Comparative effects of anesthetics on the viability and integrity of Escherichia coli ML30. Cells of Escherichia coli ML30 in a mineral salts medium were exposed to dichlorodifluoromethane (f-12), cyclopropane, halothane, or Ethrane at concentrations of 1.25, 0.2, 0.04, and 0.008 X saturation for times up to 1,200 min, and at temperatures in the range of 2 to 37 C. When any of these anesthetics were applied for 300 min at 1.25 X saturation, a substantial decrease in number of survivors occurred. Halothane was most bactericidal, cyclopropane and Ethrane were moderately bactericidal, and t-12 was least bactericidal. At saturation values of less than 1.0, none of the four anesthetics had an appreciable effect on viability of E. coli. Greatest increases in cell permeability occurred when anesthetics were used at saturation values of 1.25, and permeability changes generally decreased as the concentrations of the chemicals were reduced. In many instances, anesthetics in the vapor state caused significant increases in cell permeability but little or no loss of viability. This indicated that a close relationship did not exist between loss of viability and increased permeability. All four anesthetics caused E. coli to lose substantial and similar amounts of compounds absorbing at 260 nm. Release of compounds absorbing at 260 nm generally increased as the saturation value of a given chemical was increased. Halothane, Ethrane, and cyclopropane but not f-12 caused lysis of E. coli ML300. Considering all results, E. coli ML30 was damaged more by halothane or cyclopropane than by f-12 or Ethrane. When f-12 was applied at a saturation value of 1.25, the bactericidal effect on E. coli was much greater at 37 or 22 C than at 12 or 2 C.


Antibodies against enteric bacteria in brown bullhead catfish (Ictalurus nebulosus, LeSueur) inhabiting contaminated waters. Brown bullhead catfish were collected from sewage- and acid mine waste-polluted waters in an attempt to detect antibodies against human enteric bacteria in their sera and to investigate the association of antibody response with environmental conditions. Agglutination antigens prepared from isolates obtained from water collected at the same locations as the fish habitats were used to demonstrate such antibodies. The results showed large percentages of reactive sera for common isolates such as Escherichia coli and Enterobacter cloacae as well as lesser incidences of antibodies to other, less common isolates. In general, fish with the highest titres were collected from habitats with higher coliform counts. Acid mine drainage reduced the total coliform counts, but did not appear to affect the titers of sera from fish collected from water so affected. It was concluded that the bottom-feeding catfish might be a better subject for the study of fish as an ecological indicator of fecal pollution in acid-polluted waters.


Principal component analysis of infraspecific variation in bacteria. In certain types of ecological investigations it may be desirable to investigate infraspecific variation in bacteria. Principal component analysis is demonstrated to be satisfactory for this purpose. Hypothetical bacterial populations were used to show that such analysis can be used to compare collections of bacterial isolates taken at different times or from different sources. Alternatively, given n isolates, whether they represent a single bacterial population can be determined. The method is applied to authentic collections of bacteria in three separate analyses. The results are compatible with current taxonomic tenets.


Total rinse method for microbiological sampling of the internal cavity of eviscerated broiler carcasses. A method is described for spray-rinse sampling the entire visceral cavity of broiler chicken carcasses for microbiological analyses. The method is designed to provide a more comprehensive sample than swabbing or excision of small areas.


Immunologic phenomena in herpes gestationis. Immunofluorescence findings are of diagnostic significance in herpes gestationis. The studies in two cases demonstrated some complement components (chiefly C3 and C4, but no C(1q)), without immunoglobulins at the basement membrane zone and with no circulating anti BMZ antibodies. In one case IgG were found at the BM zone in one biopsy specimen, but were not demonstrable in several other biopsies. The IF findings seem to indicate both the alternate and classical pathways of complement activation, and speak in favor of a relation between herpes gestationis and bullous pemphigoid, in which sometimes the immunoglobulins are also not demonstrable and the immunofluorescent pattern is very much like that of herpes gestationis.


Long term studies of pancreas transplantation in experimental diabetes mellitus. Alloxan diabetes was induced in inbred rats that then were divided into four groups consisting of unoperated diabetic controls, sham-operated diabetic controls, rats given pancreaticoduodenal isografts, and rats given duct-ligated pancreas isografts. The animals were studied for from 18 months (controls) to two years (transplants) and the following important results were obtained: 1) In striking contrast to the diabetic controls, pancreas transplants of both types produced immediate and permanent relief of hyperglycemia, immediate and lasting elevation of serum insulin levels, a normal weight and growth curve, and good health for two years. Removal of the graft was followed by recurrence of severe diabetes. 2) Pancreas transplants of both types prevented the widespread and severe renal, ophthalmic and neural lesions of diabetes that were found in the diabetic controls. 3) The duct-ligated pancreas graft and pancreaticoduodenal transplant were equally effective in controlling diabetes. Ligation of the pancreatic duct was not followed by significant morphologic or clinical evidence of pancreatitis or by loss of endocrine function. 4) Portal venous drainage of the pancreas transplant was unnecessary for good endocrine function.


New surgical concepts resulting from cranio-orbito-facial surgery. The authors have defined the subspecialty of craniofacial surgery and described the organization of the multi-disciplinary team required to care for such patients. Common features of the craniofacial patient have been summarized and three major categories of patients have been proposed. These are: I. Syndromes associated with hypertelorism; II. Syndromes associated with premature synostoses or growth arrests; III. Syndromes associated with primarily mid- and lower face anomalies. Growing out of an experience with 242 operations on 106 patients, the authors have listed 9 relatively new surgical "principles." Each has led to a current surgical approach that is now being employed by the craniofacial team at The University of Virginia. A number of examples are given to show ways in which the lessons learned from the craniofacial patients are now being applied, with improved results, to patients with neoplasms, traumatic injuries, or other conditions.


Correction of the transposition of the great arteries. Inversion of the atrial flow in transposition of the great arteries is achieved in the following way: the remaining atrial septum is detached from behind, leaving it as a flap between both AV-valves. The incision is continued into the coronary sinus and the Vena magna cordis to the base of the left atrial appendage along the mitral ring. The resulting flap from the split coronary sinus is sutured to the left atrial wall in front of the left side pulmonary veins, thus forming a septum dorsal to the caval veins, diverting the left pulmonary venous blood to the right side. The atrial septal flap is reattached in front of the caval vein orifices. In 50 cases a small Dacron patch was used to complete the midportion of the new septum. Six patients died and so far we have encountered one superior Vena cava stenosis and one pulmonary vein stenosis. In 16 patients completion of the midportion of the new atrial septum was done with a partially excised flap from the right atrial wall still attached to the Vena cava inferior. This method resulted in 3 operative deaths. In 42 of 58 survivors the ECG has been repeatedly controlled 3 months to 6 years postoperatively. Thirty-six of the 42 patients have sinus rhythm, 4 vary between nodal and sinus-rhythm and 2 have a total AV-dissociation.


Complications of pulmonary resection.

Complications of pulmonary resection. The following complications of pulmonary resection are discussed with reference to their frequency of occurrence, etiology, diagnosis, and treatment: pulmonary insufficiency, arrhythmias, residual intrapleural air spaces, prolonged air leaks, postpneumonectomy empyema, bronchopleural fistula, cardiac herniation, lobar gangrene, esophagopleural fistula, pulmonary embolism, and tumor embolism.


Pulmonary artery pressure and lung water during extracorporeal circulation in experimental pulmonary insufficiency. Use of the membrane oxggenator has been advocated in the management of severe respiratory insufficiency. We have compared this method to conventional therapy in an experimental model in which 23 dogs were subjected to aspiration with 0.1 N HC1 and 18 were supported with a volume respirator and positive end-expiratory pressure or placed on partial bypass using a membrane oxygenator for 12 to 24 hours. Adequate oxygenation (POX greater than 100 mm Hg) was achieved with the membrane oxygenator. The increase in lung weight during conventional therapy was significantly greater than during membrane oxygenator support. Extracorporeal support during recovery from severe pulmonary injury allows pulmonary artery pressure to be controlled and reduces the expected increase in lung water.


Cardiovascular effects of central microinjections of apomorphin in cats. Microinjections of apomorphine (25-300 mug) were made into the lateral ventricle, dorsal nucleus of the vagus and ventromedial nucleus of the hypothalamus of cats through a stereotaxically implanted cannula-electrode. Apomorphine caused depressor and bradycardic effects without any dose-response relationship. At the above doses of apomorphine the efferent vagal discharges werr markedly increased concurrent with cardiovascular changes. Respiration was not affected except at higher doses causing depression in some experiments. Pretreatment with atropine, scopolamine or haloperidol abolished those responses. Similar results were observed with bivagotomy and midcollicular transection. Dopamine (125-100 mug), acetylcholine (10-100 mug) and norepinephrine (25-100 mug) caused similar cardiovascular changes, as in the case of apomorphine, and such effects were blocked by both specific and non-specific autonomic blackers. Thus the hypothalamus and the dorsal nucleus of the vagus appear to be inbolved in the central cardiovascular effects of apomorphine and such effects may be mediated through more than one neurotransmitter mechanisms.


Influence of temperature and cocaine on responses of the isolated mouse vas deferens to adrenergic amines. Lowering the bath temperature from 37 degrees C to 20 degrees C resulted in supersensitivity of the isolated mouse was deferens to norepinephrine. Dose-response curves to norepinephrine were shifted to the left along the log dose axis (enhanced potency) and the maximum degree of contraction that could be elicited was increased. The effect of cocaine on norepinephrine effects was qualitatively the same as hypothermia. Both treatments enhanced the potencies of other alpha receptor agonists that are substrates for the adrenergic neuronal membrane uptake mechanism. Neither treatment enhanced the potency of methoxamine, which is not taken up by adrenergic neurons, but hypothermia increased the methoxamine-induced maximum contractile height. Uptake of 14C-norepinephrine by the vas deferens was abolished by cocaine and low temperature. The data suggest that enhanced potency of adrenergic amines by hypothermia results from an inhibition of uptake by adrenergic neurons. Increased height of maximum contraction at low temperature may be partly due to decreased neuronal uptake, but other mechanisms are also likely to be involved in this effect.


Lymphocyte function in rheumatic disorders. This review discusses the immunology of rheumatic diseases, specifically rheumatoid arthritis, and systemic lupus erythematosus, and presents some known data, while suggesting what more needs to be learned. Cellular immunity occurring in rheumatoid arthritis and systemic lupus erythematosus is emphasized.


