Regulatory state of ribosomal genes and physiological changes in the concentration of free ribonucleic acid polymerase in Escherichia coli. The concept of promoter efficiency is introduced as frequency of RNA chain initiation at a given promoter normalized to the intracellular concentration of free (but functional) RNA polymerase. Previous observations from this laboratory on the synthesis of ribosomes and beta-galactosidase are used to show that during a nutritional shift-up from succinate minimal to glucose-amino acids medium (3-fold increase in steady-state growth rate) the concentration of free (active) RNA polymerase decreases to one-quarter of the pre-shift value and the promoter efficiencies of the genes for ribosomal RNA and ribosomal proteins increase 9- and 6-fold respectively. This extent of control of ribosomal genes is much greater than expected on the basis of the increase in the rate of ribosome synthesis (3-fold).


Identification of immunoglobulins and complement in rheumatoid articular collagenous tissues. Ninety-three patients with a variety of joint diseases were studied for evidence of immune complexes in articular collagenous tissues. Frozen sections of freshly obtained biopsies of hyaline articular cartilage and menisci were stained with fluoresceinated monospecific antisera for evidence of human immunoglobulins (IgG, IgM,IgA) and the beta1c component of complement. The criterion for the presence of complexes was the staining of two or more immunoglobulins and beta1c in an identical location of sequentially cut sections. Of the 42 patients with rheumatoid arthritis (RA) 83% were positive by this criterion. In those with classic RA the incidence was 92%. Sixteen patients with fresh joint trauma or nonarthritic disease had negative findings. Among 26 patients with noninflammatory disease, 4 of 8 with polyarthritis whose features suggested primary degeneration, 1 of 11 patients with secondary degenerative arthritis, and a single case of synovial osteochondromatosis had positive findings. Among 9 patients with miscellaneous inflammatory arthritides, all of 3 with psoriatic arthritis were negative; however 2 of 6 with other inflammatory arthritides were positive. The findings in classic RA suggest that immune complexes are deposited in the articular collagenous tissues. The persistence of these complexes may play a significant role in the chronicity of the synovitis.


[Centrophenoxin-induced dissolution and removal of lipofuscin. An electron microscopic study (author's transl)]. Three batches of senile guinea pigs were injected i.m. daily with 80 mg/kg, 50 mg/kg, or 30 mg/kg up to 10 weeks. With the dosage employed neurons of the central nervous system showed in all animals a significant reduction of lipofuscin pigment. A characteristic type of vacuolisation of the pigment is a preliminary sign of its reduction in size and quantity. Residuals of lipofuscin can be detected in astrocytes and in greater quantity in microglia cells and cells of the capillary wall. The dissolution of lipofuscin is not only found in the CNS but also in the myocardium and in the liver. In the latter organs the removal of pigment is carried out by phagocytes towards capillaries. The actual uptake of phagocytes is believed to occur by the process of exocytosis and pinocytosis. A destruction or an invasion of neurons by microglia has not been observed. Lipofuscin removal at a very small rate by microglia cells appears to occur normally in the ageing animal, but is greatly enhanced by centrophenoxin. Cessation of drug application does not stop the process of removal at once and the process continues as far as we can judge by our experiments for several weeks without further drug application. Even 30 mg/kg per day were found to be sufficient to initiate and sustain lipofuscin removal.


[Pathogenicity factors of gramnegative enteropathogenic bacteria and pathogenesis of intestinal diseases (author's transl)]. Enterotoxins as produced by the majority of enteropathogenic bacteria play a central role in the pathogenesis of cholera and E. coli enteritis but are obviously of little significance as to the development of Salmonella enteritis and dysentery. Pathology of Salmonella enteritis and dysentery results from multiplication of bacteria within the bowel wall and from the damage of tissue caused thereby. The lipopolysaccharides (endotoxins) of invasive enteropathogenic bacteria do not possess the character of a primary pathogenicity factor but operate as a protective agent versus bactericidal mechanisms of the macroorganism and probably also as an agent toxic for the tissue.


Current concepts on the regulation of purine biosynthesis de novo in man. In the present discussion we have presented our views on how purine biosynthesis de novo is regulated in man. The rate of the initital step unique to purine ribonucleotide biosynthesis de novo is controlled by the intracellular concentration of PP-ribose-P and purine ribonucleotides. This critical interaction of PP-ribose-P and purine ribonucleotides may be explained by a change in the physical properties of the enzyme that catalyzes this reaction. The first branch point in the pathway, following this initial step involves the utilization of IMP. Based on an in vitro analysis of the enzymes participating directly in the two biosynthetic pathways for which IMP is a substrate, we propose that the intracellular level of GTP may be more critical than previously recognized.


Halofenate. Its selection and trial as a primary uricosuric agent. In vitro binding studies on antiinflammatory and uricosuric acidic anions performed under "physiologic" conditions have demonstrated that these substances displace urate from its protein bond. The property of urate displacement appears to be a useful marker for potential uricosuric activity in vivo, and thereby a means to detect novel uricosuric drugs. One such drug, halofenate, was indeed a safe and effective uricosuric (comparable to probenecid) when used to treat hyperuricemia/gout over the long term; it did result in a modest and variable fall in serum lipid concentrations. However, used as a single fixed dose, halofenate did not produce a marked and consistent effect on the elevated serum triglyceride concentrations so commonly present in gouty patients.


[Synergism of colistin and sulfonamide in proteus species (author's transl)]. In agar diffusion testing of Proteus mirabilis strains sensitive to sulfonamide a synergistic effect of colistine (C) and sulfonamide (S) was demonstrated. By quantitative evaluation these results were confirmed in 100 strains of Proteus mirabilis and 100 strains of indol-positive Proteus spp. using broth dilution method. Combining 1 part C with 10 parts S the mean increase in sulfonamide sensitivity was enhanced fourfold. Increasing the inoculum sensitivity of Proteus against the combination of C and S was still found to range within therapeutic blood levels. Therefore in treatment of Proteus infection the positive synergistic effect of C+S should be taken into consideration.


Modified method for recording cortical and subcortical visually evoked potentials in the conscious rabbit with chronically implanted electrodes. A modified stereotaxic surgical and implantation technique for animal EEG studies in rabbits is described. The use of a stereotaxic "socklet unit" with a minimum of operational equipment, high accuracy, simple handling, and good tissue compatibility permits the repetition of EEG studies in the same animal with a large number of electrodes and over a period of several months. For example for this technique an experimental arrangement for recording visually evoked potentials is described. The importance of a careful selection of the experimental animals, of maintaining constant test conditions, and of the selecting of test parameters as a prerequisite for assessing induced alterations is emphasized and discussed.


[Coronary collateral vessels after long-term treatment with fendiline hydrochloride in a double-blind study (author's transl)]. After long-therapy with 5 mg/kg body-weight of N-(2-benzhydryl-ethyl)-N-(1-phenyl-ethyl)amine hydrochloride (fendiline hydrochloride, Sensit) in dogs the development of functioning coronary collateral vessels can be proven by filling the vessel system with synthetic resin. From earlier studies, it is known that coronary collateral vessels can protect the heart from the consequences of stenoses and tissue damage due to hypoxia.


[Administrative and juridicial problems in psychopharmacology (author's transl)]. The problem of a juridical and administrative regulation for therapeutic drugs is analyzed. There are 3 concurring factors of public interest: 1. protection of the patient against dangerous side-effects, 2. promotion of new effective therapies. 3. position and responsibility of the physician. Risks and problems of control are discussed in the light of development of a new preparation. A scheme of functions in the clinical trial under auspices of medico-ethical postulations is mentioned. Restrictive facts on behalf of a quick development of drug therapy are discussed. The necessity of intensifying biological basic research is emphasized.


Multiple sclerosis immunogenetics. A possible correlation with human leukocyte HL-A antigens. To investigate the relationships of genotype to susceptibility to multiple sclerosis, 42 patients hospitalized into Multiple Sclerosis Study Center of The Milan Medical School were typed for 14 HL-A specificities: HL-A1, HL-A2 + W 28, HL-A3, HL-A10 + W 19, HL-A11, HL-A9 of the 1st Series and HL-A5, HL-A7, HL-A8, HL-A12, HL-A13 of the 2nd Series. The increased HL-A9 frequency found in a previous study has been confirmed. Moreover in the total of multiple sclerosis patients this deviation reaches a higher degree of significance (P less than 0.01) and one antigen HL-A10 + W 19 appears to be significantly decreased (P less than 0.01). No variations were found in HL-A3, HL-A7 frequencies.


Absorption of erythromycin. A cross-over study in healthy volunteers. The absorptions of 6 erythromycin preparations were compared in a cross-over study in healthy humans. In a single-dose study, 500 mg of each preparation was, after an overnight fast, given to 10 volunteers. The two enterosoluble preparations of erythromycin base studied were absorbed slowly, and the peak serum concentration (1.5-2 mg/l) was achieved only at 4 h. The absorption of the stearates was quick, but especially one of them was poorly absorbed, the serum concentration being always below 1 mg/l. Both of the two estolates gave highest apparent concentrations, and the maximum serum level (2-2.5 mg/l) was achieved at 2 h, but the concentration of active erythromycin remains unknown. In the second part of the study, two erythromycin stearates and one base preparation were given at 6-h interval in a cross-over fashion, each for 4 days. On the 4th day, blood samples were analyzed. The erythromycin base gave higher serum concentrations than did the two stearates, which were equivalent. It seems doubtful that the erythromycin stearate at the dose of 250 mg every 6th hour would give satisfactory serum levels of erythromycin which would be effective against most bacteria during the whole treatment.


A 1,4-benzodiazepine, temazepam (K 3917), its effect on some psychological parameters of sleep and behaviour. In 30 physically and mentally healthy volunteers, 7-chloro-1,3-dihydro-3-hydroxy-1-methyl-5-2H-1,4-benzodiazepin-2-one (temazepam, K 3917) was tested for its sleep inducing action, the subjective quality of sleep and any post-medication effects. Temazepam was orally administered at doses of 15, 20 or 30 mg in hard gelatin capsules or 20 mg in soft gelatin capsules. Nitrazepam (5 mg) and amylobarbitone sodium (100 mg) were used for comparison as well as a placebo. Temazepam showed very much the same effects as they are known from conventional 1,4-benzodiazepines except for its lack of impairment in early morning behavior following night time medication.


