 125-I immune complexes or heat aggregated IgG. Bound complexes are precipitated by rabbit anti-human IgM. The binding of the rheumatoid factor to the iodinated altered IgG can be inhibited by preincubation of the rheumatoid factor with human antigen-antibody complexes. The degree to which immune complexes inhibit the binding of rheumatoid factor to heat aggregated IgG has been shown to be dependent upon the antigen-antibody ratio of the complexes, with complexes formed at or near the point of equivalence exhibiting maximum inhibitory activity. Sera from individuals suffering from certain connective tissue disorders were found to inhibit the binding of rheumatoid factor to 125-I aggregrated IgG; sera from normal individuals did not inhibit this reaction when used at comparable dilutions. This procedure can be used as a relatively simple, reproducible means of detecting immune complexes in clinical specimens. The assay is sufficiently sensitive to detect 125 ng of soluble immune complexes.


Quantitative determination of allotypic and idiotypic markers with antisera coupled to N-hydroxysuccinimide-activated sepharose. N-hydroxysuccinimide activated Sepharose beads (HAS) were used to immobilize allotypic and idiotypic antisera for use in radioimmunoassays. These HAS antisera were compared to antisera solidified with ethylchloroformate (ECF) with respect to their reactivity, specificity, and ability to detect subspecificities of the group a allotypes. The HAS antisera rated favorably in all comparisons with ECF antisera and are far superior with respect to ease of preparation and retention of antibody activity.


Antibody-dependent cytolysis of chicken erythrocytes by an in vitro-established line of mouse peritoneal macrophages. An in vitro-established line of mouse peritoneal macrophages (IC-21) was tested for its ability to mediate the cytolysis of 51chromiun-labeled chicken erythrocytes. In the presence of specific antibody, but independently of complement, the macrophages phagocytized and lysed labeled erythrocytes. The phagocytic process proved to be functionally distinct from the cytolytic reaction as demonstrated by enhanced cytolysis in the presence of iodoacetate, an inhibitor of phagocytosis. This cell line, because of its effector activity in antibody-dependent cell-mediated immune reactions, will be useful in characterizing the mechanism(s) involved in macrophage-mediated cytolysis.


Identification of macrophage-like characteristics in a cultured murine tumor line. A mouse tumor line P388D1 passaged in tissue culture for many years has been characterized morphologically and functionally as a macrophage like cell. P388D1 cells phagocytize latex particles and firmly adhere to glass and plastics. In addition they have been shown to carry cellbound receptors for immunoglobulin (Fc) and complement (C3). They fail to stain with fluorescent anti-mouse Ig or heterologous anti-mouse. Functionally, these cells exhibited high effector activity in an antibody-dependent, cell-mediated cytotoxic system. The accessibility of well characterized and pure macrophage-like cell lines, such as P388D1, will facilitate studies where cell purity is essential.


Polymorphonuclear leukocytes in antibody-dependent cellular cytotoxicity. Human polymorphonuclear leukocytes (PMN) lyse antibody-coated target cells in an immunologically specific fashion--antibody-dependent cellular cytotoxicity (ADCC). PMN-mediated cytolysis is independent of complement, de novo protein synthesis, and DNA replication. Cytolysis is rapid, detectable at low PMN:target cell ratios, and exceeds lymphocyte-mediated ADCC. The interaction appears to be mediated via an Fc receptor and is inhibited by aggregated gamma-globulin. A role for PMN in host tumor cell immunity is discussed.


Antigenic analysis of three strains of Mycoplasma arginini by two-dimensional immunoelectrophoresis. The occurrence of species-specific and strain-specific antigens in three strains of Mycoplasma arginini (G-230, leonis and 23243) was studied by two-dimensional immunoelectrophoresis. Approximately 20 antigenic components could be detected in each strain. It was possible to analyze 6 to 7 major and distinct components from each strain by two techniques: "enhancement" where antigen to an additional strain is added to the first phase of the electrophoresis which increases the size of common peaks and "suppression" where antiserum to an additional strain is incorporated in the second phase whereby peak size of components to which both sera have antibody are decreased. A total of 10 distinct antigens were recognized. Electrophoretic mobilities relative to bovine albumin ranged from 0.2 to 1.08. Three components were common to all strains; two of these represented major amounts of material. Four components represented strain-specific components. Unique fast components were found both in strains 23243 and G-230. Three antigens were distributed into only two of the three strains. The electrophoretic mobilities of some common antigens were quite different between strains.


Inhibition by epsilon-aminocaproic acid of the activation of the first component of the complement system. The influence of EACA on C1 in whole human serum and on C1 and C (see article) as isolated molecules was assessed hemolytically. There was selective inhibition of C1 without effect on the levels of C4, C2, C3, and C9 in whole human serum that was reversed by dialysis. EACA was found to inhibit the intrinsic activation of C1 without inhibiting the already active molecule. This was confirmed by the capacity of trypsin to uncover C1 activity in cellular intermediates formed by C1 treated with EACA that did not evolve in the absence of this extrinsic activating mechanism. Inasmuch as the trypsin-dependent recovery of C1 was incomplete, an effect on binding cannot be excluded.