Cryptococcal meningitis: diagnostic value of cryptococcal antigen in cerebrospinal fluid. In three previously reported cases of cryptococcal meningitis, the only laboratory evidence for this diagnosis was the presence of cryptococcal antigen in the cerebrospinal fluid (CSF). Three additional patients had chronic meningitis and repeatedly negative CSF cultures and had cryptococcal antigen demonstrated in the CSF. In our patients, the diagnosis was further supported by the complete recovery after amphotericin B therapy in two and the demonstration of Cryptococcus neoformans in the meninges at autopsy in the third. In certain patients with chronic meningitis, the detection of cryptococcal antigen in the CSF may be the only means of establishing a diagnosis during life. In such patients, if cryptococcal antigen is present in the CSF in a titer of larger than or equal to 1:8, antifungal therapy should be initiated, pending results of other diagnostic studies.


Fungal infection following renal transplantation. Twenty-seven deep fungal infections developed in 22 of 171 patients following renal transplantation. These infections included cryptococcosis (ten), nocardiosis (seven), candidiasis (four), aspergillosis (two), phycomycosis (two), chromomycosis (one), and subcutaneous infection with Phialophora gougeroti (one). Twelve infections occurred in living-related and ten in cadaveric recipients. Nineteen of the 22 patients were male. Infections occurred from 0 to 61 months after transplantation. Complicating non-fungal infections were present concomitantly in 15 patients. Thirteen patients died, eight probably as a result of fungal infection. Appropriate diagnostic procedures yielded a diagnosis in 20 of 27 infections, and therapy was begun in 18 patients. Serologic, culture, and biopsy procedures useful in making rapid diagnoses are advocated in the hope of increasing survival.


[Phagocytosis in patients with splenectomy]. Phagocytosis was evaluated in nine patients with splenctomy undergoing renal transplant, eight renal transplanted patients without splenectomy, three immunodepressed patients with terminal renal failure, four patients with splenectomy and six healthy volunteers. For phagocytosis evaluation, neutrophils adhered to glass were used. They were cover with 4% latex and albumin containing culture medium with or without phagocytosis stimulating tetrapeptide extracted from IgG in each case. Phagocytosis was decreased in all groups studied. Immunodepressed patients showed the highest diminishment of phagocytosis and patients with splenectomy and immunodepression the lesser. It was speculated that such a finding is due to a decrease of tetrapeptide in serum.


[The influence of the antivitamine pyrithiamine on the metabolism of the thiamin-autotrophic Serratia marcescens]. A new method is described for the quantitative microbiological analysis of the thiamine analog pyrithiamine present in biological material. The method uses a mutant of Lactobacillus fermenti (ATCC 9338) stimulated by pyrithiamine. By this specific method it is found that Serratia marcescens is able to consume and to phosphorylate the antivitamine (presumably to pyrithiamine monophosphate). The uptake of the analog influences neither the generation time of Serratia marcescens nor the biomass of the culture, nor the total amounts of carbohydrate and protein within the cells. On the contrary, pyrithiamin stimulates the exponential death-rate k and decreases the quantity of neutral fats. The carbon source glycerol is consumed at a higher rate, which is paralleled by an increased total amount of pyruvate as well as by an increased biosynthesis of acetoin.


Exercise electrocardiograms: Code for their clinical evaluation. This report suggests the use of a simple clinical method of codification of the most significant electrocardiographic changes as a result of the exercise stress test. It uses common abbreviations in the evaluation of the electrocardiographic findings before, during and after exercise. The five major categories to be codified include: (1) the basic electrocardiographic pattern, (2) the heart rate, (3) the pattern of ventricular conduction, (4) the pattern of rhythm and (5) the ST segment deviation. The initial sequence of five symbols indicates the findings in the preexercise electrocardiogram. This is followed by the word "to" with a  subscript number. The number in subscript indicates the peak heart rate achieved during the exercise. The sequence of symbols following "to[]" serves to indicate the electrocardiographic changes observed in the postexercise period. The data codified using this system have been found to be concise and easily comparable. The familiarity of the symbols used facilitates its learning and application.


Glucocorticoid and antibiotic effect on experimental gram-negative bacteremic shock. This study was designed to answer the three following questions: (1) Are glucocorticoids as protective in Gram-negative bacteremic shock as they are in endotoxic shock? (2) Is there any difference in efficacy between a bacteriostatic and a bactericidal antibiotic in bacteremic shock? (3) Does the combination of glucocorticoid with antibiotic potentiate the individual protective effects of both? Bacteremia was induced in male Sprague-Dawley rats by a single intravenous injection of viable Escherichia coli. The results showed that dexamethasone sodium phosphate alone afforded significant protection against Gram-negative bacteremic shock up to eight hours after challenge. The choice of a bactericidal vs a bacteriostatic antibiotic did not influence the survival rates in this study. The survival rate was maximal when dexamethasone was used with both ampicillin sodium and gentamicin sulfate.


Low-dose heparin in postoperative patients: a prospective, coded study. One hundred five patients over age 40 undergoing various major operations were randomly divided into control and treated groups; all were treated by subcutaneous injection containing either 5,000 international units aqueous heparin sodium or a placebo one hour prior to operation and every 12 hours thereafter for eight days. Deep vein thrombosis (DVT) was detected by daily -125I-fibrinogen injection and leg scanning, and confirmed by ascending phlebography. Both groups were comparably distributed by age, sex, variety of operation, incidence of previous thromboembolism, and myocardial and cerebrovascular disease. Blood loss was not increased in the treated group. Incidence of DVT was 8.6 percent for the total group, 7.5 percent in the heparin-tested group (four of 53), and 9.6 percent in the control group (five of 52), including one control patient with a normal scan (who later had a pulmonary embolus.


The leukocyte aggregation test: immunodiagnostic applications and immunotherapeutic implications for clinical renal transplantation. The leukocyte aggregation test (LAT) detects the in vitro adhesion of sensitized, but not nonimmune, recipient leukocytes onto donor kidney cell monolayers. The test specifically detects cell-mediated homograft immunity up to 15 days prior to the appearance of clinical signs or alteration of chemical indexes. The presence of a positive reaction always signified incipient homograft rejection, which was usually controlled by intravenously administered, high-dose methylprednisolone sodium succinate (Solu-Medrol) therapy. There was no instance in which methyl-prednisolone treatment effectively reversed rejection in the presence of a negative leukocyte aggregation test. One common form of homograft rejection may be characterized by positive LAT results, a cellular infiltrate on the renal biopsy specimen, and sensitivity to methylprednisolone therapy.


[Caudal vertebro-medullary dysrhaphia. Intra-sacral forms]. The mean theories of the sacral spinal dysraphisms are reviewed and 4 cases of "meningeal intrasacral diverticulum" are reported. The authors prefer this term to the more common "intrasacral occult meningocele". The proeminent symptom in their cases was late vesical incontinence, which disappeared after surgery. In the pertinent literature 24 similar cases were described till now.


Use of the Lombard response in cases of hysterical aphonia. Lombard is best known for his work on the effect of noise on vocal intensity. In 1917, he applied this concept to the treatment of hysterical aphonia and described the procedure he used in a military publication. He suggested that a masking noise will interfere with the auditory monitoring system sufficiently that the patient will not be aware he has regained use of phonation. Removal of the noise from the patient's ears confirms the return of phonation, and the aphonia disappears. Credit is given to Lombard for this pioneering effort, and suggestions for present-day clinical use are made.


Membranous glomerulopathy in a patient with sarcoidosis. We describe a case of membranous nephropathy in a patient with pulmonary, splenic and hepatic sarcoidosis. The patient was asymptomatic, and edema was absent notwithstanding the proteinuria (over 8.0 gm/100 ml daily). Prednisone cleared the pulmonary and splenic complications, but the proteinuria, although diminished, persisted. Adjunctive therapy with cyclophosphamide caused further diminishment of the proteinuria. We have reviewed the relationship between the nephropathy and the sarcoidosis and suggest that a causal relationship exists between the two diseases.


Granulomatous pneumonitis. A result of intrapleural instillation of quinacrine and talcum powder. A 73-year-old man with bilateral recurrent pleural effusions had relief of symptoms with intrapleural instillation of quinacrine hydrochloride and talcum powder. At postmortem, examination of the lungs showed granulomatous consolidation of a portion of the lower lobe of the left lung. The granulomatous reaction was in response to large numbers of talc and quinacrine crystals within the pulmonary parenchyma. A review of the literature disclosed no reports of pulmonary damage following the intrapleural administration of these two agents. This unusual complication of therapy may have resulted from the aspiration of crystals through a small, undetected bronchopleural fistula.


Changes in the composition of a nickel-base partial denture casting alloy upon fusion and casting. Three series of tensile test pieces were produced using a nickel-base partial denture casting alloy. For the first series induction heating was employed for melting the alloy, for the second a resistance crucible, and for the third an oxy-acetylene torch. In each series the same metal was cast sequentially five times, following which samples of the alloy were subjected to a ten element quantitative analysis to ascertain compositional changes associated with the three methods of fusion.


The effects of remelting on the mechanical properties of a nickel base partial denture casting alloy. Three series of tensile test pieces were produced using a nickel base partial denture casting alloy. For the first series induction heating was employed, for the second a resistance crucible, and for the third, an oxy-acetylene torch. In each series the same metal was cast sequentially a number of times and all test pieces so produced were subjected to mechanical testing. The mechanical properties were found to vary according to both the number of times the alloy was cast and the method of heating used to render the alloy molten.


The pulpless abutment.

The pulpless abutment. The use of preformed metallic anchorages inserted in satisfactorily treated teeth for increased stabilization and retention in complete and partial dentures is demonstrated. An additonal advantage is the retention of proprioceptive guidance in mastication with complete dentures. A number of clinical examples are illustrated.


Microporosity in casting alloys.

Microporosity in casting alloys. Three series of tensile test pieces were produced using a nickel base partial denture casting alloy. For the first series induction heating was employed, for the second a resistance crucible, and for the third an oxy-acetylene torch. Samples from each series were sectioned longitudinally, mounted, polished and examined microscopically for evidence of microporosity.


Periodontal treatment requirements of a group of defence forces' personnel. The periodontal treatment requirements of 187 Royal Australian Air Force men aged 18-54 years based on clinical signs and symptoms were assessed. Every man required some treatment; 60 per cent needed surgery in one or more segments of the mouth.


Papillary necrosis in renal allografts--report of 2 cases. Two patients are reported in whom early and irreversible renal allograft rejection was associated with passage of necrotic papillary material in the urine. Evidence is presented for a causal relationship between rejection and medullary necrosis. Passage of a papilla is regarded as a definite indication for graft exploration.


