Emotional complications of adolescent grand mal epilepsy.

Adolescents who have grand mal epilepsy and their parents were interviewed, and the adolescents were evaluated neurologically. Better seizure control and less neurologic disability were unexpectedly associated with less open communication between the adolescents and their families and friends, and with a poorer self-image and poorer expectations for the future. These findings were unrelated to IQ or school performance. This outcome is consistent with other studies of invisible defects and stigmatization, and suggests that youngsters who have relatively mild defects involving social disability may be more troubled than those with more apparent defects, for which denial may be operative. An incidental finding in the study was that a question more predictive of overall family, social, and academic function than the neurologic findings was simply whether or not the youngster was attending the appropriate grade in school for his or her age.


Animal infectivity of Encephalitozoon cuniculi.

Rabbits, mice, rats and Rhesus monkeys were infected experimentally with a rabbit isolate of the mammalian microsporidan Encephalitozoon cuniculi. The lesions produced were typical of those occurring in spontaneous encephalitozoonosis in rabbits, mice, and rats, respectively. Viable E. cuniculi were recovered from tissues of injected animals with and without lesions. Titration of rabbit, mouse, and hamster isolates of E. cuniculi in mice and in rabbit choroid plexus cell cultures showed that the rabbit isolate was equally infectious for mice and cell cultures. Mouse and hamster isolates were less infectious for cell cultures than for mice. The results provide further evidence that the mouse, hamster, and rabbit isolates of E. cuniculi are identical.


Glycogen synthase Hymenolepis diminuta. I. Allosteric activation and inhibition.

Glycogen synthase (UDP glucose: glycogen alpha-4-glycosyltransferase, EC2.4.1.11) of the tapeworm Hymenolepis diminuta exists in 2 forms: 1) the I-form (independent), which has significant activity in the absence of glucose 6-phosphate (G6P); and 2) the phosphorylated D-form (dependent), which has no enzymatic activity unless G6P is present. The activity of the I-form is greatly enhanced by a variety of allosteric effectors which have, as their common feature, 1 or more phosphate groups. These include inorganic phosphate (Pi), several sugar phosphates, some phosphorylated glycolytic intermediates, and nucleoside mono- and triphosphates. Competition studies suggest that while most of the positive effectors act at the same site on the enzyme (the "G6P site"), fructose 1,6-diphosphate (FDP) and 2,3-diphosphoglyceric acid (2,3DPG) act at low concentrations to stimulate the enzyme at another locus (the "diphosphate site"), while at high concentrations they competitively inhibit the binding of G6P and of the other activators. The inhibition by high uridine monophosphate (UMP) concentrations is competitive only with the activator uridine triphosphate (UTP), suggesting the existence of a third type of allosteric site (the "uridine nucleotide site"). This third site may be the locus for feedback inhibition by the product uridine diphosphate (UDP), a control mechanism which has been observed to occur in mammalian systems. The allosteric control of the D-form of the enzyme is comparatively simple, apparently involving only one site (the "G6P site") that binds a few effects with greatly reduced affinity. Pi reverses the activation of the D-form by G6P.


Experimental Babesia microti infections in non-splenectomized Macaca mulatta.

Eight non-splenectomized Macaca mulatta were inoculated intravenously with strains of Babesia microti that originally were isolated from 2 human cases of babesiosis and then were maintained in hamsters in the laboratory. Patent infections developed in 7 animals with peak parasitemias of 496 to 3,906 organisms/mm3 blood. Prepatent periods ranged from 15 to 46 days. Parasitemia persisted for at least 90 days in all animals and in one, organisms were still present 559 days after inoculation.


Hematozoa from Southern African vultures, with a description of Haemoproteus janovyi sp. n.

This study represents the first major survey of avian hematozoa from southern Africa and the only one dealing with blood parasites of vultures. Blood smears from 506 Rhodesian, Botswanan and South African vultures (Hooded, White-headed, Lappetfaced, Cape Griffon and Whitebacked Vultures) were examined for hematozoa. Haemoproteus janovyi sp. n. was observed in 35.2% of the vultures, Leucocytozoon toddi in 0.8%, Plasmodium fallax in 0.6%, Atoxoplasma sp. in 1.4% and microfilariae in 0.2%. Hematozoan prevalence increased with age of the vultures. Only 2 of 133 nestlings sampled during the dry season had patent parasitemias (L. toddi). Haemoproteid prevalence in immature vultures was depressed during the dry season, whereas it was stable throughout the year in adults. The only species which nests on cliffs (the Cape Griffon Vulture) did not harbor hematozoa whereas the other species which nest and roost in trees were infected with at least one hematozoan species.


Glycogen synthase of Hymenolepis diminuta. II. Nutritional state, interconversion of forms, and primer glycogen molecular weight as control factors.

Glycogen synthase I (UDP glucose: glycogen alpha-4-glycosyltransferase, EC2.4.1.11) of the tapeworm Hymenolepis diminuta is the form of the enzyme which is active in vivo, while the D-form represents an inactive "storage form." Utilizing the differential effect of inorganic phosphate (Pi) on the I and D-forms, the ratio of the 2 forms in vivo has been determined under conditions of starvation of the host and refeeding of the parasite with glucose. This procedure reveals that conversion of the inactive D-form to the active I-form takes place when glycogen-depleted worms are incubated in glucose. The activity of glycogen synthase I also is affected by the molecular weight of the primer glycogen. With certain molecular weight fractions, enzymatic activity is higher than with others. This specificity of the glycogen primer could explain the relatively low concentrations of those molecular weight fractions which confer the highest synthase activity.


Susceptibility of owl monkeys to Plasmodium falciparum infection in relation to location of origin, phenotype, and karyotype.

The relationship among geographic origin, phenotype, karyotype, and susceptibility of owl monkeys to 2 strains of Plasmodium falciparum was investigated. Owl monkeys from Columbia and Panama were both susceptible to fatal infections with the Asian FVO (Vietnam-Oak Knoll) strain of P. falciparum. However, when inoculated with the African FUP (Uganda-Palo Alto) strain, most Colombian owl monkeys developed fatal or potentially fatal (bled out with parasitemias of over 25%) infections, but Panamanian monkeys generally survived. Colombian and Panamanian monkeys that spontaneously recovered from malaria infection were phenotypically indistinguishable from those which died. Karyotype analysis revealed that animals considered in this study were either Karyotype II (54 chromosomes) or II (53 chromosomes). Karyotype differences between individual monkeys did not correlate with increased susceptibility or resistance to malaria. Thus, the country of origin of owl monkeys appears to play a more important role in host susceptibility to malaria infection than karyotype.


Inhibition of rat passive cutaneous anaphylaxis by 3-(tetrazol-5-yl)quinolines.

Quinoline-3-carboxylic acid (3) was found to have weak oral activity in the rat passive cutaneous (PCA) assay. In an effort to increase activity, the synthesis of structurally related compounds was initiated. This led to substituted 3-(tetrazol-5-yl)quinolines, some of which are equal in potency, when given orally, to doxantrazole. Further work resulted in the synthesis of 4-oxoquinolines, one of which, 8-chloro-1,4-dihydro-4-oxo-3-(tetrazol-5-yl)quinoline (132), is 33-fold more active than disodium cromoglycate (ip) and 32-fold more active than doxantrazole (po).


Azaprostanoic acid derivatives. Inhibitors of arachidonic acid induced platelet aggregation.

A series of 13-azaprostanoic acids (4a-h) and a 15-azaprostanoic acid (11a) have been prepared. Synthesis of the 15-aza derivative is based on a novel transformation of a ketone to an N-substituted ethylenamine using a formylmethylimino phosphate derivative. Several of the azaprostanoic acid derivatives were found to be potent inhibitors of platelet aggregation induced by arachidonic acid, whereas no effect was observed on ADP-induced primary aggregation, indicating blockade of the platelet arachidonic acid cascade. The compounds do not inhibit bovine cyclooxygenase activity and are postulated as acting beyond the synthesis of the prostaglandin endoperoxides. The inhibitory effect of the 13-aza series is highly sensitive to both stereochemistry and length of the amino side chain. Any deviation from the natural prostaglandin skeletal arrangement results in decreased biological activity.


Folate analogues altered in the C9-N10 bridge region: 11-thiohomofolic acid.

The synthesis of 11-thiohomofolic acid (2) has been accomplished by an unambiguous procedure. Reaction of 1-chloro-4-[p-(carbomethoxy)thiophenoxy]-2-butanone (10) with hydroxylamine under carefully controlled conditions gave the corresponding oxime 33. Conversion of this oxime to 1-phthalimido-4-[p-(carbomethoxy)thiophenoxy]-2-butanone oxime (4) was carried out by its reaction with potassium phthalimide using crown 18 ether as a catalyst. Hydrazinolysis of compound 4 gave 1-amino-4-[p-(carbomethoxy)thiophenoxy]-2-butanone oxime (5), which was used for the construction of the title compound 2 by modification of the Boon and Leigh procedure. An alternate synthesis utilizing 1-hydroxy-4-[p-(carbomethoxy)thiophenoxy]-2-butanone (11) and 4-hydroxy-2,5,6-triaminopyrimidine has also been carried out. Compound 2 did not exhibit any antifolate activity against Lactobacillus casei or Streptococcus faecium. The dithionite reduction product, 7,8-dihydro-11-thiohomofolic acid, was able to function as a substrate of L. casei dihydrofolate reductase. The catalytic reduction product of 2, consisting of a mixture of diastereomers, exhibited powerful antifolate activity against both these organisms.


Methotrexate analogues. 12. Synthesis and biological properties of some aza homologues.

Methotrexate analogues, in which an additional nitrogen atom is inserted between the phenyl ring and the carbonyl group of the side chain, were prepared by photochemical methods. The compounds were less inhibitory toward dihydrofolate reductase and thymidylate synthetase derived from Lactobacillus casei than was methotrexate. They were also less cytotoxic against human lymphoblastic leukemia cells (CCRF-CEM). In vivo against L-1210 leukemia in mice, the aza homologue of methotrexate showed significant antitumor activity (%ILS = 55%) compared to methotrexate (%ILS = 88%).


Synthesis of aza homologues of folic acid.

Folic acid analogues containing an additional nitrogen atom between the phenyl ring and the carbonyl group of the side chain were synthesized. None of the compounds showed significant inhibitory activity against human lymphoblastic leukemia cells (CCRF-CEM) in culture or against Lactobacillus casei (ATCC 7469) growth. Against L1210 leukemia in mice, the aza homologue of folic acid, 4, and the aspartic acid analogue, 14, showed no increase in life span over control animals. These compounds were more toxic in vivo than the corresponding methotrexate analogues. Compound 4 supported the growth of Streptococcus faecium (ATCC 8043), and its tetrahydro derivative supported the growth of Pediococcus cerevisiae (ATCC 8081). These results strongly suggest that 4 can substitute for folate derivatives as cofactors for serine transhydroxymethylase, thymidylate synthetase, and dihydrofolate reductase.