[Pharmacokinetic studies on absorption and excretion of oxazepam in Combination with alcohol (author's transl)]. The influence of 7-chloro-1,3-dihydro-3-hydroxy-5-phenyl 2H-1,4-benzodiazepin-2-one (oxazepam, Adumbran) (30 mg) on the effects produced by alcohol (0.75 g/kg) and vice versa was studied in 14 male test subjects using the following scheme: (A) alcohol; (B) placebo; (C) oxazepam; (D) oxazepam and alcohol combined; (E) alcohol given 30 min prior to oxazepam. Blood and serum levels were determined and excretion in the urine was tested during the 7-h duration of the experiment. The blood alcohol curves show a similar course, leading to the conclusion that oxazepam has no influence on the elimination of alcohol. Alcohol retrads the intestinal absorption of oxazepam but, on the other hand, does not show an effect on the specific metabolism of oxazepam.


Mutagenic activity of anthraquinone derivatives used as dyes in a textile factory. Some anthraquinone derivatives used as dyes in a textile factory have been tested for their mutagenic activity on the Ames' strains of Salmonella typhimurium. 1-amino-4-hydroxy-anthraquinone proved to be a mutagen of the frameshift type. It is active without need of metabolic activation. Studies are in progress in order to clarify if the mutagenic activity of other substituted anthraquinones, which do not act as direct mutagens, can become evident after metabolic activation. Further tests are needed to decide if substituted anthraquinones are hazardous for exposed people.


Detection of Bhanja virus in cell cultures by fluorescent antibody technique. The multiplication of Bhanja virus in CV-1 cells was studied both by staining the viral antigens with the FA technique and measuring the light intensity emitted by the fluorescent cells with a photomultiplier and by the simultaneous titration of intracellular and extracellular virus. The fluorescence appeared at 3 hrs post adsorption in the form of very small granules in the cytoplasm of the infected cells. Percentage of cells containing viral antigens reached its maximum at 18 hrs post adsorption. Later on the fluorescence slowly decreased. The virus in infected cells was demonstrated 9 hrs post adsorption and maximum titre was reached 48 hrs post adsorption. The cell sheet remained apparently normal and no sign of cytopathic effect was observed until 36 hrs post adsorption.


[Studies on the adaptation of various strains of orthomyxovirus type A, subtype A/equi 2, passaged either on bovine kidney cells or on chicken fibroblasts or on RK-13 cells or by those passages alternating with "in ovo" passages]. The Authors have researched to adapt Orthomyxovirus, subtype A/equi-2 (Naples/213/67, Andria/60/68 and Miami/63), by serial passages on kidney calf cells, chick embryo cells and RK-13 cell line or by one or two passages on these colture media alternated with one passage "in ovo". The results obtained show that only kidney calf cells are most suitable because the viruses replicate for eleven passages. On the chick embryo cells and RK-13 cell line the viruses replicate only for two passages and these results are not modified when one or two passages are alternated with one passage "in ovo". The Authors have observed the production of infectious but not haemadsorbing and haemagglutining virus. The phenomenon is discussed and integrated with collateral tests.


Search for bacterial l forms in some diseases of human pathology. The Authors report their investigations performed in the attempt to isolate bacterial L Forms or protoplasts from human materials (urine, blood) and relate their presence with an underlying pathology, and discuss the results obtained.


[Immunological aspects of bronchial asthma]. Advances in understanding of bronchial asthma are to be expected from more detailed information on the immuno-pathological mechanisms involved. Reaginic IgE antibodies are responsible for the great majority of extrinsic asthma, but in some cases immune complexes could probably be also important. A great progress has been achieved after reproducible methods for detecting cellular and serum specific IgE antibodies were introduced. Moreover, the immunological and biological changes induced by immunotherapy are better known. Standardization and purification of allergenic extracts is now under study in many laboratories, our own included. As concerns the intrinsic asthma attempts to demonstrate the importance of (auto) immune mechanisms have been performed. The results of these studies are reported.


[Effects of thymectomy on the natural bactericidal power of thymectomized mice]. T and B-dependent systems are correlated between themselves and with the macrophage system in eliciting the immune humoral response. Not all immunogens require T-B interactions to elicit the antibody response: consequently, the problem of the limits of T-dependence, with regard to different immunogen stimulations, remains open. The comparative study of the serum antibody response in mice, thymectomized at birth and in the normal ones, inferred from the values of bactericidal activity (b.a.), detected versus E. coli, S. typhi and S. albus strains did point out evident variation. Thymectomy has impaired b.a. against E. coli; it seems to have no consequence on b.a. against S. typhi; is not possible to the results of our investigation, it seems that the response to the immunogens, responsible for the so-called natural antibodies does require T-B cooperation only for some bacterial species and not for others. A higher bactericidal capacity of normal serum with regard to Gram positive as compared with Gram negative bacteria, is confirmed.


The interaction of tubocurarine and suxamethonium at different stages of recovery from tubocurarine-induced neuromuscular blockade in anaesthetized man. The interaction of tubocurarine and suxamethonium in man was studied by the use of tetanic and single twitch contractions of the adductor pollicis muscles. Suxamethonium 0.2 mg/kg was administered at the times when the recovery of the tetanic contraction from neuromuscular blockade produced by tubocurarine 0.2 mg/kg had reached 20% and 50% of the control. At the 20% recovery point, suxamethonium improved both the tetanic and single twitch contractions; at the 50% recovery point, the single twitch response was markedly enhanced but the tetanic contraction was depressed. The findings suggest that the interaction of tubocurarine and suxamethonium is both antagonistic and synergistic. The tetanic contraction is more sensitive to the synergistic action compared with the single twitch contraction. When the interaction of these two agents is being evaluated it is important to consider the doses of the agents, the stage of curarization where the interaction took place and the method of assessing the neuromuscular blockade.


Inhibition of suxamethonium relaxation by tubocurarine and gallamine pretreatment during induction of anaesthesia in man. The effect of tubocurarine and gallamine pretreatments on suxamethonium relaxation was measured in 81 patients. The blocking effect of a constant infusion of suxamethonium (0.58 mg/sec) on the recorded thumb adduction in response to supramaximal ulnar nerve stimulation was reproducible in 18 control subjects: infusion time for 50% block was 37.7 (+/- SEM 1.02) sec. Tubocurarine 3 mg and 6 mg increased the infusion time required to produce 50% block by 33.4 and 54.8% respectively. Gallamine had a similar effect. Clinical conditions for endotracheal intubation were evaluated on a blind basis. Both drugs produced impairment of clinical conditions for intubation after suxamethonium 60 mg infusion. However, when pretreatment by tubocurarine 3 mg was followed by suxamethonium infusion at 1 mg/sec the time course of neuromuscular blockade was identical to that of the controls. There were no fasciculations with this dosage and conditions for endotracheal intubation were excellent.


Evaluation of etidocaine in extradural block. Etidocaine has been evaluated in extradural block with regard to its absorption, toxicity and clinical effectiveness, which have been compared with those of bupivacaine. Etidocaine is associated with lower plasma concentrations than bupivacaine and its absorption is decreased by the addition of adrenaline 1:200,000 to a greater extent than is the case with bupivacaine. Etidocaine causes motor paralysis more frequently than does bupivacaine, especially if adrenaline is added. The duration of action of the two drugs is similar when comparing etidocaine 1% and bupivacaine 0.5%. Bupivacaine 0.75%, however, acts for a significantly longer time than etidocaine 1%. The drug is a useful addition to the range of local anaesthetics, especially with regard to the degree of motor blockade it can achieve during surgical procedures.


The skeletal muscle response to edrophonium during neuromuscular blockade by tubocurarine in anaesthetized man. Tetanic contractions of the adductor pollicis muscle in man were evaluated continuously after the administration of tubocurarine 0.2 mg/kg and during reversal by edrophonium 0.1 mg/kg. Single twitch contractions of the adductor pollicis were measured simultaneously. The response of tetanic and single twitch contractions during the reversal of tubocurarine blockade by edrophonium were compared. With the tetanic contractions edrophonium, given at recovery up to 20% of the control value, increased the peak tetanic height and restored fully sustained tetanic contractions. This reversal, however, lasted only 3-4 min. The initial peak force of the tetanus settled at a slightly greater level than the value before the administration of edrophonium. The single twitch, however, was reversed and this reversal was well maintained and showed no decline. The results indicate that tetanic and single twitch contractions of the adductor pollicis muscle in man probably represent different aspects of the responses of the neuromuscular junction to edrophonium. The recording of the single twitch alone during the reversal of neuromuscular blockade by edrophonium could lead to a false conclusion about the recovery of transmission.


Bronchial hyperreactivity.

Bronchial hyperreactivity. Bronchial hyperreactivity is characterized by increased responsiveness of bronchial smooth muscle to non-specific constrictor stimuli. Tests used in assessing airway calibre are influenced by a number of factors and should be regarded as providing a qualitative rather than quantitative index of reactivity. One important determinant of bronchial reactivity is the resting state of the airways. An increase in resting bronchomotor tone either by the direct action of spasmogens or by the autonomic nervous system may potentiate a subsequent constrictor stimulus. Bronchial challenge may help in the diagnosis of asthma in patients with normal lung function at the time of testing.


Betamethasone valerate in corticosteroid-dependent asthmatics. The integrity of the hypothalamic pituitary adrenal axis. In a single-blind trial 29 patients with corticosteroid-dependent asthma reduced their daily dose of oral prednisolone by 1 mg/week while using a placebo inhaler until an unacceptable degree of asthma occurred. Betamethasone-17-vlaerate in a dose of 800 mug/day and if necessary 1600 mug/day was then substituted for the placebo inhaler and the reduction of oral prednisolone continued until the prednisolone was withdrawn completely or an unacceptable degree of asthma recurred. In 22 patients (76%) prednisolone was withdrawn completely, 11 on 800 mug and 11 on 1600 mug betamethasone-17-valerate. The mean reduction of prednisolone was 3-8 mg on placebo, 5-4 mg on 800 mug and a further 1-8 mg in patients requiring 1600 mug of betamethasone-17-valerate. The hypothalamic pituitary adrenal (HPA) axis was assessed by tetracosactrin and insulin stress tests at the start of the study and after withdrawal of oral prednisolone. The results indicate that an HPA axis which is completely suppressed by systemic corticosteroids can regain normal integrity when the systemic steroid is replaced by betamethasone-17-valerate in a dose of either 800 mug/day or 1600 mug/day. Candidiasis was observed, but will be reported later.


Objective and subjective comparisons of terbutaline and rimiterol bronchodilator aerosols. Double-blind studies of the bronchodilator effects of rimiterol, terbutaline and a combination of rimiterol and terbutaline were performed in patients with chronic asthma. An objective assessment in 21 patients showed no significant differences between the mean improvements in FEV1 during the first 30 minutes after drug administration. At 45 minutes and thereafter terbutaline produced a significantly greater degree of bronchodilatation than rimiterol or the combination of rimiterol and terbutaline. In a subjective study of 27 patients it was found that they were able to detect the more prolonged duration of bronchodilator activity of terbutaline but not the more rapid speed of onset of action of rimiterol.