Aortic valve replacement following cadaver renal transplantation. Acute bacterial endocarditis producing aortic incompetence developed in a patient undergoin haemodialysis.  Subsequent cadaver renal transplantation and aortic valve replacement has proved successful.


Cold agglutinin formation in patients undergoing haemodialysis.  A possible relationship to dialyser re-use. Cold agglutinins with anti-N characteristics were shown to develop in as short a time as four months in patients on home dialysis.  It appeared that the development of such antibodies is related to dialyser re-use.  Those who were treated without re-using their dialyser did not develop these antibodies.  Anti-N may cause early graft failure associated with agglutination in a cold allograft.  Moreover, if these antibodies develop in patients on dialysis there may be problems with extracorporeal dialysis and blood transfusion.


The pituitary-testicular response to luteinising hormone releasing hormone administration to normal men. The response of the pituitary and testis in normal men to the injection of LH-RH has been studied using single intravenous doses of either 25 mug or 100 mug.  Serum LH levels rose with both doses but significant elevation of plasma testosterone was evident only following 100 mug of LH-RH.  Though testosterone levels were elevated within 60 minutes, peak responses occurred from 4-12 hours post-injection.  The rise in serum FSH levels was small or absent at 25 mug but was clearly present following 100 mug LH-RH, in general rising more slowly than LH levels but persisting for a longer period.


Hemodynamics, renal function, plasma renin, and aldosterone in man after 5 to 14 days of bedrest. Continous bedrest for 5 to 14 days had no significant effect on resting heart rate, blood pressure or cardiac output in six normal men. Head-up tilt induced greater tachycardia in 5 of 6 patients after bed rest than in the control period. Propranolol diminished both the tachycardia and the incidence of hypotension and faintness in upright posture. Body weight, serum electrolytes and resting renal plasma flow, and glomerular filtration rate were unchanged by bedrest. Plasma volume fell, extracellular fluid volume increased, and plasma renin activity was significantly elevated following bedrest. Unusually large increases in plasma renin followed head-up tilt or administration of isoproterenol during bedrest, and after resuming normal activity. During bedrest, plasma aldosterone was often increased in the early morning. We conclude that after bedrest, upright posture evokes strong beta-adrenergic activity, with exaggerated metabolic and circulatory responses which can be reduced or abolished by the beta-adrenergic blocker, propranolol.


Human assay of antimotion sickness drugs. The present study was undertaken to improve previous testing procedures, involving the use of a slow rotation room, for assessing the efficacy of antimotion sickness drugs which had validity for groups of subjects but not for each individual in the group. Three major changes were introduced: first, the use of an incremental increase in the intensity of the stressful stimuli of constant intensity; second, a systematic distribution of placebos, rather than a random distribution, in using a modified Latin-square design; third, categorizing the responses as "inconsquential," "beneficial," and "detrimental"--the range of the "placebo responses" was doubled to define the inconsequential range and response above or below were, respectively, beneficial or detrimental. Only drugs known to have antimotion sickness effectiveness were tested and the cardinal findings can be briefly summarized: 1) The group responses were similar to the data previously reported; 2) Great individual differences in response to antimotion sickness drugs were revealed, implying that individual assessments must be made for maximal benefits; 3) The fixed-dose combination of promethazine hydrochloride and ephedrine sulfate (25 mg each) proved to be outstanding as this combination of homergic drugs clearly exhibited a suprasummation effect; and 4) A few tests were conducted using larger than usual doses and the results supported previous findings that, for a maximal beneficial effect in response to a single dose, individuals may vary both with regard to the choice of drug and the amount administered.


Surveillance of some infectious diseases among aircrew personnel in Southeast Asia. A 2-year analysis of specimens among aircrew personnel in Southeast Asia is reported. Stool specimens were examined for intestinal parasites and enteric bacteria, blood smears for blood parasites, and sera tested for transaminases (SGOT) and antibodies to Entamoeba histolytica, Toxoplasma gondii, Chikungunya and Japanese encephalitis viruses, Rickettsia tsutsugamushi, Leptospira sp. and Pseudomonas pseudomallei. One to four specimens each were obtained from 537 adult males and 20 females. There were 56 subjects with intestinal parasites as follows: Giardia lamblia (3.8%), Endolimax nana (2.6%), Entamoeba histolytica (1.4%), Entamoeba hartmanni (0.9%), Entamoeba coli (0.7%), Trichiuris trichiura (1.7%), Ascaris lumbricoides (1.4%), hookworm (0.9%), and Clonorchis sinensis (1.2%). Two individuals had malaria, Plasmodium vivax (0.6%). Pathogenic enteric bacteria were isolated from seven stool specimens; Shigella groups B and D (0.9%), Salmonella paratyphi (0.3%), and Arizona group (0.9%). Significantly elevated SGOT levels were found in 0.9% and antibodies to Japanese encephalitis virus in 1.5%, to Rickettsia tsutsugamushi in 1.2%, to Pseudomonas pseudomallei in 0.3%, to Entamoeba histolytica in 0.9% and to Toxoplasma gondii in 10.1%. No antibodies were found to Chikungunya virus or Leptospira sp.


Effects of pyrobenzamine and plimasin on fighter pilots flying a fighter intercept mission in the F4D flight simulator. There were 26 F4D crews from a Tactical Air Force Wing which participated in a difficult intercept mission in a F4D flight simulator. They were divided into three groups, medicated with either Plimasin, Pyrobenzamine or placebo. The groups medicated with either Plimasin or Pyrobenzamine alone demonstrated decreased effectiveness in completing this intercept as compared with the nonmedicated group. These medications apparently caused impairment of mission performance and should be avoided while performing flying duties.


Induced systemic candidiasis in young broiler chickens. Systemic candidiasis was induced in broiler chickens 14 days old by intravenous injection of a suspension of viable Candida albicans cells. Injection resulted in decreased body weight, moderate mortality, swollen and reddened livers and kidneys, pancreatitis, and disturbances of the nervous system. Three types of neutral disturbances were observed: 1) extreme opisthotonus with spasmodic tremors; 2) extreme torticollis with cranial rotation of 270 degrees: and 3) extreme torticolis which resulted in the head being drawn in a medial-ventral direction. None to 50% of the inoculated birds exhibited these neural disturbances, depending on the isolant of C. albicans used. Microbiological examination of internal organs and blood revealed that C. albicans localized in the meninges of the brain. There was also a significant isolant-related effect of C. albicans on the growth rate of the inoculated birds. These easily quantitated differential effects of various isolants of C. albicans offer the prospect of correlating biochemical characteristics with virulence and obtaining information about the mechanism of pathogenicity of this microorganism.


Regulation of normal hemopoiesis in the acute leukemia L5222. Normal hemopoiesis becomes markedly depressed in rats during the development of the rat leukemia L5222. Whether this could be due to the influence of a circulating humoral factor was investigated by comparing the growth of normal bone marrow cells in diffusion chambers implanted into the peritoneal cavity of normal or leukemic hosts. Similar growth in total cell numbers was observed in both groups of hosts, and no significant difference could be detected between them. In an attempt to exclude the possibility that an inhibitor either did not penetrate to the peritoneum or was inactivated by the chamber membrane, normal serum, or serum from a highly leukemic rat was mixed with the bone marrow cell suspension in the chambers. Again, no difference in growth between the 2 groups could be detected. Therefore, the influence of a circulating humoral factor in the depression of normal bone marrow hemopoiesis in this experimental leukemia seems to be ruled out, and the decrease may be attributable to local events in the bone marrow such as short-range factors or cellular interaction between the leukemic and normal hemopoietic cell populations.


Murine and human leukemias.

Murine and human leukemias. Essentially all the drugs which are active against human leukemias and lymphomas are active against one type or another of the rodent leukemias and lymphomas. Leukemia L1210 has been generally the most successful screening tool for clinically active compounds. Leukemia P388, however, seems to be better in detecting active antibiotics and natural products and P1534 is particularly sensitive to the Vinca alkaloids, while L5178Y, EARAD, and 6C3HED are useful in detecting the activities of various asparaginase containing fractions. Cell cultures of these leukemias can demonstrate mechanism of drug action and quantitate resistance. Spontaneous AKR leukemia is a model of the advanced human disease. In these leukemias vincristine and prednisone produce a 4 log cell kill. Cytoxan and arabinosyl cytosine (Ara-C) are also effective. On the other hand drugs such as mercaptopurine (6MP) and methotrexate which are highly active in the maintenance phase of acute lymphocytic leukemia (ALL) and in L1210 have little or no activity against the AKR spontaneous system. Mouse leukemias can also detect schedule dependence, synergistic combinations, cross resistance, oral activity, and the ability of drugs to pass the blood brain barrier. A case in point is the Ara-C analog 2,2'-anhydro-arabinofuranosyl-5-fluorocytosine (AAFC) which is not schedule dependent, is active orally, is potentiated by thioguanine, and is effective against intracerebrally inoculated mouse leukemia. AAFC and its analogs might thus be a considerable improvement over Ara-C which is at the present time the most important component of the combination treatment of acute myelogenous leukemia (AML).


[Influence of indole-3-alkanecarboxylic acids on glucose utilization in rats]. The influence of various indole-3-alkane-carboxylic acids (indole-3-propionic acid, indole-3-butyric acid and derivatives of this substance) on some parameters of carbohydrate, fat and insulin metabolism were studied in normal and experimentally induced diabetic rats. Earlier it had been shown, by Schillinger and Loge (1973) using rat liver slices for in vitro studies and adrenalectomized rates for in vivo experiments that indole-3-butyric acid and its analogues investigated suppress gluconeogenesis from pyruvate and that these substances have a hypoglycaemic effect when administered orally at doses of 50-250 mg/kg. Indole-3-propionic acid, indole-3-butyric acid, 1-methyl-indole-3-butyric acid, 2-methyl-indole-3-butyric acid and 1-carboxy-6-fluor-1,2,3,4-tetrahydrocarbazol, administered orally at doses up to 1000 mg/kg, had either only a slight or no hypoglycaemic effect at all in intact and streptozotocin-diabetic rats. The suppression of gluconeogenesis observed in vitro thus only has an appreciable effect on blood glucose in vivo when the capacity of the liver to synthetize glucose is reduced. In normal rats indole-3-propionic acid and indole-3-butyric acid led to a deterioration of glucose tolerance following i.v. glucose loading and to a reduction of 14C-U-glucose oxidation. An increase in the concentration of serum free fatty acids (FFA) measured after treatment with indole-3-butyric acid and its 1- and 2-methylated derivatives was not--in accordance with a theory of an interference of the glucose and fatty acid utilization published in literature--considered to be the primary cause since the reduced formation of 14CO2 from radioactive glucose is not normalized when the increased FFA content is reduced by an inhibitor of lipolysis (5-methyl-isoxazol-3-carboxylic acid)...