N-(2-Cyanoethyl) derivatives of meperidine, ketobemidone, and a potent 6,7-benzomorphan.

The N-(2-cyanoethyl)-9alpha-ethyl-5-methyl-6,7-benzomorphan (1c) is a more potent antinociceptive and has stronger receptor binding affinity than its N-methyl analogue 1b. The N-(2-cyanoethyl)-4-phenylpiperidine compounds 2b and 3b were almost inactive compared to their N-methyl congeners 2a and 3a, respectively. It appears that the pharmacological effect of the N-(2-cyanoethyl) moiety is dependent on the opioid on which it is substituted.


A computer program for parenteral nutrition solution preparation.

The preparation of single-bottle parenteral nutrition solutions for neonates has long posed a problem for pharmacists. Frequent formulation changes and the need for rapid supply of solutions after prescription has led to a variety of preparation techniques being developed. An interactive computer program is described which enables a nonspecialist pharmacist staff to safely and quickly produce the formula, label and fully document it from a prescription for parenteral nutrition. Ordered quantities are expressed in amounts required per kilogram per 24 hours, and are input to a computer using a "prompt" system. Features of the program include automatic compensations for all non-amino acid ingredients of a variety of commercial amino acid solutions; specified or unspecified chloride anion input; and logic checks which are tied to physiologie "danger" levels, signaling to the operator when "greater than a critical level" has been requested by the prescriber, as well as the conventional check of the logic. The program has allowed infinite variability to prescribing of all ingredients of parenteral nutrition, extremely rapid and accurate production of formulate and allows the computer storage of data about nutritional input to patients. The program is extremely simple in operation, and gives significant cuts in lag time between prescription and the commencement of the new formula.


Distal decompression and proximal feeding for nutritional support during bowel obstruction.

The treatment for acute mechanical intestinal obstruction is a timely operation. A select group of patients may, however, be nutritionally supported with continual administration of elemental diet proximal to long tube decompression under two sets of circumstances: 1. while awaiting spontaneous or treatment-induced resolution of the underlying process, and 2. while reversing catabolism during evaluation prior to operation. Eleven patients with chronic intermittent bowel obstruction were studied: six with obstruction involving radiated small bowel, three with an acute exacerbation of chronic inflammatory bowel disease, one with obstruction secondary to an intra-abdominal phlegmon and one with a segmental motility problem. They received nutritional support with continual gastrointestinal administration of elemental diet proximal to long tube decompression after initial observation for signs or symptoms of altered intestinal viability and stabilization of fluid and electrolyte status. Six of the 11 patients eventually required operation. All patients maintained body weight and three gained weight. Mean nutritional input was 1,873 calories and 12.6 gm nitrogen/day. There were no complications related to the technique of proximal feeding and distal decompression because of careful patient selection and appropriate administration of elemental diet under carefully controlled guidelines.


Human granulocyte and reticuloendothelial system function during intralipid infusion.

In six patients, the phagocytic and catabolic functions of the RES (reticuloendothelial system), as measured by using 125I-microaggregated human serum albumin as a test substance, were not affected by Intralipid infusion. The nitroblue tetrazolium (NBT) reduction of granulocytes, the NBT reduction and the bacplasma, was significantly lower in eight patients during Intralipid infusion than before this treatment. When heat-killed E. coli were added for phagocytic stimulation in vitro, the same result was obtained only in the absence of plasma. The bactericidal capacity of granulocytes was significantly decreased in six patients during Intralipid infusion. When Intralipid was added in vitro to granulocytes, the NBT reduction, and the bactericidal capacity of these cells were decreased and the effect was mainly dose-dependent. Electron micrographs show that granulocytes phagocytize Intralipid. The loading of granulocytes with lipid particles may block the engulfment of bacteria and may be a reason for the decreased bactericidal capacity of these cells. An influence exerted by the lipid on the cell membrane is the most conceivable reason for the decreased NBT reduction.


Specialized nutritional support and cancer.

Over a 7 1/2-year period, 94 cancer patients presenting considerable operative risk were managed by total parenteral nutrition (TPN). All were nutritionally depleted, had obstruction to the gastrointestinal tract or had postoperative complications such as enterocutaneous fistulas, evisceration or intra-abdominal sepsis, which left TPN as the only means of achieving anabolism. When TPN was started preoperatively and continued post-operatively (28 patients--Group 1), no deaths or significant complications occurred. When TPN was first started after life-threatening complications had occurred (58 patients--Group 2), the mortality was 17.2%, so that a high incidence of recovery was achieved. When TPN was used in inoperable cachectic patients (8 patients--Group 3) to enable them to tolerate radiotherapy or chemotherapy, the mortality was 37.5%. This latter group is small, but TPN is worthwhile in selected patients where significant palliation and improvement in the quality of life can occur. With careful technique, risk of sepsis with TPN was no greater than in noncancer patients.


Elemental diet as an adjuvant for patients with locally advanced gastrointestinal cancer receiving radiation therapy: a prospectively randomized study.

Thirty patients with locally advanced, nonresectable, nonmetastatic cancer in the peripancreatic region, stomach and colorectum-anus, to be treated with radiation therapy with or without adjuvant chemotherapy, were randomized to receive standard diet and either usual between-meal feedings or 300 calories tid of a high nitrogen elemental diet. Although weight loss associated with radiation therapy was not significantly reduced in those receiving the nutritional supplement, delayed hypersensitivity skin test responses tended to improve in patients receiving the elemental dietary supplement and to deteriorate in controls. Planned radiation therapy was completed in all nutritionally supported patients. One control patient expired shortly after treatment was halted abruptly, and three other control patients required rescue by total parenteral nutrition.


Intravenous feeding of a complete diet in a child: report of a case.

A 5-month-old white infant with Hirschsprung's disease, complicated by a digestive upset and upper respiratory infection, developed a most extreme picture of marasmus which gave every indication of an impending fatal termination. The baby was given a complete intravenous feeding for five days with fats, carbohydrate, and amino acids in proportions and quantities recommended in a normal infant's diet. There was a prompt dramatic improvement in the nutrition, permitting eventual successful treatment of the Hirschsprung's disease with prostigmine. We believe that this is the first case on record of complete feeding by vein alone for a significant period of time. The success in this child suggests strongly that total feedings by vein can be a practical and lifesaving procedure, especially applicable to children who are marasmic, or are for any reason unable to handle an adequate diet by mouth, or in whom it is desirable to withhold oral feedings for therapeutic purposes.


A review of hypersomolar hyperglycemic nonketotic dehydration (HHND): etiology, pathophysiology and prevention during intravenous hyperalimentation.

The records of 200 patients, nutritionally supported by synthetic means, were reviewed for evidence of clinical hyperosmolar  hyperglycemic nonketotic dehydration (HHND). There was a 3% incidence of morbidity, with a single mortality. Laboratory values demonstrated a positive correlation between persistent glucosuria and HHND. The pathophysiology of HHND demonstrated a relative insulin lack with sufficient insulin to prevent lipolysis, but insufficient to prevent hyperglycemia, glucosuria and osmotic diuresis. The mechanism and management of the pseudodiabetes of stress is reviewed. It is concluded that HHND is an avoidable iatrogenic morbidity. Prevention of osmotic diuresis secondary to glucosuria and, therefore, prevention of HHND is achieved by providing exogenous insulin sufficient to prevent glucosuria.


Measurement of urinary kallikrein acitvity by kinin radioimmunoassay.

We have established a simplified assay system for the measurement of urinary kallikrein activity by utilizing the sensitive and specific radioimmunoassay system of kinins previously reported from our laboratory. Kinins were generated by incubating urine samples (50 microliter) with kininogen (1500 ng) in the presence of kininase inhibitors, and the generated kinins were measured by radioimmunoassay. Since the cross reactivity of kininogen in the kinin radioimmunoassay system was not recognized at dose up to 1.0 microgram, the amount of untreated kininogen in the radioimmunoassay samples did not interfere with the measurement of kinins. This eliminated the necessity for a kininogen extraction procedure. A good linear correlation (r = 0.939, p less than 0.001) was observed between the urinary kallikrein activity determined by this assay system (kininogenase activity) and that by esterolytic acitvity. Urinary kallikrein activity was 3.3 +/- 0.9 microgram/min/24 hour urine (mean +/- SEM), 1.4 +/- 0.4 microgram/min/24 hour urine and 0.25 +/- 0.06 microgram/min/24 hour urine in 6 normal subjects, 7 patients with non-complicated essential hypertension and 4 patients with chronic renal failure, respectively. Thus, urinary kallikrein activity was significantly lower in the patients with essential hypertension (p less than 0.05) and the patients with chronic renal failure (p less than 0.01) than in the normal subjects.


The effect of intravenous hyperalimentation on cell mediated immunity.

The effect of intravenous hyperalimentation (IVH) on cell mediated immunity was examined in 22 patients. Each patient received PHA and PPD skin tests before and after the performance of IVH. In this study both PHA and PPD skin reactivity showed significant increase after IVH, and serum albumin levels had positive correlation with the PPD skin reaction changes. Absence of the established delayed hypersensitivity in the surgical patient, especially those with malignant diseases, is probably secondary to generalized malnutrition, and established cell mediated immunity can be restored by proper nutritional repletion.


Phenol poisoning.

A case report of survival after severe ingestion of phenol is described. The patient developed coma, respiratory arrest 30 minutes postingestion, hypotension, ventricular arrhythmias, metabolic acidosis, seizures, selective elevation of uric acid and gastrointestinal disturbances. Treatment consisted of gastric lavage, olive oil, activated charcoal and supportive therapy. The exact amount of ingested substance was known, and ventricular arrhythmias specifically related to phenol, not its derivatives, could be described.


Effects of low-level X-radiation on 7,12-dimethylbenz[a]anthracene-induced lingual tumors in Syrian golden hamsters.