Pathological aspects of immunization of mice against influenza virus infection. Groups of mice were immunized against influenza Ao/NWS virus by a single intranasal administration of inactivated homologous virus, by 2 intranasal doses of vaccine separated by an interval of 2 weeks, or by 2 intraperitoneal doses of the same vaccine. When subjected 2 weeks later to a standard challenge of 6 x 10(5) egg infecting units Ao/NWS virus instilled intranasally, mortality fell significantly from 64% in unimmunized mice to 39% in mice given a single intranasal dose of vaccine and to 29% in animals which received double intranasal vaccine. The best protection was conferred by double intraperitoneal immunization, after which mortality was 10%. Immunity waned with time, since the mortality of mice doubly immunized by the respiratory route and challenged 30 weeks later was 49%. Intrapulmonary lymphoid tissue developed in large amounts in a proportion of mice immunized by all methods and challenged after an interval of 2 weeks. Attention is drawn to this reaction as a possible unfavourable consequence of vaccination. There were no lesions in the lungs or central nervous system after immunization without subsequent challenge. The importance of histopathology in vaccine trials in experimental animals is emphasized by the consistently higher detection rate of lesions in lungs by histological examination than by visual inspection alone.


Demonstration of immunoglobulin containing deposits in glomerular basement membrane in experimental chronic serum sickness using horseradish peroxidase labelled antiserum. Horseradish peroxidase (HRP) labelled antibodies were used in rabbit glomerulonephritis to obtain localization of immunoglobulins in the glomerulus at a light and electron microscope level. Better localization of deposits was obtained with peroxidase labelled antiserum than with immunofluorescence at the light microscope level. Ultrastructurally, immunoglobulin localization was seen to correspond to the electron dense deposits demonstrated in and on the glomerular basement membrane by conventional microscopy.


Action of histamine on endothelial cells of guinea-pig isolated hepatic portal vein and its modification by indomethacin or removal of calcium. Methods are described for preparing guinea-pig hepatic portal vein endothelium for light microscopy and electron microscopy. Histamine (100 mug/ml) caused damage to the endothelium which was visible with both the light and electron microscopes. The damage was reduced in the absence of calcium. The reduction was more apparent with the electron microscope than with the light microscope. Indomethacin (100 mug/ml) protected the endothelial cells against the damaging effects of histamine. Possible modes of action of histamine and indomethacin are discussed.


Influence of time of infection during pregnancy with coxsackievirus B3 on maternal pathology and foetal growth in mice. Coxsackievirus B3 causes foetal wastage and growth retardation in mice which may be attributed to the action of the virus in destroying the maternal exocrine pancreas. Injection of virus on Day 4 or 8 of gestation caused greater foetal wastage than injection at 12 days. Foetal and placental weights in infected animals were less than in the controls but did not vary according to the time of infection. Little support is offered for the view that the mouse foetus differs in its susceptibility to Coxsackievirus B3 according to the stage in gestation of infection.


Response of ampicillin resistant Escherichia coli to cephalosporins in an in vitro model simulating conditions of bacterial growth in the urinary bladder. Five ampicillin resistant strains of Escherichia coli were exposed to cephalosporins in an in vitro model which simulates the hydrokinetic features of the urinary bladder. Although the strains showed substantial zones of inhibition when tested against cephalosporins by the disc diffusion method, the results in the bladder model suggest that, in conditions where the antibiotic concentration is being reduced by dilution and micturition as well as enzymic hydrolysis by the organism, activity of this group of agents may be insufficient to eradicate infection. It is suggested that the results warrant a closer investigation into the efficacy of cephalosporins against ampicillin resistant Gram negative bacilli in vivo.


Failure of "thymus factor" to restore transplantation immunity in athymic mice. Implantation of cell impermeable Millipore chambers containing thymuses from normal newborn donors into athymic mice does not confer on them the capability to reject allogeneic skin grafts. The same deficiency is manifested when skin grafted female nude mice are pregnant of foetuses with normal thymuses. These data are thus in sharp contrast to those previously reported for thymic factors reconstituting neonatally thymectomized mice.


Livedo reticularis. Signs in the skin of disturbance of blood viscosity and of blood flow. The clinical sign of livedo reticularis derives from stasis of blood in the superficial venous drainage systems of the skin. Many factors can delay the flow of de-oxygenated blood away from the skin, notably hyperviscosity of the blood itself and obstruction due to disease affecting dermal arteries, capillaries, or venules.


Treatment of pemphigus with cyclophosphamide. Cyclophosphamide was ineffective in controlling acute disease activity in nine of eleven pemphigus patients, but when given during the remission phase even 50 mg cyclophosphamide per day was sufficient to prevent subsequent relapses in all the patients for quite long periods of follow-up. This small dose of cyclophosphamide could be given to outpatients and there were no side effects.


Hypothalmic-pituitary relationships in the polycystic ovary syndrome serum gonadotrophin levels following injection of oestradiol benzoate. Injection of oestradiol benzoate was found to elicit a surge of luteinizing hormone (LH) in 15 of 19 patients with polycystic ovary (PCO) syndrome; follicle stimulating hormone (FSH) was also released, in conjunction with the LH surge, in eight patients. It is concluded that the oestrogen feedback mechanisms controlling mid-cycle gonadotrophin release are functioning normally in the majority of patients with PCO syndrome. Ovulation was subsequently induced by clomiphene alone in those patients who had an LH surge, and by a combination of clomiphene and human chorionic gonadotrophin (HCG) in those who had not shown any LH release.


Endometrial washings histological and cytological assessment of material obtained with an intrauterine jet washing device. The histological and cytological assessment of material obtained with an intrauterine jet washing device from 138 patients with postmenopausal bleeding, abnormal premenopausal bleeding or infertility is presented. In the first part of the study 55 washings were examined by histological techniques and the findings compared with those in material subsequently obtained by curettage or endometrial biopsy. Only 32 (58 per cent) of the washings were satisfactory for evaluation of the endometrium compared with 46 (84 per cent) of the curettings. When, in the second part of the study the washings in 83 cases were examined by both histological and cytological methods, 76 (92 per cent) were satisfactory compared with 59 (71 per cent) of the curettings. Cytological examination of the washings in the postmenopausal women provided a significantly higher proportion of satisfactory specimens than histological examination alone or evaluation of the curettings. In the whole study, three cases of endometrial carcinoma were diagnosed by endometrial washings and by curettage, while in six cases of endometrial hyperplasia one false negative was obtained by histological examination of the washings and one by examination of the curettings. This study shows that endometrial samples obtained with the intrauterine jet washer provide information about the endometrium which is comparable with that obtained by conventional curettage, and also that in postmenopausal women endometrial lavage may be more reliable than curettage.


Mooren's ulcer. Treatment by conjunctival excision. Ten eyes with progressing Mooren's or similar ulcers were treated by excising a 3 to 4 mm ring of limbal conjunctiva adjacent to the ulcer. Eight eyes healed within 3 weeks. Seven of these eyes have remained healed, while one eye has had repeated ulcers which healed when re-treated with conjunctival exicision.


Cavernous haemangioma of the retina and optic disc. A report of three cases and a review of the literature. We report characteristics of three cases of cavernous haemangioma of the retina, bringing to 37 the number now reported in the available literature. This rare, benign, congenital malformation is non-progressive, usually unilateral, somewhat more frequent in women, and rarely a source of intraocular haemorrhage. The fluorescein angiographic features include a normal arterial and venous supply, extraordinarily slowed venous drainage, no arterio-venous shunting, no disturbances of vascular permeability, and no secondary retinal exudation. Almost always, isolated clusters of vascular globules with plasma/erythrocyte sedimentation surround the main body of the malformation. These findings differentiate the anomaly from other retinal vascular diseases. Therapeutic intervention is seldom necessary.


Lenticonus in spina bifida. A case report. A previously unreported association between spina bifida and anterior lenticonus is noted. The significance of this finding to the aetiology of some cases of spina bifida is discussed.


Utilization of 14C-labelled Escherichia coli by rats. 1. Escherichia coli was grown on 14C-labelled glucose and fractions representing "whole cells", "cell contents" and "cell walls" were administered orally to rats. 2. 14C appearing as 14CO2 in the expired air and as unidentified radioactive products in urine and faeces was measured until the cumulative recovery showed little change with time. 3. All fractions were digested. The digestibility of cell walls was less than that of the other fractions. 4. There was considerable variation among individual rats.


The induction of obesity in rodents by means of monosodium glutamate. 1. Monosodium glutamate (MSG) was administered by various methods to mice and rats of various ages and the incidence of obesity was later measured. 2. Newborn mice were injected subcutaneously with 3 mg MSG/g body-weight at 1, 2, 3, 6, 7, and 8 d of age; 16% died before weaning. Of the survivors, 90% or more became markedly obese. Mean carcass lipid content was increased by about 120% in both sexes at 20-30 weeks old. In male mice, MSG treatment increased body-weight and epididymal fat pad weight, and greatly decreased adrenaline-stimulated lipolysis in isolated fat cells. Body-eright of females was not increased significantly. Food intake was not increased in either sex from weeks 13 to 15. Blood glucose level was not generally increased by MSG but some of the male mice had abnormally high values. 3. Obesity was not detected in the offspring of female mice that had received 100 g MSG/kg diet, either from 3 weeks before mating until weaning, or from the 14th day of pregnancy until weaning. 4. Intraperitoneal injection of 10 mg MSG/g body-weight (in two doses) at weaning increased carcass lipid content in female mice by 34% by 23 weeks of age, but female rats were not affected. 5. The addition of 20 g MSG/l to the drinking-water from weaning onwards did not increase carcass lipid content in female rats or mice. 6. The addition of 20 g MSG/kg diet from weaning onwards did not alter body-weight or carcass lipid content in male and female rats by 14 weeks of age. 7. The obesity induced in mice by MSG was not associated with hyperphagia, unlike genetic obesity and obesity induced by gold thioglucose (GTG). 8. All types of mouse studied, obese and lean, had essentially the same linear relationship between carcass water content and carcass lipid content. 9. Although MSG-obese mice could not readily be differentiated from normal mice by the increase in body-weight, which was only about 10% compared to 50-120% for genetic and GTG-induced obesity, the proposed schedule of injections in the newborn was almost 100% reliable in inducing a high extent of adiposity.