[Spectral-analytical studies of the action of etifoxin on the human EEG]. Spectral analysis of EEG in healthy volunteers shows that Etifoxin, the hydrochloride of 6-chloro-4-methyl-4-phenyl-2-ethylamino-4H-3,1-benzoxazine (Hoe 36,801), in an oral dosage of 100 mg has a differential effect in function of the individual alpha organization. This differential effect appearing in the course of the quantitative evaluation can be interpreted, from a morphological point of view, as an expression of various partial phases and patterns of a process leading to reduced vigilance.


The favourable effect of adding a catalase to the treatment of syndromes due to intervertebral disc degenerations. Over a period of one year, 648 patients suffering from 4 different but very resistant syndromes due to intervertebral disc degenerations of the cervical and lumbosacral spine were treated in an identical way, but 50% of them received also catalase (Optidase¿) injections. Half of each group were males and half of them females, the age being 30--50 years. Catalase accelerates the recovery already within the first 3 weeks and the final results after 24 weeks are still significant. In addition, the study of the ischial syndrome was extended to a group of 1208 patients over a period of 5 years, confirming again the favourable effect of catalase. Catalase is generally well tolerated; dangerous reactions have not been noticed.


[Gynecological, hematological and metabolic studies during therapy with an estrogen-gestagen combination preparation]. Twenty women after menopause were submitted to substitution treatment with a biphasic hormone combination (Cyclacur¿). Nine of these had undergone hysterectomy + ovarectomy. Through 11 days each of the women was administered a daily dose of 2 mg estradiol-valerianate as estrogen. Subsequently, for another 10 days each was given the identical dose of estrogen plus 0.5 mg DL-norgestrel as gestagen. Upon this followed an intermission of 7 days without any medication. After 1 cycle of placebo, 7 cycles of medication were followed by 1 cycle of placebo again with regular controls. Climacteric signs and symptoms were eliminated entirely. In those women without hysterectomy withdrawal-menorrhagia occurred promptly 2--3 days after discontinuing medication. During the entire period of investigation there was no significant change in serum lipids, blood sugar values, or serum enzyme tests used as parameter of hepatic function. As far as hematology is concerned there was a moderate rise of hematocrit. No change of serum protein-bound iodine was observed.


Treatment of climacteric complaints with oestriol. 150 women with serious climacteric complaints were continuously treated from January, 1970, up to April, 1972, with Ovestin¿, an orally effective preparation containing 1 mg of oestriol per tablet. After one month of treatment subjective complaints were already considerably improved. Oestrogenic activity was markedly increased, according to vaginal smears and total gonadotrophin excretion in the urine. Its high tolerance and beneficial therapeutic effect make Ovestin the preparation of choice in the treatment of the climacteric syndrome.


A preliminary report on the effects of methanol extraction residue of BCG (MER) on cancer patients. Twenty-seven patients with malignant neoplasia were injected intradermally with the methanol extraction residue (MER) fraction of tubercle bacilli. Two schedules of treatment were used: every other week and once a month; 1-10 courses of MER were administered to the patients. The skin reactivity to 3 recall antigens, as well as to the injected MER itself, was used to monitor the immune response. Improvement of skin reactivity occurred in 9 of 18 patients tested with recall antigens. Five of 6 patients treated every other week improved in their immune capacity whereas only 4 of 12 patients improved on the monthly schedule. Thus, repeated injections given every other week were more effective in increasing the cutaneous reactivity than monthly injections of MER. The side-effects of MER treatment were tolerable.


Historical evidence for the genetic heterogeneity of pseudoxanthoma elasticum. Re-examination of published data from affected families with pseudoxanthoma elasticum (PXE) described in the literature between 1896 and 1968 supported the concept of clinical and genetic heterogeneity. There was clinical evidence for two autosomal dominant and one recessive type, although only a single patient with recessive type II was described. Examination of the individual pedigrees supported both autosomal dominant and recessive patterns of inheritance. The pooled genetic data were less helpful because they were so highly selected. (They had been reported in the literature.) Even so the dominant type II data fitted the hypothesis, whilst the type I and recessive groups were near the expected values. The clinical properties of the groups corresponded closely to the recent British (Pope, 1973) survey. In general, the dominant type I group showed classical cutaneous changes, vascular complications were common and the chief ophthalmic complication was a severe choroidoretinitis. The dominant type II group, in contrast, has an atypical macular rash, or sometimes no rash at all, no vascular complications and very mild retinal changes. Choroidoretinitis was rare and peau d'orange patterns and prominent choroidal vessels common. The recessive group was of intermediate severity.


Erythropoietic protoprophyria--submicroscopic events during the acute photosensitivity flare. The immediate response of erythropoietic protoporphyria (EPP) skin to long-wave ultraviolet radiation (UVR) was studied with the electron microscope. The main finding was severe vascular injury. This was confined to the superficial vessels of the dermis and consisted of endothelial cell degeneration and a pronounced leakage of vascular contents. In contrast, the epidermis showed no abnormalities. Short-wave UV irradiation of EPP skin resulted in epidermal changes typical for the usual sunburn reaction and spared the dermal blood vessels. The following conclusions are drawn: (i) Endothelial cells are the primary cellular target for the photodynamic reaction in EPP. (ii) The fibrillar material, characteristic for chronic EPP lesions, originates from the vessels and vascular contents. (iii) The multilayered basement membranes observed in such lesions reflect multiple consecutive reparative processes that follow endothelial injuries.


Reactions to aspirin and food additives in patients with chronic urticaria, including the physical urticarias. In 131 patients with chronic urticaria, including physical urticarias, oral provocation tests were done with aspirin. A total of thirty-one patients showed a reaction on aspirin challenge. Reactions were seen in 35% of patients with idiopathic urticaria, 52% of patients with cholinergic urticaria, and 43% of those with pressure urticaria. The patients with reactions to aspirin were also tested with tartrazine, sodium benzoate, 4-hydroxybenzoic acid, sodium- and phenyl salicylate and the analgesics indomethacin, paracetamol and mefanamic acid. In nineteen of twenty three aspirin sensitive patients, positive reactions to one or more of these substances were observed. Indomethacin and tartrazine had the highest scores. There was no statistically significant correlation between aspirin reactions and the presence of nasal polyposis, sinusitis, asthma or atopy.


One hundred twin pairs patch tested with primary irritants. The cutaneous sensitivity to benzalkonium chloride, sodium lauryl sulphate and potash soap was determined in 54 monozygotic and 46 dizygotic twin pairs. Comparing the intra-pair reaction strength a higher degree of concordance was found among monozygotic than among dizygotic twins, and even more so when monozygotic twins were compared with matched controls.


Puritus vulvae: treatment by multiple intradermal alcohol injections. A Preliminary report is presented on the use of multiple intradermal injections of absolute alcohol in the treatment of intractable pruritus vulvae. Twenty-five patients were followed up for over I year, and of these six (24%) were cured, and thirteen (52%) showed marked symptomatic improvement.


Simplified dithranol treatment for psoriasis. Progressive simplification of dithranol therapy has made treatment at home more acceptable. This has included ommitting ultra-violet light, talc and dressings, and substituting an ordinary bath instead of one containing tar. A stiffened dithranol 0.5% ointment was found to be slightly more effective than the best paste hitherto employed. It was also very much easier to use.


The absorption of hexachlorophane from Ultralanum ointment. The application of Ultralanum ointment to adult skin cases has produced levels of hexachlorophane in the plasma comparable with those found in infants who have undergone whole-body washing with hexachlorophane solutions. Clinical evidence of toxicity was not observed, but it is felt that the value of hexachlorophane in topical steroid preparations should be re-appraised, particularly for long-term treatment of large eczematous areas.


Effect of pindolol on intraocular pressure. Pindolol, a strong beta-adrenergic blocking agent, instilled into the conjunctival sac of normal and glaucomatous eyes, produced a significant drop in intraocular pressure. This was not, at first, accompanied by any variation in outflow facility; only after prolonged treatment did an increase in facility appear, which accounted only for one-third of the tension-lowering effect. The drug was well tolerated, and did not affect either pupil motility or corneal sensitivity. It seems suitable for a trial use in the treatment of glaucoma.


1H nuclear magnetic resonance of modified bases of valine transfer ribonucleic acid (Escherichia coli). A direct monitor of sequential thermal unfolding. Proton magnetic resonances at 220 MHz from three nucleotide residues of valine I tRNA (Escherichia coli) serve as intrinsic probes of local molecular structure. Resonances from the methyl group of ribothymidine, the methyl group of N6-methyladenosine, and the C-5 methylene of dihydrouridine monitor separate conformational transitions in the TpsiC, anticodon, and dihydrouridine loops, respectively. As the temperature is raised in a solution containing 0.23 M Na+ and no Mg2+, the dihydrouridine region melts with a Tm of 55 degrees, the anticodon region at 58 degrees, and the TpsiC region at 67 degrees. The dihydrouridine nuclear magnetic resonance (NMR) transition correlates with the major change in absorbance monitored in the uv at 330 nm which is ascribed to structural pertubations near the 4-thiouracil moiety. On the NMR time scale slow exchange is seen throughout the temperature range for dihydrouridine and below the apparent Tm for the ribothymidine methyl group. Chemical shift and line width differences between folded and unfolded forms of the polynucleotide indicate that, in the native structure, ribothymidine is in a highly structured region and N6-methyladenosine is in a somewhat less restricted environment. Narrow line widths for the C-5 methylene triplet are found over the whole temperature range indicating that this base is undergoing rapid internal reorientation relative to the overall macromolecule.


Fluorescence detected circular dichroism study of the anticodon loop of yeast tRNAPhe. Fluorescence detected circular dichroism (FDCD) measurements have been used to study the conformations of the anticodon loop of yeast phenylalanine tRNA. To our knowledge this is the first application of fluorescence detected circular dichroism. Much smaller amounts of tRNA are needed for the measurement of FDCD than for the conventionally measured circular dichroism. Furthermore, FDCD is specific for conformational changes near the anticodon loop. The FDCD measurements suggest a transition in the anticodon loop near 20 degrees in 0.01 M MgCl2-0.1 M NaCl (pH 7). This is followed by a broad transition from 30 to 60 degrees and finally a sharp melting at 75 degrees consistent with the absorbance detected melting of the entire tRNA. Removal of Mg2+ from the tRNA at 1 degrees causes nearly a factor of two decrease in the FDCD near 230 nm. This indicates a decrease in conformational rigidity in the anticodon loop on removal of Mg2+.


