Paradoxical lithium neurotoxicity: a report of five cases and a hypothesis about risk for neurotoxicity. There have been many reports of probable lithium-induced organic brain syndromes occurring when serum lithium levels are within or close to the therapeutic range. The authors report on five patients who developed clinical syndromes suggestive of severe neurotoxicity during lithium treatment. In all cases lithium levels were between .75 and 1.7 mEq/liter. The patients who developed neurotoxicity had markedly higher global ratings of psychotic symptomatology and anxiety in the pretoxic period than did patients who never deveoped neurotoxicity. When the acute manic state is characterized by marked psychotic symptoms and intense anxiety, it may be associated with increased vulnerability to the development of severe lithium neurotoxicity.


Controlling the cost of dental care.

Controlling the cost of dental care. Methods for controlling dental care expenditures are taking on greater importance with the rapid increase in prepaid dental plans. The use of regulatory systems to monitor provider performance are necessary to prevent gross over-utilization but are unlikely to result in net savings of more than five per cent of total gross premiums. Theoretically, prepaid group dental practice (PGDP) may reduce expenditures by changing the mix of services patients receive. The modest estimated savings and the small number of PGDPs presently in operation limit the importance of this alternative for the next five to ten years. If substantial reductions in dental expenditures are to be obtained, it will be necessary to limit dental insurance plans to cover only those services which have demonstrated cost-effectiveness in improving health for the majority of people. The concept that richer benefit plans may have small marginal effects on improving oral health may not be easy for the public to accept but, until they do, expenditures for dental care will be difficult to control.


Control of schistosomiasis: report of a workshop. Nineteen scientists, field workers, and representatives of funding agencies active in schistosomiasis research and control met in Bellagio, Italy in October 1977 to attempt to evaluate the effectiveness of current control methods and what might be accomplished with available technology. The deliberations included summaries of knowledge on the biology, transmission, and control of schistosomiasis and assessment of major control programs and methodologies. The groups concluded that in the major endemic areas considerable gains in control of schistosomiasis could be made with current technology. However, maintenance of control in most countries, and establishment of serious control programs in countries in which schistosomiasis is a less severe public health problem, would require development of less expensive modalities which would need little monitoring and possibly have benefits extending beyond schistosomiasis control.


Immunological study of hydatidosis. I. Evaluation of immunoelectrodiffusion tests and enzyme-linked immunoelectrodiffusion assay (ELIEDA) in human hydatidosis. The comparative sensitivity, specificity and rapidity of immunoelectrodiffusion (IED) on cellulose acetate membranes and of immunoelectrophoresis (IEP) on agarose gel, were evaluated in the diagnosis of hydatidosis. Pooled crude fluid from several cattle hydatis cysts was used as antigen in tests on 1,750 non-hydatis and 400 hydatis sera obtained from patients with hepatic, pulmonary, splenic, peritoneal and cerebral hydatidosis before and after surgery. Coupled to a specific human reference serum for arc 5, IED shows a specificity comparable to that of IEP but its sensitivity is slightly higher and the amount of antigen needed is very small. The appearance of a typical "gloved finger" pattern in sera from patients with ruptured cysts emphasizes the interest of quick results (3 hours) obtained by this method. In order to increase the sensitivity of IED and to define the class of immunoglobulins involved in the antigen-antibody reaction, we have coupled this method to an enzymatic technique. The immune complexes precipitated by IED were treated with peroxidase-labelled antibodies specific to each class of human immunoglobulins. The specificity of this enzyme-linked immunoelectrodiffusion assay (ELIEDA) permits one to follow the immunologic evolution of hydatidosis and to identify IgM in ruptured hepatic cysts and IgA in pulmonary cysts.


Host selection patterns of some Pakistan mosquitoes. The host selection patterns of 18 species of Pakistan mosquitoes are described, emphasizing the presumed malaria vectors, Anopheles culicifacies, An. fluviatilis, and An. stephensi, and the probable vectors of West Nile virus, Culex pipiens fatigans and Cx. tritaeniorhynchus. All species tested, with the exception of Cx, p. fatigans, were considered to be essentially zoophilic. Few human feeds were recorded for An. annularis (0.7%), An. culicifacies (0.5%), An. fluviatilis (1.1%), An. nigerrimus (14.3%), and Cx. bitaeniorhynchus (2.8%), while Cx. p. fatigans (37.6%) commonly fed on man. No human positive feeds were recorded for An. stephensi. Temporal or spatial changes in host selection patterns were not discerned with the exception of Cx. p. fatigans, whose feeding patterns varied opportunistically with host availability. Cx. p. fatigans females collected from houses fed more commonly on man than those resting in cattle sheds or in agricultural fields. Cx. p. fatigans resting in cattle sheds during winter fed mostly on birds and bovids, changing to man and bovids during the spring and then to man and birds during summer. The relationships between these results and vector-borne disease transmission in Pakistan are discussed.


The gonotrophic cycle in Simulium ochraceum. Oogenesis in Simulium ochraceum is characterized in terms of distinct stages of follicular development following emergence, sugar feeding, and completion of a blood meal. Changes in the ovary following the first oviposition are also described. This black fly species was found to be anautogenous and exhibited gonotrophic concordance. The formation of follicular dilatations after oviposition followed a distinct pattern with traces of cellular debris evident up to 144 hours. The presence of fructose, as detected by the cold Anthrone test, indicated that both parous and nulliparous flies routinely imbibe nectars as part of their foraging behavior. The availability and frequent use of nectar may contribute significantly to longevity of parous flies, hence increasing their vector potential.


Chemotherapy with ethidium bromide-DNA complex in established Chagas' disease. Two different strains of mice (AKR and NMRI-IVIC) were inoculated intraperitoneally with the virulent Y strain of Trypanosoma cruzi, and then treated with the lysosomotropic ethidium bromide-DNA complex, according to several different treatment schedules. When animals were treated 48 hours after intraperitoneal inoculation with three intraperitoneal doses of EB-DNA no parasitemia was detected, even after 11 weeks, confirming previous results. However, when infection was allowed to become fully established, that is 3-4 weeks after inoculation, and then challenged with several different treatment schedules (with varied doses and timing of administration) we failed to cure established Chagas' disease, suggesting that the claim of effectiveness for this EB-DNA complex is limited to early Chagas' disease.


Prevalence of parasitemia and seroreactivity to Trypanosoma cruzi in a rural population of Northeast Brazil. Age-specific prevalence rates of parasitemia and seroreactivity to Trypanosoma cruzi were determined in a rural area endemic for Chagas' disease in Northeast Brazil. Parasitemia was detected by blood cultures and xenodiagnosis, and serum antibodies to the parasite were measured by the complement fixation (CF) and indirect immunofluorescence (IFA) tests. Of the 116 persons examined, 39 (33.7%) had antibodies and 23 (19.8%) had parasitemia. Ninety-six percent of parasitemic individuals were seropositive and 56% of seropositive individuals were parasitemic. The percentage of seropositive individuals with detectable parasitemia declined with age; all seropositive children in the 1- to 4-year age group and two-thirds of seropositive persons 5-19 years old had parasitemia while only one-third of seropositive adults above 19 years had parasitemia. CF and IFA tests were equally sensitive in detecting persons with parasitemia. Xenodiagnosis was more sensitive than culture for detecting parasitemia, but the two methods together were more sensitive than either method alone. Using the age-dependent relationship of parasitemia to seropositivity determined in this study, the prevalence rate of T. cruzi parasitemia was estimated in a much larger adjacent population in which seropositivity rates and the demographic structure were already known.


Enteral and parenteral feeding to evaluate malabsorption in intestinal parasitism. General evidence of malnutrition such as loss in body weight associated with intestinal parasitism has been attributed to decreased food intake, to intestinal malabsorption, and to change in host basal metabolism. To establish the relative importance of these factors in this regard, rats with trichinosis were studied. The weights of infected and uninfected animals were followed after being placed on one of three feeding regimens for 1 week--stock diet ad libitum, intraduodenal nutrition, and intravenous nutrition. Infected rats on a stock diet lost weight whereas those on the other two regimens maintained the same weight pattern as uninfected counterparts. The maintainance of body weight occurred despite alterations at the level of the intestinal brush border as indicated by a depression of intestinal disaccharidase activities (sucrase and lactase) and by reduction of monosaccharide absorption (measured as accumulation of beta-methyl glucoside) in the proximal, heavily infected region of the small intestine. There was no compensatory increase in enzyme activity nor in the absorptive capacity in the distal gut. Results support the conclusion that inadequate oral food intake rather than changes in basal metabolism or intestinal pathophysiology accounts for weight loss during the intestinal phase of infection.


Schistosomiasis in Omo National Park of southwest Ethiopia. Schistosomiasis mansoni infection was found in more than 50 tourists who had visited Omo National Park, Ethiopia, and bathed and swum in the Mui River. A survey revealed Schistosoma mansoni infection in 41% of Park residents and in 33% of the neighboring Suri people. Eggs were found in stools and adult worms at autopsy of wild Papio anubis and Cercopithecus aethiops. Trematode larvae were found in 27% of Biomphalaria pfeifferi snails found in the Mui River. The source of the disease and the implications of its spread with the future development of the Omo Valley are discussed.


Epidemiologic studies among Amerindian populations of Amazônia. I. Pyoderma: prevalence and associated pathogens. Pyoderma was studied among a representative sample of the residents of four remote Amerindian villages, Amazonas State, Brazil, during July-August 1976. The overall prevalence among the 775 inhabitants examined was 11%, with little intervillage variation. When the attack rates for the entire sample population were calculated by 5-year age intervals, the 0- to 4-year-olds had the highest rate, 31%. The highest prevalence, 38%, was found among 3-year-olds. Attack rates were not apparently related to sex. Cultures which were taken from representative pyoderma lesions from people in the four survey villages and from three additional villages were studied by a modified delayed culture technique for recovery of gram-positive pathogens from silica-gel desiccated swabs. Group A and group G B-hemolytic streptococci, coagulase positive Staphylococcus aureus, and Corynebacterium diphtheriae were isolated. Group A S. pyogenes was most commonly found, occasionally as the sole pathogenic species. No nephritogenic M-types were found, although most isolates were not M-typable. The T-types found corresponded to those previously reported as being pyoderma-associated. Most pyoderma-associated C. diphtheriae isolates were non-toxigenic. Biotypes gravis and mitis were equally represented.


The continuous cultivation of Plasmodium fragile by the method of Trager-Jensen. Using the Trager-Jensen method, a second malaria species, Plasmodium fragile, a simian counterpart to the human malaria P. falciparum, has been cultivated successfully. The average growth rate every 3--4 days was 5-fold and the average number of merozoites observed was 14. To date, only rhesus monkey red blood cells (RBC) would support the long-term cultivation of this parasite. Short-term observations indicate that RBC from the squirrel monkey (Saimiri sciureus) may support growth but human RBC of each of the four major AB-O types failed to support growth of the parasite. Availability of the P. fragile-rhesus monkey model would allow for a second parasite-host system for the in vitro and in vivo study of the immunologic responses of the falciparum-like parasite in a more natural host.