The effects of repeated low-dose-rate, high-dose-rate X-radiation of the head and neck on lingual tumor induction by 7,12-dimethylbenz[a]anthracene (DMBA) in Syrian golden hamsters were studied. Animals received either topical application as 0.5% DMBA in acetone on the lateral middle third of the tongue three times a week for 15 consecutive weeks, 20-R X-radiation exposures of the head and neck once a week for 15 consecutive weeks, or concurrent radiation and DMBA treatments for 15 consecutive weeks. Animals were examined visually at regular intervals, and all were killed 35 weeks after the start of treatments. All tissues were than examined histopathologically. Animals receiving radiation alone had no detectable changes. Animals receiving DMBA plus radiation had an excess of papillomas compared to animals receiving only DMBA (35% vs. 15%). In addition, an excess of nonlingual oral tumors (lip, gingiva, and floor of mouth) was found in DMBA-treated plus radiation-treated animals versus DMBA-treated animals. These results suggest that repeated, localized, low-level X-radiation exposures enhance chemical tumorigenesis in a variety of oral tissues of Syrian golden hamsters.


Aspects of the control of feeding: application of quantitation in psychobiology.

In rhesus monkeys equipped with indwelling gastric cannulae and studied in the unanesthetized state, it is possible to demonstrate that their feeding is a precisely regulated behavior functioning to maintain caloric intake quite constant. The infusion of nutrients into the stomach through the cannula results in a reduction in their feeding by an amount that is equivalent to the caloric value of the infusion. A similar precise recovery from caloric deficits by overeating can also be demonstrated. One means of controlling intake of food may derive from a remarkable change in gastric function that occurs with feeding. If the stomach is filled with non-nutrient saline it functions like a physiological "pump" expelling its contents into the small bowel in an exponential fashion--more rapidly with increasing volume. If the stomach is filled with nutrients then it functions like a precise "valve," delivering its contents at a constant and linear rate of 0.4 kcal/min to the duodenum regardless (within limits) of the volume, concentration or character (carbohydrate, fat, protein) of the nutrient meal. This change in gastric activity is provoked by the calories that are passed into the duodenum and sustained for a period depending on the amount of calories in the duodenum. The relation of these physiological events to the control of feeding is discussed.


[Prostaglandins in cardiovascular and renal function. Biochemical, physiological and clinical findings (author's transl)].

Prostaglandins (PG) are highly unsaturated, cyclic fatty acids with 20 carbon atoms which are biosynthesized from dihomo-gamma-linolenic, arachidonic and eicosapentaenoic acids. These fatty acids are either ingested or are biosynthesized from linoleic and linolenic acids, respectively. The PG-precursor fatty acids are liberated from membrane phospholipids by phospholipase A and are converted to prostaglandins by the multienzyme complex PG-synthetase. The activity of the PG-system is influenced by extracellular hormonal, neural and mechanical stimuli and by intracellular factors such as ion-concentration and activity of the enzymes adenyl- and guanylcyclase. Prostaglandins are tissue hormones or autacoids which act on their receptors near their site of synthesis and degradation. The prostaglandin family constitutes a group of more than 10 natural occurring compounds showing a variety of biological actions. In arteries and veins the different PG:s have vasodilating as well as vasoconstricting effects. In addition, they are involved in the regulation of vascular smooth muscle proliferation. Within the kidney PG:s have vascular and tubular actions. They antagonize the effect of ADH, mediate renin secretion and are involved in the control of electrolyte balance. In the regulation of platelet aggregation and platelet adhesion PG:s have opposite functions: Prostacyclin which is synthesized in the vascular wall antagonizes the aggregating action of Thromboxane A2 which is formed in the platelets. A defect or an imbalance in the production of PG:s in the vascular wall, in platelets or in the kidney is assumed to play a pathogenetic role in a variety of cardiovascular and renal diseases such as in hypertension, atherosclerosis, persistent ductus arteriosus and Bartter's syndrome.


Drinking of ethanol by adult and infant rhesus monkeys (Macaca mulatta).

Levels of voluntary consumption of ethanol were obtained which were sufficient for use in developing a nonhuman primate model of heavy drinking.


Combined chemoimmunotherapy and radiation therapy of inflammatory breast carcinoma.

Thirty-two patients with a diagnosis of primary inflammatory breast carcinoma were treated with combination chemoimmunotherapy consisting of 5-fluorouracil, doxorubicin hydrochloride (Adriamycin), and cyclophosphamide (FAC) plus BCG followed by radiation therapy. This group of patients was compared to a group of 32 consecutive historical controls treated with radiation therapy alone. The estimated mean disease-free interval for the FAC-BCG group was 16 months, compared to 9 months for the radiation therapy alone group (P less than 0.01). The estimated median survival for the FAC-BCG treated group was 24 months, compared to 18 months for the radiation therapy alone group (P = less than 0.03). The combined modality approach consisting of chemoimmunotherapy and radiation therapy significantly improved the disease-free interval and survival of patients with inflammatory breast carcinoma.


Comparison of intramural 5-fluorouracil and more conventional routes of drug administration on concentrations in gastric regional lymph nodes: a potential for trans-endoscopic adjuvant chemotherapy.

Intramural injection of 20 muCi (2-14C) 5-fluorouacil (5-FU) into the gastric submucosa of dogs and baboons was employed to evaluate the kinetics of 5-FU distribution following this route of administration and to compare these results with those following intraluminal, intravenous, and local intraarterial injections. Measurements of radioactivity taken over a six-hour period after injections demonstrated that the greatest concentration of isotope in samples of gastric wall and perigastric lymph nodes was found after administration of the drug directly into the stomach wall. Clinical application of intramural injections of 5-FU into the stomach wall under direct gastroscopic visual control, may be worthy of trial in patients with gastric cancer.


Lymphocytes subpopulation in normal family members of patients with alpha-chain disease.

Normal family members of eight diagnosed patients suffering from alpha-chain disease had been investigated by immunoelectrophoresis for abnormal alpha-chain protein in their serum. The pattern was demonstrated in four families only, of different members, but all are of the first-degree relationship. The aim of the study was to determine any possible hereditary defect of the immune system in such patients, as compared to the immune system of the normal members of the family who had abnormal protein in their serum. It was found that in both patients and normal members, the proportion of circulating B-lymphocytes was much higher than normal, whereas that of T-lymphocytes was lower than normal. Neither could be sensitized to DNCB, and their skin tests to tuberculin were negative. From all these findings, it was concluded that the disease was a B-cell disease of IgA type transmitted by a hereditary factor associated with a low level of cellular immunity. Further studies are required to support this hypothesis.


Transfer RNA in aging Drosophila: I. Extent of aminoacylation.

Transfer RNA and aminoacyl-tRNA synthetases have been isolated from 5 day and 35 day old Drosophila melanogaster males. Transfer RNA isolated from old males cannot be aminoacylated to the same extent as tRNA from young flies. The average reduction is 16% and 53% for tRNALeu. The aminoacylation ability of some aminoacyl-tRNA synthetases isolated from 35 day old flies is reduced by more than 50%.


Transfer RNA in aging Drosophila: II. Isoacceptor patterns.

Transfer RNA and aminoacyl-tRNA synthetases have been isolated from 5 and 35 day old Drosophila melanogaster males. The isoacceptor pattern of the following tRNAs is not changed irrespective of the source of the tRNA and amino-acylating enzymes: tRNAAla, tRNALeu, tRNAMet, and tRNASer. However, the Q base containing tRNAAsp, tRNAAsN, tRNAHis, and tRNATyr show dramatic changes in their isoacceptor patterns during the aging process. This probably reflects a change in the activity of a tRNA modifying enzyme. These findings are discussed in relation to Strehler's theory of aging and development.


The biology of Encephalitozoon cuniculi.

Encephalitozoon cuniculi is a widespread and often sub-clinical microsporidian parasite of homeothermic animals, including man. The biology, pathology and taxonomy of the organism is reviewed and the available diagnostic methods discussed. Transmission is almost invariably via the oral route either by ingestion of contaminated tissues and other foods or by ingestion of infected urine, perhaps on food, or when animals lick the coats of others. Transplacental transmission does not seem common but can probably occur when susceptible animals are infected during pregnancy. It has been demonstrated once in mice and once in rabbits. The possibility of arthropod vector transmission awaits thorough investigation but this is unlikely to be as important as the oral route. No drugs have yet been found to be effective against E. cuniculi but control of the spread of encephalitozoonosis in laboratory animals, at least, can probably be achieved by maintaining laboratory hygiene.


Air caloric testing in otitis media. (preliminary studies).

On certain occasions it becomes important to evaluate vestibular function in a patient with otitis media. The potential application of the air caloric test in evaluating such patients was examined. Patients with unilateral otitis media before and after surgery were studied to answer certain clinical questions. More questions were raised than answers provided. The preliminary conclusions from this study are: 1. Patients with tympanostomy tubes or small perforation of one ear may show a caloric response in the perforated ear equal to that of the intact ear. 2. Patients with a large tympanic membrane perforation on one side may show hyperactive caloric responses on the perforated side. 3. Patients with a moist ear may show inverted horizontal nystagmus to warm air caloric testing. This applies to patients with a large perforation or mastoidectomy cavity. 4. Patients with a dry open mastoid or fenestration cavity are likely to show a hyperactive caloric response on the side of previous surgery, accompanied by vegetative symptoms. This size of the cavity appears to be less important than the presence of the cavity per se. 5. Patients may be safely tested in the early postoperative period.


Nutritional concepts in the management of the head and neck cancer patient. I. Basic concepts.

Proper nutritional management has assumed an increasingly important role in the care of cancer patients in recent years. Head and neck cancer creates disturbances in host nutritional balances. Profound nutritional depletion and cachexia are induced by local and distant effects of head and neck carcinoma. Radiation, chemotherapy and surgery tend to compromise the host further by compounding these nutritional deficits. This paper reviews the factors responsible for the malnutrition associated with head and neck malignancy, and discusses the current methods available for identifying and following the nutritionally depleted patient.


Nutritional concepts in the management of the head and neck cancer patient. II. Management concepts.

Essential to the management of the head and neck cancer patient is carefully monitored nutritional support. Traditionally, enteral alimentation, using the nasogastric feeding tube, has been the mainstay of treatment. Tube feedings should provide ample amounts of essential nutrients, minerals, vitamins, and adequate calories and protein. Knowledge of the tube feedings available and problems associated with their administration helps to avoid the pitfalls which limit their effectiveness. The inadequacies of enteral alimentation preclude its use in selected circumstances of severe nutritional depletion. Parenteral hyperalimentation, as a primary or adjuvant mode of therapy, may be capable of rapidly reversing deficits, improving postoperative morbidity and increasing tolerance to radiation and chemotherapy. Postoperative deglutition abnormalities may prolong the nutritional problems of head and neck cancer patients as well.


Adrenocortical function in experimental protein malnutrition.