Interaction of actin with N-ethylmaleimide modified heavy meromyosin in the presence and absence of adenosine triphosphate. N-Ethylmaleimide modified heavy meromyosin in only 3-fold activated by actin rather than 200-fold as is normal heavy meromyosin  (Silverman, R., Eisenberg, E., and Kielley, W. W. (1972), Nature (London) 240, 207). Ultracentrifuge studies demonstrated that in the absence of ATP the N-ethylmaleimide modified heavy meromyosin binds to actin at a ratio of 2 actins to 1 N-ethylmaleimide modified heavy meromyosin. However, it was found that most of the N-ethylmaleimide modified heavy meromyosin was not bound to actin during ATP hydrolysis. Ultracentrifuge studies demonstrated that in the presence of 25 or 50 mM KCl under conditions where the ATPase is maximally activated by actin, less than 5% of the N-ethylmaleimide modified heavy meromyosin was bound to actin. In the absence of KCl there was limited binding but even this binding did not appear to correlate with the N-ethylmaleimide modified heavy meromyosin ATPase rate. Turbidity and viscosity studies also indicated that in the presence of ATP under conditions of maximal actin activation the N-ethylmaleimide modified heavy meromyosin and actin are almost completely dissociated, whereas there is a marked increase in turbidity and viscosity after all of the ATP is hydrolyzed. These results suggest that in the presence of ATP and actin N-ethylmaleimide modified heavy meromyosin exists most of the time in a refractory state unable to bind to actin and only a small part of the time in a nonrefractory state which can interact with actin. It follows that the major rate-limiting step during actin activation is the transition from the refractory to the nonrefractory state. Since the actin activation of N-ethylmaleimide modified heavy meromyosin is lower than that of normal heavy meromyosin this transition may be slower for N-ethylmaleimide modified heavy meromyosin than for normal heavy meromyosin.


Reversible inactivation of vectorial phosphorylation by hydroxybutynoate in Escherichia coli membrane vesicles. The acetylenic hydroxy acid 2-hydroxy-3-butynoate causes irreversible inactivation of the Escherichia coli membrane-bound flavoenzyme D-lactic dehyrogenase, and thus blocks D-lactate dependent active transport in isolated membrane vesicles [Walsh, C. T., Abeles, R. H., and Kaback H. R. (1972), J. Biol. Chem. 247, 7858]. The inactivator is a suicide substrate for the dehydrogenase, undergoing a small number of turnovers before partitioning between oxidation and inactiviation. It is now demonstrated that reactive product molecules of 2-keto-3-butynoate can diffuse in the membranes to a component of the phosphotransferase system and cause time-dependent and covalent inactivation of phosphoenolpyruvate-dependent hexose uptake. Membrane vesicles from double mutants with low levels of both D- and L-lactic dehydrogenase lose only 30 percent of their hexose uptake capacity on exposure to hydroxybutynoate under conditions sufficient to fully inactivate hexose transport in wild type vesicles. Transport of 1-[14C]hydroxybutynoate into vesicles is followed by rapid covalent labeling of membrane proteins by the reactive, enzymatically generating keto acid oxidation product. Incubation of hydroxybutynoate-inactivated vesicles (5% residual activity) for 20 min in buffer with 10 mM dithiothreitol results in reactivation of 63% of the hexose transport activity, a 12-fold increase in activity. No reactivation occurs if the vesicular phosphotransferase system is inactivated by keto acid derived from membrane oxidation of the olefinic congener 2-hydroxy-3-butenoate. In contrast to thiol reactivation of acetylenic-blocked glucose transport, blockage of D-lactate-stimulated proline uptake is not alleviated, stressing different modes of inactivation of the phosphotransferase system compared to the membranous lactate dehydrogenases.


Limited trypsinolysis of native Escherichia coli elongation factor G. Native elongation factor G possesses three peptide bonds or limited regions of the molecule which are especially sensitive to trypsinolysis. Cleavage at these sites occurs in sequence. Initially, the protein, a single peptide chain of 74,000 daltons, is rapidly split into a fragment of 71,000 daltons and one or more small peptides totaling 3000 daltons. This initial scission does not alter the gross three-dimentional structure of the protein, as the native and trypsin-cleaved protein have the same Kd on Sephadex gel filtration. Although cleaved, the modified elongation factor G retains full activity as measured by its ability to form complexes with the ribosome and guanine nucleotides. A single peptide bond (or region) in the 71,000-dalton fragment is then completely cleaved, yielding fragments of 47,000 and 29,000 daltons. These fragments do not remain associated under native conditions, as they are clearly resolved on gel filtration. Loss of activity is concomitant with the scission of the 71,000-dalton fragment. The third cleavage is complete under the conditions employed here and occurs in the 47,000-dalton peptide generating a fragment of 45,000 daltons and one or more small peptides totaling 2000 daltons. Prolonged treatment with higher levels of trypsin ultimately reduces the protein to small peptides but without generating further discrete fragments visible on sodium dodecyl sulfate gel electrophoresis.


Raman spectra and structure of yeast phenylalanine transfer RNA in the crystalline state and in solution. Laser Raman spectra of yeast phenylalanine transfer RNA have been obtained in solution and in orthorhombic and hexagonal crystals. So far as one can tell from the spectra, which are identical in the two crystal forms, the molecular structure of the tRNA is not altered by differences in molecular packing in these two unit cells. In addition, the spectra of the two crystal forms show the same characteristic Raman frequencies and intensities as those of the tRNA in aqueous solution. Thus the structure of the tRNA molecule appears to be the same in the crystals and in aqueous solution. From the spectroscopic changes that result when Mg2+ ions are removed from the native tRNA, it is concluded that the removal of Mg2+ produces a partial disordering of the ribophosphate backbone of the molecule and a lowering of its melting temperature. The melting is shown to be a complex process in that the vibrations specific for adenine indicate a slightly lower melting temperature and those specific for guanine a slightly higher melting temperature than that of the ribophosphate backbone.


Escherichia coli stringent factor binds to ribosomes at a site different from that of elongation factor Tu or G. The binding of Escherichia coli stringent factor to ribosomes has been studied; the reaction depends on 50S and 30S ribosomal subunits and poly(U) as messenger RNA. The ribosome-stringent factor complex is formed at 5-10 mM magnesium acetate; NH4 ions are inhibitory. Binding of the stringent factor to the 70S-mRNA complex is not stimulated by uncharged tRNA. The ribosomal binding site(s) for the stringent factor does not overlap with the one known for the elongation factor Tu (EF-Tu) or G (EF-G). Ribosomes carrying either EF-Tu or EF-G are active in binding the stringent factor; however, they are inactive in synthesizing guanosine 5'-triphosphate 3'-diphosphate (pppGpp) and guanosine 5'-diphosphate 3'-diphosphate (ppGpp). The latter result is due to the blockage of the ribosomal acceptor site by the aminoacyl-tRNA and/or elongation factors. That stringent and elongation factors do not compete for identical ribosomal region(s) is supported by: (1) the reverse experiments where ribosomes charged with the stringent factor are fully active in EF-Tu or EF-G dependent functions; (2) ribosomes that lack the two ribosomal proteins L7 and L12 known to be essential for EF-Tu and EF-G functions bind the stringent factor and are active in synthesizing pppGpp and ppGpp.


Cobalt(III) labeled aspartokinase-homoserine dehydrogenase of Escherichia coli. The kinase activity of the threonine-sensitive aspartokinase-homoserine dehydrogenase enzyme complex of Escherichia coli was selectively inactivated by Co(III) incorporation. Incubation of the enzyme with Co(II) in the presence of oxygen or H2O2 resulted in incorporation of one Co(III) per subunit. The cobalt(III) bound to the enzyme was not removable by dialysis and presumably results from formation of "inert" coordination complexes with ligands contributed by the enzyme. Cobalt was released from the enzyme by incubation with dithiothreitol but not by metal chelating agents. The Co(III)-labeled enzyme was aspartokinase inactive but still retained 60% of its original homoserine dehydrogenase activity. Studies of the time course of inactivation showed aspartokinase inactivation paralleled Co(III) incorporation. The residual dehydrogenase activity of aspartokinase inactive enzyme was still inhibited by threonine Thus, Co(III) incorporation seems to result in a specific inactivation of kinase activity which permits enumeration of the number of aspartokinase sites. Limited alpha-chymotrypsin digestion of Co(III)-enzyme produced homoserine dehydrogenase-active fragments devoid of Co(III), further confirming the specificity of the labeling procedure. Aspartokinase inactivation obtained without concomitant desensitization of homoserine dehydrogenase to threonine inhibition suggests that kinase active site integrity is not required for threonine binding and inhibition of homoserine dehydrogenase.


Fragments of bovine serum albumin produced by limited proteolysis. Isolation and characterization of peptic fragments. Five new fragments of bovine serum albumin have been isolated following limited peptic hydrolysis. These fragments, and the two peptic fragments previously described by King (King, T.P. (1973), Arch. Biochem. Biophys. 156, 509), were positioned within the albumin sequence published by Brown (Brown, J.R. (1975), Fed. Proc., Fed. Am. Soc. Exp. Biol. 34, 591) on the basis of molecular weight, amino acid composition, and amino- and carboxyl-terminal sequences. The fragments correspond to residues 1-385, 1-306, 307-581, 49-185, 186-306, 307-385, and 353-503 in the albumin sequence. These peptides are likely to be native in structure since disulfide bonds were not cleaved during their preparation. In each case the amino acid composition and terminal sequences have confirmed the bovine serum albumin sequence and disulfide bridging pattern proposed by Brown, and the offer further proof that bovine albumin is composed of a series of nine independent loops. These fragments should be useful in elucidating the structure-function relationships of albumin.


DNA-binding chromosomal non-histone proteins. Isolation, characterization, and tissue specifity. A fractionation schedule is described which allows the isolation of a group of chromosomal non-histone proteins (NP) with affinity for DNA. In polyacrylamide gel electrophoresis these proteins isolated from rat liver are represented principally by a group of low molecular weight polypeptides. The NP fraction comprises about 2-4% of the total chromatin protein content in rat liver or Novikoff hepatoma. Experiments in vivo and in vitro revealed that the NP proteins do not incorporate significant amounts of 32P. Complexes of the chromosomal proteins NP with homologous DNA are immunologically tissue specific and the specificity can be transferred by reconstituting the NP proteins from one tissue to the residual chromatin from another.