Substitution of cellular fatty acids in yeast cells by the antibiotic cerulenin and exogenous fatty acids. Cell growth of Saccharomyces cerevisiae ATCC 12341 inhibited by the antibiotic cerulenin, a specific inhibitor of fatty acid and sterol syntheses, was reversed by various exogenous fatty acids. Myristic acid (14 : 0), pentadecanoic acid (15 : 0), palmitic acid (16 : 0), and oleic acid (18 : 1) reversed effectively the growth inhibition by cerulenin. When these cells were reversed by adding pentadecanoic acid, over 90% of native even-numbered fatty acids was substituted by odd-numbered fatty acids. Those in the cells reversed by adding oleic acid were almost all unsaturated fatty acids. Cerulenin did not inhibit either elongation or desaturation systems in S. cerevisiae.


Abundant species of poly(A)-containing RNA from Saccharomyces cerevisiae. Hybridization experiments using uniformly labeled poly(A) RNA derived from Saccharomyces cerevisiae strains carrying the "killer character" showed that (1) these molecules appear to be transcribed from repetitive DNA sequences. (2) there are approximately 35 DNA template sequences that are transcribed into poly(A) RNA. It is concluded that under the RNA extraction procedure used, most of the poly (A) RNA represents killer-RNA as judged by the dependence of the kinetic complexity of poly(A) RNA on the genomic complexity of killer-RNA.


Activities of guanosine triphosphate analogues in reactions catalyzed by elongation factor Tu and initiation factor 2 of Escherichia coli. In earlier studies two natural analogues of GTP, guanosine 3'-diphosphate 5'-triphosphate (pppGpp) and dGTP, were found to substitute for GTP in reactions catalyzed by initiation factor 2 (IF-2) and elongation factor Tu (EF-Tu), while only dGTP could replace GTP with elongation factor G. These observations with IF-2 and EF-Tu have been extended to two analogues of GTP modified at the 3' ribose hydroxyl position, 3'-deoxyguanosine 5'-triphosphate (3'dGTP) and 3'-deoxy-3'-aminoguanosine 5'-triphosphate (3'dNH2GTP). These compounds were found to be similar to GTP, dGTP, and pppGpp in IF-2-dependent formation of N-formylmethionyl-puromycin and EF-Tu-dependent formation of N-acetyl-Phe-Phe-tRNA. The apparent Km values for the five guanosine nucleotides were 2 - 10(-6)-4 - 10(-6)M in the former reaction and 2-10(-7)--6-10(-7) M in the latter. These reactions did not have an absolute requirement for either an intact pentose ring or for the guanine base in the nucleotide. Although substantially less active than the guanine nucleotides, ITP and the dialcohol derived from GTP by periodate oxidation and borohydride reduction (ox-redGTP) were partially active in both the IF-2 and EF-Tu-dependent reactions, with apparent Km values about 40-100 times those of GTP.


Evidence for the participation of a host enzyme in the activation of poly (A)-Qbeta RNA as an infectious agent. It has been reported earlier that phage Qbeta RNA (Gilvarg, C., Bollum, F.J. and Weissmann, C. (1975) Proc. Natl. Acad. Sci. U.S. 72, 428-432) elongated at its 3' terminus with up to 100 or more AMP residues retained its full infectivity for Escherichia coli spheroplasts, and that the resulting progeny did not inherit the poly (A) appendage. We now show that while poly (A)-Qbeta RNA appears to function normally as messenger for the synthesis of virus-specific proteins it has lost its capacity to serve as template for Qbeta replicase. Template function could be restored by phosphorolysis with polynucleotide phosphorylase. Taken in conjunction, these results imply that after poly (A)-Qbeta RNA enters the spheroplast a host enzyme (perhaps polynucleotide phosphorylase) removes part or all of the adenylate residues prior to replication of the RNA.


Isolation and partial characterization of Chlamydomonas reinhardtii chromatin. The use of a cell wall less mutant of Chlamydomonas reinhardtii allowed the isolation of the nuclei of this organism. The study of the extracted desoxyribonucleoprotein shows that a set of established properties of the chromatin of higher eucaryotes do not apply to this material. This is particularly the case for the failure to sediment in  a good yield. This could be due to a low content in basic proteins of the chromatin of Chlamydomonas.


The determination of the primary structure of the 16S ribosomal RNA of Escherichia coli. III. Further studies. In this paper, we describe in detail the recent progress in the nucleotide sequence analysis of the 16S ribosomal RNA from E. coli. The sequence which has been partially or completely determined so far encompasses 1520 nucleotides, i.e. about 95 percent of the molecule. Possible features of the secondary structure are suggested on the basis of the nucleotide sequence and data on sequence heterogeneities, repetitions and the location of modified nucleotides are presented.


Further analysis of acyl-CoA-ACP-transacylases of mycobacterium smegmatis. Identification of a long chain alkyl malonyl-CoA-ACP-transacylase. Homogenates were prepared from three sources, Mycobacterium smegmatis Saccharomyces cerevisiae and Escherichia coli and tested for docosyl malonyl-CoA-ACP transacylase activity, using ACP purified from E. coli strain B and [2R, 2S, 1, 3-14C2] docosyl malonyl-CoA synthesized chemically, as substrates. Only homogenates of M. semegmatis showed positive transacylase activity. Successive chromatographies on Sephadex G-150 and then on DEAE-Sephadex A-50 prove that neither the palmityl-CoA-ACP-transacylase nor the malonyl-CoA-ACP-transacylase of M. smegmatis are responsible for this activity. The question concerning the identity of the enzyme with one of the two entities exhibiting acetyl-CoA-ACP-transacylase activity, previously identified in homogenates of this microorganism (1973 this journal, 55, 1381-1394), remains open for further experimentation. The physiological significance of the presence of a long chain alkyl malonyl-CoA-ACP-transacylase in homogenates of M. smegmatis, a representative of the Actinomycetales, is discussed in relation to the mechanism of the biosynthesis of mycolic acids. Chromatography on Sephadex G-100 showed that the substrate of the enzyme, docosyl malonyl-CoA, exists, in 50 mu molar aqueous solution, mostly in an aggregated state. A factor has been identified in the homogenates, which in the presence of radioactive docosyl malonyl-CoA, leads to the formation of a radioactive material showing an apparent molecular weight less than 10000. The nature of this material is discussed.


[Effect of ultraviolet light on sterol-containing membranes in nystatin solutions]. The nystatin induced conductivity of artificial phospholipid membranes containing ergosterol or 7-dehydrocholesterol drops sharply under the action of ultra-violet light as a result of the transformation of these provitamins into adequate D2 and D3 vitamins. The UV-light does not change the conductivity of the membranes containing cholestrol in nystatin solution. Nystatin slightly effects the membrane resistance in the presence of D2 and D3 vitamins.


[Kinetic equation for the process of accumulation of paramagnetic centers in proteins exposed to UV-radiation]. The process of accumulation of paramagnetic centres in UV-irradiated solutions of simple proteins at 77 degrees K has been studied. A kinetic equation describing the accumulation of radicals in protein is obtained. Experimentally obtained curves of radical accumulation coincide with the theoretical ones.


[Stimulation of rna synthesis by chloramphenicol]. The influence of different concentrations of chloramphenicol on the RNA synthesis was studied in E. coli strains CP 78 (rel+) and CP 79 (rel-). In cells growing in the medium with succinate as a carbon source the maximal stimulation of RNA synthesis was obtained in the presence of 5-10 mug/ml of chloramphenicol-that is when the protein synthesis in the cells is decreased to 50-30%. Concentrations of antibiotic 50-100 mug/ml only slightly stimulate the incorporation of 14C-uracil in TCA-insoluble fraction. The high level of RNA synthesis once achieved in the presence of small doses of chloramphenicol was not affected by higher concentrations of antibiotic up to 105 mug/ml. It is suggested that at low doses of chloramphenicol some proteins of positive control of RNA synthesis are synthesized. Under amino acids starvation in the presence of succinate only rel+ cells stopped RNA synthesis. The phenotypic suppression of RNA synthesis with chloramphenicol in these cells is achieved only by high concentrations of antibiotic (50-100 mug/ml). It is proposed that under amino acid starvation RNA synthesis is independent on the protein formation and is determined by relative contents of programmed ribosomes in cells.


[Conformational aspects of peptide interaction with proteolytic enzymes. Effect of amino acid residue configuration on binding of stereoisomeric N-acetylleucyltyrosine and N-acetyltyrosylleucine methylamides with pepsin]. Ki, Km and kcat. constants which characterize the interaction with pepsin of a complete series of Ac-Leu-Tyr-NHMe and Ac-Tyr-Leu-NHMe stereoisomers are determined. In compounds, containing residues of different configuration, Ki was found to have higher values than Ki or Km in DD- and LL-dipeptides. The data obtained show that not only the rate of peptide bond hydrolysis but also the efficiency of the binding with the enzyme depend on the configuration of the amino acid residue. Differences in the efficiency of pepsin binding of diastereomers is a manifestation of differences in their conformational characteristics. Data on "secondary" binding sites of pepsin are used to account for a mixed type inhibition observed in some cases.


[Isolation and characteristics of homopolyribonucleotide synthesized in toluene-treated E. coli cells]. Optimal conditions of homopolyribonucleotide (poly-A-14C) synthesis in toluene-treated E. coli cells under incubation with ADP-14C, Mg2+ and tris. HCl buffer (pH 8.0) are studied. Optimal Mg2+ concentration was 0.75.-10(-3) M. Heterogeneity of the isolated poly-A-14C from E. coli cell was demonstrated by means of sucrose density gradient (5-20%) centrifugation and polyacrylamide gel electrophoresis. Actinomycin D was found not to affect the reaction rate of polymerization of ADP-14C, UDP-14C and GDP-14C, catalyzed by polynucleotide phosphorylase in toluene-treated E. coli cells.


[Inactivation of yeast inorganic pyrophosphatase by sodium dodecyl sulfate and cetyltrimethyl ammonium bromide]. The kinetics of the inactivation of yeast inorganic pyrophosphatase by sodium dodecyl sulfate (SDS) and cetyltrimethyl ammonium bromide were studied. Micellar forms of detergents were shown to be an active reagetns under conditions tudied. The possible scheme of the inactivation including reversible formation of the micellar-protein complex and subsequent penetration of the bound detergent molecules in the protein is proposed. The enzyme ionogenic group with pK 7-8 by 25 degrees C is found to be responsible for conformational changes of the enzymes. The influence of the specific ligands on inactivation of yeast pyrophosphatase by SDS is studied, and dissociations constants of corresponding enzyme-ligand complexes are calculated.