Surgical technic for treatment of splenic rupture. In nine baboons and one dog, a technic for suturing the injured spleen and also providing immediate control of hemorrhage is described. By placing Silastic strips along the wound edges of the transverse incision into the spleen and along the surface and bottom edges of the wound after resection, it was possible to control bleeding with deep U sutures passing through the Silastic bolsters and the spleen tissue. The procedure appears to provide a reliable method of salvaging splenic tissue and thus avoiding the hazard of postsplenectomy sepsis.


Operative and postoperative choledochofiberoscopy. Choledochofiberoscopy is effective for both diagnosis and therapy. It is a valuable diagnostic technic because it permits a definite diagnosis and it allows the surgeon to determine the operative method by direct viewing and by biopsy of the biliary tract wall. Choledochofiberoscopy is therapeutically valuable because it permits cleaning of the bile duct, the removal of retained or remnant stones in the common and intrahepatic ducts, and the removal of granuloma and benign tumor with forceps under direct vision.


An SEM AND TEM study of suppression of eye development in eyeless mutant axolotls. Morphology of the primary optic rudiment of normal eyed and mutant eyeless (e/e) axolotl embryos was studied at light. Tem and SEM levels. The presumptive eyeforming region of eyeless embryos differs from that of normal embryos in several important respects including premature formation of basal lamina, separation from overlying ectoderm by mesenchyme cells and persistence of granules in the interspace surrounding the optic anlage into relatively late developmental stages. These differences suggest that the gene that causes failure of eye formation in the mutant axolotls produces structural differences that interfer with normal physical and/or biochemical inductive interactions between neurectoderm and mesenchyme cells due primarily to the precocious development of basal lamina over the extermal surface of the optic primordia.


Mutagenicity studies with halothane in Drosophila melanogaster. Halothane was investigated for its mutagenic effects in Drosophila melanogaster. The induction of sex-linked recessive lethals was used as an indicator of genetic damage. Adult male flies were exposed to halothane either for 14 days at 1,000 or 1,600 ppm (v/v) or for one or two days in 2,100 or 20,000 ppm. In several experiments slight increases of the mutation frequency were observed. For the pooled data of the 14 day-exposure experiments, the increase amounted to a doubling of the spontaneous rate, just reaching the level of 5 per cent significance. The authors consider this a borderline result, indicating, with a fair degree of probability, that halothane has weak mutagenic activity, under the conditions studied.


Prolongation of pancuronium-induced neuromuscular blockade by intravenous infusion of nitroglycerin. Based upon clinical observation of undue prolongation of pancuronium-induced blockade in the presence of intravenous infusion of nitroglycerin, neuromuscular blockades produced by pancuronium, succinylcholine and d-tubocurarine were studied in 51 cats using the sciatic-gastrocnemius nerve-muscle preparation. Pancuronium-induced blockade was found to be significantly prolonged (P less than 0.1) in the presence of a nitroglycerin infusion of 1 microgram/kg/min (65 vs. 127 min). Less, but still significant, prolongation occurred when nitroglycerin, 0.5 microgram/kg/min, was infused. The intravenous infusion of nitroglycerin must be started prior to the pancuronium injection for the block to be prolonged. Neuromuscular blocks produced by succinylcholine and d-tubocurarine were not altered by nitroglycerin. In experiments using the isolated rat diaphragm preparation, the depth of pancuronium-induced block was found not to be changed by nitroglycerin, suggesting an effect of nitroglycerin on the process of recovery from blockade. These findings indicate a selective pancuronium-nitroglycerin interaction.


Cromolyn DSG effects in hyperkinetic and psychotic children with allergies. Preliminary findings of an open trial of cromolyn in hyperkinetic and psychotic children indicates that the medication is useful, safe and well tolerated. Further controlled studies are suggested to determine the efficacy and indications of cromolyn in child psychiatry. Positive findings could clarify the biochemical mechanisms of hyperkinesis.


Prophylactic drugs in the management of childhood asthma. Recurrences of explosive, severe asthma in children can often be prevented by immunotherapy and constant surveillance of environmental and dietary control. In addition, judicious and preserving use of pharmaceuticals may be employed for prophylaxis in childhood asthma. These include cromolyn sodium by inhalation, beclomethasone dipropionate aerosols, round-the-clock methylxanthines and beta 2 adrenergic bronchodilator agents. Their application is discussed.


Elevated concentrations in serum immunoglobulins due to infection by ovine progressive pneumonia virus. Sixty-seven serum samples were obtained from 2 sheep flocks. Agar gel immunodiffusion (AGID) was used to separate progressive pneumonia virus (PPV)-infected sheep from noninfected sheep by the presence of precipitating antibodies. Immunoglobulin (Ig), total protein, and albumin concentrations were then measured from all 67 sera to determine whether differences existed between PPV-infected and non-infected sheep. A significant difference (P less than 0.0005) was found in both total protein and Ig concentration between PPV-infected and noninfected sheep. This corresponding difference was absent in albumin measurements. The significant differences (P less than 0.0005) in Ig and total protein concentrations were then used to evaluate a field test for diagnosing progressive pneumonia. The possibility of using either total protein or Ig concentrations as a field test was found to be highly unlikely due to variation in individual values.


U.S. Public Health Service Cooperative trial of three rifampin-isoniazid regimens in treatment of pulmonary tuberculosis. A total of 822 patients with newly diagnosed pulmonary tuberculosis were assigned randomly to one of 3 daily rifampin-isoniazid (RIF-INH) regimens: 450, 600, or 750 mg of RIF in combination with 300 mg of INH. After an initial 20 weeks of therapy with RIF-INH, patients recieved 300 mg of INH and 15 mg of ethambutol (EMB) per kg of body weight for either 12 or 18 months after their sputum cultures became negative. The rate of bacteriologic conversion of sputum among the 3 RIF-INH regimens was compared for 552 patients who completed the 20 weeks of RIF-INH therapy. Apporximately 60 per cent of these patients also completed their assigned INH-EMB therapy and were examined for relapse for at least one year after therapy was stopped. There was no significant difference in the rate of sputum conversion or rate of relapse between the group of patients who received 600 mg of RIF and those who received 750 mg of RIF. However, the 450-mg RIF regimen was significantly less effective than the other 2 regimens, as manifested by a lower rate of sputum conversion and a higher rate of treatment failures. Further analysis showed that RIF dosages of less than 9 mg per kg of body weight per day may be inadequate for treatment of pulmonary tuberculosis. The acceptability of these regimens was high, and the incidence of adverse reactions requiring discontinuation of RIF-INH therapy was quite low (3.3 per cent). A large proportion of patients (44 per cent) developed increased concentrations of transaminase during therapy with RIF-INH. These abnormalities were usually transient and, in most cases, of no clinical significance. In the relapse analysis, 12 months of chemotherapy after sputum conversion was shown to be as effective as 18 months of therapy after conversion of these RIF-containing regimens.


Sodium cromoglycate-induced changes in the dose-response curve of inhaled methacholine and histamine in asthmatic children. The effect of inhalation of 2 per cent solution of sodium cromoglycate compared to that of saline on the bronchial response to methacholine and histamine was studied in 30 asthmatic children. Seven of 17 children challenged with methacholine showed decreased sensitivity after pretreatment with sodium cromoglycate. In 4 of 13 children, sodium cromoglycate attenuated the response to inhaled histamine. We conclude that in some asthmatic children, sodium cromoglycate provides significant protection against these nonallergenic challenges. This may have some therapeutic implications in the management of these patients. Our findings raise the possibility that sodium cromoglycate might have an action on cholinergic or irritant receptor sites in addition to inhibition of mast cell degranulation.


Pulmonary alveolar proteinosis: analysis of airway and alveolar proteins. Lavage effluents from the lungs of patients with pulmonary alveolar proteinosis were analyzed for soluble constitutents. Antiserums monospecific for normal plasma components, C-reactive protein, and secretory piece were used to investigate the presence of these proteins in lavage effluent, wheras 2-dimensional polyacrylamide gel electrophoresis provided a comprehensive map of the major proteins that were present. The proteins of lung washing obtained from normal subjects and of patient and normal serums were similarly analyzed. Most of the soluble-phase proteins from lavage of patients with pulmonary alveolar proteinosis were also found in patient serum, and were present in amounts consistent with a theory that they originate from the plasma by passage through channels of approximately normal size and selectivity. This findings suggests that the abundant soluble protein found in the alveoli of the lungs of these patients does not arise by leakage through a serverly damaged blood-air barrier. Patients had in their lung lavage effluents 2 soluble proteins of molecular weights 47,000 and 52,000 daltons not found in their serum and not present in lung washings from normal subjects. Uniformly increased concentrations of immunoglobulins in patient lavage effluent, abnormal immunoglobulin concentrations in patient serum, and the presence of C-reactive protein in the serum from 4 of 5 patients indicate a response of the immune system to the disease process and suggest that an atypical hypersensitivity reaction may be involved.


Possible correlation between induction modes of hepatic enzymes by PCBs and their toxicity in rats. Acute toxicity of individual PCBs, which were categorized as either phenobarbital (PB)- or 3-methylcholanthrene (MC)-type inducers, was examined in young male Wistar rats, comparing their effects on growth rate, organ weight and liver lipid content, 5 days after a single i.p. injection. PB-type PCBs (2,4,3',4'- and 2,5,2'5'-tetrachlorobiphenyl), which slightly increased a content of cytochrome P450, did not show any significant toxicity at a dose of 100 mg/kg. On the contrary, MC-type PCBs (3,4,5,3',4'-pentachloro- and 3,4,5,3',4',5'-hexachlorobiphenyl), which markedly increased a content of cytochrome P448, strongly reduced growth rate and weights of thymus and spleen at a dose of 10 mg/kg. Liver enlargement accompanied by fatty liver was also observed only with MC-type PCBs. 3,4,3',4'-Tetrachlorobiphenyl was also toxic at a dose of 50 mg/kg, in keeping with its weak MC-type-inducing ability. Pretreatment with MC affected neither growth rate, spleen weight, nor liver lipid content. These results suggest that the toxic potency of PCBs is related to their MC-type inducing ability, but the toxic characteristics are different from those of MC itself.