Impaired macrophage function in Friend virus leukemia: restoration by statolon. Phagocytic and migratory functions of peritoneal macrophages from Friend virus (FV) leukemic mice are significantly depressed as compared with normal controls. Leukemic macrophages exposed in vivo and in vitro to statolon, an extract of the mold Penicillium stoloniferum, shown previously to suppress FV erythroleukemia, regain normal function and release reduced amounts of FV. Statolon's in vivo restoration of leukemic macrophage function is paralleled by restoration of humoral immune competence. Statolon induces interferon in vitro but its effects on leukemic macrophages are probably direct, since restoration of macrophage function occurs at dosage levels far below those that induce interferon. These studies suggest that macrophages play an integral role in both the pathogenesis and the statolon-induced suppression of FV disease.


Cytolytic lymphocytic cells with complement receptor in human blood. Induction of cytolysis by IgG antibody but not by target cell-bound C3. Human blood lymphocytes were fractionated on glass bead columns charged with sheep erythrocyte (Es) membranes-bearing human C3b (7,000-10,000 molecules/Es). In the passaged cells the proportion of C receptor lymphocytes was strongly reduced, in parallel with the capacity to lyse chicken erythrocytes (Ec) in the presence of IgG-rabbit anti-Ec antibody. In other experiments, lymphocytes forming rosettes with Es bearing activated rabbit complement [C(ra)] from C6-deficient rabbits were removed by centrifugation through human serum albumin-gelatine mixtures. This procedure also depleted the lymphocyte preparations of antibody-dependent cytolytic effector cells. The results suggest that rations of antibody-dependent cytolytic effector cells. The result suggest that such effector cells have receptors for human C as well as for C(ra). Lymphocytes were not able to lyse erythrocytes bearing either human C3b (similar to 30,000 molecules/Ec) or activated C(ra) in the absence if IgG antierythrocyte antibodies. Under the same experimental conditions these target cells were efficiently lysed in the presence of small amounts of IgG antitarget cell antibodies. This suggests that the interaction between the cellular Fcreceptors and the Fc part of the inducing antibodies is of special significance for the triggering of the cell-mediated lytic reaction. However, although target cell-bound C did not trigger cytolysis, it seemed to potentiate antibody-dependent cytolysis, probably by enhancing effector cell-target cell contacts.


Antibody-mediated inhibition of immune responsiveness in baby chicks. An experimental model for the analysis of antibody-mediated regulation of specific immune responsiveness in chickens is described. Chick embryos and day-old chicks were passively administered homologous serum with high anti-mouse erythrocyte (MRBC) activity and then tested for immune responsiveness to MRBC several weeks later. Compared to the untreated controls, the hemagglutinin titers of experimental birds were obviously depressed for 2 to 3 weeks but near normal by the 4th week. The inhibitory property of homologous serum appeared directly related to the anit-MRBC activity; serum samples collected on day 1 and 2 after immunization were ineffective and low in specific activity, whereas those collected after the 3rd day were progressively more inhibitory as the anti-MRBC activity increased.


Biosynthesis of lysine in Rhodotorula glutinis: role of pipecolic acid. Glutamate-alpha-ketoadipate transaminase, saccharopine reductase, and saccharopine dehydrogenase activities were demonstrated in extracts of Rhodotorula glutinis but alpha-aminoadipate reductase activity could not be measured in whole cells or in extracts. Lysine auxotroph lys1 grew in the presence of L-lysine or DL-alpha-aminoadipate and incorporated radioactivity from DL-alpha-amino-[I-14C]adipate into lysine during growth. Growing wild-type cells converted L-[U-14C]lysine into alpha-amino-[14C]adipate, suggesting both biosynthetic and degradative roles for alpha-aminoadipate. Lysine auxotrophs lys1, lys2 and lys3 of R. glutinis, unlike lysine auxotrophs of Saccharomyces cerevisiae, satisfied their growth requirement with L-pipecolate. Moreover, extracts of wild-type R. glutinis catalysed the conversion of L-pipecolate to alpha-aminoadipate-delta semialdehyde. These results suggest a biosynthetic role for L-pipecolate in R. glutinis but not in S. cerevisiae.


Rapid onset of hyporesponsiveness to interferon induction on re-exposure to polyribonucleotide. Only doses of complexed polyriboinosinic: polyribocytidylic acids sufficient to induce interferon, stimulated hyporesponsiveness to re-induction in rabbit kidney cells. Results of experiments designed to delineate the earliest time of appearance of hyporesponsiveness suggested the time of onset was less than i h after the end of a i h exposure to the inducer. The duration of hyporesponsiveness was about 24 h. Interferon itself did not produce hyporesponsiveness. Hence hyporesponsiveness must be related to an event or a substance synthesized before interferon production. The best candidate is a control protein inhibiting interferon production which is rapidly synthesized following exposure to an inducer.


Micromethod for the titration of lymphocytic choriomeningitis virus in cell cultures. A quantal microassay for the titration of LCM virus strains is described. It is based on the detection of virus-specific complement-fixing antigen in the medium of infected L cell microcultures.


Identification of self-complementary virus-specific ribonucleic acid in chick kidney cells infected with chicken embryo lethal orphan virus. RNA was extracted from primary chicken embryo kidney (CEK) cells infected with chicken embryo lethal orphan (CELO) virus and exposed to a pulse of (5-3H)-uridine late in infection. When this RNA was self-annealed, 4.5% became resistant to pancreatic ribonuclease digestion. The ribonuclease-resistant RNA was isolated by chromatography on Sephadex G-100, and the RNA was found to have the characteristics of a double-stranded molecule of sedimentation coefficient 8S. Half of the column-isolated RNA hybridized to CELO DNA with equal amounts of virus RNA binding to the heavy or light stands of the CELO DNA, indicating the presence of complementary RNA species late in the infectious cycle of CELO.