[Conformational aspects of peptide interaction with proteolytic enzymes. Pepsin-catalyzed hydrolysis of cyclic peptides, containing leucyl-tyrosine fragments]. Pepsin was found capable of splitting cycloota- and cyclodecapeptides cyclo (-L-leucyl-L-tyrosyl-glycln-), n=6 or 8. The 18-membered peptides cyclo(-L-leucyl-L-tyrosyl-glycyl4-) and cyclo(-L-leucyl-L-tyrosy-delta-aminovaleroyl2-) were found stable to the effect of pepsin. To study the kinectics of hydrolysis for tyrosine-containing substrates of pepsin, a method of isolation of the unsplit substrate using ion-exchange resins and quantitative spectrophotometric estimation by absortion of tyysis of cycoocta and cyclodecapeptides was characterized by Km and kcat values.


[Application of the automatic Edman procedure to establishing the primary structure of swine pepsinogen and its fragments]. The automatic Edman procedure was applied to elucidate N-terminal sequences of swine pepsinogen, pepsin, and the fragments of its degradation by BrCN, i. e. B-1 and B-5. A "Beckman"  model 890 instrument was used in experiments. 50 amino acid residues were split off the pepsinogen molecule and identified and 55 amino acid residues-off the pepsin molecule by means of gas chromatography. A continuous N-terminal sequence of pepsinogen was 119 amino acids, in which the overlapping of the known peptide sequences with enzymic hydrolysers was taken into account. In B-1, B-4, B-5 fragments 22, 31 and 38, residues, respectively were analyzed with the sequencer.


Comparison of batch and semicontinuous cultures for production of protein from mesquite wood by Brevibacterium sp. JM98A. The production of protein by a Brevibacterium sp. JM98A using mesquite wood as the substrate was compared in batch and semicontinuous cultures. A 14 liter glass fermentor with automatic pH, temperature, and foam control was used for the study. A pH range of 6.6 to 7.2 was optimum for the growth of JM98A. The batch and semicontinuous cultures were compared on the basis of viable cell counts, protein production, CMC-Ase (beta-1,4-glucanase) activity, and filter paper cellulase (beta-1,4-glucan cellobiohydrolyase) activity. Total hexose, cellulose, and reducing sugar consumption were measured. The semicontinuous process yielded 2.97 times as much protein in 72 hr as the batch cultures. Most of the biomass resulted from the utilization of soluble sugars rather than from the degradation of cellulose during the semicontinuous process.


Engineering analysis of continuous production of L-aspartic acid by immobilized Escherichia coli cells in fixed beds. The reaction mechanism and decay behavior of aspartase activity for immobilized Escherichia coli cells were investigated by using a sectional packed column. Reaction within the immobilized cell column proceeded at zero-order on substrate solutions ranging in concentration from 0.1 to 1.0M, and the initial reaction rate was found to be 1.556 X 10(-2) mol/min/liter of immobilized cells. The effect of temperature on the reaction rate constant was investigated. The Arrhenius plot was a straight line at temperatures below 43 degrees C, and the activation energy for immobilized cells was calculated to be 12.36 kcal/mol. Aspartase activity in the immobilized cell column decayed exponentially and uniformly in all sections of a column. Its half-life was approximately 120 days. The rate of formation of L-aspartic acid was shown to be independent of column dimensions.


The heterogeneity and properties of folate binding proteins from chronic myelogenous leukemia cells. Previous studies have demonstrated that some chronic myelogenous leukemia cells contain a macromolecular binding factor for folic acid. This binder, which previously was believed to be a single factor, has now been resolved into two distinct binding proteins. Separation of each binder was obtained by DEAE chromatography of the partially purified lysate of chronic myelogenous leukemia cells. One binder has a molecular weight of 30;000-35,000, and the second binder has a molecular weight of 40,000-45,000. Both proteins bind the mono-, di-, and triglutamates of folic acid, N10-methyl-folate, dihydro-folate, and N5-methyltetrahydrofolate. Neither binder has determinants for N5-formyltetrahydrofolate or methotrexate. The preferred substrates for both binders appear to be the fully oxidized and partially reduced folates rather than the fully reduced folates. The lower-molecular-weight folate binding protein shows reversible binding with partially and fully reduced folates but irreversible binding with oxidized folates. This property suggests that this binder may have some function in the transport and storage of folate. The higher-molecular-weight folate binding protein, however, has only slight reversibility of binding with the partially and fully reduced folates, and it is therefore more difficult to postulate a physiologic function for this binding factor.


Central nervous system complications in patients with diffuse histiocytic and undifferentiated lymphoma: leukemia revisited. Fifteen of 52 patients (29%) with diffuse histiocytic and undifferentiated pleomorphic lymphoma developed central nervous system (CNS) complications, primarily leptomeningeal lymphoma. Lumbar puncture with cerebrospinal fluid cytology was the most useful test for diagnosis, and for following the response to therapy. Leptomeningitis developed during all stages of the patients' clinical course: at time of diagnosis, during progression of systemic disease, and most importantly as the initial site of relapse within 7 mo of attaining a complete clinical remission. Patients with bone marrow involvement are at high risk for the development of leptomeningeal lymphoma. Pathologic findings suggest that entry into the leptomeninges involves extension from the medullary bone marrow cavity along perforating vessels through dura into the arachnoid space. The leptomeningeal lymphoma has been successfully controlled in all patients receiving intensive central nervous system therapy consisting of a combination of intrathecal drug administration and radiotherapy. The high frequency of this syndrome and the success in its control suggest that a controlled trial of prophylactic CNS therapy be instituted in patients with these histologic types of non-Hodgkin's lymphomas.


Bone marrow transplantation in children with aplastic anemia and acute lymphatic leukemia. A boy with severe Aplastic Anemia (AA) and a girl with Acute Lymphoblastic Leukemia (ALL) in relapse have been grafted with marrow from HL-A identical, mixed leukocyte culture (MLC) negative siblings after appropriate immunosuppressive and antileukemic therapy. Both of them are well 7 and 2 months after transplantation respectively. Bone marrow transplantation should be considered in children with AA and ALL in relapse, if HL-A identical, MLC negative siblings are available.


A comparison of EMG feedback and progressive muscle relaxation training in anxiety neurosis. Two similar groups of adult psychiatric patients carrying the diagnosis of anxiety neurosis were compared in their response to different methods of training in deep muscle relaxation. One group received EMG feedback and the other a modification of the Jacobson Progressive Relaxation method. The frontalis muscle was chosen as the target for feedback training and for the measurement of tension reduction in both groups, for the reason that this muscle has been shown to reflect the general muscle tension level in anxious patients. Training was carried out under controlled laboratory conditions, and objective muscle tension levels were obtained. Overall changes in the status of the anxiety symptoms, as determined by global ratings from patients and from primary therapists, were also compared in the two groups at the end of training. The results of the study indicated that both EMG feedback and the progressive muscle relaxation training produced significant reductions in frontalis tension levels. However, EMG feedback was found to be generally superior in producing larger reductions in muscle activity, with a concomitant relief in anxiety symptoms, for a greater number of the patients.


Recognizing faces.

Recognizing faces. Following a review of the stimulus and subject factors which have been found to affect recognition faces, the question of whether this process can be considered a special one is dealt with. Evidence from studies involving the development of face recognition, the recognition of inverted faces, and the clinical condition prosopagnosia is considered, and in each case found to be inadequate for the unequivocal conclusion that the processes underlying face recognition are qualitatively different from those employed in recognizing other pictorial material.


How tight should tension sutures be tied? A controlled clinical trial. In a prospective randomized trial there was no advantage in tying tension sutures flush with the wound; by tying them loosely (over three fingers of the assistant's hand) there was less wound pain and less cutting in of the tension suture into the ridge of the wound and along the suture tracks.


Does the choice of suture material affect the incidence of wound infection? A comparison of dexon (polyglycolic acid) sutures with other commonly used sutures in an accident and emergency department. One hundred and four patients with superficial lacerations were sutured with either Dexon (polyglycolic acid), silk, polyethylene or nylon sutures. The incidence of postoperative tissue reaction and wound infection was compared. There were few complications and these occurred with almost the same frequency in each suture group, although there were slightly more cases of infection in the patients who were sutured with silk. Dexon was seen to possess certain advantages in that it caused as little tissue reaction as the other sutures but did not have to be removed subsequently. This could clearly be of benefit to the patients and hospital staff alike.


Comparison between ibuterol hydrochloride and terbutaline in asthma. The bronchodilating effect, circulatory effects, and subjective side effects of ibuterol hydrochloride, the di-isobutyric acid ester of terbutaline, at two dose levels (2 and 4 mg) were compared with those of 5 mg terbutaline sulphate in a double-blind cross-over study on 12 patients with asthma. Both drugs were given by mouth. The 2-mg dose of ibuterol had the same bronchodilating effect during the first three hours as 5 mg terbutaline. The 4-mg dose, however, produced a significantly greater increase in the peak expiratory flow rate between the 30th and 120th minutes than terbutaline. No significant changes in heart rate or pulse amplitude were noted, and there was no difference in the incidence of subjective side effects between ibuterol at either dose level and terbutaline.


Saccharomyces cerevisiae petite mitochondrial DNA of suppressive and neutral haploids and of [rho-] diploids obtained from crossing [rho+] to a neutral petite. An unusual property of GR25a [rho+] was the production of 20 to 30 percent [rho-] zygote colonies when crossed to a tester strain lacking mitochondrial DNA. Spontaneous [rho-] isolates of GR25a [rho+] were observed to be highly suppressive and to contain mitochondrial DNA of a parental buoyant density (1.685 g/cm3). Three ethidium bromide induced neutral petites of GR25 a [rho+] did not have detectable mitochondrial DNA and were neutral in crosses to [rho+] strains. Seven [rho-] zygote colony isolates obtained from crossing GR25a [rho+] to a neutral peptite were shown to contain abnormal mitochondrial DNA. Six zygote colony isolates had mitochondrial DNA of a buoyant density less than, or equal to, GR25a (1.682 - 1.685 g/cm3), whereas one isolate had a buoyant density greater than GR25a (1.688 g/cm3). It was suggested that abnormal mitochondrial DNA is generated during the mating reaction.


Action of "colonie lisse" mutation on cell morphology of Saccharomyces cerevisiae. The mutation of one of the genes of the series PLi in Saccharomyces cerevisiae Hansen brings about a reduction in the size of the cells. Besides, the mutation of some genes (PLi 5 and PLi 7) induces a change in the scanning microscopy appearance of the cell wall.


An account of the radium collar and the Finzi Harmer implant. Dr. J. G. Stewart of the Christie Hospital and Holt Radium Institute in Manchester gave an anecdotal account of two methods which were previously used to radiate laryngeal carcinomata and neck nodes. Both techniques have been superseded by newer irradiation techniques, but in view of their considerable success and historical interest, they are included here. The first paper concerns the radium collar which was used for the treatment of late cases of laryngeal carcinoma, and in particular for cases with neck nodes. The second is a brief account of the Finzi Harmer radium needle implant technique, which was at one time widely used.