Decrease in vital capacity in PCB-exposed workers in a capacitor manufacturing facility. Pulmonary function was evaluated in 243 workers exposed to PCB in the manufacture of capacitors. Mean employment was greater than 15 years. Thirty-four of the workers (14%) were found to have a reduced Forced Vital Capacity (FVC less than 80% of Morris' predicted). Of the 34 with reduced FVC, 27 (80%) demonstrated a restrictive pattern of impairment (FEV1/FVC greater than 0.7). Only one of these 27 workers had an abnormal chest roentgenogram (greater than or equal to 1/0 by ILO UC Classification of Radiographs of Pneumoconioses). These findings are of interest in view of recent experimental data indicating the accumulation of PCBs and PCB metabolites in lung tissue (Brandt and Jansson). Restrictive spirometric impairment with no radiographic change is unusual in occupational exposure.


Neuropathology of "spinning syndrome" induced by prenatal intoxication with a PCB in mice. A striking motor dysfunction, "spinning syndrome," developed with a high frequency in weaning mice whose dams received oral 3,4,3',4'-tetrachlorobiphenyl (4-CB) during gestation (day 10 through day 16). The syndrome is permanent and is characterized by swift circling movements sustained in one direction at a minimal rate of 40 turns/min (usually 50 to 150 turns/min), restlessness, and hyperkinesia. Twenty-four spinners and 4-CB nonspinners and age-matched controls were subjected to histopathologic, histofluorescent, histochemical, and electron microscopic studies. The most reliable histopathologic marker for prenatal 4-CB injury to the CNS is the presence of cylindrical CNS peninsulas (CCPs) in the spinal and cranial nerve roots. The CCPs consist of either CNS-type myelinated fibers, unmyelinated fibers, or astroglial bundles in varying proportions, and are enclosed by a basement membrane. The CCPs are also observed in 4-CB nonspinners but in none of 12 controls studied. A selective defect in synaptogenesis induced prenatally by 4-CB is proposed as the primary event pursuant to the development of the CCPs, while interference with synaptogenesis may have occurred selectively in the striatonigral dopaminergic system. This is suggested by electron microscopy on the nucleus accumbens and also by the responses to administration of dopaminergic agonists and antagonist. The 4-CB induced clinico-pathologic anomaly may serve as a singular model for understanding human neurologic disorders, in particular, Werdnig-Hoffmann disease and minimal brain dysfunction syndrome.


Developmental toxicology of the halogenated aromatics: effects on enzyme development. Our studies were designed to define structure-activity relationships for the effects of pure PCB congeners and 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) on hepatic enzyme development. The organohalogens were usually administered to pregnant rats and enzyme activities measured in fetuses and offspring (6, 20, and 55 days after birth). Offspring of pregnant rats exposed to TCDD had elevated levels of benzpyrene hydroxylase and nonsteroid UDP glucuronyltransferase (UDPGT). Postnatal inductive actions of TCDD were caused primarily by newborn exposure to TCDD in milk. 3,4-3',4'-Tetrachlorobiphenyl (4-CB) induced P448-dependent enzymes in 6- and 20 day-old rats whereas 2,4,5-2',4',5'-hexachlorobiphenyl (6-CB) induced P450-dependent enzymes. 4-CB was a potent teratogen (behavioral and kidney abnormalities) but 6-CB produced no such developmental defects. 4-CB might also alter the normal imprinting of sexual differentiation of hepatic metabolism as evidenced by a feminization (decrease) of UDPGT in adult male rats which were exposed neonatally to 4-CB. Therefore, the organohalogens might alter the ontogeny of hepatic metabolism in two ways: induction and imprinting.


Quantification of total IgM and IgG and specific IgM and IgG to a thymus-independent (LPS) and a thymus-dependent (tetanus toxoid) antigen in the rat by enzyme-linked immunosorbent assay (ELISA). Rat IgM and IgG was determined by mechanized "sandwich" enzyme-linked immunosorbent assay (ELISA) using peroxidase labeled anti-rat-IgM and -IgG. Linear ranges in standard curves of a reference rat serum had a slope similar to the slopes found with sera of 25 rats of various age. IgM and IgG measurements by ELISA in these sera correlated well with results obtained by single radial immuno-diffusion (SRID). In addition, the precision of the enzyme immunoassay was the same as obtained with the SRID. Compared with SRID, ELISA is less time consuming and the amount of antiserum used in the macro-ELISA is one order of magnitude lower; and again 10 times lower in the mechanized micro-ELISA that is currently being developed. In conclusion, the ELISA is a specific, reliable, sensitive, and economic method for routine measurement of rat serum IgG and IgM e.g. in toxicity studies. In the second part of this study, ELISA and the passive hemagglutination test were compared to determine the primary and secondary antibody response to E. coli lipopolysaccharide (LPS) and tetanus toxoid in rats. In the ELISA, the antigens were bound to the wells of polystyrene microplates. Tetanus toxoid was coated directly, LPS after complexing with methylated bovine serum albumin. After incubation with dilutions of the rat sera, the amount of antibody bound to the solid phase was quantified by means of peroxidase-labeled antiimmunoglobulin. The specificity of the enzyme immunoassay was tested by absorption of the sera with the respective antigens. ELISA proved to be more sensitive than the hemagglution reaction, except when titers were determined during the secondary response to tetanus toxoid. Besides its specificity and sensitivity, ELISA is a convenient method for measuring both IgM and IgG antibodies. Finally, evidence is presented that in the rat, the humoral immune response to LPS is a thymus-independent phenomenon. Thus, by using the antibody response to LPS and tetanus toxoid in function studies of the immune system of the rat, insight can be obtained in the thymus-independent and thymus-dependent humoral immune response.


PCB removal from the Duwamish River estuary: implications to the management alternative for the Hudson River PCB cleanup. The results of the Duwamish River PCB cleanup and recovery project are discussed as a basis for comparison in selecting potential management alternatives for removing PCB residues in the Hudson River. Application of the Pneuma dredge system at the Duwamish River spill site removed approximately 92% of the PCBs spilled. No significant changes in the PCB load of suspended matter and water column were observed. These results suggest that a carefully planned dredge/disposal operation can be effective in minimizing environmental impacts in the Hudson River. On the basis of observations of the Duwamish case, the existing baseline data on the distribution of PCBs in the Hudson River, and the removal alternatives and associated environmental tradeoffs, hot spot dredging is recommended as the best practical alternative.


World PCBs map: storage and effects in man and his biologic environment in the 1970s. PCB residues are found in biota all over the world. A biologic magnification of PCBs has been demonstrated in the food chain: plankton-fish-fish eating birds. A world map of the PCBs residues in biota and some of their biologic effects are given in this study. The biologic effects of PCBs are varied and may generally be explained by the induction or inhibition of the activity of a large number of enzymes, which upsets quantitatively, normal biochemical processes. Harm to reproduction, growth, development, defense systems, tissues, and organs appeared in susceptible organisms as a result of such changes or as a chain reaction to heterostases. The adverse health effects, observed in persons occupationally exposed and in those accidentally poisoned, point to the risk for the general population of an ever-increasing environmental pollution by PCBs. There is need for an integrated approach, consisting of epidemiologic studies on high risk groups in the general population and in persons occupationally exposed, as well as periodic assessment of PCB residues in man, his food, and feed supplies.


Congenital nonprogressive facial hemiatrophy with ipsilateral eye abnormalities and juvenile glaucoma. Congenital nonprogressive facial hemiatrophy is reported in association with anisometropia and ipsilateral cornea, iris, angle abnormalities, and juvenile glaucoma. The importance of complete ocular examination in patients with this uncommon abnormality is emphasized.


Bacillus cereus panophthalmitis after intravenous heroin. Two healthy young black men developed panophthalmitis after intravenous heroin injections. Bacillus cereus, considered to be a relatively noncommon pathogen for man, was found to be the causative agent as it was recovered from the anterior chamber and viterous cavity of both cases. The ocular findings were unilateral in each case, and neither patient had any sistemic involvement from the bacteremia. The onset of visual symptoms varied from 24 to 36 hours after the last intravenous injection with the eye becoming rapidly blind. Photographs of the early fundus lesions included preretinal hypopyon-like lesions and peculiar changes in the blood vasculature. Intracameral gentamicin and steroids did not alter the cause, and treatment was enucleation.


Are changes in serum immunoglobulin and complement levels following open heart surgery influenced by oxygenator type or postoperative parenteral nutrition? The serum IgG and IgA concentrations significantly decreased through the seventh day and the serum IgM and complement (C3 and C4) concentrations through the second day after elective open heart surgery for heart valve disease in a series of 16 patients. A significant increase over the preoperative concentration was seen, however, in the serum IgA, IgM, C3 and C4 on day 14 postoperatively. The serum IgG and IgM concentrations declined significantly only in patients perfused by a bubble oxygenator, although the two small series of patients perfused by bubble and membrane oxygenator did not differ significantly from each other. Parenteral nutrition had no effect on postoperative changes in serum immunoglobulin and complement concentrations.


Atenolol once daily in angina.

Atenolol once daily in angina. Both atenolol 50 mg daily (A-50) and atenolol 100 mg daily (A-100) significantly reduced the angina attacks per week compared with placebo (p less than 0.05). Nitroglycerine consumption was less on A-50 (p less than 0.05) and on A-100 (p less than 0.025) than on placebo. There was no difference between the atenolol dosages in these respects. In comparison with placebo, atenolol gave a significant increase in total work performed until the appearance of 1 mm ST segment depression in bicycle exercise tests made between 3--4 p.m. (A-50: p less than 0.05; A-100: p less than 0.0017). The rate-pressure products (systolic BP X heart rate X 10(-2) at 6' at 30 W) was 163.5 +/- 12.5 (S.E.) on placebo. It diminished to 129.8 +/- 7.9 on A-50 (p less than 0.01) and to 113.9 +/- 6.6 on A-100 (p less than 0.001). The effect of A-100 on the rate-pressure product was stronger than that of A-50 (p less than 0.05). Relative heart volume did not change on A-50 compared with placebo but A-100 resulted in a slight enlargement of 22 ml/m2 BSA when compared to placebo (p less than 0.01). Side effects were minimal.


Reconstruction of the audiogram using brain stem responses and high-pass noise masking. Contributions to the brain stem electrical responses (BSER) presumably initiated from specific frequency regions of the cochlea with center frequencies similar to the major audiometric frequencies (0.5, 1, 2, 4, and 8 kHz) are derived by the application of a high-pass noise masking technique utilizing click stimuli. In normal hearing subjects, these derived narrow-band responses from the midfrequency regions (4, 2, and 1 kHz) can be recognized at click levels as low as 10 dB HL. For the frequency regions around 8 kHz and 0.5 kHz, these derived responses can be discerned at click levels of 30 dB HL and higher. When one uses the lowest click level at which these derived responses can be obtained from a given frequency region, the differences between a patient with a hearing loss and a normal hearing subject correlate well with the amount of hearing loss (air conduction) recorded by conventional pure tone audiometry. Use of the high-pass noise masking technique to reconstruct the audiogram may be of great potential value in assessing young children and other individuals who cannot or will not respond to conventional audiometry.