Phospholipid metabolism of stimulated lymphocytes. Composition of phospholipid fatty acids. Lymph node lymphocytes and thymocytes from different species were isolated. Rabbit and calf thymocytes were stimulated in vitro with concanavalin A. Phospholipid fatty acids of these cells were analyzed and their positional distribution was determined. When compared with liver, phosphatidylcholine of unstimulated lymphocytes was found to contain relatively high amounts of palmitic acid in position 2 and oleic acid in position 1. After stimulation of rabbit thymus cells, the content of polyunsaturated fatty acids (linoleic and arachidonic acid) increased. Thus the ratio of polyenoic acids (18:2 + 20:4) to saturated fatty acids was doubled when compared to control cells. Similar results were obtained after in vivo stimualtion with Mycobacterium Calmette Guerin. The correlation of these findings with the activation of acyl-CoA:lysolecithin acyltransferase, and their relevance for changes of membrane fluidity during lymphocyte stimulation is discussed.


Formation of lipid-bound oligosaccharides in yeast. Incubation of a membrane fraction from Saccharomyces cerevisiae with UDP-N-acetyl [14C]glucosamine catalyzes the transfer of N-acetylglucosamine to an endogenous lipid fraction as well as a methanol-insoluble polymer. The glycolipid was shown to separate into three compounds by thin-layer chromatography. The biosynthesis of two of them could clearly be stimuated by the addition of dolichol monophosphate to the incubation mixture. Evidence is presented that the substances are dolichol pyrophosphate derivatives: dolichol pyrophosphate N-acetylglucosamine and dolichol pyrophosphate di-N-acetylchitobiose. The formation of the chitobiose-containing lipid was increased by reincubation of the glycolipid with non-radioactive UDP-N-acetylglucosamine. The same particulate preparation transferred mannose from GDPmannose to dolichol pyrophosphate di-N-acetylchitobiose, giving rise to a lipid-bound oligosaccharide. Molecular weight determination of the oligosaccharide moiety gave a value of 780, which is consistent with a tetrasaccharide containing two mannose subunits attached to di-N-acetylchitobiose. The methanol-insoluble radioactive product obtained in the presence of UDP-N-acetyl[14C]glucosamine was transformed by pronase treatment to a large extent into dialyzable material. It is suggested that the glycolipids described serve as intermediates in the glycosylation of yeast mannoproteins.


Inhibition of thiamine transport in anaerobic baker's yeast by iodoacetate, 2,4-dinitrophenol N,N'-dicyclohexylcarbodiimide and fatty acids. 1. [14C]Thiamine uptake by baker's yeast (Saccharomyces cerevisiae) was strongly inhibited by 0.2 mM iodoacetate, 0.2 mM 2,4-dinitrophenol and 0.1 mM N,N'-dicyclohexylcarbodiimide under anaerobic conditions. 2. The inhibition of anaerobic [14C]thiamine uptake by these inhibitors was accompanied by almost parallel decreases in the ATP level of the yeast cells. 3. On the other hand, the short-chain fatty acids inhibited [14C]thiamine uptake to a large extent, without greatly affecting the intracellular ATP level. This suggests that the acids primarily block the use of energy from ATP for the transport rather than the fermentation process. 4. Caproate, which has a most pronounced inhibitory effect on [14C]thiamine uptake, significantly prevented the dissipation of an energized membrane state of yeast cells necessary for the active transport of thiamine. 5. Possible ways in which the inhibitors may affect thiamine uptake were discussed.


The effects of D- and L-glyceraldehyde on glucose oxidation, insulin secretion and insulin biosynthesis by pancreatic islets of the rat. D-glyceraldehyde stimulated insulin secretion from isolated rat pancreatic islets in static incubation and perifusion systems. At low concentrations (2-4 mM) D-glyceraldehyde was a more potent secretagogue than glucose. The insulinotropic action of 15 mM D-glyceraldehyde was not affected by D-mannoheptulose, was potentiated by cytochalasin B (5 mug/ml) and theophylline (4 mM), and was inhibited by both adrenalin (2 muM) and somatostatin (10 mug/ml). D-glyceraldehyde at a concentration of 1.5 mM produced a 10-fold increase of L-[4,5-3H]leucine incorporation into proinsulin and insulin without a significant increase into other islet proteins. Glucose at 1.5 mM did not stimulate proinsulin biosynthesis. D-Glyceraldehyde at concentrations higher than 1.5 mM, in marked contrast to glucose, progressively inhibited incorporation of labelled leucine into proinsulin + insulin and other islet proteins. D-Glyceraldehyde also inhibited the oxidation of glucose. L-Glyceraldehyde did not stimulate proinsulin biosynthesis and had less effect than the D-isomer on insulin release and glucose oxidation. The results strongly suggest that metabolites below D-glyceraldehyde-3-P are signals for insulin biosynthesis and release. Interaction of D-glyceraldehyde with a "membrane receptor" cannot, however, be excluded with certainty.


Glucosamine-labelled envelope proteins of Escherichia coli K-12. I. Electrophoretic studies and partial fractionation of phenol-soluble proteins. Hydrophobic envelope proteins were extracted by phenol from a glucosamine- and leucine-requiring mutant of Escherichia coli K-12 (E-110). Three protein fractions labelled with D-[1-14C]glucosamine and L-[4,5-3H]leucine were obtained by electrophoretic separation. Envelopes were isolated from cells labelled with D-[1-14C]glucosamine--HCL and acid hydrolyzed. At least 68% of the radioactivity was recovered as glucosamine and glucose with no random distribution of label. Fingerprinting of pronase digests of glucosamine-labelled proteins showed four radioactive spots associated with peptides. The glycoproteins were pronase- and trypsin-sensitive and had apparent molecular weights of 11 000 (fast mobility), 35 000 (intermediate mobility) and 62 000 (slow mobility) as estimated by sodium dodecyl sulfate-polyacrylamide disc electrophoresis. The two heavier fractions were labelled with meso-diamino[1,7-14C2]pimelic acid, while orth[32P]phosphate was not incorporated into any fraction. The glucosamine radioactivity of the fast fraction underwent rapid changes upon a chase with non-radioactive glucosamine. Using a Sephadex LH-20 column, the radioactive proteins were separated from the phenol and subsequently fractionated on a DEAE-cellulose column. The DEAE-cellulose fractions were distinct from each other in the number and composition of protein bands, when analyzed by sodium dodecyl sulfate-polyacrylamide disc electrophoresis. Radioactive bands with intermediate and fast electrophoretic mobilities were found in separate DEAE-cellulose fractions.


Glucosamine-labelled envelope proteins of Escherichia coli K-12. II. Location in inner and outer membranes. Outer and inner membranes were prepared from a culture of a glucosamine- and leucine-requiring mutant of Escherichia coli K-12, which was grown on L-[4,5-(3)H2]leucine and D-[1-(14)C]glucosamine and "chased" with unlabelled medium. Phenol-soluble glycoproteins were obtained from these membranes after phospholipid extraction. Sodium dodecyl sulfate-disc electrophoresis of the outer-membrane glycoproteins separated two components which were labelled with both isotopes and had a fast (F) and intermediate (I) migration. Dodecyl sulfate electrophoresis of the inner-membrane glycoproteins showed that only the F band incorporated both 14C and 3H labels. The 3H to 14C ratio in the F band from the outer membrane was completely different from the isotopic ratio in the F band from the inner membrane, indicating that these components were not identical despite their similar molecular weights. The F bands from both membranes lost their label during the chase with unlabelled medium, while the I band remained relatively stable. Chloroform-soluble 14C label extracted from the outer membrane decreased during the chase. In contrast, the chloroform-soluble 14C from the inner membrane, accumulated during the chase.


Effect of activated nitrofurans on DNA.

Effect of activated nitrofurans on DNA. Enzymically activated nitrofurazone reacts with co-valently closed circular DNA (derived from Escherichia coli minicells carrying lambdadv) to give at least two kinds of damage: breaks which are detected on neutral sucrose gradients and alkali-labile lesions in DNA which are converted to breaks when the DNA is subsequently Treated with alkali. DNA, isolated from nimicells exposed to the drug, also contains lesions which are converted to breaks upon treatment with endonuclease preparations obtained from Micrococcus luteus. Minicells repaired both breaks and nuclease-susceptible lesions within 2 h but did not repair alkali labile lesions within that time. Experiments with three other nitrofurans show that there are considerable differences in the degree to which DNA is damaged by activated metabolites of various derivatives and that the potency of the compounds as mutagens and carcinogens is correlated with the amount of damage caused to minicell DNA.


Temperature and salt effects on the formation of preinitiation complexes between RNA polymerase and phage DNA. The influence of temperature and KCl concentration on the formation of rifampicin-resistant preinitiation complexes by holo RNA polymerase has been compared for T4 DNA and Azotobacter phage A21 DNA. The sharp transition with respect to temperature between an inactive complex of polymerase and DNA and a preinitiation complex reflects an equilibrium between the two complexes, the position of which depends on the temperature and the salt concentration. The transition is shifted to higher temperatures by increasing the KCl concentration. The position of this transition is characteristically different for T4 and A21 DNA. The midpoint for A21 DNA is about 15 degrees C above that for T4 at 0.006 M KCl. At 0.15 M KCl the transition for A21 DNA cannot be observed below 37 degrees C. This difference is responsible for the apparent inhibition of a21 dna transcription by KCl and for the low template activity of A21 DNA under the conditions of the standard assay. Both holo and core RNA polymerases are able to form complexes with A21 DNA that are resistant to attack by rifampicin. The second-order rate constant for the inactivation  of the complex with the core enxyme is three times greater than that for the complex with the holoenzyme.


Ribosome run through of the termination codon in the absence of the ribosome releasing factor. The ribosome releasing factor (RR factor) which releases ribosomes from mRNA at the termination codon has been examined for its effects on the amino acid incorporation programmed by wild type R17 Phage RNA and amB2 R17 RNA. When RR factor was added at the beginning of the incorporation, there was no effect on the initial rate of incorporation but it reduced the final level of incorporation. The reduction of the final level of incorporation was more pronounced for histidine incorporation than for valine incorporation suggesting that the translation of the RNA polymerase cistron was more influenced by RR factor. These experiments were carried out under conditions where no reinitiation of protein synthesis occurred. In the presence of RR factor, suppressor tRNA functioned better for the incorporation of amino acids into proteins with amB2 R17 RNA than did wild type tRNA. No such differential effect of suppressor tRNA was observed in the absence of RR factor. This suggests that the ribosome has to be released from mRNA by RR factor in order for the amber mutation to be effective.