Delayed primary closure in colon operations. Secondary closure of incisions by tape is a simple, safe procedure. In a retrospective series of 179 contaminated surgical wounds (incisions for colon operations) delayed wound closure resulted in a lower incidence of wound infection (5.8%) than did primary closure of similar wounds (11.8%). Although wounds left open do become infected, the acute swelling and systemic signs typical of an infection in a closed wound never develop. Delayed closure facilitated wound healing in hospital: only 3.8% of patients thus treated left hospital with open wounds, but 9.5% of patients whose wounds were closed primarily left hospital with wounds that were partially or completely open.


Delayed primary closure of the skin and subcutaneous tissue in abdominal surgery. Infection-prone abdominal incisions in 143 patients were managed by delayed primary closure of the skin and subcutaneous tissue of the abdominal wall. The overall wound infection rate was 1.4% and the rate for 74 open, large bowel operations was 2.7%. These rates compare favourably with a rate of 3.1% for clean inguinal hernia repairs done by the same surgical team over the same period. Delayed primary closure, carried out on the ward 3 days after operation, was simple, easy and did not prolong hospital stay. Its use is recommended in closing all abdominal wounds associated with a special risk of infection.


Dexon: an excellent suture for midline incisions. In a period of 18 months, 100 midline incisions of 93 patients were closed with interrupted 0 Dexon sutures. The Smead-Jones far-and-near stitch was used. There were no early disruptions and no incisional hernias in follow-up periods ranging from 5 days to 14 months. Four wound infections healed readily without prolonged drainage. Incisional pain was minimal in all patients. A group of 18 patients with potential for poor healing because of hemorrhage, sepsis, malignancy and steroids had no wound complications apart from an acceptable rate of infection. It is concluded that interupted 0 Dexon with wide fascial placement provides an excellent closure for midline incisions.


Involvement of the central nervous system in non-Hodgkin's lymphoma. Twenty-four patients with non-Hodgkin's lymphoma and CNS involvement are presented. There were 7 cases with diffuse histiocytic lymphoma (HL), 9 with diffuse poorly differentiated lymphocytic lymphoma (PDLL-D), of whom 6 patients were in leukemic conversion, 5 patients with nodular poorly differentiated lymphocytic lymphoma (PDLL-N), and 3 cases with undifferentiated lymphoma (UL). CNS complications were noted only in Stage IV lymphoma; the prognosis was generally poor. Histiocytic lymphoma was associated with widespread parenchymatous infiltration, whereas PDLL was usually associated with leptomeningeal seeding. The clinical course and the neuropathologic findings are discussed.


1,3-bis(2-chloroethyl)-1-nitrosourea(BCNU), cyclophosphamide, vincristine- and prednisone-(BCOP). A new therapeutic regimen for diffuse histiocytic lymphoma. Sixty-seven evaluable patients with mixed cellular and histiocytic lymphomas, both nodular and diffuse, were treated with a combined drug regimen of BCNU, cyclophosphamide, vincristine, and prednisone administered for six monthly courses. Of 28 previously untreated patients with diffuse histiocytic lymphoma, 14 (50%) achieved a complete remission, and an additional 7 (25%) had a good partial response for an overall remission rate of 75%. Complete remissions were also obtained among the small number of mixed and nodular histiocytic lymphomas which were treated. The median survival for previously untreated patients with diffuse histiocytic lymphoma who achieved a complete remission is nearly two years and 42% (8/19) of these patients remain in their initial unmaintained remission (range 2-119 weeks). Hematologic toxicity, although acceptable, was the limiting factor. Granulocytes were more often and more severely depressed than platelets.


Differential uptake of alkylating agents by normal and leukemic lymphocytes. When 14C-labeled cyclophosphamide and nitrogen mustard were incubated separately with normal lymphocytes and lymphocytes from patients with chronic lymphocytic leukemia, the amount of radioactivity associated with the normal cells far exceeded that detected on the leukemic lymphocytes. This comparative diminution may be analogous to the impaired PHA response and excess surface immunoglobulin which serve as identifying markers of the malignant B cell. Cytotoxicity and neuraminidase experiments indicated that drug uptake by lymphocytes is not capricious and may occur in an optimum, predetermined fashion. Although surface uptake and therapeutic response are not necessarily directly interrelated, initial peripheral contact with an antineoplastic agent may be an essential step which modifies tumor sensitivity or resistance.


Metastatic reticulum cell sarcoma and lactic acidosis. The clinical course of a woman with metastatic reticulum cell sarcoma and intractable lactic acidosis is described. Although her illness was dominated by a myelopathy, she developed severe lactic acidosis which could not be related to decreased tissue oxygen delivery. Necropsy showed extensive hepatic replacement with tumor and widespread disease. This and other possible pathogenetic factors causing lactic acidosis are discussed.


Clinical evaluation of a method of quantitative determination of homovanillic acid for the estimation of degree of tumor dissemination process in melanoma of the skin. Homovanillic acid (HVA) was determined both in 45 patients with different forms of melanoma and those hospitalized on the suspicion of melanoma and in 15 healthy persons. Excretion rate of homovanillic acid within the normal limits was observed in patients with benign melanotic neoplasms of the skin (4.4 +/- 0.56 mg/24 hr) and in local forms of melanotic melanoma of the skin (4.5 +/- 0.24 mg/24 hr). In patients with metastases to regional nodes as well as in patients with locally disseminated forms of melanotic melanoma of the skin the concentration of homovanillic acid increased up to 11.1 +/- 0.93 mg/24 hr. Especially strong concentration of homovanillic acid was registered in patients with disseminated malanotic melanoma of the skin, and it made up 15.7 +/- 2.04 mg/24 hr. There was evident tendency to the elevation of HVA excretion level as generalization of the process which was taking place. The data obtained enable the authors to recommend the determination of HVA excretion as an additional diagnostic test as well for prognosis of melanoma development together with other clinicomorphological data.


Breast thermography as a screening technique. An evaluation of performance data. The concepts of relative operating characteristics (ROC-curves) and detectability index (d') are introduced for the purpose of evaluating performance in breast thermography. In assessing published information on the subject we have found that much of it is anecdotal and lacks sufficient data to determine performance. We have also found that for those published findings which had sufficient data it was possible to reconcile conflicting conclusions as to the efficacy of thermography, and that performance could be quantitated with the use of the detectability index. Conclusions for optimizing available clinical thermographic techniques are given together with ways for future improvement.


Interaction of Rhodium(II) carboxylates with molecules of biologic importance. Rhodium(II) acetate, propionate, and butyrate showed a considerable variation in their antitumor activity against Ehrlich ascites tumor cells in mice, with the butyrate complex being the most active. The three complexes markedly inhibited DNA synthesis of Ehrlich ascites tumor cells in vivo. Rhodium (II) butyrate was the most potent inhibitor followed by the propionate complex. One hour after administration, rhodium(II) propionate and butyrate induce more uridine-5-3H incorporation into RNA than is seen in the controls. Equilibrium dialysis studied showed that rhodium(II) acetate-1-14C binds to single stranded DNA, poly-A, ribonuclease A, and bovine serum albumin but not to highly polymerized native calf thymus DNA, poly-G, or poly-C. In these cases binding occurred at the two axial positions of rhodium(II) acetate to a nitrogen donor in the ligands. The formation constants of the rhodium(II) acetate and propionate complexes with 5'-adenosine monophosphate were determined. The rhodium(II) propionate complex was more stable. Sedimentation and viscosity measurements of poly-A and poly-A/rhodium(II) acetate complexes indicate a high degree of intramolecular crosslinking in the rhodium(II) acetate/poly-A complex. The rhodium(II) carboxylate complexes were also found to be potent inhibitors of purified DNA polymerase I and RNA polymerase from Escherichia coli.


Review of the current clinical status of platinum coordination complexes in cancer chemotherapy. During the past 3 years, eight hospitals and one cooperative study group have reported their initial clinical results with cis-dichlorodiammineplatinum (II). The most popular clinical schedule was 15-25 mg/m2/day for 5 days repeated every 3-4 weeks. Almost all patients had extremely advanced disease. Of 323 patients in whom response could be evaluated, there were 12 complete responses, 25 partial responses (greater than 50% decrease in tumor size), and 23 improvements (greater than 50% decrease in tumor size) for a 19% overall response rate. The tumor most sensitive to cis-dichlorodiammineplatinum (II) was testicular carcinoma in which seven complete responses, three partial responses, and three improvements were observed in 16 patients treated at Roswell Park Memorial Institute. Other sensitive tumors were lymphoma (63% response and improvements), squamous cell carcinoma of the head and neck (41% response and imporvements), and ovarian carcinoma (40% response and improvements). Complete responses were also seen in one patient with thyroid carcinoma and two with bladder carcinoma, while partial remissions were recorded in two patients with breast carcinoma and one patient each with acute myelogenous leukemia, endometrial carcinoma, renal carcinoma, malignant thymoma, neuroblastoma, adenocarcinoma of the lung, and an undifferentiated tumor of unknown origin. Five major types of toxicity were encountered: gastrointestinal, hematopoietic, immunosuppressive, otologic, and renal, with the last two generally the most serious. Serial audiometry testing can generally warn of the otologic toxicity and thus prevent permanent acoustic damage. Renal toxicity, which is similar to that seen with heavy-metal poisoning, appears to be dose related, cumulative, and only partly reversible, thus, severely limiting the repeated administration of cis-dichlorodiammineplatinum (II). Recent laboratory studies suggest that combination chemotherapy with this drug may be rewarding. Studies of this nature should be pursued along with attempts to find more effective less toxic platinum compounds.


Clinical phase I trial of inosine dialdehyde (NSC-118994). An initial clinical phase I trial of inosine dialdehyde has been carried out in 40 patients at dose levels of 30-4000 mg/m2 for 5 days given intravenously (iv) monthly. At 1.5 g/m2, noncumulative dose-related toxicity occurred in all patients which consisted of nausea and vomiting, local pain, alterations in coagulation mechanism, elevated partial thromboplastin time, and positive Coombs' test. No dose-limiting leukopenia, thrombocytopenia, anemia, or bleeding occurred; however, depression of the leukocyte and platelet counts, and decreased hemoglobin value were observed. The dose-limiting toxic effect was renal tubular damage with reversible acute renal failure in one of four patients who received 3000 mg/m2 iv for 5 days. Refractory hypercalcemia was controlled in three of three patients without tumor effect. Responses occurred in patients with seminoma, oat cell carcinoma, and melanoma. A starting dose of 2 g/m2 for 3 days monthly is recommended for phase II trials and a trial in lung carcinoma is now being conducted.