Adrenocorticoid activity was investigated in experimentally induced protein malnutrition in Rhesus monkeys. Control studies were carried out in the same animals before inducing protein malnutrition. Plasma cortisol levels were elevated in the protein malnourished state. There was a total abolition of the diurnal rhythm of cortisol secretion. Fasting hypoglycemia was also observed in the protein malnourished state. It is concluded that increased adrenocorticoid activity and sustained steroidogenesis result from protein deficiency. Hypoglycemia may be an important stimulus, in addition to the metabolic stress imposed by the protein deprivation. The recognition of increased adrenocorticoid activity is important in a protein-deficient host, since the defense against infections might be impaired in such a situation.


Treatment of diabetes mellitus: the future.

For more than half a century the management of hyperglycemia in diabetes mellitus has included rigid diets and intermittent subcutaneous insulin administration. These methods have been totally unsuccessful in restoring glucose homeostasis to normal in most diabetic patients. This review focuses on techniques that offer promise as alternatives or adjuncts to the current modalities of treatment. Specific areas discussed include pancreatic transplantation, islet cell transplantation, artificial beta cell devices, and the glucagon-suppressing agent somatostatin. Although many of these show promise for the future, a cure for the metabolic abnormalities of diabetes is not imminent.


The genome of B. subtilis phage SSP1: the topology of DNA molecules.

DNA molecules of B. subtilis phage SPP1 exhibit terminal redundancy and are partially circularly permuted. This was established by the hybridization of selected EcoRI restriction fragments to single strands of SPP1 DNA and by an analysis of the distribution of denaturation loops in partially denatured SPP1 DNA molecules. Deletions in SSP1 DNA are not compensated by an increase in terminally repetitious DNA. This finding, which is unique to SPP1, is discussed in terms of a modification of the Streisinger/Botstein model of phage maturation.


Evidence that induction and suppression of mutations and recombinations by chemical mutagens in S. cerevisiae during mitosis are jointly correlated.

Mutagen-induced intergenic and interallelic recombination as well as forward mutation were studied in one and the same strain of S. cerevisiae. In nontoxic dose ranges, the induction of mutants and recombinants was parallel after treatment with ethyl methanesulfonate (EMS), methyl methanesulfonate (MMS), N-methyl-N'-nitro-M-nitrosoguanidine (MNNG), triethylene melamine (TEM), 4-nitroquinoline 1-oxide (4-NQO), sodium nitrite (NaNO2), and 1-fluoro-2,4-dinitrobenzene (2,4-DNFB). Acridine orange (AO) after treatment without light induced recombinants, but reduced the frequency of spontaneous mutations. In combination with TEM, AO exerted the same effect, i.e., reduced its mutagenic effect and enhanced its recombinogenic effect. 4,5,6-Trichloro-2-(2,4-dichlorophenoxy) phenol (Cl5-predioxin) induced mutants and intergenic recombinants, but specifically reduced the spontaneous frequency of interallelic recombinants. In combination with TEM, it enhanced its mutagenic and intergenic recombinogenic effects but reduced its interallelic recombinogenic effect. The main conclusions of the present study, that is 1. Essentially similar lesions can lead to different genetic consequences, and 2. Induction of mutation and recombination are jointly correlated, i.e., suppression of mutations leads to an enhancement of recombinations, while suppression of recombinations leads to an enhancement of mutations, are used to set up a speculative concept for mutation and recombination induction in the diploid yeast cell during mitosis.


The genome of Bacillus subtilis phage SPP1: the arrangement of restriction endonuclease generated fragments.

SPP1 DNA was cleaved by the restriction endonucleases, BglI, BglII, EcoRI, KpnI, SmaI, and SalI. The molecular weights of the DNA fragments obtained by single enzyme digestion or by consecutive digestion with two enzymes were determined by electron microscopic measurements of contour length and by gel electrophoresis. The major fragments from the six digests could be ordered to give a consistent restriction map of SPP1. The electropherograms of several digests indicated that certain fragments occurred in less than stoichiometric amounts or were heterogeneous in size. Such bands carried a major part of radioactivity, when SPP1 DNA was terminally labelled with P32 prior to degradation by restriction enzymes. These results, and studies of the effect of exonuclease III treatment on restriction enzyme patterns define the terminal restriction fragments. All data obtained support the conclusion drawn in the preceding paper (Morelli et al., 1978 b) that the SPP1 genome is terminally redundant and partially circularly permuted.


Different behaviour of Xbb+ and Xbb chromosomes in D. melanogaster with only one nucleolus organizer.

We have examined the rDNA content of male and female adult flies having only one nucleolus organizer (NO), using X chromosomes carrying wild or partially deleted bobbed loci (Xbb+/O, Xbb+/XNO- and Xbb/O, Xbb/XNO-). The results show that in Xbb+/O and Xbb+/XNO- flies, where only somatic gene compensation is supposed to occur, the rDNA increase, althought less pronounced than previously reported, is directly proportional to the number of rRNA genes initially present in the nucleolus organizer. In Xbb/O and in Xbb/Xbb/XNO- flies the rDNA increase is relatively much higher than that observed in flies carrying bb+ instead of bb. It is suggested that this may be due to rDNA premagnification and somatic gene compensation occurring simultaneously in the former flies.


Mapping of the loci involved in the catabolic oxidation of L-hydroxyproline in Pseudomonas aeruginosa PAO.

Genes specifying the oxidative utilization of hydroxyproline (Hyp) in P. aeruginosa PAO were located on the chromosome, around 19th minute by conjugation experiments. A map order of his-68-his-07-Hyp was assigned. Confirmation of this gene order was also demonstrated by transductional mapping studies. All the genes determining the enzymes of Hyp dissimiliatory pathway were closely linked.


Ultrastructure of the capsule of Bacillus megaterium ATCC 19213.

The ultrastructure of the firmly adherent capsule produced by Bacillus megaterium cultured on fructose mineral salts medium was examined using thin sectioning, freeze-etching, and critical point drying by transmission and scanning electron microscopy. The capsule material was shown to be fibrillar, with most fibrils containing bulbous protrusions. Two types of fibres were resolved. These were termed primary and cross-linking fibres. Primary fibres originated at the cell wall and had a diameter of 34-50 nm. They also contained bulbous protrusions and enlarged areas where branching occurred. Cross-linking fibres connected the primary fibres. The cross-linking fibres were much smaller, usually 15 micro m in diameter, and contained few enlarged areas. The primary fibres originated at sites on the cell wall approximately equidistant and 0.26 micro m apart.


Spasticity due to phenytoin toxicity.

A young epileptic presented with spasticity as well as ataxia, diplopia and nystagmus; his serum phenytoin level was very high. All the abnormal signs disappeared after withdrawal of phenytoin. Spasticity, hyperreflexia, and clonus are features of phenytoin intoxication, present in this case, which are not commonly seen, and which have rarely been mentioned previously in the literature.


[Possible role of macromolecular components in the functioning of photosynthetic reaction centers of purple bacteria].

The temperature dependencies of the photoconversion of pigments P870--P890 were studied using isolated chromatophores and photosynthetic reaction centres (RC's) of purple bacteria. The samples were prepared by extraction with organic solvents (light petroleum and a combination of light petroleum and methanol) and modified through cross-linking the functional groups of proteins by treatment with glutaraldehyde or denatured by various physical and chemical treatments. The data provide further evidence that the pool of RC secondary acceptors is formed by the compounds of quinone nature located in the hydrophobic surrounding. Similar molecules localized in a more polar medium act as primary acceptors. The findings indicate on the essential role of macromolecular components in the RC's functioning and also suggest that the photochemical charge separation is conformation-controlled.


[Heterochromatin and chromosomal polymorphism].

The structure and sizes of different short deletions of Drosophila melanogaster X-chromosome: left break in the region of locus y-ac-sc, right break in different parts of the heterochromatic region (HR) near the centromere, of the nuclei of salivary glands and metaphases of oogonia were compared. It is suggested, that the development of large blocks of deoxynucleoproteins by a small number of loci of the HR, observed in mitotic chromosomes, is due to their ability to undergo additional replication, as it previously has been shown for locus bobbed. HR in human chromosomes provide high polymorphism of the chromosome sets. The same chromosome can appear in several varieties within a population. The varieties differ in sizes of the HR and their ability for intensive fluorescence. The appearance of these heredical varieties are conditioned by deletions, duplications and inversions in the HR. Studies on revealing boundaries between normal and pathological HR polymorphism of human chromosomes are in progress.


[Spectral position of the principal absorption band of pigment complex P870 and the kinetics of photo-induced oxidoreductions in the reaction centers and chromatophores of purple bacteria with preparations at different temperatures and having different degrees of hydration].

In isolated photosynthetic reaction centres of Rps. spheroides and chromatophores R. rubrum the spectral position of the longest wavelength absorption band of P870, effectiveness of electron removal from the photochemical pair (P870 -- primary electron acceptor, A1) and the rate constant for recombination of photooxidized P870 with photoreduced A1 undergo marked and fully reversible changes over the temperature interval from +20 to -70 degrees. Dehydration of the samples has the effect similar to that induced by temperature lowering. The data suggest that the spectral position of the main maximum of pigment complex P870 absorption band may be regarded as a sensitive inner probe of the structure-functional state of the investigated preparations.


A radioimmunoassay method for the rapid detection of Candida antibodies in experimental systemic candidiasis.

Rabbits were employed as experimental models to evaluate a solid-phase radioimmunoassay (RIA) method for the diagnosis of systemic candidiasis. Ten rabbits were inoculated subcutaneously to mimic superficial candidiasis and were found to produce no antibodies to Candida as determined by both immunodiffusion and RIA procedures. However, 94 per cent of 18 rabbits systemically infected by intravenous injection of Candida cells were observed to produce antibody as assessed by the RIA technique. These data encourage further tests with human sera and the continued development ofthis RIA procedure as a useful tool in the early serodiagnosis of systemic candidiasis.


t-Haplotypes of the mouse may involve a change in intercalary DNA.

The naturally occurring t-haplotypes of the mouse exhibit a set of peculiar genetic properties, including strong suppression of crossing over in the segment of chromosome 17 between the loci of T and H--2. Study of the genetics of mutant haplotypes suggests that the observed effects on meiosis and embryonic development may be due to an altered form of intercalary DNA (iDNA) in the relevant chromosomal region (band 17B).


Autonomic nervous dysfunction in patients on long-term hemodialysis.

The level of plasma dopamine-beta-hydroxylase (DBH) activity in subjects at rest was found to be significantly lower in 12 patients on long-term hemodialysis than in a healthy 8-member control group: 28.3 +/- 7.2 and 13.6 +/- 7.6 IU/1, respectively (p less than 0.01). Following immersion of one hand of each subject into cold water (4 degrees C) for 1 min, a significant rise was observed in both groups, 6.1 +/- 4.8 IU/1 for the control and 1.6 +/- 1.4 IU/1 for the patient group (p less than 0.01). Upon tilting up the head of all subjects, activity in both groups increased significantly, but a markedly smaller rise was found in the patient group: 5.8 +/- 4.8 and 1.1 +/- 1.6 IU/1 for the two groups, respectively (p less than 0.01). The data suggest an autonomic nervous dysfunction in patients on long-term hemodialysis.