[Biochemical mutants of Actinomyces griseus, a producer of the antibiotic grisin. The isolation of the mutants and a study of the level of grisin antibiotic synthesis]. Biochemical mutants of Actinomyces griseus producing grisin were obtained under the action of chemical mutagens. The mutants were divided into 2 groups. The mutants with impaired synthesis of amino acids of the aspartic acid family, i.e. lysine, homoserine and methionine were included into the 1st group. The 2nd group included the mutants with impaired synthesis of the other amino acids, i.e. histidine, arginine, tyrosine, phenylalanine and valine. The antibiotic production level in the biochemical mutants was studied. It was found that the level of the antibiotic production was decreased in most of the biochemical mutants. A necessity for increased fonds of lysine and arginine for biosynthesis of grisin was shown.


[Comparative antiviral and interferonogenic activity of natural and synthetic interferon inducers in vitro and in vivo]. Biological activities of the RNA replicative form of phage f2, a natural interferon inductor and poly-I -- poly-C, a synthetic polyribonucleotide complex were studied comparatively. Differences in the comparative interferonogenic and antiviral activity of the inductors were as dependent on the type of the cell system. It was shown that DEAE-dextran increased the interferon-inducing activity of RFf2 in the cell culture by 4 to 8 times. The dynamics of the interferonogenic and antiviral activity of RFf2 in the L-929 cell culture was studied. Interferon appeared in the culture fluid in 6--8 hours and reached its maximum titers (128 IU50/ml) by the 24th hour, the maximum protection of the cells being also developed by the 12th--24th hour, reaching on an average 51 g PFU/ml. It was shown in the experiments with green marmosets that administration of RFf2 in the form of aerosol in a dose of 2.3 mg/kg induced interferon production in the blood serum the titers of which amounted to 80--160 IU50/ml 24 hours after the administration.


[Effect of penicillin series antibiotics on the properties of Brucella]. Investigation of the properties of antibiotic-resistant variants obtained experimentally from various species of Brucella indicated that benzylpenicillin, ampicillin and ceporin induced significant changes in the main properties of Brucella, i.e. phagolysability, agglutinability, production of brucellacin and hydrogen sulfate, attitude to dyes. Only the nutrient requirements of Brucella did not change when it was grown on synthetic media which is of great importance for investigation of changed Brucella strains.


Iron requirement and chelator production of staphylococci, Streptococcus faecalis and enterobacteriaceae. The effect of iron deprivation on growth of 101 aerobic strains of gram-positive and gram-negative bacteria was studied on agar media in the presence of various concentrations of the synthetic iron chelator ethylene diamine diorthohydroxyphenyl acetic acid (EDDA) and the iron binding protein transferrin. Growth of Staphylococcus epidermidis was inhibited by 15 mM EDDA and 1.5 mM transferrin. Staphylococcus aureus was only inhibited by 44 mM EDDA and not by transferrin. None of the strains of S. faecalis was inhibited. The majority of the enterobacteriaceae (E. coli, Salmonella spp, Klebsiella spp) was inhibited by 44 mM EDDA and 1.5 mM transferrin. The relation between susceptibility and concentration of EDDA and transferrin was expressed as S-value for each species. Iron supply with various iron compounds could restore the effects of inhibition. In all species except in S. faecalis iron chelator production could be demonstrated, using indicator plates of media containing EDDA and flooded with 10(4)--10(5) colony forming units of indicator organisms. The iron chelator of both S. epidermidis and S. aureus could stimulate growth of S. epidermidis, but not that of enterobacteriaceae. Iron chelators from all gram-negative bacteria were functionally interchangeable, but did not stimulate growth of gram-positive bacteria.


A glucan from the cell wall of the cyanobacterium Spirulina platensis. A polysaccharide was isolated from the cell wall of the cyanobacterium Spirulina platensis. Hydrolysis of the polysaccharide only yielded glucose. Curie-point pyrolysis mass spectrometry of the polysaccharide resulted in a spectrum very similar to that of beta-1,2-glucan. Probably the glucan originates from the fibrillar inner layer of the cell wall.


Impairments and recovery of the spatial and temporal components of a visuomotor pointing movement after unilateral destruction of the dentate nucleus in the baboon. In baboons trained to perform visuomotor pointing movements, unilateral electrolytic lesions are performed in the dentate nucleus. The consequences of these lesions on the following movement parameters are then studied:--temporal parameters: reaction time and movement time;--spatial parameters: pointing area and directional errors. From the observations taken 3 months after the dentate nucleus exclusion, a distinction can be made between parameters showing recovery phenomena (reaction time and pointing area) and components definitively affected (movement time and directional error).


Conditions for induction of bacteriophage from lysogenic Bacillus megaterium with aflatoxin B1. The present study was conducted to determine whether or not aflatoxin B1 was an effective inducing agent for lysogenic bacteria and to characterize some of the parameters involved in induction. A lysogenic strain of Bacillus megaterium (NRRL-B-3695) and an indicator strain of this species (NRRL-B-3694) were used. Cultures of the lysogenic strain were incubated for various periods of time in the presence of aflatoxin B1. Plaque-forming units as well as colony-forming units were then determined. Results of the present study indicated that bacteriophage lysogenizing B. megaterium could be induced with aflatoxin B1. The optimum concentration for induction was 25 micrograms of toxin per ml of early-log-phase culture. Evidence suggested that: (i) higher concentrations of aflatoxin B1 formed hydrophobic complexes which would not efficiently induce B. megaterium; (ii) the toxic effect of aflatoxin B1 severely limited the number of cells which could be induced prior to killing action of the toxin; and (iii) concentrations less than 25 micrograms of aflatoxin B1 per ml were not efficient inducers of bacteriophage production nor did they demonstrate the toxic effect observed at higher concentrations.


Degradation of polychlorinated biphenyls by mixed microbial cultures. Three different enriched mixed cultures capable of degrading polychlorinated biphenylas were isolated from two soil samples and a river sediment, respectively. The predominant organisms found in all three mixed cultures were Alcaligenes odorans, Alcaligenes dentrificans, and an unidentified bacterium. The polychlorinated biphenyl isomers that were more water soluble and had lower chlorination were not only degraded at a faster rate than those that were less water soluble and had higher chlorination, but were also more completely utilized by these mixed cultures. This resulted in the presence in the environment of polychlorinated biphenyl residues consisting mainly of higher-chlorinated isomers. A form of cometabolism of polychlorinated biphenyls was also found with these cultures in the presence of acetate as the cosubstrate.


Contact dermatitis due to nitroglycerin ointment. Topical nitroglycerin ointment is among the most effective long-acting vasodilators currently available. We report two cases of contact dermatitis in which patch testing showed allergy to nitroglycerin-containing compounds. The severity of this dermatitis necessitated the discontinuation of topical nitroglycerin therapy in both patients. A potent corticosteroid cream applied once to previous nitroglycerin application sites prevented or minimized this dermatitis and enabled continuation of topical nitroglycerin therapy. Contact dermatitis due to topical nitroglycerin preparations may be more common than is presently realized.


Cutaneous melanoma in a patient with neurofibromatosis. Although neurofibromatosis and cutaneous melanoma are both diseases of neuroectodermal origin, reports of their association are rare. The case history of a patient with histologically documented neurofibromatosis and a nodular melanoma unrelated to a cafe-au-lait spot or congenital nevus is reported, and the literature reviewed. The appearance of only one patient with neurofibromatosis in a series of 900 patients with melanoma suggests that these diseases are probably not associated with greater frequency than that predicted by chance alone.


Drug metabolism in malnourished children: a study with antipyrine. The effect of malnutrition on hepatic drug-metabolising enzymes was investigated in 8 Sudanese children aged between 9 and 12.5 years using as a model the drug antipyrine. Antipyrine half-life and clearance were measured in the malnourished state and after 3 or 4 weeks of good nutrition. Associated with the improvement in nutritional state was a shortening of antipyrine half-life and an increase in its clearance. There was also a rise in serum triiodothyronine. It is concluded that poor nutrition is associated with impairment of drug metabolic capacity and that many factors are responsible.


Intrapartum sensitisation of human infants to the Rhesus-D antigen. A study is reported of the numbers of nonimmunoglobulin-bearing lymphocytes specifically binding to Rhesus-D positive erythrocytes in the blood of Rhesus-dd infants born to either Rhesus-dd or Rhesus-Dd mothers, at birth and at 6 months of age. There was a significant increase in the number of such antigen-binding cells by age 6 months in babies born to Rhesus-Dd mothers. It is concluded that the Rhesus-negative offspring of Rhesus-positive mothers may be sensitised against the Rhesus-D antigen at birth and that protection of such female infants by the injection of antiRhesus-D antibody should be considered.


Adequate anticoagulation during cardiopulmonary bypass determined by activated clotting time and the appearance of fibrin monomer. The adequacy of anticoagulation during 2 hours of cardiopulmonary bypass at 30 degrees C in 9 rhesus monkeys was determined by measuring the whole-blood activated clotting time (ACT) and by noting the appearance of thrombin-altered fibrin (fibrin monomer) and the relative consumption of clotting factors. Factor V and VIII, the heparin cofactor, antithrombin III, prothrombin time, partial thromboplastin time, ACT, platelets, hematocrit, fibrinogen, and fibrin monomer were determined prior to heparinization and after protamine. In 6 of 9 experiments, fibrin monomer became positive in the plasma during cardiopulmonary bypass (CPB), indicating that active coagulation was occurring. In 5 of the 6 animals, initial ACT was less than 400 seconds, and fibrin monomer appeared within the first 30 minutes of bypass. In 1 animal with an initial ACT of 439 seconds, fibrin monomer appeared after 60 minutes of bypass, at which time the ACT was less than 400 seconds. An abnormal level of fibrin monomer was not detected in 5 pediatric patients with an ACT greater than 450 seconds during CPB. Our experimental study and clinical data suggest that the lower limit, as measured by the ACT, for anticoagulant effect to provide coagulation-free CPB is at least 400 seconds.


Diverticula in association with the proximal left subclavian artery: a cause of mediastinal mass lesions. The case reports of 5 patients with an aortic arch diverticulum associated with the proximal left subclavian artery are presented. In 3 patients, the left subclavian artery arose from a diverticulum off a right aortic arch and traversed the retroesophageal area to the opposite arm. In 1 patient, the left subclavian artery arose from a diverticulum off a left aortic arch; this represents the second instance of this anomaly to be reported in the literature. In 1 patient, the left subclavian artery was not attached to the aorta, and a diverticulum was present in the expected area of attachment. Confusion of these anomalies with solid mediastinal mass lesions and with aortic aneurysms may occur. Treatment is predicted on symptomatology, and resection is usually not indicated. It is necessary to remain cognizant of aortic diverticula and to differentiate them from other mediastinal mass lesions.