The relationship between emotionality and dominance in the hooded rat. Hooded rodents were tested in an emotionality condition and two competitive "dominance" conditions. It was found that stable rankings of subjects within the test groups could be established for each of the three conditions. Significant negative correlations were also found between a subject's "emotionality" ranked in a group and his ranking in the group in relation to either condition of competitive dominance, while a significant positive correlation was found between a subject's ranking in both of the competitive dominance conditions. These data are taken as evidence for a debilitating effect of high emotionality on one's success in competition.


Malignant metastasizing angioblastic meningiomas. The authors report and discuss the unusual clinical and roentgenographical features of three cases of angioblastic meningioma with extracranial metastases. The primary lesions occurred in the cervical canal, the parasagittal region, and the posterior fossa. Metastases were to lungs, liver, pancreas, and kidneys.


Maduromycosis of the brain. Case report. A rare case of a gas-containing brain abscess due to verified maduromycosis is described. Blackish material over the abscess cavity revealed the fungal elements. The pus from the abscess contained Streptococcus hemolyticus and Klebsiella organisms.


Liver scan showing intense lung uptake in neoplasia and infection. Two cases of intense lung activity uptake during routine liver imaging are presented. One patient died 6 days after uptake was seen, and had Kupffer cell tumor, or liver angiosarcoma, at autopsy. The second patient with an acute infection superimposed on alcholic hepatitis showed intense lung uptake on the tenth day of a sustained course of very high fever. A repeat liver scan after the patient became afebrile showed no lung uptake.


Computer-based method for calculation of the available energy of proteins. An aspect of protein nutrition that has not been resolved in a satisfactory manner is the utilizable energy equivalence of proteins. The existing methods overestimate the utilizable energy of proteins. A more accurate method fo calculating the utilizable energy of proteins is to calculate the moles of adenosine triphosphate formed during the complete oxidation of a given amount of proteins. The moles of adenosine triphosphate formed can be calculated from knowledge of the amino acid composition and knowledge of the metabolic pathway for each amino acid. A computer program is described that provides the bookkeeping required for these calculations. The available energy of a protein is calculated in this computer-based method by adjusting the energy value of the proteins so that it is equivalent to that of carbohydrates and fats in providing the energy for adenosine triphosphate formation. The computer-based method was used to calculate the available energy of a group of proteins of known amino acid composition. The available energy varied from 3.02 keal/g for colagen to 3


Iron availability to rats from soybeans. Experiments were carried out to determine the effects of endogenous phytic acid in soybean seeds (Glycine max, var. "Amsoy") on the bioavailability of 59Fe to iron-depleted rats. Soybean plants were grown in nutrient solutions labeled with 59Fe supplied at either 0.4 or 1.0 ppm iron in solution. Immature and mature soybean seeds were harvested and fed to male rats as a single dose. The phytic acid content of the immature and mature seeds averaged 0.61 and 1.71% dry weight, respectively. Varying the nutrient solution iron concentration in the mature soybean seeds. When the 59Fe-labeled seeds were fed to iron-deficient rats, 59Fe from the mature seeds was more available than that from immature seeds even though the mature seeds contained approximately three times as much phytic acid. Autoclaving the mature seeds did not affect the absorption of 59Fe by rats from the seeds. Immature soybean seeds contained much more insoluble 59Fe than did mature soybean seeds. It was concluded that the directly correlated to the phytate content of the seeds; also immature soybean seeds apparently contain a factor or factors (other than phytic acid) that depresses iron availability.


Role of hand contamination of personnel in the epidemiology of gram-negative nosocomial infections. Gram-negative organisms, responsible for a high rate of colonization and infection in an intensive care nursery, were recovered frequently from hands of personnel, despite regular hand antisepsis. In sequential hand cultures contamination of hands by gram-negative organisms was transient in some nurses, but active multiplication of the organisms occurred on the hands of others and in a few nurses a temporary carrier state appeared to exist. The results of this study suggest that the hands of hospital personnel may serve not only as a passive vehicle in the nosocomial transmission of gran-negative bacteria, but also may constitute a reservoir of nosocomial organisms.


Dystocia and broad ligament hematoma secondary to hip pinning. A case of dystocia and broad ligament hematoma after hip pinning is presented. Abdominal section was performed after a secondary arrest of labor and failure of descent.


Intramitochondrial bodies in a human cell line carrying oncornavirus-like particles. Electron-dense particles 80-100 nm in diameter were observed in electron micrographs of mitochondria in transformed mouse fibroblasts and in a human amnion cell line infected with supernatants of a human mammary carcinoma cell culture. Intramitochondrial particles were more numerous in the human cell cultures early after subculture. These particles were morphologically similar to those reported in mitochondria of Rous sarcoma virus-infected cells.


Therapy with antibody-coated immune and hyperimmune peritoneal cells in a murine lymphoma system. Goat-antimouse (DBA/2) SL2 lymphoma immunoglobulin was specifically cytotoxic to DBA/2-derived sl2 lymphoma cells. This anti-SL2 immunoglobulin, or a part of it, was cytophylic for peritoneal macrophages as shown by target cells adhering to macrophages incubated in vitro with the immunoglobulin. The target cells became free in the medium after incubation with 0.1% trypsin for 1 hour. In in vivo experiments, the incubation of immune or hyperimmune peritoneal cells with immunoglobulin from normal goat serum or from goat-anti-SL2 serum before use in immunotherapy decreased the number of DBA/2 mice surviving for more than 35 days an intraperitoneal injection with SL2 cells compared to the number of survivors inoculated with immune or hyperimmune cells only. These results show that, in immunotherapy with immune cells, we must consider the possibility that specific antitumor antibodies and antibodies not directed against the tumor cloud the therapeutic potnecy of the immune cells.