Use of combination adriamycin (NSC-123127) and DTIC (NSC-45388) in children with advanced stage IV neuroblastoma. Adriamycin and DTIC were used in combination because of their reported effectiveness in neuroblastoma when administered as single agents and because of the poor survival rate of patients with this malignancy in its disseminated stage. Eighteen patients with previously treated stage IV neuroblastoma received this combination chemotherapy every 21 days. Two to eight courses were administered. Two partial and no complete remissions were seen. Mild to moderate gastrointestinal and hematologic toxicity was observed.


Structural studies on the O-specific side-chains of the cell-wall lipopolysaccharide from Escherichia coli O 75. The structure of the O-specific side-chains of the cell-wall lipopolysaccharide of Escherichia coli O 75 has been investigated, using methylation analysis and Smith degradation as the principal methods. The O-specific side-chain was found to be composed of a tetrasaccharide repeating-unit of the following structure: (see article)


Haemodynamic studies during short-term preservation of the autoperfusing heart-lung preparation. Preservation of the donor heart in a viable state for even a few hours would greatly facilitate clinical cardiac transplantation. The autoperfusing heart-lung preparation has been investigated for periods of 2 to 6 has a means of short-term preservation of the heart. Its functional capability has been assessed by observations of a number of haemodynamic parameters, including those parameters believed to give an indication of the state of myocardial contractility. Although conversion of the normal circulation to that of the preparation was accompanied by considerable falls in max dP/dt and its derivative max dP/dt/P, there was a subsequent stabilization or improvement in these parameters during the period of autoperfusion. Haematoxylin-basic fuchsin-picric acid staining of the myocardium showed no evidence of myocardial ischaemia during the perfusion period. These observations, together with other haemodynamic and histological observations, suggested that myocardial perfusion by this technique was satisfactory. The effects of a coronary dilator such as dipyridamole have been observed, and its value in approximating the coronary and pulmonary blood flows in such a preparation is discussed. These studies would suggest that the autoperfusing heart-lung preparation can provide a simple and satisfactory form of short-term storage of the heart and, with further experience, may prove of value in clinical cardiac transplantation.


Effects of catecholamines on the coronary circulation in the Langendorff-type transplanted dog heart. Intracoronary administration of isoprenaline, adrenaline, and the noradrenaline in the Langendorff-type transplanted dog heart transiently decreased coronary blood flow measured from the inflow vessel; flow then increased. The reduction in coronary flow coincided with increased myocardial contractility and was prevented by keeping the left ventricle empty. Propranolol prevented changes in flow and contractility. It is concluded that positive inotropic changes result in the ejection of accumulated Thesbesian flow from the ventricle and affect coronary flow measurements and that the interpretation of flow changes requires a steady state.


Alternating administration of adriamycin (NSC-123127) and vincristine (NSC-67574)-actinomycin D (NSC-3053) in advanced sarcomas. Fifteen patients with surgically incurable, advanced, and metastatic sarcomas were treated with courses of adriamycin and vincristine-actinomycin D alternating within a 7-week cycle. Three patients had objective partial responses for more than 3 months (liposarcoma, 4 months; fibrous histiocytoma, 15+ months; desmoid tumor, 19+ months) while two other patients (liposarcoma, leiomyosarcoma) had lesser responses. No beneficial effect could be attributed to therapy in two patients with leiomyosarcoma, two patients each with synovial cell sarcoma, fibrosarcoma, and chondrosarcoma, and one patient each with rhabdomyosarcoma and mesenchymoma. No additive effect of alternating full doses of these agents could be demonstrated over the published data on response to adriamycin or actinomycin D alone.


Dibromodulcitol (NSC-104800) compared with cyclophosphamide (NSC-26271) as remission maintenance therapy in previously treated children with acute lymphoblastic leukemia or acute undifferentiated leukemia: possible effectiveness in reducing the incidence of central nervous system leukemia. Dibromodulcitol and cyclophosphamide are both alkylating agents. In this study, these two drugs were compared for their effectiveness as remission maintenance therapy for childhood acute lymphoblastic leukemia or acute undifferentiated leukemia. Toxic effects were similar in both groups of patients although cystitis did not occur with the dibromodulcitol treatment. The duration of remission was slightly shorter for dibromodulcitol than for cyclophosphamide (P = 0.04). There was, however, a lower incidence of CNS leukemia in the patients treated with dibromodulcitol, which did not seem to be related to a basic difference in the patient groups.


Phase II study of cis-dichlorodiammineplatinum(II) (NSC-119875) in combination with cyclophosphamide (NSC-26271) in the treatment of human malignancies. The effectiveness of cis-dichlorodiammineplatinum(II) in the treatment of human malignancies is evaluated. The first stage of our investigation consisted of a phase I study to determine toxicity. In the second stage attempts were made to reduce toxicity by varying the modes of administration, and the third stage comprised studies of combination chemotherapy including cis-dichlorodiammineplatinum(II). A total of 74 patients have been treated, 20 of whom received the combination of cis-dichlorodiammineplatinum(II) and cyclophosphamide. Major toxic effects included vomiting, mild leukopenia and thrombocytopenia, decreased creatinine clearance, audiologic toxic effects, hyperuricemia, and nephrotoxicity. Measurable regression of tumors was seen in 18 of the 74 patients and ten of the 18 patients who responded had been given the combination of cis-dichlorodiammineplatinum(ii) and cyclophosphamide.


The surface structure of isolated pancreatic islet cells. The surfaces of isolated pancreatic islet cells were studied with the scanning and transmission electron microscopes. Islets were isolated from the pancreas of Wistar rats by collagenase treatment and were incubated either in glucose-free medium or in 300 mg% glucose for one hour. Immunoreactive insulin (IRI) in the media of both control and experimental preparations was assayed. Islets were then transferred to 4% glutaraldehyde, buffered with cacodylate, pH 7.4, and prepared for scanning and transmission electron microscopy. Cell masses average 200 mu in diameter. Alpha cells appear pyramidal in shape, are about 8 mu in diameter and appear in groups. Beta cells are round or oval in shape and have an average diameter of 10 mu. Glucose stimulation raised the IRI value tenfold and increased the number of blebs and other surface irregularities per unit area of beta cell surface. Comparison with transmission electron micrographs suggests that the blebs are related to the process of emiocytosis.


Changes in myosin distribution in dedifferentiating and redifferentiating smooth muscle cells in tissue culture. Isolated smooth muscle cells and fibroblasts from the newborn guinea-pig vas deferens were grown in culture. In the first 2 days, all cells characterized as smooth muscle by phase-contrast microscopy reacted intensely with fluoresceinated antibodies against smooth muscle myosin. The fluorescence was in the form of particles (termed here "myosin aggregates"), which were often aligned to give the cell a striated appearance. After 3-5 days, coarse fluorescent fibrils were also visible. These were termed "attachment fibrils" ("A-fibrils") since they were thought to represent myosin in microfilament bundles. Between 6 and  days in culture, the smooth muscle cells began to dedifferentiate morphologically. At this time, the "myosin aggregates" became clumped and less intensely fluorescent. "A-fibrils" also decreased in fluorescence intensity. By 8 days in culture, the dedifferentiated cells had undergone intense proliferation and gave only a minimal reaction with myosin antibodies. However, when a confluent monolayer of cells formed on day 9 or 10, they immediately began to redifferentiate ultrastructurally and to regain immunofluorescence in both "myosin aggregates" and "A-fibrils". Throughout the entire culture period, cells characterized as fibroblasts by phase contrast microscopy gave only a weak reaction with fluoresceinated antibodies to myosin showing "A-fibrils" but no "myosin aggregates".


Lung water and urea indicator dilution studies in cardiac surgery patients. Comparisons of measurements in aortocoronary bypass and mitral valve replacement. We measured transpulmonary indicator dilution curves of 51Cr-erythrocytes, 125I-albumin, 14C-urea, and 3H-water before and six and 24 hours after operation in seven patients undergoing aortocoronary bypass (ACB) and eight patients undergoing mitral valve replacement (MVR). We calculated cardiac output (CO), extravascular lung water (EVLW), the difference between 125I-albumin and 51Cr-erythrocyte distribution volumes (EV albumin), the difference between 14C-urea and 51Cr-erythrocyte distribution volumes (EV urea) and 14C-urea extraction (E) and permeability -surface ares (PS) products. Comparisons between 16 ACB studies and 17 MVR studies showed the MVR group to have a higher EVLW (P less than 0.01). Extravascular lung water decreased after operation. The ratio of EV urea to EV albumin averaged 1.35 in the MVR group and 0.91 in the ACB group (P less than 0.001). 14C-urea E was also higher in the MVR group (P less than 0.05), but PS was similar in the two groups. None of the differences was related to the time that studies were done. We showed that EVLW, calculated using both 125I-albumin and 51Cr-erythrocytes as intravascular indicators and measured blood water content, had a constant relationship to EVLW calculated using only 125-I-albumin as the intravascular indicator and neglecting blood water content, over a broad range of cardiac outputs, hematocrist, and lung water volumes. We conclude that patients with mitral valve disease have an increased distribution volume and E for urea, probably due to hemodynamic changes but possibly due to increased vascular permeability. Extravascular lung water decreases after cardiac surgery regardless of the type of operation. A single intravascular indicator is adequate for estimating extravascular lung water in humans.


Electrophoretic amylase fractionation as an aid in diagnosis of pancreatic disease. Six alpha-amylase (EC 3.2.1.1) isoenzymes have been resolved electrophoretically on cellulose acetate membranes in a discontinuous buffer system. The fastest migrating isoenzymes are of salivary origin (S1, S2, S3), the slower ones of pancreatic origin (P1, P2, P3). We determined the amylase isoenzyme distribution in the sera of 240 subjects. A specific pancreatic isoenzyme (P3) was observed in all clinically diagnosed cases of acute or chronic pancreatitis as well as in 15 of 40 renal-transplant patients. Moreover, P3 isoenzyme activity declined during apparent recovery from pancreatitis. The P2 isoenzyme appeared in 95% of all specimens, P1 in only 2%. The pancreatic isoenzymes were preferentially excreted in the urine of both renal-transplant patients and normal individuals. The major salivary isoenzyme, S1, was observed in 95% of all serum and urine samples; however, the S2 and S3 appeared less consistently. Our method is simple and rapid, and quite applicable for use in clinical evaluation of patients with pancreatitis or with certain nonpancreatic dysfunctions.