Selective nonoperative management of contained intrathoracic esophageal disruptions. Eight patients with intrathoracic esophageal disruptions were managed nonoperatively and without pleural drainage. Criteria for nonoperative treatment included the following: disruption contained in the mediastinum or between the mediastinum and visceral lung pleura; drainage of the cavity back into the esophagus; minimal symptoms; and minimal signs of clinical sepsis. Cause of the esophageal perforation was pneumostatic dilatation (1 patient), vomiting (2), and a leak following esophageal operation (5). Antibiotics were administered intravenously to all patients; hyperalimentation was accomplished intravenously in 5, and nasogastric suction was used in only 1. The cavities contracted and the esophageal leaks sealed in all instances. Time before oral intake was resumed ranged from 7 to 38 days (average, 18 days). Days until discharge ranged from 15 to 52 days (average, 28 days).


Esophageal atresia and tracheoesophageal fistula: preoperative assessment and reduced mortality. One hundred twenty-nine infants with esophageal atresia and tracheoesophageal fistula were reviewed from 1955 to 1978. The overall mortality was 38%. Factors associated with the increased mortality include prematurity, pneumonia, and congenital defects other than this anomaly. A classification based on these factors in introduced, which provides the clinician with a prognostic survival rate greater than 90% with only physical examination, chest and abdominal roentgenography, and intravenous pyelography. Postoperative mortality was reduced to 11% in the last five years of the study; this is attributed to the exclusive use of the retropleural approach to the esophagus, more intensive postoperative ventilatory support, and routine use of parenteral nutrition.


Thyroid axis in patients with Cushing's syndrome. Thyroid hormone values and serum thyrotropin (thyroid-stimulating hormone [TSH]) responses to the intravenous administration of 400 micrograms of protirelin were determined in ten patients with Cushing's syndrome and in ten matched normal subjects. In patients with Cushing's syndrome, the serum thyroxine (T4) level was mildly depressed and free T4 level was normal. The mean (+/- SD) concentrations of serum triiodothyronine (T3) and free T3 were both reduced in patients compared with normal subjects (P less than .001). At 20 and 60 minutes after protirelin administration, serum TSH levels were, respectively, 3.3 +/- 2.7 microU/mL and 2.6 +/- 2.3 microU/mL in patients with Cushing's syndrome and 12.3 +/- 5.4 microU/mL and 10.7 +/- 5.4 microU/mL in normal subjects (P less than .001). The reduced serum T3 and free T3 levels are due to a glucocorticoid suppressive effect on the peripheral conversion of T4 to T3. The protirelin test is of limited value in assessing the thyroid status because the response of TSH is frequently blunted or absent due to glucocorticoid excess.


Reversible adrenocorticol insufficiency in fulminant meningococcemia. A 13-year-old girl who had been well previously was admitted with fulminant meningococcemic purpura. Her plasma cortisol level was only 3 micrograms/dL and failed to rise after cosyntropin administration. Maintenance doses of corticosteroid were administered. As her infection cleared her adrenal glands were restimulated and shown to have near normal response. Replacement doses of corticosteroids should be given to patients with fulminant meningococcemia until adrenal cortical insufficiency can be excluded because of the high incidence of adrenal failure in these patients and the possibility that excessive adrenal stimulation might contribute to adrenal abnormality.


[Preparation and testing of polyvalent EDTA-Na-extract vaccine from Escherichia coli strains pathogenic to Swine]. Monovalent vaccines were prepared of E. coli strains with pathogenicity to swine by means of a technique described elsewhere. A polyvalent vaccine was obtained by mixing the monovalent vaccines. This polyvalent vaccine was tested by criteria usually applied to vaccine of E. coli strains with pathogenicity to man, and it exhibited the same quality characteristics.


Cushing's syndrome and autoimmunity.

Cushing's syndrome and autoimmunity. Is Cushing's syndrome ever caused by adrenal stimulating autoantibodies? Specific antiadrenal antibodies were found in the serum of three of seven patients with Cushing's syndrome. The immunologic techniques that showed positive results were complement fixation, tanned RBC hemagglutination, and immunofluorescence. Lymphocytic infiltration of the adrenal was present in two cases, in one of which nodular hyperplasia seemed to be of primary adrenal origin with autoimmunity as a possible cause. However, in cases with clear-cut pituitary origin, the lymphocytic adrenalitis and circulating adrenal antibodies are likely secondary to antigen leaking from damaged tissue.


Waldenström's macroglobulinemia associated with a mixed cryoglobulin. Report of a case with partial precipitation in vitro at 37 degrees C. A 68-year-old man had Waldenström's macroglobulinemia associated with a mixed cryoglobulin (monoclonal IgM kappa and polyclonal IgG, 0.96 g/dL) manifested by purpura, weight loss, hepatosplenomegaly, and proteinuria. On cooling of serum warmed to 50 degrees C, the cryoglobulin begins to precipitate above 40 degrees C, with substantial precipitation at body temperature. Incubation at 37 degrees C (after equilibration at 50 degrees C) causes approximately two thirds of the cryoglobulin to precipitate in 30 to 60 minutes; the precipitate dissolves on rewarming the serum to 50 degrees C. The ability of this cryoglobulin to precipitate at 37 degrees C in vitro indicates that the temperature spectrum of cyroglobulin precipitation can extend to body temperature or above it, and suggests that some serum samples that contain cryoglubulins must be separated quickly at a temperature of 37 degrees C or higher.


Formaldehyde toxicity.

Formaldehyde toxicity. The presence of formaldehyde in biologically active concentrations in several occupational and environmental atmospheres has created concern about the hazards of exposure to these contaminated atmospheres. A review of the clinical and animal toxicologic data demonstrate both primary irritant and sensitizing actions of the compound. The minimal concentration of formaldehyde that can be detected by odor is similar to the level that produces minimal irritant effects on the eyes and in the pulmonary airway, thereby serving as a warning of exposure to the compound. The irritant effects are reversible when exposure is discontinued. Some persons show allergic type responses to concentrations of the compound that are well below the odor threshold. The very limited amount of toxicologic data concerned with effects from prolonged and chronic exposures suggest that the compound does not produce specific delayed types of toxicity. I discuss the factors that influence establishment of "safe" conditions or levels of exposure to formaldehyde.


Judgments of the potential benefits of rehabilitation research. A critical step in a project to develop a benefit-cost approach to evaluating proposed rehabilitation research has been to specify the potential benefits of such research in a reasonably systematic and comprehensive manner. The purposes of this study were: (1) to assess the contemporary relevance of 46 benefit factors yielded by a 1973 survey: (2) to acquire judgmental data to reduce these factors to fewer and more generic 2nd-order factors: (3) to compare 7 different rehabilitation role groups in terms of how the potential benefits of rehabilitation research are valued. "Hierarchical clustering" analyses of similarity judgments by 96 participants yielded 18 2nd-order benefit factors. A number of reliable differences were identified among the role groups regarding the value placed upon specific potential benefits of rehabilitation research.


Spinal cord injury: effect of thyrocalcitonin on periarticular bone formation in three subjects. Sodium fluoride 18F scintimetry was performed before and after 1 month of salmon thyrocalcitonin treatment of 3 spinal cord injured patients with periarticular ossification of the hips and knees. Thyrocalcitonin therapy caused a marked diminution of 18F uptake in 1 patient with long-standing periarticular bone of both hips. Clinically, the range of motion in this subject increased by 25 degrees and there was a marked decrease in pain locally. The results were, however, not duplicated in the 2 patients with periarticular bone formation of short duration.


Maintenance of home hyperalimentation in patients with high-output jejunostomies. We studied four patients with high-output end jejunostomies (greater than 3,000 mL/day) who were receiving home hyperalimentation. With knowledge of urine and stoma output, standard total parenteral nutrition fluid was infused and titrated according to individual needs, first during the hospital stay and then on follow-up office visits. Mean daily stoma output for the group was 3,556 mL/day. With oral fluid intake restricted to 1,500 mL/day, average daily stoma output dropped to 2,892 mL. The mean daily volume of nutrient fluid required to achieve stabel biochemistries and weight was 3,550 mL. Daily caloric requirements ranged from 23 kcal/kg to 44 kcal/kg, with a mean of 32.5 kcal/kg. Amino acid requirements averaged 1.6 g/kg. With 65 patient months of continuous overnight catheter infusion, there have been no serious metabolic derangements or deaths. The program of home hyperalimentation can maintain a satisfactory metabolic state in patients with high-output jejunostomies.


Effects of 4-dimethylaminophenol and Co2EDTA on circulation, respiration, and blood homeostasis in dogs. The effects of intravenously injected 4-dimethylaminophenol and Co2EDTA on peripheral circulation, respiration, acid-base balance, and several other physiological and biochemical parameters were studied on dogs. DMAP increased the respiratory minute volume and mean arterial pressure, diminished the lactate-to-pyruvate ratio, and induced an increase in arterial oxygen pressure caused by liberation of oxygen from oxyhemoglobin during the formation of ferrihemoglobin. A study in vitro of the fate of the oxygen during the reaction between DMAP and oxyhemoglobin showed that only 30--40% of the oxygen released by the formation of ferrihemoglobin appeared in the gas phase. Co2EDTA caused circulatory depression, hyperventilation, and metabolic acidosis resulting in a decrease in base-excess and pH. The concentrations of lactate, pyruvate, potassium, and urea nitrogen and the hemoglobin content were increased by Co2EDTA. The side effects of Co2EDTA in therapeutic doses were more serious than those of DMAP. Thus the latter is superior in the therapy of cyanide poisoning, all the more since it detoxifies more cyanide.


Photoreactivation of ultraviolet irradiated blue-green alga: Anacystis nidulans and cyanophage AS-1. Ultraviolet (UV) inactivation and photoreactivation of Anacystis nidulans and cyanophage AS-1 was studied at different wavelengths. UV inactivation of free phage particles and one and two hour host-phage complexes (intracellular phages) were exponential. UV resistance of plaque forming units was attained at the latter phase of latent period. Black, blue and white lights were able to photoreactivate the UV irradiated A. nidulans whereas green, yellow and red lights were not. However, incubation of A. nidulans for more than 2 hours in black light resulted in loss of viability but shift to red light caused significant recovery. This suggests the involvement of two types of photoreactivation, i.e. of photoenzymatic repair of DNA and of the repair of the photosynthetic apparatus of A. nidulans.


Messenger ribonucleoprotein complexes containing in vitro-synthesized 26S and 42S Semliki Forest virus RNA. An extract derived from Semliki Forest virus (SFV) infected cells is described which catalyzes the synthesis of virus-specific RNAs. The newly-synthesized 26S and 42S RNAs are found complexed with protein as messenger ribonucleoproteins (mRNPs). These mRNPs either are non-membrane bound or are associated with large cytoplasmic lipoprotein membranes, and they are found as free mRNPs as well as mRNPs bound to ribosomal subunits, ribosomes, and polysomes. Following treatment with Tween 40 and deoxycholate, membrane-bound mRNPs containing in vitro-synthesized 26S RNA are dissociated and sediment at 33S. These membrane-dissociated mRNPs contain relatively little protein. In contrast, the free or ribosome-bound mRNPs, which are isolated as 30S to 160S particles, remain heterogeneous after detergent treatment and have a much higher protein content. Addition of purified, native 40S ribosomal subunits to the extract leads to the formation of complexes between the added ribosomal subunits and the newly-synthesized viral mRNA. The in vitro-synthesized 26S and 42S RNAs participate in the assembly of translational initiation and elongation complexes.