Diamine oxidase activity in plasma and urine in uremia.

Diamine oxidase activity was measured in plasma or urine in 12 normal men, 4 men with chronic liver or heart disease, 13 men with chronic renal failure, and 12 men undergoing maintenance hemodialysis. Also in five studies in 4 patients, plasma diamine oxidase activity and total amine levels were measured at hourly intervals during a hemodialysis treatment. Plasma diamine oxidase activity was normal in patients with liver or heart disease and was at least three times normal in chronically uremic patients and in patients undergoing maintenance hemodialysis. Plasma diamine oxidase activities before and after a hemodialysis therapy were similar and did not change during dialysis until the 4th hour when they fell transiently; plasma total amine levels, which were elevated initially, tended to rise during the 4th hour of dialysis. Urinary diamine oxidase activity was reduced in the chronically uremic patients as compared to normal subjects. These observations are consistent with three alterations in diamine oxidase in patients with renal failure: activity (a) is increased in plasma of chronically uremic patients and those undergoing maintenance hemodialysis, (b) does not increase normally in response to heparin administration during dialysis therapy, and (c) is reduced in urine of chronically uremic patients.


Brain region differences and some characteristics of monoamine oxidase type A and B activities in the vervet monkey.

Monoamine oxidase in the vervet monkey showed greater variations in activity in six brain regions when tyramine or phenylethylamine was used as the substrate (3.8- to 4.1-fold differences) than when serotonin was the substrate (1.8-fold differences). With phenylethylamine and tyramine as substrates, the highest MAO specific activities were found in the hypothalamus and the lowest in the cerebellum and cortex. With serotonin as the substrate, the highest specific activities were in the mesencephalon and cortex. The inhibition of tyramine deamination by clorgyline and deprenyl yielded biphasic plots indicative of the presence of MAO-A and MAO-B enzyme forms in the vervet brain. On the basis of these inhibitor curves, the vervet brain could be estimated to contain approximately 85% MAO-B and 15% MAO-A, in contrast to rat brain which contains 45% MAO-B and 5% MAO-A. The inhibition of serotonin deamination by deprenyl in vervet brain yielded a biphasic plot, suggesting that some serotonin deamination in the vervet is accomplished by the MAO-B enzyme form. Estimations of the relative amounts of MAO-A and MAO-B based on inhibitor curves or based on substrate ratios yielded proportionate results which were in close agreement across the different brain regions, supporting the validity of these approaches to estimating MAO-A and MAO-B activities.


Myasthenia gravis and monoclonal IgG gammopathy.

Monoclonal IgG gammopathy of the lambda light-chain type was detected in a 51-year-old woman who had unexplained fever, muscle fatigue, and myalgia. One year later, myasthenia gravis was diagnosed. There was no evidence of myelomatosis or other malignancy. On the assumption that her M-component (gammopathic paraprotein) was related to myasthenia, she was treated with melphalan and cyclophosphamide, but her clinical condition was not improved. Despite therapeutic trials of other agents and a time course of 6 years, the quantity of the M-component remained unchanged. Serum AChR antibody activity was not located in the paraprotein peak. The findings do not support a relationship between the M-component and myasthenia gravis.


Effects of phenobarbital and 3-methylcolanthrene treatment on microsomes of Morris hepatoma 3924-A, tumour-bearing and normal rat liver.

Microsomal cytochromes and some oxidative activities were determined in normal rat liver, tumour-bearing rat liver and Morris hepatoma 3924-A. Except for a moderate lowering of cytochromes and enzymes in host livers, the relation between TPNH-cytochrome c reductase activity and cytochrome P-450 TPNH reduction, both increased by phenobarbital (PB) and decreased by 3-methylcolanthrene (3-MC) treatment, is noteworthy. In tumour cytochromes b5 and P-450 are absent and TPNH-cytochrome c reductase is unmeasurable and not induced by PB or 3-MC treatment. Aminopyrine demethylase activity, instead, is comparable with normal or host liver and it is modified by PB or 3-MC treatment in the same way, despite the microsomal enzymes pathway disorganization. Microsomal enzymatic defect selectivity in tumours may be due to a deranged microsome-linked growth control.


The laboratory diagnosis of plasma cell myeloma and related disorders.

Plasma cell myeloma constitutes the commonest primary malignant tumor affecting the skeleton of the adult. It is also one of several forms of monoclonal gammopathy that include Waldenström's macroglobulinemia, the heavy chain diseases, "benign" (nonprogressive, asymptomatic) monoclonal gammopathy, and primary amyloidosis. The diagnosis of classic or advanced myeloma rests on the findings of sheets of plasmacytic cells in a bone marrow biopsy sample or aspirate; a serum monoclonal immunoglobulin usually of the IgG or IgA class or Bence Jones protein in the urine; and evidence of multiple lytic lesions in x-ray views of the skeleton. Pitfalls in the diagnosis of earlier or unusual forms of myeloma require a careful application of classic diagnostic criteria or of criteria established by clinical cooperative groups.


Traumatic pharyngeal pseudodiverticulum in neonates and infants. Two case reports and review of the literature.

Thirty one cases of pharyngeal pseudodiverticulum have been reported in the literature; twenty nine were diagnosed during the neonatal period. Respiratory distress, increased oral secretions, difficulty with feeding and the impossibility of passing a nasogastric catheter were the most common symptoms and/or signs. Pneumomediastinum, pneumothorax, cervical emphysema and ectopic location of a feeding catheter, alone or in combination, were identified in the chest roentgenograms of 16 patients. Esophagography and/or endoscopy were the diagnostic methods of choice. The exact location of the perforation was identified in 18 patients. Most of the perforations were in either the posterior pharyngeal wall or in the pyriform sinuses. The survival rate was as good amongst the medically treated patients as in those who underwent surgery.


Fatal tuberculosis in young children.

Three young children with advanced cavitary pulmonary tuberculosis died between 1974 and 1978. Two of the patients had tuberculous meningitis as well. All diagnoses were verified at autopsy; however, all tuberculin skin tests were negative. These instances emphasize the difficulty of establishing the diagnosis of tuberculosis in young children. Tuberculosis should be considered in the differential diagnosis of cavitary chest disease in young children, even those with negative tuberculin skin tests.


The use of rifampicin to evaluate tRNA transcriptional organization in Escherichia coli.

The antibiotic rifampicin, which in prokaryotes inhibits the initiation of RNA synthesis but not the completion of nascent strands, was used to explore tRNA gene transcriptional organization in Escherichia coli. Cultures were grown in [32P] orthophosphate to constant specific radioactivity and labeled with [3H] uridine in the presence of rifampicin. Numerous tRNA species then were isolated by polyacrylamide gel electrophoresis and their 3H/32P ratios determined; these ratios, following correction for the base compositions of the tRNAs, should reflect the distances of the corresponding tRNA genes from their promoters. Individual tRNA species were identified, where possible, by oligonucleotide fingerprint analysis. Observed isotopic ratios were correlated with promoter-gene distances, measured in nucleotides, using the nucleotide sequence of the 16S ribosomal RNA gene as a reference. The protocols developed should be applicable to most prokaryotes.


Sequence of 1000 nucleotides at the 3' end of tobacco mosaic virus RNA.

The sequence of 1000 nucleotides at the 3' end of tobacco mosaic virus RNA has been determined. The sequence contains the entire coat protein cistron as well as regions to its left and right. Sequence characterization was by conventional methods for use with uniformly 32P labeled RNA complemented by newer methods for in vitro 5' and 3' 32P end-labeling of RNA and its subsequent rapid analysis. The noncoding region separating the coat protein cistron from the 3' terminus is 204 residues long and may be folded into a clover-leaf-type secondary structure. The distribution of termination codons to the left of the coat protein cistron suggests that the end of the adjacent cistron is separated from the beginning of the coat protein cistron by only two nucleotides. The subgenomic viral coat protein mRNA was isolated from infected tissue and shown to be capped. The nontranslated sequence separating the cap from the AUG initiation codon is 9 residues long and thus overlaps a portion of the adjacent cistron on the genome RNA.


Cooperative interactions among protein and RNA components of the 50S ribosomal subunit of Escherichia coli.

Copperative interactions among constituents of the 50S ribosomal subunit of Escherichia coli have been analyzed in order to elucidate its assembly and structural organization. Proteins L5 and L18 were shown to be necessary and sufficient to effect the association of the 5S and 23S RNAs into a quaternary complex that contains equimolar amounts of all four components. Measurement of diffusion constants by laser light scattering revealed that integration of the 5S RNA induced the 23S RNA to adopt a somewhat more open conformation. An investigation of relationships among proteins associated with the central and 3' portions of the 23S RNA demonstrated that attachment of L5, L10 + L11, and L28 depends upon the RNA-binding proteins L16, L2, and L1 + L3 + L6, respectively, and that L2 interacts with the central segment of the 23S RNA. These data, as well as the results of others, have been used to construct a scheme that depicts both direct and indirect associations of the 5S RNA, the 23S RNA, and over two-thirds of the subunit proteins. The 5' third of the 23S RNA apparently organizes the proteins required to nucleate essential reactions, whereas a region within 500 to 1500 bases of its 3' terminus is associated primarily with proteins involved in the major functional activities of the 50S ribosomal particle.


The regulatory region of MS2 phage RNA replicase cistron. III. Characterization of fragments resulting from S1 nuclease digestion.

The functionally active fragments MS2 R(-53 leads to 6) and MS2 R(-53 leads to 3) comprising the regulatory region for the replicase cistron have been isolated from MS2 RNA-coat protein complex following T1 RNase digestion. In order to obtain shorter fragments, active in coat protein binding and initiation of translation, MS2 R(-53 leads to 6) was cleaved with S1 nuclease. The results indicate that S1 nuclease attacks the most susceptible loop regions of the two hairpin helices of MSZ R(-53) leads to 6). Among the three fragments which have been isolated, only MS2 R(-35/33 leads to 6) containing the intact hairpin (b) region with initiation codon AUG is active in the coat protein binding. Functional activity exerted by another polynucleotide MS R(-17 leads to 6) supports the assumption that specific binding with the coat protein is determined by the hairpin (b) region prior to the replicase cistron.


Initiation of translation with Pseudomonas aeruginosa phage PP7 RNA: nucleotide sequence of the coat cistron ribosome binding site.