Differential oncogenic effects of methylnitrosourea. We investigated differences in the oncogenic effects of methylnitrosourea (MNU) which were induced by varying dose schedules and changing administration routes. The nervous system represented the target organ when MNU was given intravenously to rats. Carcinogen by other routes resulted in decreased numbers of neurogenic tumors and the appearance of neoplasms at the injection site.  Increased oral doses of MNU caused shorter survival times, a decreased incidence of neuroglial tumors, and increased numbers of thymic lymphomas and mesenchymal tumors of the nervous system. The results suggest that many tissues are susceptible to the oncogenic effects of MNU, but the degree of exposure necessary for neoplastic transformation varies.


1-beta-D-arabinofuranosylcytosine-induced chromatid breakage: effect of inhibition of DNA synthesis. The frequency of chromatid breakage induced by 1-beta-D-arabinofuranosylcytosine was decreased when hamster cell fibroblasts were treated with chemical agents that interferred with DNA synthesis immediately before addition of the analog. This phenomenon occurred in both the S and G2 phases of the cell cycle.


Host-cell antigen potentiated by incomplete growth cycle of influenza virus. Lysis of Ehrlich ascites tumor cells in mice was induced with the Hong Kong influenza A-strain HKH virus not previously adapted to the tumor. Despite high pathogenicity of HKH, mice not genetically resistant to the lethal action of myxoviruses survived the actue phase of oncolysis. Virus infection of tumor cells resulted in high titers of hemagglutinin with low infectivity which indicated incomplete virus growth. Serial passages of HKH in Ehrlich ascites tumors failed. HKH oncolysates induced solid antitumor immunity in several mouse strains, including those fully susceptible to the virus. The immunizing power of HKH oncolysates could be abolished by mouse antibody against egg-grown HKH (H3, N2) but not by antiserum raised aganist TUR virus (Havl, Nav3).


Transcription of the genome of adenovirus type 12. I. Viral mRNA in abortively infected and transformed cells. In baby hamster kidney (BKH-21) cells abortively infected with adenovirus type 12, polysome-associated, virus-specific RNA could be detected starting 5 to 7 h after infection. The amount of this RNA reached a maximum between 10 to 12 h after infection and continued to be synthesized at a reduced level until late in infection (48 to 50 h.). In BHK-21 cells transformed by adenovirus type 12 (HB cells), 0.26% of the polysome-associated mRNA was virus specific. The size of the virus-specific mRNA isolated from polysomes of BHK-21 cells abortively infected with, or transformed by adenovirus type 12 was determined by electrophoresis in polyacrylamide gels in 98% formamide, i.e., under conditions which eliminated secondary structure or aggregation of RNA. In abortively infected hamster cells viral mRNA size classes of molecular weights 0.9 times 10-6 and 0.65 times 10-6 to 0.67 times 10-6 were predominant. A minor fraction of 1.5 times 10-6 daltons was consistently found and increased with time after infection. Late after infection (24 to 26 h), viral mRNA of 1.9 times 10-6 daltons was also observed. The size distribution of adenovirus type 12-specific mRNA from transformed hamster cells (HB line) was very similar to that in abortively infected cells, except that the relative amount of the viral mRNA fraction of 1.5 times 10-6 daltons was much higher. It is uncertain whether the viral mRNA of high-molecular-weight represents mixed transcripts derived from integrated viral genomes and adjacent host genes.


RNA polymerase associated with virions of pike fry rhabdovirus. The association of an RNA-dependent RNA polymerase activity with virions of pike fry rhabdovirus has been demonstrated by both in vitro and in vivo studies. The temperature optimum for the in vitro assay is around 20 C, although enzyme activity can be observed at 4 C. Preparations of pike fry virus possess a glycoprotein, a membrane protein, a nucleoprotein, an L protein, and a phosphoprotein, as well as an RNA of about 3.8 times 10-6 mol wt. A protein kinase activity has been found associated with virus preparations. In vitro RNA product analyses indicate that the virus-associated enzyme functions principally as a transcriptase synthesizing viral-complementary, heteropolymeric RNA.


Ribonucleoproteins of Uukuniemi virus are circular. The internal ribonucleoprotein (RNP) of Uukuniemi virus was released with Triton X-100 and analyzed on sucrose gradients. Three species of RNP sedimenting at 140 to 150, 105 to 120, and 85 to 90S could be separated. All of them contained the same ratio of core polypeptide (mol wt, 25,000) to RNA. Eelctron microscopy using rotatory shadowing showed that all three species were circular. Free ends were rarely seen. Measurements of the strands revealed three distinct length classes of about 2.8, 1.4 and 0.7 mu m. Polyacrylamide-agarose gels showed that the largest RNP contained the L RNA, the medium-sized RNP the M RNA, and the smallest RNP the S RNA.


Alterations in Sindbis viral enbelope proteins by treating BHK cells with glucosamine. Addition of d-glucosamine to BHK cells infected with Sindbis virus inhibited the formation of the E-2 viral envelope from its precursor PE-2. Release of virus was blocked, and two new viral protein bands replaced the normal envelope protein bands detected in SDS-gel electropherograms of infected cell extracts.