A novel plaque method for attenuated rubella virus in Vero cell cultures. Brief report. Under defined plaquing conditions, three attenuated strains of rubella virus showed different plaque morphology and plaque size when tested in Vero cell cultures. The Cendehill strain of rubella virus formed distinct, ring-shaped plaques, while strains HPV77 and RA27/3 produced clear plaques that could be distinguished by their different size. These distinguishing characteristics remained preserved even after the vaccine strains were passaged five consecutive times in Vero cell cultures.


[Morphological analysis of spontaneous hemoblastoses in dogs and cats]. Information of the frequency and morphological characteristics of spontaneous hemoblastoses in dogs and cats observed in 1966-1978 are presented. Hemoblastoses were diagnosed in 90 dogs and 12 cats. Lymphoproliferative diseases were most frequent: lymphosarcomas (80 cases) and lymphocytic leukemias (11 cases). Ten dogs had mastocytomas and one--myeloid leukemia. The cell composition of the organs and tissues involved permitted to distinguish 3 cellular variants of lymphosarcoma: poorly differentiated, lymphoblastic, and prolymphocytic; in both species the poorly differentiated type of lymphosarcoma was predominant (47 cases). It is recommended to use spontaneous hemoblastoses of dogs and cats as an experimental model for investigations of the pathogenesis and therapy of human leukemias.


Valproic acid. Review of a new antiepileptic drug. Valproic acid is a new antiepileptic drug. It has a marked effect on generalized spike-wave discharges. The exact mechanism of action is uncertain; however, some evidence suggests an effect on the metabolism of gamma-aminobutyric acid. It is rapidly absorbed from the gastrointestinal (GI) tract. Concurrent administration with phenobarbital may result in elevated phenobarbital plasma concentrations. Administration with phenytoin sodium may transiently result in lower total phenytoin plasma levels. Side effects are generally mild and include fatigue, GI disturbances, weight gain, a fine postural and resting tremor, mild thrombocytopenia, and an increase in hepatic enzymes. Platelet counts and liver function monitoring should be done during valproic acid therapy. Drowsiness may be seen in patients receiving other antiepileptic drugs concurrently.


Absence status (petit mal status) with focal characteristics. Two patients, aged 23 and 74 years, manifested prolonged episodes of mildly impaired consciousness in conjunction with rhythmical spike waves or spikes (mostly 3/s). This paroxysmal EEG activity was consistently accentuated unilaterally over the superior frontal regions. The first patient showed ictal aphasia and occasional right hemiparesis during these episodes, and partial left frontal lobectomy resulted in temporary freedom from seizures. The classification of these ictal episodes is difficult. They apparently fall into the category of absence status (petit mal status), but the focal neurological signs do not fit the presently valid definitions of absence status, nor does the lack of symmetrical bilateral-synchronous paroxysmal discharges. Perhaps a special category of status epilepticus should be established.


Effect of Salmonella enteritidis 11RX infection on two-stage skin carcinogenesis in mice. S. enteritidis 11RX infection inhibits the growth of a number of transplantable tumours in mice. In addition, oral infection of mice with S. enteritidis 11RX inhibits colon carcinogenesis by 1,2 dimethylhydrazine. This study has examined the effect of S. enteritidis 11RX infection on two-stage skin carcinogenesis in mice using 7,12 dimethylbenz(a)anthracene (DMBA) as initiator and croton oil as promoter. No protection was observed in either LACA or (BALB/c x C57Bl/6J)F1 mice when live 11RX was repeatedly administered i.v. during promotion. When a protein antigen extract from S. enteritidis 11RX was administered i.v. to previously immunised mice during skin carcinogenesis, significant protection was observed both in terms of the number of mice with papillomas and the number of papillomas per mouse. However, the protection was weak and transient. LACA mice were much more susceptible to skin carcinogenesis by DMBA and croton oil than were (BALB/c x C57B1/6J)F1 mice. A preliminary study indicated that BALB/c, C57B1 and CBA mice were also relatively resistant to skin carcinogensis.


Biochemical properties of esterase 6 in Drosophila melanogaster. Biochemical properties of esterase 6 in Drosophila melanogaster were investigated using partially purified preparations from three genotypes, 1/1, 1/2, and 2/2. The molecular weight of the enzyme is estimated to be about 90,000, and treatment with sodium dodecylsulfate cleaves the enzyme into four units with a molecular weight of about 22,000. The activity toward 28 naturally occurring esters was assayed and shown to vary considerably with substrate, the 1/1 preparation having in general higher activity than 1/2 and 2/2, which were very similar. Heat sensitivity, the effect of metal ions, and the effects of the presence or absence of an end product were also studied. The differences demonstrated between allozymes would allow considerable scope, under appropriate conditions, for differential selection to operate between genotypes.


Isolation and biochemical analysis of a temperature-sensitive scarlet eye color mutant of Drosophila melanogaster. Six new ems-induced scarlet mutants were selected. Four of these were partially pigmented, with xanthommatin levels ranging from 12% to 45% of normal. In one (st754ts), pigment production was temperature sensitive; the level of xanthommatin changed from less than 10% of normal at 29 C to more than 70% at 18 C. In all of the new mutants tested, the level of early pupal 3-hydroxykynurenine was as low as that in st1. Thus reduced larval accumulation of this metabolite also appears to be a characteristic feature of scarlet mutants. Temperature-pulse and temperature-shift experiments were carried out with st754ts to determine the temperature-sensitive period for the scarlet gene during development. The major sensitive period commenced prior to the onset of pigmentation and was over before adult emergence. Thus the initiation of xanthommatin synthesis is not brought about by the activation of the scarlet gene. In similar experiments carried out with a temperature-sensitive white mutant (Wbl), a similar temperature-sensitive period was obtained.


Studies of esterase 6 in Drosophila melanogaster. I. The genetics of a posttranslational modification. A locus has been found, an allele of which causes a modification of some allozymes of the enzyme esterase 6 in Drosophila melanogaster. There are two alleles of this locus, one of which is dominant to the other and results in increased electrophoretic mobility of affected allozymes. The locus responsible has been mapped to 3-56.7 on the standard genetic map (Est-6 is at 3-36.8). Of 13 other enzyme systems analyzed, only leucine aminopeptidase is affected by the modifier locus. Neuraminidase incubations of homogenates altered the electrophoretic mobility of esterase 6 allozymes, but the mobility differences found are not large enough to conclude that esterase 6 is sialylated.


Developmental changes of sepiapterin synthase activity associated with a variegated purple gene in Drosophila melanogaster. A variegated position effect on the autonomous gene, purple, has been studid enzymologically in Drosophila melanogaster. Sepiapterin synthase, the enzyme system associated with pr+, was examined for activity in different developmental stages of the fly. The results indicate that T(y:22)prc5, cn/prc4 cn flies (flies in which pr+ has been translocated and which exhibit variegation) have a reduced amount of enzyme activity as compared with both Oregon-R and pr1 flies. This reduction in activity was not found in larval stages, which suggests that the inactivation process probably occurs in late larval or early pupal stages. The phenotype of the variegated adult has white eyes with red-colored spots and patches where drosopterins occur. The phenotype of the fly carrying the translocation is modified by the presence of additional Y chromosomes. This extends the observation from other systems that extra heterochromatin acts to suppress the variegated position effect. The advantages of studying the variegation by measuring enzyme activity, as well as the phenotypic expression, are several; for example, the developmental time at which variegation occurs may be estimated even though drosopterin synthesis is not occurring.


Form II DNA-dependent RNA polymerase from Drosophila melanogaster: general in vitro catalytic properties and template interactions. Several in vitro properties of partially purified form II RNA polymerase from Drosophila melanogaster embryo nuclei are described. The enzyme preparation is free from contaminating RNase, protein kinase, and polyphosphate kinase activities and can be used to study the incorporation of gamma-32P-labeled nucleoside triphosphates. The enzyme exhibits a biphasic heat inactivation pattern which is probably related to differential lability of its two subforms. However, a considerable protection against heat inactivation is provided by the nucleoside triphosphates present in the in vitro reaction system such that the enzyme catalyzes RNA synthesis in a nearly linear mode for over 2 hr at 30 C. Two initiation inhibitors, rifamycin AF/013 was found unsuitable for critical studies because of the high concentrations necessary for total inhibition (200 micrograms/ml) and particularly because of the obligate use of solvents which secondarily have a destabilizing effect on native DNA. Poly[I] was found to effectively block initiation at very low concentrations (1 microgram/ml). The enzyme rapidly forms poly[I]-resistant preinitiation complexes on both double- and single-stranded DNA. These complexes decay with a half-life of 2.5--3 min. RNA synthesis from poly[I]-resistant complexes amounts to 10% of the total potential synthesis on both double- and single-stranded DNA. Enzyme-DNA saturation experiments indicate that the form II enzyme discriminates two types of sites on Drosophila DNA, tight binding and weak binding, from which RNA synthesis proceeds slowly and rapidly, respectively. The tight-binding sites appear to be analogous to those sites with which the enzyme is able to form poly[I]-resistant complexes.


Enumeration of drosophila form II DNA-dependent RNA polymerase initiation sites on Drosophila DNA. The in vitro incorporation of gamma-32P-labeled nucleoside triphosphates into RNA by Drosophila melanogaster form II RNA polymerase from template sites which afford protection from the initiation inhibitor, polyriboinosinic acid (poly [I]), is used as a method for enumerating a specific class of transcription initiation sites on D. melanogaster DNA. Such sites number about 4000 per haploid genome for D. melanogaster. This value is in good agreement with the number of functional genetic units in the D. melanogaster genome as determined by classical cytogenetics.