Initiation complex formation between PP7 RNA and ribosomes of Pseudomonas aeruginosa and Escherichia coli has been investigated. The PP7 RNA fragments protected by both species of ribosome have been isolated, and their sequences have been determined. Only one binding sites is available on the intact PP7 RNA strand, and this site is recognized by ribosomes of both species. The PP7 RNA binding site is approximately 38 nucleotides long. It contains two AUG sequences and a purine-rich segment near the 5'-end that is complementary to segments near the 3'-ends of the 16S ribosomal RNA's of both P. aeruginosa and E. coli. In order to establish which of the AUG codons acts as the initiator, the H2N-terminal amino acid sequence of PP7 coat protein was determined. This sequence is compatible with the codon sequence following the second AUG codon. The extent of the reaction of PP7 RNA with E. coli ribosomes is greater than with P. aeruginosa ribosomes, but our results do not indicate a qualitative difference in the initial interaction between intact PP7 RNA and the ribosomes of either species.


Nucleosome periodicity in HeLa cell chromatin as probed by micrococcal nuclease.

When HeLa cell nuclei were treated with micrococcal nuclease (nucleate 3-oligonucleotidohydrolase, EC 3.1.4.7), lysed, and centrifuged, the supernatant from early digests contained two predominant classes of polynucleosomes of repeat size 8N and 16N. With increasing digestion time, the 16 N polynucleosome appeared to be cleaved to the 8N species and finally to the basic subunit of chromatin. The size of the polynucleosomes has been determined by DNA analysis and on polyacrylamide electrophoretic gels of native chromatin particles. The 16N polynucleosome appears to be a unique higher ordered structural component of HeLa cell chromatin. Our recent report, showing that the nuclear protein-modifying enzyme poly(ADP-ribose) polymerase increases in specific activity progressively with increasing nucleosome repeat size up to 8-10N, has been extended in the present study. Activity was also elevated in the polynucleosomes of the 16N structure preferentially cleaved by micrococcal nuclease, although specific activity of the enzyme was highest in octanucleosomes. Acceptors for poly(ADP-ribose) have also been determined in these particles.


Positive control of lac operon expression in vitro by guanosine 5'-diphosphate 3'-diphosphate.

Maximal expression of the Escherichia coli lactose operon in a coupled in vitro transcription-translation system from a Salmonella typhimurium relA mutant was strongly dependent upon addition of guanosine 5'-diphosphate 3'-diphosphate (ppGpp). Without added ppGpp, at saturating 3',5'-cyclic AMP (cAMP) concentrations, synthesis of beta-galactosidase (beta-D-galactoside galactohydrolase, EC 3.2.1.23) was reproducibly only 5-7% of that which can be obtained with 0.5-0.8 mM ppGpp. Experiments in which transcription was uncoupled from translation indicated that this 14- to 20-fold stimulation by ppGpp occurred at the level of transcription. When coupled beta-galactosidase synthesis was primed with a template containing a well-characterized mutant lac promoter (lacP(r)L8UV5), the dependence on ppGpp was greatly reduced. This result provides an important experimental control previously unavailable for verifying the significance of ppGpp effects on gene regulation in vitro; it indicates that activation of lacP(+) expression by ppGpp is specifically an effect of increased transcription initiations. Furthermore, the large ppGpp stimulation of lacP(+) DNA enabled the level of expression of this template to approach that of lacP(r)L8UV5 DNA, an observation expected from results in vivo but not obtained with other transcription-translation systems in vitro. The importance of these results is considered with respect to previous ideas on the physiological role of ppGpp as a supercontrol molecule in bacterial regulation.


Tryptic dissection and reconstitution of translocation activity for nascent presecretory proteins across microsomal membranes.

The ability of microsomal membranes to translocate nascent presecretory proteins across their lipid bilayer into the intravesicular space was investigated by using trypsin as a proteolytic probe. We found that under defined conditions trypsin is able to dissect the translocation activity of microsomal membranes into components that can be separated into two fractions, one soluble and the other membrane bound. The trypsinized membrane fraction has lost its translocation activity. Addition of the trypsin-generated soluble fraction, however, results in reconstitution of translocation activity. These results are compatible with the notion proposed in the signal hypothesis that the translocation activity of the microsomal membrane resides in transmembrane protein(s). We propose that trypsin effects solubilization from the membrane of cytosol-exposed domain(s) involved in recognition of the signal sequence or ribosome or both, leaving behind membrane-integrated domain(s) that provide the environment for the passage of the nascent chain across the membrane. Signal peptidase activity was unaffected by trypsinization of microsomal vesicles consistent with a localization of the active site of this enzyme on the cisternal side of the vesicles.


Selective growth of some rodent epithelial cells in a medium containing citrulline.

We have defined a medium (called Sun's modified Waymouth medium) that selectively cultures some rodent epithelial cells that are capable of using citrulline in place of arginine. A growth-response study of the ability of 47 different mammalian cell cultures (of mouse, rat, Syrian hamster, Chinese hamster, guinea pig, rabbit, monkey, and human origin) to use arginine or its biosynthetic precursors, ornithine, citrulline, or argininosuccinate, showed that all epithelial cells and some fibroblasts are capable of growing in citrulline medium; however, primary embryo fibroblasts and 12 established fibroblast cell lines derived from Syrian hamsters failed to grow. The citrulline medium also allowed selective outgrowth of epithelial cells, without contaminating fibroblasts, from Syrian hamster tracheal explants. This absolute nutritional difference between Syrian hamster epithelial and fibroblast cells allows citrulline medium to be used for selective cultivation of epithelial cells, which should be valuable for study of growth, differentiation, and malignant transformation of mammalian epithelial cells.


Lateral and vertical displacement of integral membrane proteins during lipid phase transition in Anacystis nidulans.

Alterations in membrane structure as a result of lipid phase transitions have been studied in Anacystis nidulans, a blue-green alga. Cells grown at 38 degrees C were subjected to temperature transitions of 38 degrees C leads to 21 degrees C and 38 degrees C leads to 10 degrees C, previously shown to produce substantial changes in photosynthetic activities, and examined by freeze-fracture electron microscopy. As a result of these treatments, large particle-free regions appeared on the fracture faces of both the plasma and thylakoid membranes. Particle density measurements suggest that the displacement of the integral membrane protein complexes occurs in both lateral and vertical directions. Returning the cells to 38 degrees C resulted in the restoration of normal membrane morphology, indicating that the proteins were not lost from the membrane. Such displacement of the integral membrane protein complexes could contribute significantly to the temperature-dependent alterations in the functional activity of membrane-bound enzymatic complexes.


Rapid evolution of animal mitochondrial DNA.

Mitochondrial DNA was purified from four species of higher primates (Guinea baboon, rhesus macaque, guenon, and human) and digested with 11 restriction endonucleases. A cleavage map was constructed for the mitochondrial DNA of each species. Comparison of the maps, aligned with respect to the origin and direction of DNA replication, revealed that the species differ from one another at most of the cleavage sites. The degree of divergence in nucleotide sequence at these sites was calculated from the fraction of cleavage sites shared by each pair of species. By plotting the degree of divergence in mitochondrial DNA against time of divergence, the rate of base substitution could be calculated from the initial slope of the curve. The value obtained, 0.02 substitutions per base pair per million years, was compared with the value for single-copy nuclear DNA. The rate of evolution of the mitochondrial genome appears to exceed that of the single-copy fraction of the nuclear genome by a factor of about 10. This high rate may be due, in part, to an elevated rate of mutation in mitochondrial DNA. Because of the high rate of evolution, mitochondrial DNA is likely to be an extremely useful molecule to employ for high-resolution analysis of the evolutionary process.


Bilirubin diglucuronide synthesis by a UDP-glucuronic acid-dependent enzyme system in rat liver microsomes.

Incubation of rat liver homogenate or microsomal preparations with bilirubin or bilirubin monoglucuronide with (BMG) resulted in formation of bilirubin diglucuronide (BDG). Both synthesis of BMG and its conversion to BDG were critically dependent on the presence of UDP-glucuronic acid. Pretreatment of the animals with phenobarbital stimulated both reactions. When 33 microM bilirubin was incubated with microsomal preparations from phenobarbital-treated rats, 80-90% of the substrate was converted to bilirubin glucuronides; the reaction products consisted of almost equal amounts of BMG and BDG. When phenobarbital pretreatment was omitted or when the substrate concentration was increased to 164 microM bilirubin, proportionally more BMG and less BDG were formed. Homogenate and microsomes from homozygous Gunn rats neither synthesized BMG nor converted BMG to BDG. These findings in vitro suggest an explanation for the observations in vivo that, in conditions of excess bilirubin load or of genetically decreased bilirubin UDP glucuronosyltransferase (EC 2.4.1.17) activity, proportionally more BMG and less BDG are excreted in bile.


Selection and characteristics of a Vibrio cholerae mutant lacking the A (ADP-ribosylating) portion of the cholera enterotoxin.

After mutagenesis with nitrosoguanidine and selection by immuno-halo techniques, an avirulent mutant, designated Texas Star-SR, which produces no detectable A (active; ADP-ribosylating) region of the cholera enterotoxin (choleragen) but produces the B region (choleragenoid) in amounts similar to the hypertoxinogenic wild-type parent Vibrio cholerae (biotype E1 Tor serotype Ogawa), has been isolated. The mutant retains the colonizing ability, motility, prototrophy, and serologic characteristics of the parent. In relevant intestinal experimental models, it has been shown to be avirulent and to induce protection against challenge with virulent cholera vibrios. The mutant appears to be suitable for further evaluation in volunteers as a candidate living enteric vaccine against cholera and related enterotoxic enteropathies.


A genetic approach to analysis of transposons.

Integration of the tetracycline resistance transposon Tn10 into lacI of a lacI-lacZ gene fusion permits the isolation of deletions that excise DNA from one end of Tn10 and fuse Tn10 genes with lacZ in such a manner that chimeric proteins with beta-galactosidase activity are produced. The synthesis of the chimeric proteins is under the same control as the transposon genes. Thus, regulation of expression of Tn10 genes can be investigated by measuring beta-galactosidase activity. Analysis of Tn10-lacZ fusions revealed different deletion endpoints within Tn10; lacZ has been fused to at least three different Tn10 genes or operons. Two of these genes are under the control of a tetracycline repressor.


Reevaluation of level of genic heterozygosity in natural population of Drosophila melanogaster by two-dimensional electrophoresis.