Complementary strands of CELO virus DNA. When alkali-denatured DNA from CELO virus (an avian adenovirus) was annealed for 15 min at 37 C in 0.1 M NaCl, 70% of the molecules formed single-stranded circles. This is probably due to base pairing of complementary sequences not more than 110 nucleotides long at the ends of the single strands and implies an inverted terminal repetition in the duplex DNA similar to that reported for the DNA from human adenoviruses. The circular molecules had a uniform length that was approximately the same as that of linear single-stranded molecules. The complementary strands of CELO virus DNA were separated on a preparative scale, and at least 40% of the heavy strands and 56% of the light strands were found to be intact as judged by the formation of single-stranded circles.


Effect of fatty acids on growth of Japanese encephalitis virus cultivated in BHK-21 cells and phospholipid metabolism of the infected cells. Growth of Japanese encephalitis virus (JEV) in BHK-21 cells was stimulated in the presence of 20 to 40 mug of the sodium salt of oleic acid (cis-9-octadecenoic acid, 9-18:1) per ml supplemented in Waymouth medium. The stimulatory effect of the salt was highest when 9-18:1 was added after adsorption of the virus. Study of the effect of other fatty acids on growth of JEV showed the following results: the longer the chain length of the saturated fatty acid salt, the higher the stimulatory effect on viral growth. In contrast, polyunsaturated fatty acids had an inhibitory effect on viral growth. The effect of isomeric cis-octadecenoic acids on viral growth was variable, depending upon the position of the double bond. The cis-6-octadecenoic acid had the highest inhibitory effect on growth of JEV compared to other isomeric octadecenoic acids. The sodium salt of (1-14C) cis-9-octadecenoic acid (9-18:1, 20 mug/ml) was rapidly incorporated into control and JEV-infected cells. Specific radioactivity in phosphatidylcholine dropped 12 to 24 h after virus inoculation, whereas synthesis of phosphatidylethanolamine increased 12 to 24 h after virus inoculation in infected cells compared to uninfected cells. Results from these studies suggest that phospholipid metabolism of infected cells is markedly changed, which can be associated with altered fatty acid metabolism when using labeled 9-18:1 fatty acid as a marker.


Complete duplication of the urethra with vaginal stenosis. Complete urethral duplication, clitoral hypertrophy and vaginal urethra in a girl with normal adrenal function are described. These anomalies caused severe micturitional disturbance by several mechanisms. Reported cases of complete duplication form a continuous spectrum of abnormalities and were classified according to anatomical pattern.


Genitourinary aspects of disseminated coccidioidomycosis. Genitourinary involvement is common in disseminated coccidioidomycosis. Urologic evaluation should include careful examination, cultures and radiograms. The radiographic features of renal coccidioidomycosis parallel those of renal tuberculosis, with feathery, moth-eaten calices, infundibular constriction and caliceal ballooning, and eventual calcification of granulomas. Coccidioidal epididymitis may be chronic, with long asymptomatic intervals. A patient with a 15-year interval between episodes of right and left coccidioidal epipidymitis is described. Complement fixation titers should be used to determine the activity of genitourinary coccidioidomycosis. In 3 of 12 patients dissemination of coccidioidomycosis occurred during treatment with immunosuppressive medications. Renal transplant patients should be evaluated for active coccidioidomycosis prior to transplantation.


Lymphangiography in genitourinary cancer. The rationale for pedal lymphangiography in the management  of genitourinary cancer is discussed. The comparative studies between lymphangiography and subsequent histologic section of the nodes in the literature and our own instituiion are reviewed. Itis concluded that while lymphangiography is a safe procedure, it is unlikely to play a major role in the management of cancer of the kidney and penis. If more such studies were performed in cases of prostatic, testicular and bladder malignancies, the diagnostic accuracy might be improved and the lymphangiogram could then help significantly in planning treatment for these patients.


Rhabdomyosarcoma of the spermatic cord: report of 3 cases with review of the literature. Rhabdomyosarcoma of the spermatic cord is a rare, highly malignant lesion, for which the optimum treatment has yet to be ascertained. Results of treatment of 3 patients are presented, making the total reported experiences 53 cases. Two of our 3 patients are alive. One survivor underwent retroperitoneal lymphadenectomy and the other received radical orchiectomy only. The third patient had metastasis at the time of diagnosis and subsequently died. Because of a paucity of knowledge concerning the disease, its propensity for youthful patients and its highly malignant characteristics, all treatment modalities must be considered and tailored to the needs of the individual patient.


Endoparasites of black-bellied whistling ducks in southern Texas. Endoparasites detected in the black-bellied whistling duck (Dendrocygnaautumnalis) include one nasal mite, four nematodes, three trematodes, five cestodes, and two acanthocephalans. Each of these represents a new host record. Overall, 47% of the birds were parasitized with an average of 12 helminths per infected bird. Helminths were significantly more common in juveniles than in adults; more females than males were parasitized, but the difference was not statistically significant. No correlation existed between the parasite load and the physical condition of the host. Tissue damage in infected birds was minimal and limited to nematode cysts in the proventriculi. Parasite loads were light, apparently reflecting the food habits of the host.


