Immunologic evidence for addition of oviductal components to the hamster zona pellucida. An indirect immunofluorescent procedure was used to determine whether zonae of unfertilized and fertilized eggs would bind antibodies specific for hamster reproductive tract antigens. Appropriate controls indicated that the fluorescent staining was probably due to antigen-antibody interactions. Antigens were not detected in zonae of ovarian eggs. However, components in zonae of unfertilized eggs collected from the oviduct at metestrus, mid-diestrus, and late diestrus bound antibodies specific for oviductal and uterine antigens. The antigens appeared to be concentrated in a peripheral band within the zona. An outer area of the zona around the eggs obtained from the uterus at proestrus also bound the antibodies, although the fluorescent staining was relatively faint. Antigens were observed in the peripheral area of zonae of fertilized eggs collected from oviducts on days 1 and 2 postcoitum. However, the antibody-binding component of the zona was noticeably reduced in embryos taken from the uterus on day 3 of pregnancy. The results are consistent with the idea that the zonae of both unfertilized and fertilized hamster eggs are composed of layers and that the outer region contains specific antigens which are dereived from the oviduct. Furthermore, loss or alteration of these components in the zona of an embryo (but not of an unfertilized egg of the same postovulation age) may reflect some chemical or structural change which is a prerequisite for zona shedding. Anti-uterus sera (unabsorbed and absorbed) did not interfere with zona removal by trypsin, indicating that the antigens detected in the present study are different from the receptor sites which bind anti-ovary antibodies and certain plant agglutinins (as reported by other workers).


[Stationary exitation according to A.A. Ukhtomskiĭ and its role in the mechanism of completion of temporary connections]. When recording potentials form the cortical surface, an increase in the processes of local excitation was shown to occur at the moment of revealing the first signs of the conditioned reflex: appearing of slow waves, increasing amplitude of evoked potentials, negative shift of the cortical steady potential.  When recording evoked potentials from deep cortical layers, successive stages of transition of the local excitation into a spread one which carries out the conditioning, were shown.  Stationary depolarization occurring in conditioning and revealed in gross forms as slow waves, increasing amplitude of evoked potentials, and negative shift of steady potential, at the cellular level is revealed as formation of neuron's polysensory acitvity and is usually followed by increase in the action potential (AP) duration in neurons of the cortical ends of analysers participating in conditioning.  Etimizol is known to improve memory processes in man and in animals (2).  When acting on the mollusk neurons, etimizol increases AP duration by 144 plus or minus 64 percent.  The data obtained suggest that the observed in conditioning increase of AP durration of neurons in the cortical representation of conditioned and unconditioned stimuli is associated with alterations in ion exchange and with trace processes in neurons, which apparently is related to the fine mechanisms underlying the retention in memory.


[Role of A.A Ukhtomskiĭ in the development of the problems of work, working capacity and fatigue (peculiarities and characteristics of these states in various age periods)]. The data of research which has been carried out for years, are summarized.  They are presented in a succession conforming to the A.A. Ukhtomsky views on the gradual enlargement of amplitude of muscles contraction, on capacity for work, and on fatigue.  In the early postnatal period the phenomenon of gradual enlargement of amplitude of muscles contraction does not yet exist.  It only appears after realizaiton of standing posture, when skeletal muscles transit from the type of tonic activity to the phasic--tetanic type.  This phenomenon is due to that the first contractions induce hyperrelaxation which leads to the subsequent higher amplitude of contraction.  The steady state of work capacity of neuromuscular system without any changes of amplitude, in the conditions of undisturbed circulation in the organism as a whole, can be maintained in all the periods of age for a long time without any signs of fatigue.  This phenomenon takes place during the intervals between the contraction activity, the anabolic process having ample time to be completed.  Such a form of work capacity is called stability.  If the anabolic processes are unable to be completed during the intervals between the contractions, their amplitude gradually decreases reflecting the beginning of fatigue.  The contractile activity which is going on during the fatigue state, is called endurance.


Insulin biosynthesis in isolated pancreatic islets of fetal and newborn rats. The effect of glucose and glucose plus glucagon on the incorporation of H-3-L-leucine into proinsulin and insulin was examined in isolated islets of twenty-one-day old fetal and five- and ten-day old newborn rats. Maximal stimulation of (pro-) insulin biosynthesis was achieved with 100 mg. per cent of glucose in isolated islets of twenty-one-day old fetal rats. No additional effect was observed with 300 mg. per cent of glucose. On the other hand, in islets of five- and ten-day old newborn rats the incorporation of H-3-L-leucine into proinsulin and insulin was gradually augmented by glucose up to concentrations of 300 mg. per cent. Addition of glucagon to the various glucose concentrations only enhanced the synthesis of insulin in ten-day old newborn islets, whereas it had no effect on the islets of the younger age groups. The results show a different pattern of insulin biosyntheses in fetal and newborn islets, which may be related to the varied plasma glucose concentrations of the perinatal period.