We have found the two-dimensional electrophoretic technique of O'Farrell to be highly efficient in the detection of charge-change substitutions in a large number of proteins. We have applied this method to determine the level of heterozygosity of the most abundant proteins in Drosophila melanogaster adults from a natural population. The estimate of per-locus heterozygosity obtained from approximately 54 loci screened was 4% with 6 loci polymorphic. This is much lower than overall estimates obtained by standard gel electrophoresis but is not different from estimates for "Group I" enzymes--i.e., those utilizing a narrow spectrum of substrates of intracellular origin. We consider these data to throw open the question of the level of genetic variability in nature.


Mutation generating a fragment of the major heat shock-inducible polypeptide in Drosophila melanogaster.

Drosophila melanogaster tissues carrying a third chromosome with the deletion Df(3R)Kar(D2) make a 40,000-dalton (Dal) heat shock protein not made by wild type. The unusual polypeptide was inducible in every tissue examined. Tryptic peptide fingerprints showed it to include part of the 70,000-Dal major heat shock protein. Mapping experiments placed the mutation responsible for the 40,000-Dal protein at or close to the kar(D2) deletion. One break point of the deletion is in subdivision 87A, close to or at a heat shock locus that codes for the 70,000-Dal protein. The results are consistent with the possibility that this break point is within a gene for the 70,000-Dal protein, leaving only the initial portion of its coding sequence. This would specify the direction of transcription of the mutant gene as proximal to distal on the normal chromosome. The 87A heat shock locus should contain at least two genes for the 70,000-Dal protein, because embryos homozygous for the kar(D2) deletion and lacking the heat shock locus at 87C, which also codes for the 70,000-Dal protein, nevertheless produced both the 40,000-Dal and the 70,000-Dal proteins upon temperature elevation. Using the presence of the 40,000-Dal protein to monitor chromosome segregation, we found that embryos homozygous for deletions of the heat shock puff site at 93D exhibited a normal electrophoretic pattern of heat shock proteins.


The effect of metal complex of thyrotropin-releasing hormone on locomotor activity of neonatal chicken.

Neonatal chickens were injected intraventricularly with Ni(II), Pd(II), Cu(II) or Zn(II) complex of TRH and the potencies of stimulating locomotor activity were compared with that of TRH, Ni(II)-TRH was more potent than the ligand while Pd(II)-TRH was inert. Cu(II)- and Zn(II)-TRH induced the response to the same extent of the ligand. These results indicate that the action of TRH in the CNS resulting in the locomotor hyperactivity is dependent on the tertiary conformation of molecule which is modified by chelate formation.


Interactions of acetylmethadol or methadone with other drugs in rhesus monkeys.

Behavioral effects and blood or plasma levels of d-amphetamine, ethanol, cocaine, and diazepam were examined in rhesus monkeys treated chronically with alpha-l-acetylmethadol (LAAM), methadone, or vehicle. Chronic treatment with the opiates failed to alter blood or plasma levels and behavioral effects of d-amphetamine or ethanol. LAAM-maintained monkeys were somewhat less sensitive to rate-decreasing effects of cocaine on schedule-controlled responding, but cocaine plasma levels and half-lives generally did not differ across the chronic treatment conditions. Behavioral depression after diazepam was prolonged substantially in LAAM- and methadone-maintained monkeys, but blood levels of diazepam and metabolites were not increased prolonged in those animals. Naloxone partially antagonized the residual depression LAAM- and methadone-maintained monkeys 24 hr after diazepam, but had no effect on the weaker sesidual depression in vehicle-maintained aniamals. Thus, diazepam appeared to interfere with the metabolic inactivation of the opiates. One LAAM-maintained monkey showed recurrent episodes of LAAM overdose and eventually died during the course of the study.


Effects of the neuropeptide DG-AVP on morphine and food self-administration by dependent rhesus monkey.

Pretreatment with the neuropeptide DG-AVP (desglycrinamide9-arginine8-vasopressin) at two dose levels (25 and 125 mcg/kg) did not reduce intravenous morphine self-administration (0.25 mg/kg/inj) by morpine dependent monkeys, in comparison to pretreatment with saline or DG-AVP vehicle placebo. Food self-administration was also unaffected by DG-AVP pretreatment in comparison to control conditions. These data do not confirm previous reports of a dose-dependent suppression of heroin self-administration in rat following DG-AVP pretreatment [14].


Synthesis of quaternary 8-hydroxyquinoline and 8-hydroxyquinaldine-carbamates structurally related to cholinesterase.

The quaternary carbamates were synthesized by reaction of 8-hydroxyquinoline and 8-hydroxyquinaldine with alkylisocyanates, then quaternizing the carbamates produced with alkylhalides, aralkylhalides or sulphate. The UV spectra of the compounds obtained showed lambdamax at 285 nm. The new quaternary carbamates were found to be of high toxicity and parasymapathomimetic activity in white mice.


Skin necrosis from extravasation of intravenous fluids in children.

From 16,380 administrations of I.V. fluid in children during a 6-month period, there were 1,800 extravasations (11%) with 40 skin sloughs. All that resulted in either partial or full-thickness skin loss were treated by one of 3 conservative protocols, and no skin grafts were needed in this series. Most of the extravasations resulting in skin loss were associated with hypertonic fluids and the use of infusion pumps. Careful hourly monitoring of such cases seems highly desirable. We found no discernible differences in the healing among the 3 treatment regimens used. The importance of systematic monitoring of children receiving I.V. fluids by nursing personnel, the elevation of an extremity involved in an extravasation, and the care of any resulting wounds are discussed.


Effects of piracetam on brain monoamine metabolism and serum prolactin levels in the rat.

Levels of monoamine metabolites in three different regions of the rat brain were determined following treatment with piracetam (0.5 and 5 g/kg, i.p.). The concentration of prolactin in serum was also measured. Piracetam, at 5 g/kg, increased the levels of dihydroxyphenylacetic acid, homovanillic acid and 3-methoxy-4-hydroxyphenylethylene glycol, whereas 5-hydroxyindoleacetic acid was unaffected. The drug also increased prolactin concentrations in serum. The level of dopamine was unchanged in the olfactory tubercle and the striatum. These effects are different from those obtained with amphetamine-like drugs. The results would seem to indicate that piracetam accelerates brain catecholamine (CA) turnover via a blockade of CA receptors, as suggested for neuroleptic drugs. This effect could be responsible for the therapeutic action of piracetam on psychotic symptoms in psycho-organic disorders of old age. A blockade of brain CA receptors by piracetam is not compatible with facilitated learning, which seems to be mediated via other neuron systems than CA pathways.


Cholinergic involvement in ethanol intoxication and withdrawal-induced seizure susceptibility.

The enzymes of the cholinergic system have been investigated in discrete brain areas in alcohol-dependent rats, which were still intoxicated or were undergoing withdrawal. The ethanol intoxication resulted in a slight, but significant increase in choline acetyltransferase (CAT) activity in the caudate nucleus both 1 and 7 h after the last dose of ethanol. We also found a significant decrease in CAT activity in the temporal limbic cortex while rats were highly intoxicated. All other brain regions investigated, e.g., cerebellum, pons-medulla, frontoparietal cortex, hypothalamus and septum showed unchanged CAT activity. Rats were also analysed immediately following the onset of a withdrawal-induced audiogenic convulsive seizure where, in addition to the striatum, depressed CAT activity was observed in the hippocampus. In all the analysed situations acetylcholinesterase activity remained unchanged. These results show that ethanol intoxication leads to a perturbation in the synthetic capacity of acetylcholine in certain defined brain structures and that this may have some correlation to the observed behavioural impairments.


Serotonergic function in mouse head twitches induced by lithium and reserpine.

We examined the relationship between lithium-induced head twitches and serotonergic neurons. Head twitches were elicited by combined treatment with lithium chloride (2 or 5 mEq/kg x 5, s.c. administered hourly) and rauwolfia alkaloids, i.e., reserpine (5 mg/kg, s.c.), tetrabenazine (20 mg/kg, s.c.), and syrosingopine (10 mg/kg, s.c.). Neither lithium nor the alkaloid alone induced the twitches; nor did combined administration of lithium with methamphetamine or p-chloroamphetamine. The head twitches induced by lithium in combination with reserpine were strongly inhibited by antiserotonin drugs, methysergide and cyproheptadine, and also by a serotonin synthesis inhibitor, p-chlorophenylalanine (PCPA), when administered between lithium and reserpine. When PCPA was administered before lithium for 3 days, the head twitches were potentiated. In addition, the head twitches were potentiated by a serotonin receptor stimulant, 5-methoxy-N,N-dimethyltryptamine. The results imply that lithium can induce head twitches in the presence of rauwolfia alkaloids and may exert its effect in part by acting on the serotonergic neuron system.


[Influence of lithium and rubidium on exploratory behaviour and locomotor activity in isolated male mice (author's transl)].

The effect of lithium and rubidium on curiosity rearing, and locomotor activity in isolated and group-housed male mice (DBA/2 Han) was investigated. LiCl and RbCl were given with the drinking water (30 mol/l) during 3 weeks. The behaviour of the animals was tested before and after the chronical application of either lithium or rubidium. The results show that lithium was able to enhance the decreased curiosity of the isolated mice which now was no longer different from that of normal group-housed animals. The locomotor activity of the isolated animals was also increased by LiCl, while rearing was not altered. Rubidium even more decreased the already diminished curiosity of the 'fighting mice' but had no influence on rearing and locomotor activity. Three weeks after cessation of treatment the behaviour of the isolated mice returned to predrug levels. Neither lithium nor rubidium had any influence of the behaviour of animals kept in groups.


Facilitation of responding for rewarding brain stimulation by a high dose of amphetamine when hyperthermia is prevented.

Rats given a 5 mg/kg injection of d-amphetamine did not respond for brain stimulation reward when tested under normal laboratory temperatures. In addition to the usual manifestations of stereotypy the rats were markedly hyperthermic. If the hyperthermia was prevented, however, by initially placing the rats in a cold room (10 degrees C) and subsequently testing for brain stimulation under a cool chamber temperature (14--16 degrees C), the rats responding for brain stimulation was facilitated. Thus, the occurrence of hyperthermia appears to be a critical factor responsible for this behavioral dysfunction produced by a high dose of amphetamine.


L-Dopa feeding suppression: effect on catecholamine neurons of the perifornical lateral hypothalamus.