The role of valve replacement in the treatment of fungal endocarditis. The role of valve replacement in the treatment of fungal endocarditis has not been defined. The diagnosis of fungal endocarditis was made in 11 patients, 8 before death and 3 at autopsy. Nine patients were infected with Candida species, one with Torulopsis glabrada and one with Aspergillus. None of the 8 patients whose disease was diagnosed during life was cured despite up to 160 days of amphotericin B therapy. All required valve replacement for heart failure (1 patient), emboli (1 patient, or both (6 patients). We recommend early valve replacement for fungal endocarditis before the onset of heart failure and emboli.


Glucagon secretion in unaffected monozygotic twins of juvenile diabetics. Glucagon secretion during a 50-g oral glucose tolerance test has been investigated in 16 nondiabetic monozygotic twins of juvenile diabetics and compared with the results in 10 normal controls and 10 untreated, newly diagnosed, maturity-onset diabetics. Normal subjects showed a significant mean fall in glucagon at 15, 30, and 60 min, with a return to the baseline at 120 min. Maturity-onset diabetics showed a significant mean rise 15 min after oral glucose. The mean of the twin group was intermediate between normals and diabetics, although there was considerable individual variation with some showing suppression and others stimulation of glucagon release. When the twins were divided according to the length of discordance it was found that the mean response in the 8 twins who had been discordant for a mean of 19 years was indistinguishable from that of normal subjects, whereas the mean response of twins discordant for a mean of only 4.5 years was similar to that of the diabetic patients. It is possible, therefore, that hypersecretion of glucagon may occur in some subjects predisposed to develop diabetes mellitus, and the finding of lack of suppression of glucagon in the identical twin of a juvenile diabetic may be of prognostic significance. Identical twins who have been discordant for over 10 years are thought on other grounds to be unlikely to develop diabetes, and the finding of a normal glucagon response is further confirmation of their normal metabolic status and reinforces the suggestion that they are not prediabetic.


[The behavior of the hospitalized child awaiting a surgical operation]. The literature and personal experience are referred to in an assessment of psycho-affective situations in hospitalised children awaiting non-emergency surgery (appendicectomy, adenotonsillectomy, inguinal hernia, phymosis, hypospadias, cryptorchidism). Psychological and somatic damage resulting from an absence of preparation before hospitalisation and during admission immediately prior to surgery is described. A method whereby this may be avoided as far as possible with the aid of relatives is described.


PSRO: an educational force for improving quality of care. The major purpose of the PSRO program is to improve the quality of medical care.  Peer review is expected not only to detect problems but to analyze causes, and to develop, implement, and evaluate corrective programs.  Such peer-review efforts are in themselves educational.  In addition, educational programs focusing on identified deficiencies must be organihe educational efforts of local health facilities and organizations of all types is urged.  Although PSRO's will not specifically underwrite continuing health-care education, they should provide a community-wide means for integrating patient-care review with continuing medical education.


The subunit structure of the eukaryotic chromosome. New strucutral data have been obtained from neutron scattering studies of chromatin. The concentration-dependent meridional peak at 10-11 nm comes from the interparticle spacing of a subunit structure. Peaks at 5.5 and 3.7 nm have a different contrast behaviour to those at 11.0 and 3.7 nm showing that histones and DNA have a different spatial arrangement in the subunit. A globular model in which apolar segments of histones from the core surrounded by DNA complexed with the basic segments of histones agrees with the data.


Computer simulation of protein folding.

Computer simulation of protein folding. A new and very simple representation of protein conformations has been used together with energy minimisation and thermalisation to simulate protein folding. Under certain conditions, the method succeeds in "renaturing" bovine pancreatic trypsin inhibitor from an open-chain conformation into a folded conformation close to that of the native molecule.


Arteriographic correlates of handedness. Bilateral intracranial arteriograms of 123 right-handed patients and 38 left-handed patients were evaluated for: (1) the angulation of the branches of the right and left middle cerebral arteries as they leave the Sylvian fissure, (2) the position of the posterior saggital sinus with respect to the anatomic saggital midline, and (3) the dominant right-sided and left-sided superficial cortical venous drainage (Labbé, Trolard, and Sylvian). Arteriographic evaluations of these points may prove useful in assessing cerebral dominance.


Estrogen-withdrawal migraine. I. Duration of exposure required and attempted prophylaxis by premenstrual estrogen administration. The minimum exposure to estrogen required to cause estrogen-withdrawal migraine has been studied by giving long-acting estradiol valerate to four women and short-acting estradiol benzoate to two women. It was found that several days of exposure to high estrogen levels were needed to cause migraine on estrogen withdrawal. Oral administration of estrogen supplements in the form of estradiol valerate or as conjugated equine estrogens during the premenstrual phase in four women did not significantly affect plasma levels of estradiol, nor was it effective in preventing menstrual migraine.


Estrogen-withdrawal migraine. II. Attempted prophylaxis by continuous estradiol administration. Prophylaxis of menstrual migraine was attempted in five women, using estradiol implants. The treatment was successful in suppressing ovulation and in producing high, although fluctuating, levels of estradiol in the plasma. Clinically this was associated with severe menstrual disturbance. The regular periodicity of the headaches was lost, but clinical benefit was unpredictable, with some patients actually experiencing more headaches than before the treatment. The administration of progesterone against a background of prolonged exposure to high estrogen levels did not provoke migraine, nor in any case did its subsequent withdrawal result in migraine. These findings cast further doubt on the importance of progesterone withdrawal in the etiology of menstrual migraine.