Pituitary responsiveness to luteinizing hormone-releasing hormone in insulin-dependent diabetes mellitus. Impaired fertility and menstrual disturbances are common in diabetes mellitus. In order to study the effect of diabetes on gonadotropin secretion by the pituitary gland, twenty premenopausal diabetic females and seven diabetic males were investigated using luteinizing hormone-releasing hormone (LH-RH). Although the gonadotropes responded to LH-RH in the patients studied, a relative impairment was apparent when compared to normal matched control groups. In the female diabetic patients there was no difference in the gonadotropin responses obtained in those with oligomenorrhea or amenorrhea when compared to those with normal menses. A significant inverse relationship was found between the maximum rise in plasma luteinizing hormone and the fasting plasma glucose. These findings suggest an influence of glucose metabolism on pitiutary gonadotropin function. However, the reduced gonadotropin response is unlikely to be the sole cause of the abnormal gynecologic function.


Sodium valproate in the treatment of intractable childhood epilepsy. Sodium valproate has been used, in combination with a variety of standard anticonvulsant drugs, to treat 24 children with intractable epilepsy.  A reduction of fit frequency greater than 90 percent was obtained in six patients, and a reduction of over 50 per cent in another six, with an improvement in alertness and school performance in the majority of cases.  Grand mal, myoclonic and petit mal epilepsy gave the best response, while infantile spasms responded least well.  No serious side-effects were encountered.


Correction of equinus in cerebral palsy by the Murphy procedure of tendo calcaneus advancement: a preliminary communication. The biomechanics of the Murphy procedure for correction of equinus in cerebral palsy are presented.  A series of 79 surgical procedures on 48 patients, with a follow-up period of from one to four years, is reviewed.  Correction of equinus was succeswful in 89-9 per cent of the procedures.  Using the Murphy procedure the following advantages over other methods of correction are anticipated: (1) no loss on 'push-off' in gait; (2) no loss of correction (recurrence) during longitudinal growth of the child, necessitating repeated surgery; and (3) no need for continual night bracing during the growth period.


Factors affecting the concentration of combustible gases in the colon during colonoscopy. The colonic concentrations of the combustible gases, H2 and CH4, were well below hazardous levels in 60 consecutive patients at the time of colonoscopy. Independent analysis of the effect of a low residue liquid diet, a 12-hr fast, and bowel cleansing on the pulmonary excretion of these gases suggest that the low colonic concentrations encountered were largely the result of our patient preparation procedure. Both ingestion of the liquid diet and fasting decreased the pulmonary excretion of H2 markedly but caused only a slight fall in CH4 excretion. Thus H2, but not CH4, production appears to depend on the delivery to the colonic bacteria of exogenous fermentable substrate. Bulk removal of bacteria from the colon resulted in about a 10-fold reduction in the excretion of both gases. The results of these studies do not support the need for routine CO2 insufflation prior to colonoscopic electrosurgical polypectomy.


Depression of antral and serum gastrin concentration by food deprivation in the rat. In rats fasted 4 days, immunoreactive gastrin concentrations decreased to one-third of the fed levels in antral tissue and to one-eight of the fed levels in serum. The number of antral cells that reacted with fluorescent antigastrin antiserum was also correspondingly decreased. After refeeding, serum gastrin returned to normal levels in 6 days, whereas antral gastrin concentration recovered after 9 days. Normal gastrin levels were maintained in rats fed a nutritious liquid diet over a 6-day period, whereas tissue and serum hormone concentrations decreased to low levels in rats placed on a high bulk non-nutritive diet over the same period. These results suggest that food in the gastrointestinal tract is necessary for the maintenance of normal serum and antral gastrin concentration in rats. The effect of food is most likely attributable to chemical constituents and not distention by bulk.