Energy-dependence of calcium accumulation during sporulation of Bacillus megaterium KM. Ca2+ accumulation and endogenous respiration of sporulating Bacillus megaterium are inhibited to the same extent by electron-transport of inhibitors and the uncoupler carbonyl cyanide p-trifluoromethoxyphenylhydrazone, suggesting that Ca2+ is accumulated by an active transport process. Forespores isolated in stage V of sporulation demonstrated Ca2+-specific carrier-mediated Ca2+ uptake, consistent with downhill transfer [Hogarth & Ellar (1978) Biochem. J. 176, 197-203]. In the present studies forespore Ca2+ uptake was unaffected by carbonyl cyanide p-trifluoromethoxyphenylhydrazone and by concentrations of respiratory inhibitor that inhibited forespore endogenous respiration by 85%. These data suggest that Ca2+ enters the isolated forespore by facilitated diffusion. Ca2+ uptake into sporulating protoplasts was completely inhibited by concentrations of respiratory inhibitors that had no effect on either Ca2+ uptake or respiration of stage-V forespores, but which resulted in inhibition of mother-cell membrane NADH oxidase. These results indicate that the mother-cell membrane is a site for active transport of Ca2+ into the sporulating cell. The effects of the adenosine triphosphatase inhibitor dicyclohexylcarbodi-imide on mother-cell membrane adenosine triphosphatase, NADH oxidase and protoplast Ca2+ uptake were examined.


C3 activation by monosodium urate monohydrate and other crystalline material. Monosodium urate monohydrate (MSUM) is a potent activator of the complement system as measured by electrophoretic conversion of beta1C to beta1A. Activation of C3 in human serum by MSUM is both time- and dose-dependent. The sensitivity of the assay allows detection of C3 activation by as little as 0.2 mg/ml of MSUM. It was observed that C3 activation is calcium dependent and elimination by both EDTA and EGTA, a finding that demonstrated the major role of the classic pathway of complement activation. Excess calcium or magnesium alone inhibited C3 activation by MSUM in accord with the inhibitory effect of these cations on sensitized sheep cell hemolysis by complement. Heating of MSUM at 200 degrees C for 2 hours removes the water of crystallization such that heated crystals may no longer be considered MSUM. Such treatment has a variable effect on C3 activation. Of the crystals and other material studied, only zymosan was more potent than MSUM in activating C3. Calcium prophosphate dihydrate and hydroxyapatite activated significant amounts of C3. Asbestos, glass wool, or a variety of microcrystalline steroids activated little or no C3.


Acute metabolic effects of nitrogen mustard and thiotepa on rabbit articular cartilage and synovium. Metabolic alterations in immature rabbit joint tissue were examined following in vitro and in vivo exposure to the alkylating agents Thiotepa and nitrogen mustard. Brief exposure in vitro to either agent resulted in marked suppression of incorporation of radiolabeled precursors of protein, RNA, and glycosaminoglycan synthesis in articular cartilage, which was partially reversible after Thiotepa exposure. In vivo, nitrogen mustard has little effect on synovium and transient inhibitory effects on cartilage vital processes, whereas Thiotepa caused a prolonged inhibition of synovial metabolism with little effect on cartilage. Autoradiographic localization of labeled agents indicated that synovial tissue and cartilage were readily penetrated by nitrogen mustard, but only a few synovial lining cells and superficial chondrocytes were labeled with 35S-Thiotepa. Furthermore, trypsin significantly reduced labeling of cartilage with 14C-nitrogen mustard. These data suggest that alkylating agents differentially affect metabolic processes in joint tissues in vivo and that with Thiotepa, this interference occurs primarily in the synovium. The degree of interference is apparently dependent upon the time of exposure to the agents and the relative DNA-RNA synthetic activity of the joint tissue.


Changes in thylakoid structure associated with the differentiation of heterocysts in the cyanobacterium, Anabaena cylindrica. The thylakoids of vegetative cells of the filamentous cyanobacterium, Anabaena cylindrica, are capable of oxygen-evolving photosynthesis and contain both Photosystems I and II (PSI and PSII). The heterocysts, cells specialized for nitrogen fixation, do not produce oxygen and lack Photosystem II activity, the major accessory pigments, and perhaps the chlorophyll a associated with PSII. Freeze-fracture replicas of vegetative cells and of heterocysts reveal differences in the structure of the thylakoids. A histogram of particle sizes on the exoplasmic fracture face (E-face, EF) of vegetative cell thylakoids has two major peaks, at 75 and 100 A. The corresponding histogram for heterocyst thylakoids lacks the 100 A size class, but has a very large peak at about 55 A with a shoulder at 75 A. Histograms of protoplasmic fracture face (P-face, PF) particle diameters show single broad peaks, the mean diameter being 71 A for vegetative cells and 64 A for heterocysts. The thylakoids of both cell types have about 5600 particles/micrometers2 on the P-face. On the E-face, the density drops from 939 particles/micrometers2 on vegetative cell thylakoids to 715 particles/micrometers2 on heterocyst thylakoids. The data suggest that the 100 A E-face particle of vegetative cell thylakoids is a PSII complex. The 55 A EF particle of heterocysts may be part of the nitrogenase complex or a remnant of the PSII complex. The role of the 75 A EF particle is unknown. Other functions localized on cyanobacterial thylakoids, such as respiration and hydrogenase activity, must be considered when interpreting the structure of these complex thylakoids.


Photosynthetic vesicles with bound phycobilisomes from Anabaena variabilis. Photosynthetically active vesicles with attached phycobilisomes from Anabaena variabilis, were isolated and shown to transfer excitation energy from phycobiliproteins to F696 chlorophyll (Photosystem II). The best results were obtained when cells were disrupted in a sucrose/phosphate/citrate mixture (0.3 : 0.5 : 0.3 M, respectively) containing 1.5% serum albumin. The vesicles showed a phycocyanin/chlorophyll ratio essentially identical to that of whole cells, and oxygen evolution rates of 250 mumol O2/h per mg chlorophyll (with 4 mM ferricyanide added as oxidant), whereas whole cells had rates of up to 450. Excitation of the vesicles by 600 nm light produced fluorescence peaks (-196 degrees C) at 644, 662, 685, 695, and 730 nm. On aging of the vesicles, or upon dilution, the fluorescence yield of the 695 nm emission peak gradually decreased with an accompanying increase and final predominant peak at 685 nm. This shift was accompanied by a decrease in the quantum efficiency of Photosystem II activity from an initial 0.05 to as low as 0.01 mol O2/einstein (605 nm), with a lesser change in the Vmax values. The decrease in the quantum efficiency is mainly attributed to excitation uncoupling between phycobilisomes and Photosystem II. It is concluded that the F685 nm emission peak, often exclusively attributed to Photosystem II chlorophyll, arises from more than one component with phycobilisome emission being a major contributor. Vesicles from which phycobilisomes had been removed, as verified by electron microscopy and spectroscopy, had an almost negligible emission at 685 nm.


Differential perturbation of erythrocyte membrane-associated transport and enzyme activities by structurally related lipophilic compounds. The alteration of two erythrocyte plasma membrane functions, acetylcholine hydrolysis and glucose exchange, by a series of structurally related small lipophilic compounds which exhibit antihemolytic behavior was studied. 2-Methyldimethylaminoazobenzene is a more potent inhibitor of acetylcholinesterase than the 3'-methyl analogue, while the unsubstituted compound fails to inhibit. Esterase inhibition by the 2-methyl compound is non-competitive and dependent on the anion composition of the assay buffer. The temperature dependence of acetylcholinesterase activity in the presence of the 2-methyl compound suggests that interaction with inhibitor is influenced by the state of lipids tightly bound to the enzyme. Glucose exchange is inhibited to the same extent by both methyl derivatives but not by the unsubstituted dye, and the temperature dependence in the presence of inhibitor is not grossly altered. The lack of correlation between inhibition of membrane function adn stabilization of erythrocytes against osmotic hemolysis is discussed.


Hydrophobic membrane protein from chromatophores of Rhodospirillum rubrum. Structural and spectroscopic studies of monolayers and multilayers. A hydrophobic, lipid- and pigment-free polypeptide from the chromatophore membrane of Rhodospirillum rubrum was spread from chloroform/methanol, pyridine and formic acid solutions at an air-water interface. Surface pressure versus area isotherms of the monolayers formed at the interface were partially dependent upon the spreading solvent used. From the surface area at 20 dynes/cm compression, an average molecular area of 12.9 nm2/molecule was calculated for a polypeptide monolayer spread from chloroform/methanol. Multilayers built up on germanium plates at different surface pressures were subjected to attenuated total reflection infrared spectroscopy. In all cases the amide I and II absorption bands were typical of alpha-helical and random conformations. Electron microscopy of transferred monolayers replicated by rotary platinum shadowing revealed domains of regular texture in specimens prepared at 20 dynes/cm. Such domains were virtually absent in specimens prepared at 10 and 30 dynes/cm. Light optical diffractometry of the ordered arrays yielded a smallest repetitive area of 13.5 nm2 which agrees well with the molecular area obtained from the monolayer surface. Although no drastic changes in secondary structure were detected in the course of this study, some conformational changes are indicated by solvent-dependent differences in the surface pressure versus area isotherms.


Sugar hydrolases of the infant rat intestine and their arrangement of the brush border membrane. Lactase and maltase, the predominant sugar hydrolases associated with the intestinal brush bordermembrane of the suckling rat, were purified essentially free of the other to near homogeneity (lactase at specific activity 23, maltase at specific activity 58), and their specific physiocochemical properties determined. Antisera prepared to each showed by immunodiffusion a single common precipitin line with pure enzyme and solubilized proteins of the brush border membrane. Brush border membranes were purified 26--35-fold from infant rat intestine. Membranes prepared from 10-day-old rats contained 32% protein, 43% lipid and 25% carbohydrate with lactase and maltase estimated to comprise in excess of 10% and 2%, respectively, of the membrane protein. Immunotitration curves of lactase and maltase showed equivalent antibody binding by the membrane-bound and free enzyme forms. Furthermore, antibody binding to one enzyme did not affect the immunotitration curve or the extractability (by papain or Triton X-100) of the other membrane-bound enzyme. It was concluded that the lactase and maltase molecules are attached singly on the external membrane surface in a spatially independent manner with their antigenic sites as freely available to antibody binding as exhibited by their papain-solubilized counterparts.


Replication of the linear mitochondrial DNA of Tetrahymena pyriformis. 1. Electron micrographs of the linear mtDNA from Tetrahymena pyriformis strain GL show linear molecules with a duplex 'eye' of variable size in the middle. This indicates that replication of this DNA starts near the middle of the molecule and proceeds bidirectionally to the ends, as previously shown for the mtDNA of strain ST (Arnberg, A.C., Van Bruggen, E.F.J., Clegg, R.A., Upholt, W.B. and Borst, P. (1974) Biochim. Biophys. Acta 361, 266-276). The mtDNAs of these two strains have little base sequence homology beyond the ribosomal RNA cistron (Goldbach, R.W., Bollen-De Boer, J.E., Van Bruggen, E.F.J. and Borst, P. (1978) Biochim. Biophys. Acta 521, 187-197). 2. Electron micrographs of mtDNA from strain ST, spread under non-denaturing conditions, contain only molecules with fully duplex ends. mtDNA spread under conditions of early denaturation contains duplex loops on one end (40% of all molecules) or both ends (37%). The loops are stable to partial denaturation and vary in size from 0.15 to approximately 1.0 micron, most loops measuring 0.25--0.40 micron. No loops are formed with single-stranded DNA under analogous conditions and we conclude from this result that loop formation is based on the presence of straight, rather than inverted, duplications near the ends. 3. When full-length 3H-labelled mtDNA from strain ST, 32P-labelled at the 5'-termini with T4 polynucleotide kinase, was sedimented in alkaline sucrose gradients, greater than 70% of the 3H and less than 30% of the 32P cosedimented with full-length molecules; the remaining 32P sedimented heterogeneously and predominantly with the DNA less than 10% the size of intact single strands. Brief incubations of full-length mtDNA with DNA polymerase I from Escherichia coli and labelled dNTPs at 15 degrees C did not lead to preferential labelling of terminal EcoRI fragments of the DNA. From these results we infer that the DNA contains nicks or gaps near the termini and that these are not bordered by free 3'-OH groups. 4. A model is presented in which straight sequence repetitions at the termini of Tetrahymena pyriformis mtDNA are involved in the later stages of replication. This model can also account for the pronounced terminal heterogeneity previously observed in this DNA.