Leukocyte and lymphoid cell counts in myasthenia gravis. Serial leukocyte counts and lymphoid cell counts were compared before and after thymectomy via sternal-splitting incision in 50 patients with myasthenia gravis. Leukocyte counts and lymphoid cell counts of the thymectomized patients also were compared to those of 30 non-thymectomized myasthenic patients and 150 non-myasthenic patient controls. While mean leukocyte counts did not change significantly before and 5.5 plus or minus 5 years after thymectomy, mean lymphoid cell counts were slightly lower during this observation period post-thymectomy (t equals 1.918, p equals 0.03). Lower lymphoid cell counts appeared to correlate with a favorable response of myasthenia gravis in patients with thymic hyperplasia and mainly involuted thymus.


The effect of alcohol on essential tremor. Five patients with essential tremor had a dramatic diminution in tremor amplitude within 15 minutes of ingesting small doses of ethyl alcohol. The same patients were given equivalent amounts of ethyl alcohol infused into a brachial artery, and there was no decrease in tremor amplitude in the perfused limb. It is concluded that, in patients with essential tremor, ethanol acts in a specific fashion on sensitive structures within the central nervous system and has no effect on peripheral tremorogenic mechanisms. This provides additional evidence for a central mechanism in essential tremor, distinguishing it from other tremors arising primarily from oscillation in peripheral servo-loops.


Effect of chronic levodopa treatment on pyridoxine metabolism. Pyridoxine metabolism was investigated in eight chronic levodopa-treated parkinsonian patients and in eight matched dopa-naive controls by measurement of plasma and erythrocyte pyridoxal-5-phosphate concentrations over a 24-hour period, following intravenous administration of pyridoxine. After both a 10 mg and 100 mg pyridoxine dosage, plasma and erythrocyte pyridoxal-5-phosphate concentrations were significantly higher in the chronic levodopa-treated group than in the controls. The percentage change in pyridoxal-5-phosphate concentrations as a function of the administered doses of pyridoxine was greater in the dopa-treated group. Chronic administration of levodopa may result in an adaptive alteration in the decarboxylase system with an increased cellular ability to synthesize pyridoxal-5-phosphate from pyridoxine, possibly through enzyme induction. This metabolic alteration would protect against pyridoxine deficiency states and may explain in part the progressive loss of levodopa effectiveness sometimes observed.


Downbeat nystagmus as a sign of brainstem involvement in acute meningoencephalitis. Two cases of acute meningoencephalitis, presumably viral in origin, with brainstem involvement are reported. Common to both patients during the course of their illness was coma, downbeat nystagmus, and ataxia. Although both reached what appeared to be a terminal state, recovery began following treatment with corticosteroids. The significance of the neurologic signs, particularly the unusual occurrence of downbeat nystagmus, is discussed, as is the appropriate management of the disorder.


Predicting the duration of Guam amyotrophic lateral sclerosis. During the years of study of amyotrophic lateral sclerosis on Guam we have observed a wide range in clinical signs and rate of progression of the disease. Some patients died within 6 months of onset, while others have lived for 20 years. It was our assumption that some aspects of the early neurologic involvement would be related to length of survival, and hence be of prognostic value. We found that an early age at onset and male sex were associated with longer survival. The detailed analysis of degree of involvement of four major neurologic components of amyotrophic lateral sclerosis (progressive muscular atrophy, lateral sclerosis, bulbar paralysis, and pseudobulbar palsy) showed no meaningful pattern of association with duration of illness that could be useful in predicting the course.


Internuclear ophthalmoplegia and associated abnormalities in eye motion timing (differential delays). Studies of the dynamic characteristics of horizontal saccadic eye motion give velocity, acceleration, and timing information. Average time differences between the motion of the eyes on left and right gaze, termed "differential delays," are statistical measures of the average differences of right-going and left-going velocity waveforms, and can be determined to within 0.5 msec. These relative time measurements give information that is not necessarily apparent from the velocity or acceleration data, and provide a sensitive measure of early oculomotor dysfunction. Patients with internuclear ophthalmoplegia have been shown to give characteristic abnormal values, and two such patients are described in detail to illustrate how these measurements may be used both to help in diagnosis and to characterize the presumed lesions.


Midbrain deafness following head injury. A patient is described who became deaf following a head injury. Postmortem examination revealed bilateral lesions of the lateral lemnisci and inferior colliculi. The clinical pattern of midbrain deafness is examined and compared with syndromes of cortical and peripheral auditory impairment.


Deanol in the treatment of levodopa-induced dyskinesias. Levodopa-induced dyskinesias frequently limit the clinical efficacy of levodopa therapy in Parkinson's disease. Deanol recently has been reported to be of value in relieving the dyskinesias by acting through a central cholinergic mechanism. Seventeen outpatients with levodopa-induced dyskinesias were given deanol in dosages of 300 to 900 mg per day. The dyskinesias improved in four of the patients, remained unchanged in five, and worsened in eight. In all four patients who showed improvement after institution of deanol therapy, the improvement continued after the patients were switched to a placebo. One of these patients also demonstrated improvement in his parkinsonism, while two others experienced a worsening in their parkinsonism. Deanol does not appear to be effective in the treatment of levodopa-induced dyskinesias.