Injection of L-Dopa (0.8--200 nmoles) into the perifornical hypothalamus produced a dose-dependent suppression of feeding in hungry rats. This effect was positively correlated in magnitude with the same effect produced by the catecholamine agonists dopamine and epinephrine, and by the catecholamine-releasing drug amphetamine. L-Dopa's action was partially antagonized by separate injections of the dopaminergic blocker haloperidol (58% blockade) and the beta-adrenergic blocker propranolol (38% blockade). Combined injections of these two antagonists produced a 90% blockade of L-Dopa's effect. Perifornical administration of the dopa decarboxylase inhibitors Ro 4-4602 and MK-486 was also shown to reverse L-Dopas feeding suppression, at doses that enhanced the effect of injected dopamine and epinephrine. On the basis of these findings, L-Dopa appears to suppress food consumption in part through increased catecholamine synthesis, specifically within dopaminergic and adrenergic neurons of the perifornical hypothalamic region.


Differential effects of chlordiazepoxide on conditioned and unconditioned behavior in mice with septal lesions.

The time course of action of repeated daily doses of chlordiazepoxide (CDP) was examined in mice with septal lesions. Bilateral septal lesions produced changes in both conditioned and unconditioned behavior. These lesions increased rate of operant responding on a VI-40 s schedule and produced hyperreactivity to tactile stimuli. The effects of repeated administration of CDP on these two classes of behavior were observed to follow different time courses. In a dose dependent fashion, CDP first reduced VI response rates in septal mice, but after several daily doses the response rate increased above post-surgical baseline levels. The suppressant effects of CDP on septal hyper-reactivity were quite different. Hyper-reactivity was persistently attenuated by all CDP doses tested during the eight-day drug regimen. The pattern of drug effects suggested that serotonin antagonism was involved in CDP's effects on lesion produced hyper-reactivity.


Tolerance to amphetamine-induced impairment of rotarod performance in rats.

Acute administration of d-amphetamine sulfate (30 mg/kg, i.p.) caused muscle weakness and 80% failure in rotarod performance in rats. The incidence of performance failure progressively decreased to 30% in animals receiving d-amphetamine and tested daily for 8 days. A similar progressive reduction in performance impairment was observed in animals receiving d-amphetamine for 3, 5, or 7 days (12.5%) and evaluated only once after the last dose. The tolerance is attributable to a physiological or biochemical rather than learning phenomenon.


The four basic types of evaluation: clinical reviews, clinical trials, program reviews, and program trials.

Four basic types of evaluation, each appropriate in a distinctive situation, are the clinical review and the clinical trial, which are concerned with the care of the individual patient, and the program review and program trial, which deal with programs or services directed at groups or populations. Evaluative reviews are primarily motivated by concern with the welfare of the specific population served, and they appraise specific activities in specific settings as a basis for decisions concerning these activities. Clinical and program trials aim to generate knowledge of more general applicability, especially concerning causal relationships between care and outcomes. The types of evaluation differ in the questions they pose and in the methods used to answer them. Failure to draw a distinction between program reviews and program trials is a frequent cause of wasteful or unhelpful evaluative studies.


Radiation therapy in bronchogenic carcinoma.

Response of intrathoracic symptoms to thoracic irradiation was evaluated in 330 patients. Superior vena caval syndrome and hemoptysis showed the best response, with rates of 86% and 83%, respectively, compared to 73% for pain in the shoulder and arm and 60% for dyspnea and chest pain. Atelectasis showed re-expansion in only 23% of cases, but this figure increased to 57% for patients with oat-cell carcinoma. Vocal cord paralysis improved in only 6% of cases. Radiation therapy has a definite positive role in providing symptomatic relief for patients with carcinoma of the lung.


The inverse compensating filter.

When a single-field technique is used in megavoltage x-ray therapy, a uniform entrance dose is often required. If there is a large variation in source-to-surface distance over the field, an inverse type of compensating filter is necessary to achieve entrance dose uniformity. The production and utilization of these filters is discussed.


Relative biological effectiveness of low- and high-LET radiotherapy beams for jejunal crypt cell survival at low doses per fraction.

In order to determine the relative biological effectiveness (RBE) of various low- and high-LET radiotherapy beams of low coses, LAF1 mice were exposed to one dose and 10 equally fractionated doses of 60Co gamma rays, 137Cs gamma rays, 4-MeV x rays, 300-kVp x rays, helium ions in the plateau and Bragg-peak region, and 15-MeV neutrons; survival of jejunal crypt cells with the microcolony assay of Withers and Elkind was determined using 60Co as the standard. RBE values for the survival of 10 cells/circumference for 10 fractionated exposures (2.4 Gy [240 rad] per fraction of 60Co) were: 1.07 for 137Cs, 1.06 for 4-MeV x rays, 1.16 for 300-kVp x rays, 1.10 for helium ions in the plateau, 1.29 for helium ions in the peak, and 3.02 for 15-MeV neutrons. As LET increased, RBE increased with decrease of dose/fraction.


[EEG changes induced by long-term lithium therapy].

The effect of longstanding therapy using lithium salts compared to other psychotropic drugs in combination with lithium salts or other psychotropic drugs alone, on the human EEG is examined and compared with the literature. We found in our sample of 56 patients under lithium therapy: (1) that lithium salts - like other psychotropic drugs - evoke a significant increase in paroxysmal dysrhythmic activity and EEG abnormalities collectively; (2) that lithium salts cause an increase in rhythmical patterns and abnormalities in vigilance; and that (3) the combination of lithium salts with other psychotropic drugs leads to a significant augmentation of focal abnormalities of the left brain area and epileptic potentials, while EEG recordings without abnormalities are significantly diminished under this therapy. It results from this investigation that the risk for central decompensation in relation to the individual disposition seems to be especially high for the combination therapy.


Plasma and tissue levels of furosemide in dogs and monkeys following single and multiple oral doses.

35S-Furosemide was administered to beagle dogs and rhesus monkeys as an oral solution on a single and a 20 repeated 5 mg/kg/day dosing regimen. In both species, furosemide is rapidly but incompletely absorbed with peak plasma levels being achieved within one hour after dosing. The plasma level versus time profiles do not appear to be significantly altered as a result of the repetitive dosing regimen employed, although the profiles themselves are quite different for the two species studied. Following single oral doses, the observed peak plasma levels achieved in dogs are approximately eight-fold higher than in monkeys. In dogs, oral administration of furosemide results in a biexponential disposition curve while in monkeys the profile appears monoexponential. The major disposition phase in dogs has a half-life or approximately 30 minutes and is followed by a slow elimination phase with a half-life of approximately 7 hours. Only a small percentage of the absorbed dose is affected by the slow disposition phase and consequently accumulation of furosemide on the once-daily dosing regimen is slight. In the monkey studies, the rapid disposition phase was not observed and the slow disposition phase had a half-life of approximately 11 hours. On the repetitive dosing regimen, the monkey appeared to accumulate furosemide to a slightly greater extnet than the dog and showed a lesser degree of fluctuation within a dosing interval. In both species, liver and kidney radioactivity levels were not detectable three days after a single dose although low levels of furosemide and/or metabolites were observed for six days following the last dose of the multiple dosing regimen. At the dosage level and regimen employed, no unusual or "typical" lesions associated with furosemide were found in the dog and monkey tissues examined histopathologically.


Antagonism of vasopressin-induced water flow by somatostatin.

The ability of somatostatin to modify the water premeability of the toad bladder was examined. Somatostatin had a small effect on basal water flow and antagonized the hydrosmotic effect of vasopressin. Water flow induced by cyclic AMP was enhanced. These results may explain the diuretic and hyposthenuric effects of somatostatin in vivo.


V. Efforts at stabilization of rat liver phenylalanine hydroxylase.

The best from a series of matrices for the covalent coupling of rat liver phenylalanine (Phe H) was activated thiol-Sepharose 4B. An average of 0.38 mumoles of enzyme thiol was coupled per mumole of matrix thiol, representing 21 mg of protein per g of carrier. After thiolation with N-acetylhomocysteine thiolactone the enzyme, bound to the same matrix, was thermally more stable with a broad pH range of optimal activity. Thiolation with S-acetylmercaptosuccinic anhydride markedly reduced hydroxylase activity whereas succinylation with succinic anhydride had little or no effect. Treatment of the thiolated enzyme with various oxidants to form disulfide bridges did not increase the thermal stability. Efforts to improve stability by formation of new linkages while the enzyme was still bound to the matrix were unsuccessful. The untreated or thiolated enzyme was inactivated upon reaction with glutaraldehyde.


Is there a place for chemical and radiation synovectomy in rheumatic diseases?

Synoviorthesis has the same indications as surgical synovectomy. But it is easy to perform, can be repeated and does not leave any scars. It should be tied first. The choice of the product to be injected depends on the joint, the number of injections and the age of the patient. In all cases a long-acting local steroid should be tried first. In combination, osmic acid seems to potentiate the effect of the steroid. If osmium tetroxide damages the cartilage, it has been demonstrated that the damage is limited and superficial and as far as we know without clinical consequence. The fear of side-effects has limited the use of synoviortheses in the United States. Sledge et al. (1977), in animals, have used a new rare earth isotope, dysprosium-165, a beta-emitter with a maximum tissue penetrance of 5.7 mm and the advantage of a very short half-life of 140 minutes. In so far as systemic therapy is only partly successful there is still a place for chemical and radiation synovectomy in rheumatic diseases.


Retention in glass bead columns as a measure of leucocyte adhesiveness. I. Influence of sample size and erythrocyte sedimentation.

Erythrocyte sedimentation significantly influenced test results when leucocyte retention in glass bead columns was used as a measure of leucocyte adhesiveness. In blood with elevated ESR, this influence was especially evident. Erythrocyte sedimentation highly increased leucocyte retention in vertical columns with a downward flow, whereas in slightly tilted columns with an upward flow, the retention was reduced. The error in measurements caused by erythrocyte sedimentation could be corrected by means of the haematocrit values in samples and eluates. Correction was better for results obtained with slightly elevated than with vertical columns. The granulocyte retention rates increased with increasing sample size, whereas lymphocyte retention remained constant.


Erythrocyte and leucocyte enzymes in a case of paroxysmal nocturnal haemoglobinuria.

In a patient with paroxysmal nocturnal haemoglobinuria (PNH) enzymatic activities of erythrocytes and leucocytes were studied. Studies of autohaemolysis were also performed. The following erythrocytary enzymes were measured: Glucose-6-phosphate dehydrogenase (G-6-PD), pyruvate kinase (PK), glutathione reductase (GR), and acetylcholinesterase (AcChE). The following enzymes were measured in leucocytes: Adenosine deaminase, purine nucleoside phosphorylase, adenine phosphoribosyltransferase, hypoxanthine phosphoribosyltransferase and adenosine kinase. Normal activity of G-6-PD, GR and PK in erythrocytes was found. In leucocytes and lymphocytes activity of purine nucleoside phosphorylase was reduced. Auto-haemolysis in vitro was increased, which could not be compensated by addition of glucose or ATP.