DNA synthesis in vitro in lysates of Escherichia coli. A system using crude lysates of Escherichia coli PolA- for in vitro synthesis of DNA has properties similar to those described previously for washed DNA - membrane complexes, i.e. short-lived synthesis of DNA, 2-3-fold stimulation by ATP, semiconservative form of synthesis, and most of the product in the size distribution of Okazaki fragments. The proximate substrate is shown to be the deoxynucleoside triphosphate. The lysate system can be fractionated into a particular membrane - DNA complex and a soluble portion, each of which is inactive by itself, but which upon mixing restores activity. The particulate fraction provides template as well as essential protein(s). The soluble fraction contains DNA polymerases II and III both of which are capable of the necessary polymerase function, and a factor that is required for the stimulation of DNA synthesis by ATP. The stimulatory activity can also be assayed by restoration of activity to an aged lysate, which has lost the ATP-dependent synthesis; the activity is not a kinase, and is not an ATP-dependent nuclease, at least of the type currently known. The ATP-dependent stimulatory factor has been partially purified but further purification or characterization of it has been limited by its extreme instability. Both the stimulatory factor and ATP are required for semiconservative synthesis in the lysate, and for synthesis of the short fragments. Similar to results in vivo and in permeable cells the new DNA appears to contain RNA, as judged by the evidence for RNA - DNA junctions from modified nearest neighbor experiments. However, the nucleotides at the RNA - DNA junctions in the lysate system are not specific, in contrast to results in permeabilized Escherichia coli.


Wall mannan of Saccharomyces cerevisiae. Metabolic stability and release into growth medium. Selective labelling of cell wall mannan with radioactive precursors in growing Saccharomyces cerevisiae showed that this polysaccharide is metabolically stable during exponential growth. Mannan once inserted into the wall is not subject to turnover or release into the growth medium. However, about 10% of the amount of mannan incorporated into the cell wall fraction can be recovered in the non-dialyzable material isolated from the growth medium. Therefore, the mannan escaping from the cell must be either a mannan de novo synthesized, not trapped in the growing wall structure, or a mannan with a non-structural role. Radioactivity was also retained in the wall fraction of cells pre-labelled with [14C] glucose which pointed to metabolic stability of all cell wall polysaccharides in growing S. cerevisiae.


Levels and turnover of the proteinase B inhibitors in yeast. The ratio of the proteinase B inhibitors IB1 and IB2 from baker's yeast was shown to depend on the yeast strain by specific immunoprecipitation from boiled yeast extract and subsequent electrophoresis of the heat-dissociated precipitates on polyacrylamide gels. Bothe IB1 and IB2 were found, IB2 being by far predominant. Saccharomyces carlsbergensis NCYC 74 contained IB1, whereas in Saccharomyces cerevisiae X 2180 only IB2 was present. When cells of the latter strain were labelled with [14C] leucine from the beginning of growth and pulsed with [3H] leucine during the stationary phase, no short-lived IB1 could be detected. However, the peak of IB2 resolved on the gel showed an increased 3H/14C ratio in comparison to the majority of the other cellular proteins. The increased 3H/14C ratio was found to be the result of catabolite repression of inhibitor synthesis during exponential growth: cells growing on glucose as carbon source contain high inhibitor levels only during the stationary phase of growth, whereas during growth on acetate high amounts of inhibitor are present even in exponentially growing cells. During the stationary phase of growth the inhibitor is degraded with the same half-life as the total cellular proteins (about 50 h).


The use of computed activity coefficients in Raoult's reference system, for n-alcohol/water mixtures, and their application to the study of proteins. Activity coefficients of alcohols in Raoult's reference system have been calculated. Using data from papers dealing with transition temperatures in the presence of n-alcohols, lipase deactivation and competitive inhibition by n-alcohols of pepsin and alpha-chymotrypsin, it is shown that the conversion of concentrations into activities (using Pierotti's equation) profoundly affects the original data. Within experimental error, if inhibition constants by alcohols keep the same value, lupase deactivation becomes independent of the length of the alcohol chain, and except for butanol, transition temperature is no longer dependent on the number of methylene groups. Interpretations which take into account the thermodynamic reference system and final environment are related to the fact that alcohol molecules bind to some inner parts of the proteins.


A novel electrophoretic fractionation of Escherichia coli envelopes. Particulate fractions of Escherichia coli have been submitted to electrophoretic fractionation in a buffer stabilized by sucrose gradient. Inner membrane and outer membrane were readily resolved. A combination of electrophoresis, fractional centrifugation and gel filtration can remove remaining contamination by ribosomes and cytoplasm. The presence of particles containing no phospholipids was detected after differential centrifugation. The nature of this fraction is unknown. The inner membrane exhibited heterogeneity on electrophoresis.


Inducible gluconate permease in a gluconate kinase-deficient mutant of Escherichia coli. Gluconate-resistant mutants were isolated from Escherichia coli strain DF 1070 deficient in phosphogluconate dehydrogenase (EC 1.1.1.44) and in phosphogluconate dehydrogenase (EC 4.2.1.12) which is inhibited by gluconate. Among the resistant mutants, AR 13 has been identified as a gluconate kinase (EC 2.7.1.12)-deficient strain. This mutant exhibits an inducible gluconate transport system capable of concentrating gluconate in the cytoplasm against a concentration gradient. The accumulated gluconate is subject to permanent turnover, and is not chemically modified. The kinetics of induction and deinduction indicate a single inducible component, rate limiting for the transport function, and the distribution of transport capacity among non-induced progeny of induced parents indicates that the inducible protein is membrane bound.


Heterogeneity of schizophrenia.

Heterogeneity of schizophrenia. One of the difficulties in defining schizophrenia is the possibility of its heterogeneity. Schizophrenia may be divided into organic and idiopathic on the basis of precipitating factors; the difference between the two is supported by evidence from family studies and differences in symptomatology. A further division of idiopathic schizophrenia may be made into typical and atypical schizophrenia based on differences in clinical features, family history, mode of inheritance, and prognosis. There is suggestive evidence that within typical schizophrenia, paranoid and nonparanoid subtypes may be idstinguished due to clinical and family differences. Within nonparanoid schizophrenia, simple, catatonic, and hebephrenic schizophrenia may also prove to be distinct entities. The available evidence for differentiating the subtypes of typical schizophrenia is inconclusive to date. However, several recent developments in the methodology of family research may lead to more convincing findings for differentiating subtypes within typical schizophrenia.


Sequential treatment assignment with balancing for prognostic factors in the controlled clinical trial. In controlled clinical trials there are usually several prognostic factors known or thought to influence the patient's ability to respond to treatment. Therefore, the method of sequential treatment assignment needs to be designed so that treatment balance is simultaneously achieved across all such patients factor. Traditional methods of restricted randomization such as "permuted blocks within strata" prove inadequate once the number of strata, or combinations of factor levels, approaches the sample size. A new general procedure for treatment assignment is described which concentrates on minimizing imbalance in the distributions of treatment numbers within the levels of each individual prognostic factor. The improved treatment balance obtained by this approach is explored using simulation for a simple model of a clinical trial. Further discussion centers on the selection, predictability and practicability of such a procedure.


Cost evaluation of a two-stage classification procedure. A two-stage classification rule is suggested for classifying an individual into one of two multivariate normal populations. An individual may be classified using measurements on just the first subset; otherwise the second subset is measured and he is classified using the combined set. On the basis of misclassification costs and costs of collecting observations, the two-stage rule is compared with the usual rules based on the first subset and on the combined set. Comparisons are made for the case of known population parameters and also for the case of unknown population parameters which are estimated from samples. It is assumed throughout that a priori probabilities, cost of misclassification, and cost of making observation are known.


A generalized modified-chi2 analysis of categorical bacteria survival data from a complex dilution experiment. A well-known method of estimation for the density of bacteria in a sample solution is the most probable number (MPN) procedure. This paper considers the change in density through time of bacteria populations which are undergoing extinction. The MPN with its estimated variance for a fixed time point is a basic module of these investigations and is treated as a general implicit function of the cell proportions in linear categorical data analysis. These quantities are then used to fit exponential decay models over time by weighted least squares. When such models are supported by the data, comparisons of decay rates between populations under possibly different experimental conditions can be undertaken. This methodology is illustrated with an example pertaining to the survival experience of Leptospira autumnalis.


Analysis of data from multiclinic experiments. The analysis of data from multiclinic experiments is put in the framework of a mixed model in which the treatment effects are assumed to be fixed, and clinic and clinic by treatment effects are assumed to be random. The analysis in the unbalanced case is developed under the usual mixed model assumptions. The maximum likelihood estimates and likelihood ratio tests of the parameters are derived following the approach of Hemmerle and Hartley (1973). The T2 test is shown to be asymptotically equivalent to the analysis of variance F test in the balanced case. The distribution of the test statistic in the case of proportional cell frequencies is also discussed. Some investigation is made of the distribution of the test statistic by applying the method of Box (1954).


Statistics and medical experimentation.

Statistics and medical experimentation. Starting from a review of four major experimental areas in medicine (Polio, Coronary Surgery, Diabetes, Breast Cancer), some of the special statistical problems arising in the design and analysis of clinical experiments are reviewed, and the limitations of current formulations are emphasized. Particular attention is given to the ethical dilemma and an alternative formulation of the inference/decision problem is proposed.


Randomization and the analysis of variance. This paper attempts to strengthen faith in randomization, first by defining it in general and second by showing how it is the fundamental means, in many experiments, of generating the probability space. It is defined by the natural structure of the experimental units (E.U.'s) not by a particular experimental design imposed on that structure. Three examples are given, with the same design but different E.U. structures and randomizations, to show that models developed from experimental designs are superficial, but that their randomization is fundamental.


Efficiency of case-control studies with multiple controls per case: continuous or dichotomous data. The statistical efficiency of a study design which matches each case with k controls is compared with that of the standard matched-pairs setup. For the situation in which the distribution of measurements on the factor under investigation is continuous, the comparison criterion used is the reciprocal of the variance ratio of the corresponding case-control difference estimators; for dichotomous observations, the Pitman efficiency of Miettinen's test [1969] or of Pike and Morrow's test [1970] relative to McNemar's test is used. When the variances for continuously distributed case and control measurements are equal to the same result, 2k/(k + 1), is obtained for both situations and, more generally, the efficiency of k1 controls relative to k2 controls is given by k1(k2 + 1)/k2(k1 + 1). The Pitman efficiency is compared with the "practical efficiency" for large-sample dichotomous situations.