The thymus gland in elderly patients with myasthenia gravis. This study was undertaken to determine the amount and histologic appearance of the persistent thymic tissue removed post mortem from 20 patients over 60 years of age with myasthenia gravis. One patient died several days after thymectomy. No recognizable thymic tissue on gross examination was seen in any patient. On microscopic examination, 11 patients had no thymic tissue. The other nine patients, including the one with thymectomy, all showed marked involution of the thymus. No germinal centers were seen. The thymus glands of two additional patients, still alive after thymectomy at ages 62 and 70, showed similar findings. Marked involution of the thymus also was found in each of six elderly controls. Thymectomy is not likely to be effective treatment of the elderly myasthenic patient.


Short-chain organic acidemia and Reye's syndrome. Short-chain fatty acids were determined prior to therapy in seven patients with Reye's syndrome. Elevated concentrations of propionate, butyrate, and isobutyrate were found in all patients. Isovalerate concentrations were high in three patients. In view of the fact that the administration of certain short-chain fatty acids to experimental animals results in coma, electroencephalographic changes, and fatty accumulation in the viscera, the elevations of short-chain fatty acid concentrations observed in the present study suggest that these fatty acids may play a role in the clinical manifestations of Reye's syndrome.


Autonomic dysfunction in botulism B: a clinical report. Nine cases of botulism B with preponderant effects upon cholinergic autonomic innervation are presented. Blurred vision and dry mouth were constant symptoms. Impairment of salivary and lacrimal secretion were detectable for months. In the absence of clinical signs, electromyographic studies did not reveal neuromuscular involvement. Administration of antitoxin in the late course of such cases is not recommended, but guanidine can be used. Doubt is cast upon a new clinical entity called "acute autonomic neuropathy."


Effects of ketamine in epilepsy.

Effects of ketamine in epilepsy. Ketamine, a rapid-acting general anesthetic, was administered intravenously to 26 epileptics. The effects of ketamine on the patients' clinical seizures and electroencephalograms were compared with similar periods during alert and sleep states. Epileptic discharges were present in the alert electroencephalogram of 17 (65 percent) of the patients. Epileptic discharges were precipitated or aggravated by sleep in 15 patients (58 percent) and by ketamine in eight patients (31 percent). No siezures were recorded during ketamine anesthesia. Ketamine neither precipitates nor aggravates seizures and is less effective than natural sleep as an activator of epileptic discharges.


Brief, small, abundant motor-unit action potentials. A further critique of electromyographic interpretation. The concept that there is a constellation of electromyographic (EMG) findings diagnostic of myopathy is criticized. The EMG pattern generally referred to as "myopathic" is one of brief-duration, small-amplitude overly-abundant (for the amount of power being exerted) motor-unit action potentials on slight voluntary effort. For it, the author prefers the descriptive and diagnostically noncommittal acronym "BSAP". In addition to myopathy, three hypothetical neurogenic mechanisms could equally well cause a BSAP EMG pattern: (1) Acquired functional or structural loss of a fraction of the axonal twigs in many individual motor-unit trees, (2) congenital motor-unit hypoplasia (too few muscle fibers per motor unit), and (3) acquired (atrophic) or congenital (hypotrophic) smallness of many muscle fibers. The term and concept of a diagnostic "myopathic EMG" thus is considered invalid.


Patterns of clinical response and plasma dopa levels in Parkinson's disease. Serial determinations of plasma dihydroxyphenylalanine (dopa) in 16 Parkinson's disease patients receiving levodopa showed a negative correlation between plasma dopa levels and disability scores among patients who exhibited daily fluctuations of signs and symptoms. This suggests that the amount of levodopa delivered to the brain from the periphery is of major importance in the production of the "on-off" phenomenon. A close relationship between plasma dopa levels and abnormal involuntary movements was present in six patients. In three a striking dissociation between control of Parkinson's disease and abnormal involuntary movements was present, suggesting that in some patients these two effects are mediated through different underlying mechanisms. Administration of levodopa in such a way as to prevent both high and low levels of dopa in plasma minimizes disability in Parkinson's disease and may lessen abnormal involuntary movements in patients with the "on-off" effect.


Cognitive and motor-sensory performances in toxic and nontoxic epileptic subjects. Psychometric, adaptive ability, motor proficiency, and sensory discrimination measures were administered to adult seizure patients within 36 hours of blood serum determinations of diphenylhydantoin, phenobarbital, and primidone. Thirty-five subjects in whom at least one of the three drug levels fell within stated mug per milliliter toxicity ranges were compared with 28 subjects of comparable age and IQ who were identified by the same procedure to be nontoxic. The toxic group consistently obtained poorer mean test scores, and significant intergroup differences were found on several measures of attention and concentration and on tests of motor coordination and static tremor.


Cerebrovasculopathy following irradiation in childhood. Symptomatic intracranial vasculopathy developed in four patients following irradiation for central nervous system tumors. All the patients presented with a stroke-like picture from 2 to 22 years after the completion of radiotherapy. Two of the patients showed abnormalities on arteriography consisting of narrowing of the supraclinoid portion of the internal carotid artery and the adjacent proximal anterior and middle cerebral arteries. Although the risks of radiotherapy for central nervous system tumors of malignant potential are outweighed by potential benefit, the risks should be carefully considered in cases of tumors with little or no malignant potential.


Infantile spasms associated with histidinemia. A case of infantile spasms associated with histidinemia is presented. Histidinemia was well-documented through biochemical assays. The patient was treated with the standard anticonvulsant regimen for infantile spasms, as well as an elimination diet for histidinemia. Despite low levels of histidine and adequate anticonvulsant therapy, the child continues to have seizures and is markedly retarded. The natural history of infantile spasms and its possible association with histidinemia is discussed.