Improved microscale assay for purine phosphoribosyltransferase activities. We describe an improved and rapid filter-type assay for purine phosphoribosyltransferase activities. 14C-labeled purine bases are used as substrates, and the equipment includes a sampling manifold and glass-fiber filters coated with polyethyleneimine-cellulose. The method is especially suited for assay of a large number of samples, and may be useful for other enzyme activity measurements.


The immunological consequences of antigen overload in experimental mycobacterial infections of mice. Mice infected in the tail with M. ulcerans developed transient cell-mediated immunity which disappeared as the bacterial load increased. Lymph node cells from animals in this late phase of the disease transformed spontaneously in vitro. This transformation was inhibited by mycobacterial antigen. Lymph node cells from mice injected intravenously with 10(8) or 10(9) BCG also transformed spontaneously in vitro. Such animals did not become foot pad test-positive. Evidence is presented that the spontaneous transformation may represent an accumulation of specifically sensitized cells due to trapping in nodes overloaded with persistent antigen. The relevance of such a phenomenon to 'desensitization' in human and animal disease is discussed.


Rapid quantitation of changes in macrophage volume induced by lymphokine in vitro. A new machine, the Coulter Channelyzer P128, has been used to measure the mean cell volume of suspensions of normal guinea-pig macrophages. Macrophage volume, expressed in cubic microns, has been examined after various times of storage and subsequently after contact with varying concentrations of lymphokine. It is suggested that this machine improves the speed and accuracy of quantitating one in vitro assay of lymphokine activity.


Guinea-pig nephrotoxic nephritis. I. The role of complement and polymorphonuclear leucocytes and the effect of antibody subclass and fragments in the heterologous phase. In guinea-pig nephrotoxic nephritis induced by a sheep antibody there was minimal glomerular capillary deposition of C3 and accumulation of polymorphonuclear leucocytes (PMN) in the heterologous phase. The C4-deficient strain developed the same injury as normal Duncan-Hartley animals. Complement depletion with cobra venom factor, polymorph depletion with nitrogen mustard or anti-PMN serum and treatment with antihistamines provided no protection. The relationship between the dose of nephrotoxic antibody and the proteinuria was similar for gamma1 and gamma2 subclasses and the F(ab')2 fragment of gamma1 antibody. However, the F(ab') and F(ab) antibody fragments, though fixing on the glomerular basement membrane, did not cause proteinuria. It is concluded that the development of proteinuria in this system: is largely independent of the complement-polymorph system; is due to the fixation of the F(ab')2 fragment of the antibody molecule; and does not depend on an intact Fc piece.


A paraprotein in severe combined immunodefeciency disease detected by immunoelectrophoretic analysis of plasma. A qualitative study was made of the plasma immunoglobulins of a child with severe combined immunodeficiency. By immunoelectrophoresis an immunoglobulin with an abnormal electrophoretic mobility was detected. This protein possessed mu heavy chain determinants, gave no detectable reaction with antisera specific for light chains, was of a relatively small molecular size, and was probably not composed of subunits held together by easily reduced disulfide bonds. The light chains that were present in this patient's plasma had a homogeneous electrophoretic mobility. The patient's plasma also contained at least two other immunoglobulins whose antigenic identity could not be established. One of these was abnormal in its electrophoretic mobility. The presence of the abnormal protein with mu determinants in the plasma of the second unrelated child with a similar disease suggests that the detection of this protein may have implications for the diagnosis or classification of immunodeficiency diseases.


An indirect immunofluorescent test for human antibodies to tetanus toxoid using an insoluble toxoid as antigen. An indirect fluorescent antibody (FA) test for detection of human antibodies to tetanus toxoid is described using an ethylchloroformate-prepared polymer of tetanus toxoid as the particulate slide test antigen. Titres of the FA test were compared with those obtained with the toxin neutralization (TN) test in mice. No antisera were FA-positive at less than 0-0025 AU/ml. Positive correlation of the FA test with the TN test was 50% between 0-0025 and 0-01 antitoxin units/ml (AU/ml) and 100% between 0-02 and 160 AU/ml. In general, an increase in FA titres correlated with an increase in TN titres beginning at about 0-08-0-16 AU/ml.


Clinical applications of the continuous flow blood separator machine. The NCl/IBM or Aminco Continuous Flow Blood Separator Machine is a safe apparatus for the selective removal or exchange of either packed red blood cells, leucocyte-rich or platelet-rich layers or plasma. Abnormal fractions from any of these layers may be collected and discarded. Normal constituents may be collected for therapeutic uses. The wide scope of its applications includes important uses in clinical immunology: temporary provision of good leucocytes or platelets; harvesting of immune leucocytes (preparation of transfer factor at up to 10 units per harvest); removal of cryo- or macro-globulins, immune complexes or blocking factors; replacement therapy for antibody or complement deficiencies. Examples are given of such uses together with some of the medical problems so far encountered.


Binding of the complement intermediate C56 to zymosan in acute phase human sera. C56 is known to appear in the fluid phase when zymosan is incubated at 37 degrees C with certain acute phase 'reactor' sera. In the present study, C56 was detected bound to the zymosan particle prior to its appearance free in solution. In reactor sera C56 was formed and released with kinetics similar to that of the generation and decay of a C56-binding site formed when zymosan was incubated with normal serum. Bound and fluid phase C56 was detected only in reactor sera, and was generated only by agents known preferentially to activate the properdin pathway. Elution of C56 from zymosan in hypertonic salt solutions proved to be a convenient step in the partial purification of large amounts of this haemolytically active bimolecular complex.


Increased antibody production following depression of hepatic phagocytosis. The influence of Kupffer cell blockade on the humoral immune response to suboptimal doses of intravenous sheep red blood cells has been measured in mice. Hepatic phagocytosis was suppressed using dextran sulphate. Direct (IgM) and indirect (IgG) plaque-forming cells were measured in spleens from treated and untreated mice at varying times after the antigen. The results show that after Kupffer cell blockade both IgM and IgG responses correspond to the responses seen after a 10-fold greater dose of cells in control animals. The implications of this are discussed.


Ultrastructural and serological studies on the resistance of activated B cells to the cytotoxic effects of anti-immunoglobulin serum. Patch and cap formation of surface immunoglobulin on mitotic B lymphocytes. Previous studies have shown that rabbit anti-mouse immunoglobulin sera (anti-Ig) which kill non-activated B lymphocytes in the presence of complement, are incapable of doing so when the cells are activated by antigen or mitogen into mitosis. Results reported here indicate that the resistance is not dependent on either the source of antiserum or complement, or on the presence of a mitotic inhibitor, colcemid. Immunoperoxidase staining-electron microscopy techniques were applied to assess whether there was any conspicuous difference between unstimulated versus mitogen-stimulated, mitotic cells with respect to density or distribution of cell surface Ig. No such differences were found; furthermore, mitotic cells showed rapid classical 'patch and cap' formation of cell surface Ig when incubated with anti-Ig at room temperature, indicating the retention of fluid membrane dynamics by lymphocytes in this stage of the cell cycle. In contrast to this cytotoxic resistance, T or B lymphocytes in mitosis were found to be as sensitive, or more so, to lysis by various other antisera when compared to non-mitotic cells. Thus the resistance of mitotic B cells to the cytotoxic effects of anti-Ig serum seems unique and appears independent of any conspicuous quantitative or qualitative change in cell surface Ig.


Effects of oestrogens and pregnancy on the distribution of sheep erythrocytes and the antibody response in mice. Mice pre-treated with three oestrogenic preparations showed increased hepatic and reduced splenic uptake of 51Cr-labelled sheep erythrocytes (SRBC). The anitbody response to SRBC wahe number of antibody-forming cells in the spleen parallels the amount of SRBC localizing in this organ and that both can be depressed or enhanced by appropriate pre-treatments with oestrogens or colloidal carbon. The effects of these agents are mediated through stimulation or 'blockade' of the phagocytic activity of liver macrophages. Changes in localization of SRBC in pregnant mice were similar to those found after treatment with oestrogens. These changes were, however, rather small and the antibody response of pregnant animals was not affected.


Human sperm antigens and antisperm antibodies I. Studies on vasectomy patients. This study documents the types and incidence of antisperm antibody, detectable by indirect immunofluorescence, in 114 patients before vasectomy, 112 at 2 months and 71 patients at 6-9 months after vasectomy. Indirect immunofluorescence techniques revealed antibodies to seven distinct sperm antigens. Five of these antigens were readily accessible to antibody in vitro, and the remaining two were accessible only after treatment of spermatozoa with dithiothreitol and trypsin. Antisperm antibodies were detected in 61% of patients before vasectomy. The incidence rose to 77% at 2 months and 90% at 6-9 months after vasectomy. These antibodies were distinguishable into two groups based on their incidence before vasectomy. The first group included antibodies to antigens in the acrosome with a diffuse distribution, the equatorial region, the postacrosomal region and the midpiece of the tail. Its incidence was 61% before vasectomy; increased to 73% at 2 months and 80% at 6-9 months after vasectomy. The second group included antibodies to the sperm nucleus, the tail and to discrete antigens over the acrosome. They were found rarely (3%) in patients before vasectomy; increased in incidence to 25% at 2 months and 55% at 6-9 months after vasectomy. Antisperm antibodies of both groups existed as IgG and IgM classes; an exception being antibodies to sperm nucleus which were almost exclusively IgG. Of the antibodies, 14% were found to fix complement in vitro. Other autoantibodies, including antinuclear, antimitochondrial and antismooth muscle antibodies, did not develop following vasectomy.


Humoral and cell-mediated immunity in experimental progressive thyroiditis in rabbits. Rabbits immunized over a long period of time with serial injections of aqueous preparations of either bovine thyroglobulin or chemically altered rabbit thyroglobulin develop progressive thyroiditis. As is short-term thyroiditis in rabbits and mice, this thyroiditis is characterized by lesions and cellular infiltration similar to that observed in Arthus reactions. Once the progressive thyroiditis is established, the rabbits respond readily to subsequent injections of native rabbit thyroglobulin. No significant reduction of lesions or circulating antibody is observed when injections of native rabbit thyroglobulin are substituted for the preparations used to induce the disease. Cell-mediated hypersensitivity to rabbit thyroglobulin, as evidenced by MIF activity, develops in rabbits after prolonged immunization with altered or cross-reacting thyroglobulin. It is suggested that this activity develops as a result of a loss in the unresponsive state in T lymphocytes. The data indicate that it is the persistence of circulating antibody to autologous thyroglobulin which sequesters autologous thyroglobulin from peripheral lymphoid tissue, and thus, results in the loss of the unresponsive state in lymphocytes of these tissues. It is suggested that similar events may be involved in the development of cell-mediated hypersensitivity in thyroiditis in humans.