The relationship between separation time and genetic distance based on angular transformations of gene frequencies. Expressions are constructed for the theoretical expectation and the variance of the statistic chord2 used in connection with the genetic distances introduced by Cavalli-Sforza and Edwards. The gene frequency probability distribution is taken from Kimura's solution of the diffusion equation for pure random drift. The expression for the expectation is used for discussing whether distances are independent of original gene frequencies, and whether they are proportional to separation times. A modified method for estimating such separation times is proposed, and an example is given with cattle breeds. The accuracy of procedures based on genetic distances is evaluated using the variance of chord2.


F-cells in the adult: normal values and levels in individuals with hereditary and acquired elevations of Hb F. Specific antibodies to human fetal hemoglobin were prepared and, after conjugation with a fluorescent dye, were used to determine the distribution of Hb F-containing cells in blood smears from normal adults and individuals with hereditary and acquired conditions associated with abnormal levels of Hb F. The mean proportion of F-cells in normal persons was 2.7% +/- 1.4%, with a range of 0.5%-7.0%. Proportions of F-cells were increased in persons with several acquired and inherited disorders that are associated with an increased percentage of Hb F in hemolysates. A strong linear correlation between the amount of Hb F and proportion of F-cells was observed. This technique may prove useful in studies of a variety of disorders associated with Hb F elevations and also in investigations of the mechanisms controlling the transition from fetal to adult hemoglobin during the course of human development.


Adrenergic mechanisms in the rabbit ear artery. A review. A number of factors which have been implicated in the response of the rabbit ear artery to adrenergic agents and stionstriction in the arterial smooth muscle; (b) the interrelationship between the intramural distribution of sympathetic nerves and muscle, and the influence of uptake and enzymic inactivation on the response to norepinephrine, and (c) the influence of presynaptic adrenergic and cholinergic receptors on the rwsponse to sympathetic nerve stimulation.


The NBT test: Erratic behaviour in acute leukaemia. Non-stimulated and stimulated nitroblue tetrazolium (NBT) dye test was studied in sixteen adult patients with acute leukaemia in sequence, viz. at the time of diagnosis, during and after treatment. It was found that the capacity of the neutrophilic granulocytes to reduce NBT (non-stimulated) was below, within or above the range for the normal group. Two of the patients had a constant negative non-stimulated NBT test. The stimulated NBT test was below the normal range. Generally, there was no correlation between the percentage of NBT positive cells and fever of bacterial origin. All sixteen patients but four were found to be within the normal range concerning phagocytic ability.


Effect on platelet counts and fever of platelet transfusion in leukemia. The efficacy of platelet transfusions in pretransfused HL-1 sensitized patients has been doubtful, particularly where repeated platelet transfusions were required. The efficacy of one hundred and nine ABO-compatible, cross match negative platelet transfusions in 26 adult patients with acute leukemia was therefore studied. Although most of the patientswere old and pretransfused and although no HL-A typing could be made, a statistically significant average increase of about 11 000 platelets/mul was seen not only after the first but also after subsequent (up to 8) platelet transfusions. Moreover, significant reduction in temperature followed platelet transfusions. However, platelet recoveries were worse in severely infected patients.


Depolarizing effects of the ionophores X-537A and A23187 and their relevance to secretion. The ionophore X-537A depolarized frog skeletal muscle fibres about 10 mV in 10 min and a further 30 mV or so over the next 50 minutes. With A23187, depolarization was relatively feeble and delayed in onset but was hastened when calcium was removed from the bathing medium. The results support conjecture that some functional responses to these ionophores are attributable, in part, to depolarization.


Studies on the hypotensive action of alpha-methyldopamine. 1. Intraventricular alpha-methyldopamine (50-200 mug) produced a dose-related fall in blood pressure in conscious spontaneously hypertensive rats. Pretreatment with intraventricular 6-hydroxydopamine prevented this hypotensive effect of alpha-methyldopamine. 2. The hypotensive effect of alpha-methyldopamine was prevented by intraventricular injection of phentolamine or desmethylimipramine, but not by intraperitoneal injection of haloperidol. 3. Pretreatment with U-14,624, a selective central dopamine-beta-hydroxylase inhibitor, prevented the hypotensive effect of alpha-methyldopamine. 4. Alpha-methyldopamine was considerably less potent than noradrenaline as a pressor agent in the pithed rat, but noradrenaline and alpha-methylnoradrenaline were found to be equipotent. 5. Alpha-methyldopamine (1-5 mg i.c.v.) reduced pressor responses elicited by electrical stimulation of the midbrain reticular formation in cats anaesthetized with chloralose. 6. It is concluded that the hypotensive action of alpha-methyldopamine in conscious animals involves intact central alpha-adrenergic neurones and a central adrenergic uptake mechanism for the formation of alpha-methylnoradrenaline.


Bile salts and gallstone disease.

Bile salts and gallstone disease. Bile salts play an important role in maintaining cholesterol in aqueous solution in bile. There is evidence that in some patients at least gallstones arise as a result of bile salt deficiency. The evidence to date suggests that although oral bile salts may be of use in treating some gallstone patients they are unlikely to replace cholecystectomy in the foreseeable future. The instillation of bile salts via an indwelling T-tube may well be of great use in the management of stones retained in the common duct after choledocholithotomy. There is no doubt that in this branch of medicine as in all others prevention is better than cure and our aim should be to perfect our technique so that we do not leave stones behind. However, should the situation occur we may soon have safe effective physiological solvents to dissolve our mistakes.


Orthotopic liver transplantation: an experimental study on the prevention of infections with Gram-negative organisms. Infection with Gram-negative bacteria is frequently the cause of death in human liver transplantation. In experimental liver transplantation in mongrel dogs we detected Gram-negative septicaemia in 73% of our first 15 cases; maximal survival was 26 days. In a second series of 27 orthotopic liver transplantations in tissue-typed littermate dogs and littermate pigs the standard surgical technique and aftercare were changed with regard to four factors: an end-to-end common bile duct anastomosis was made instead of a gallbladder to duodenum anastomosis; the continuous postoperative bacteriostatic antibiotic therapy was changed to a single 2-day course of bactericidal antibiotics; preoperative selective bowel decontamination combined with postoperative protective isolation was introduced; the dosage of azathioprine used for immunosuppression was reduced..


Gastric acid secretion following renal transplantation. Gastric acid secretion studies were performed in 10 patients on regular dialysis and again following transplantation. While on dialysis, the acid output was high, with mean peak values after pentagastrin of 37-2 mEq/h in males and 26-3 mEq/h in females. These compared with values in normal subjects of 21-6 mEq/h in males and 12-3 mEq/h in females. Following transplantation continuous steroid therapy was given. Repear gastric acid secretion studies showed no significant change, with values of 37-1 mEq/h in males and 24-6 mEq/h in females. The high incidence of gastro-intestinal haemorrhage and perforation following transplantation is probably due mainly to steroid therapy, but the ulcerogenic effect of steroids would not seem to be mediated by increased gastric acid secretion.


Analysis of treatment in childhood leukaemia. I. Predisposition to methotrexate-induced neutropenia after craniospinal irradiation. Report to the Medical Research Council of the Working Party on Leukaemia in Childhood. The degree of drug-induced neutropenia resulting from a controlled trial (UKALL I) of treatment in acute lymphoblastic leukaemia was analysed. The main agent associated with severe neutropenia was methotrexate, and methotrexate-induced neutropenia was significantly greater in patients who had received craniospinal irradiation. The synergistic toxic effect of irradiation followed by methotrexate treatment seems to have contributed to three of the five deaths which occurred in complete remission in this trial; all deaths in remission occurred in patients who had received central nervous system prophylaxis. Analysis of patients who subsequently relapsed compared with those still in remission after 18 months of treatment indicated that the former, on average, had slightly lower neutrophil counts. This suggests that the children who relapsed did not receive any less aggressive treatment than those who remained in remission.


Serum digoxin in patients with thyroid disease. Serum digoxin concentrations were measured by radioimmunoassay in 17 hyperthyroid and 16 hypothyroid patients after a seven-day course of oral digoxin. The significantly higher levels of serum digoxin in patients with hypothyroidism and lower levels in those with hyperthyroidism were closely related to the measured changes of glomerular filtration rate and digoxin serum half time in these two groups. Differences in serum digoxin concentration contribute to the altered sensitivity to digoxin shown by patients with thyroid disease.


Use of bacteriological investigations by general practitioners. The numbers of vaginal swabs, faecal specimens, throat swabs, and urine specimens submitted by 104 city general practitioners to the bacteriology laboratory at Aberdeen differed widely. The doctors who made most use of the laboratory service rarely did so equally for all four investigations but usually for only one or two. Similarly, those who used the service least often made frequent use of one particular investigation. Further studies are needed to identify and evaluate the reasons for the apparently substantial disagreement among general practitioners concerning the value of these diffferent bacteriological investigations in general practice.


Agonal phase, ischaemic times, and renal vascular abnormalities and outcome of cadaver kidney transplants. A retrospective study of 250 cadaver kidney transplants was carried out to determine the effects of the agonal period, the warm and cold ischaemic times, and the use of kidneys with vascular anomalies on the primary success and failure and the subsequent level of function of the transplants. Kidneys with vascular anomalies or from non-ventilated donors had a primary failure rate of over 30%, whereas those with normal vasculature or from ventilated donors had a rate of 17%. An initial warm ischaemic time of more than 60 minutes was associated with a primary failure rate of 57% and a cold ischaemic time of over 550 minutes with a primary failure rate of 47%. The interrelationship between the warm and cold ischaemic times in the primary success or failure of the transplants was examined and criteria defined for selecting potentially viable cadaver kidneys for transplantation, as follows: (1) The donor should be (a) ventilated, (b) aged 6-50 years, and (c) have normal ante-mortem renal function and have secreted more than 1-5 1 of urine in the 24 hours before death (or an equivalent volume if the urinary output was recorded for less than 24 hours before death); (2) the kidney should have normal renal vasculature enabling single arterial and venous anastomoses to be performed; (3) kidneys with I.W.I.T.s of longer than 60 minutes should not be used; (4) for kidneys with I.W.I.T.s of less than 20 minutes the C.I.T. is not critical but should not exceed 12 hours; (5) for kidneys with I.W.I.T.s of 20-60 minutes the C.I.T. should not exceed 450 minutes.