Regular, rodlike, macromolecular structures in Artemia salina. Artemia salina of different origins and in different developmental stages contains regular, rodlike, macromolecular structures. The particles were visualized by electron microscopy after negative staining. They can be purified from the postmitochondrial supernatant by sucrose gradient centrifugation in the presence of EDTA. Their size distribution has been measured from electron micrographs. The particles are probably nucleoprotein complexes.


Kinetic mechanism of the aliphatic amidase from Pseudomonas aeruginosa. The kinetic constants for hydrolysis and transfer (with hydroxylamine as the alternate acceptor) of the aliphatic amidase (acylamide amidohydrolase, EC 3.5.1.4) from Pseudomonas aeruginosa were determined for a variety of acetyl and propionyl derivatives. The results obtained were consistent with a ping-pong or substitution mechanism. Product inhibition, which was pH dependent, implicated an acyl-enzyme compound as a compulsory intermediate and indicated that ammonia combined additionally with the free enzyme in a dead-end manner. The uncompetitive activation of acetamide hydrolysis by hydroxylamine and the observation that the partitioning of products between acetic acid and acetohydroxamate was linearly dependent on the hydroxylamine concentration substantiated these conclusions and indicated that deacylation was at least partially rate limiting. With propionamide as the acyl donor apparently anomalous results, which included inequalities in certain kinetic constants and a hyperbolic dependence of the partition ratio on the hydroxylamine concentration, could be explained by postulating a compulsory isomerisation of the acyl-enzyme intermediate prior to the transfer reaction.


Differential biosynthesis of polyunsaturated fatty acids by Tetrahymena supplemented with ergosterol. Tetrahymena grown with foreign sterols such as ergosterol incorporate them into cellular membranes at the expense of the native compound, tetrahymanol. It is shown that cells grown with ergosterol have a lessened capacity to produce the polyunsaturated linoleic and gamma-linolenic acids from [14C]oleic acid. However, the same cells have normal capacities to introduce double bonds at C-6 into linoleate, alpha-linolenate, or cis-vaccenate. Thus, a presumed 12-desaturase is inhibited in the presence of ergosterol, while desaturation at C-6 is unaffected.


Rabbit skeletal muscle F-actin can be stable at low ionic strength, provided trace amounts of Ca2+ are absent. Addition of low concentrations (0.2--2.0 mM) of EGTA to rabbit skeletal muscle G-actin in the presence of ATP caused increase in viscosity. The effect is probably due to chelation of Ca2+. EGTA-polymerized actin was sedimented in the ultracentrifuge as a pellet which could be depolymerized in the presence of Ca2+ and then repolymerized. Electron microscopy indicated that formation of filamentous actin which appears to be somewhat more flexible than F-actin obtained by polymerization with KCl. The EGTA-polymerized actin was dissociated by DNAase I faster than KCl-polymerized actin. F-Actin can thus be stable also in very low ionic strength media if Ca2+ is removed whereas for G-actin to be the only form of the protein in such media, micromolar concentrations of Ca2+ must be present.


Raman spectra and conformational structures of Fab mu and (Fc)5 mu fragments of cryoglobulin IgM-kappa McE. Raman spectra have been obtained on aqueous solutions of the following human immunoglobulins: IgM-kappa McE, IgG-kappa Ger, IgM-kappa WSm and IgG-lambda Gui. The former two species exhibit the property of cryoprecipitation. Comparison of the spectra shows that all immunoglobulins have similar secondary structures, predominantly of the beta-sheet type. Fab mu and (Fc)5 mu fragments of IgM-kappa McE also yield Raman spectra which indicate closely similar secondary structures. Minor differences among the spectra can be explained by differences in amino acid compositions of the respective proteins.


Solution conformation of virginiamycin S. II. The conformation of allohydroxy- and deoxyvirginiamycin S. The experimental results of 1H- and 13C-NMR studies of allohydroxy-, and of deoxyvirginiamycin S strongly confirm the conformation that was proposed earlier for the parent virginiamycin S (Anteunis, Callens and Tavernier (1975) Eur. J. Biochem. 58, 259--268). The changing nature of dipole-induced dipole interaction is responsible for the specific gradually increasing libration of the N-MePhe side chain along the series virginiamycin S, allohydroxy-, deoxyvirginiamycin S. Previous methods for the estimation of rotameric populations around the alpha, beta bonds are critically evaluated and compared to the present results obtained from interpretation of geminal 2J (beta) coupling constants.


Solution conformation of virginiamycin S. III. Patricin A: A further model for cooperative effects of the Pro ring conformation and backbone. Although the main characteristics of the parent virginiamycin S conformation i.e. a bend of type VI (Lewis, P.N., Momamy, F.A. and Scheraga, H.A. (1973) Biochim. Biophys. Acta 303, 211--229) formed by the Pro-N-MePhe-X-PhGly sequence is still present in patricin A, the substitution of X = pipecolic acid by proline in the latter results in a destabilization of the tertiary structure of the depsipeptide, since two isomeric states of a peptidic bond appear in C2HC13 solution. Addition of +/- 30% (v:v) (C2H3)2SO totally shifts this equilibrium in favor of the major parent isomer. These results completely fit with what is known up to now on the occurrence and structure of turns (Chou and Fasman (1977) J. Mol. Biol. 115,135--175).


The presence of a Ca2+ bridge within the gamma chain of human fibrinogen. The presence of Ca2+ increased the mobility of fragment D, and the gamma chain from fibrinogen on polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulphate, suggesting that a Ca2+ was associated with these fibrinogen derivatives. The mobilities of the constituent chains from fragment D produced under various conditions, indicate that Ca2+ bound to fibrinogen form an intrachain bridge towards the C-terminus of each gamma chain.


Manual solid phase sequence analysis of polypeptides using 4-N-N,-dimethylaminoazobenzene 4'-isothiocyanate. A manual solid-phase method for sequence analysis of polypeptides is described. The immobilized polypeptide was subjected to stepwise degradation by Edman-type reagent, using the 4-N,N-dimethylaminoazobenzene 4'-isothiocyanate phenyllisothiocyanate double coupling method. The N-terminal amino acids were released (after conversion reaction) as 4-N,N-dimethylaminoazobenzene 4'-thiohydantoin (identified by thin layer chromatography) and phenylthiohydantoin derivatives. The method required 2--10 nmol polypeptide.


Immunological comparison of proline-rich proteins from human and primate parotid secretion. Antisera raised in response to proline-rich proteins purified from parotid secretions of man and the primate Macaca fascicularis were employed to investigate the interrelationships of these proteins by immunodiffusion, immunoelectrophoresis and the combined use of disc gel acrylamide electrophoresis with radial immunodiffusion. The major human proline-rich proteins, PRP I, PRP II, PRP III and PRP IV as well as several minor proline-rich proteins cross-react with antiserum to PRP I or PRP III. Similarly primate parotid saliva contains several components cross-reacting with antiserum directed against a purified primate proline-rich protein, MPRP. Antiserum to PRP I or PRP III cross-reacted with MPRP and primate parotid saliva protein, whereas antiserum to MPRP cross-reacted only with human parotid saliva protein and not with the isolated human proline-rich proteins. The immunological relationships of these salivary proline-rich proteins within and between species suggest their origin from a common precursor molecule.


Characterization of an inhibitor of the intracellular protease from Bacillus subtilis. A specific inhibitor of intracellular serylprotease from Bacillus subtilis has been isolated from both growing and sporulating cells. Like other protease inhibitors isolated from eukaryotic cells, the inhibitor from B. subtilis is a thermostable protein. A purification method is described. The molecular weight estimated by Biogel filtration and SDS gel electrophoresis is about 15,500. Both proteolytic and esterolytic activities of intracellular protease are equally sensitive to inhibition. With azocoll or Z-tyrosine p-nitrophenylester as substrates, noncompetitive inhibition patterns are observed. The inhibitor has no effect on the proteolytic or esterolytic activities of the extracellular serylprotease. A similar thermostable inhibitor is also present in Bacillus megaterium.


[Subtilisin 72: a serine protease from Bac. subtilis strain 72 - an enzyme similar to subtilisin Carlsberg]. Subtilisin 72, a serine proteinase secreted by Bac. subtilis strain 72 was purified by covalent chromatography on Sepharose sorbent containing p-(omega-aminomethyl)phenylboronic acid as a ligand. The homogeneity of subtilisin 72 was confirmed by isoelectrofocusing in a thin layer of polyacrylamide gel (pl 8.6). The amino acid composition of this enzyme is different from that of other subtilisins, e. g. subtilisin Carlsberg. The N = terminal amino acid sequence of subtilisin 72 traced up to the 35th residue turned to be the same as that of subtilisin Carlsberg with the exception of the 21st (Tyr) and the 30th (Ile) residues. This very pronounced extent of homology shows that subtilisin 72 is very similar although not identical to subtilisin Carlsberg.


Porous polysulfone as an attachment vehicle for orthopedic and dental implants. The mechanical properties of porous polysulfone (PSF) were determined and its performance as a surface coating on orthopedic and dental implants evaluated. Ten coated femoral prostheses were implanted in nine dogs. A second series of four uncoated "control" prostheses and all of the acetabular cups were implanted using conventional bone cement techniques. Six porous PSF coated tooth roots were implanted in healed mandibular premolar extraction sites in three Rhesus monkeys. The shear strength of porous PSF (6 MPa) was comparable to that of trabecular bone. Pushout tests of 1cm. thick sections of the prostheses yielded interfacial shear strength values over 1.4 MPa for the PSF coated hips after 14 weeks and mean values under 0.7 MPa for the bone cemented specimens after 3 and 36 weeks. Bone and fibrous tissue was identified in the pores of coated specimens. Preliminary clinical evaluation of the functioning dental implants revealed zero mobility and other favorable clinical and radiographic indications after 2 months.