Significance of elevated liver alkaline phosphatase in serum. The serum alkaline phosphatase was fractionated by polyacrylamide gel electrophoresis in 317 patients with elevated serum alkaline phosphatase activity. In 253 patients the source of the elevation was the isoenzyme of presumed liver origin, band L. In 87 of these patients, there was either no obvious liver disease or the alkaline phosphatase elevation was inappropriately high. In 19 of the 87, liver disease was further excluded by liver biopsy or by laparotomy. Because of this, biochemical studies were done to verify the hepatic origin of band L. Band L and alkaline phosphatase extracted from human liver migrated together on polyacrylamide gel electrophoresis before and after digestion with Vibrio cholerae neuraminidase. They had identical pH optima, sedimentation coefficients, Michaelis constants, and rates of inactivation at 55.5 degrees C. They had different rates of inactivation in 3 M urea. Over-all, the data indicate that band L is of liver origin, and that elevation of the hepatic alkaline phosphatase isoenzyme may be a nonspecific finding in certain patients.


Evaluation of radiographic lucency or opaqueness of gallstones as a means of identifying cholesterol or pigment stones. Correlation of lucency or opaqueness with calcium and mineral. A major criterion for the selection of patients with gallstones for treatment with chenodeoxycholic acid is the radiographic demonstration of lucent gallstones. In this study, we sought to evaluate the degree of selectivity of that criterion for distinguishing patients with cholesterol stones from those with pigment stones and to define the determinants of stone lucency or opaqueness. Of 92 lucent stones, 14% were pigment stones; and of 18 of opaque stones, 33% were cholesterol. Thus, the criterion of stone lucency allows inclusion of a significant number of subjects (14%) with lucent pigment stones, which may account for about one-half of the reported 33% incidence of treatment failures with chenodeoxycholic acid. Conversely, of patients with opaque stones, the one-third with cholesterol stones would be excluded from chenodeoxycholic acid treatment. Calcium is the major metal of both types of stones. However, opaque stones contain 6 times more calcium than lucent stones, which accounts for the difference in radiographic appearance.


Recombination-deficient deletions in bacteriophage lambda and their interaction with chi mutations. We have isolated a new class of deletion mutants of phage lambda that extend from the prophage attachment site, att, into the gam and cIII genes. In this respect they are similar to certain of the lambda pbio transducing phage, but they differ in having a low burst size and in forming minute plaques. Lytically grown stocks of the deletions contain a variable proportion of phage that produce large plaques. These have been shown to carry an additional point mutation. Similar mutations, called chi, have been described by Lam et al. (1974), who showed that they result in a hot-spot for recombination produced by the host recombination system (Rec). We show that chi mutations can occurat several sites in the lambda genome and produce a Rec-dependent increase in the burst size of the one deletion tested.---In addition to reducing burst size, the one deletion tested reduces synthesis of DNA  and emdolysin but increases production of serum blocking protein. A chi mutation partially restores DNA synthesis and endolysin production and reduces serum blocking protein to normal levels. Our results are consistent with the hypothesis put forward by Lam et al., that chi enhances the frequency of Rec-promoted recombination, which provides the only pathway for production of maturable DNAin a red gam infection. The mechanism of the differential effect on protein production is, however, unclear.---Chi mutations are found to occur in DNA other than that of lambda. We show that, as has been suggested elsewhere (McMilin, Stahl and Stahy 1974), the lambda pbio transducing phages carry a chi mutation within the E. coli DNA substitution. A chi mutation also arose in a new substitution of unknown origin isolated in the course of this work.


A new method for mutant selection in Saccharomyces cerevisiae. A new method for the selection of auxotrophic, antibiotic- and temperature=sensitivemutants in Saccharomyces cerevisiae is reported. The technique is based upon the observation that certain fatty acid auxotrophs of yeast die when deprived of fatty acid only under conditions supporting growth. When macromolecular synthesis is blocked, the fattyacid-starved cells survive. By appropriate manipulation of a fatty acid-requiring strain enrichment as great as 75-fold was achieved for certain classes of auxotrophic mutants. An enrichment of approximately 100-fold is possible for some antibiotic-sensitive mutants. Selection for temperature-sensitive mutants, however, resulted in less than a 2-fold increase in the frequency of such mutants, probably because of the heterogeneity ofthis mutant category. It is likely that only that fraction of temperature-sensitivemutations which rapidly and reversibly blocks macromolecular synthesis is selected by this technique.


Recessive lethal amber suppressors in yeast. Recessive lethal amber suppressor mutations have been isolated in a diploid strain of Saccharomyces cerevisiae. Diploids carrying these suppressors upon sporulation yield asci with only two live spores, both lacking the suppressor. At least two classes of recessive lethal suppressors exist. Aneuploid strains carrying one wild type and one suppressor locus have been isolated and used in mapping studies; one suppressor maps on chromosome III, the other does not.


Evidence of preferential pairing of chromosomes at meiosis in aneuploid yeast. Meiotic pairing in homothallic S. cerevisiae was studied by tetrad analysis, using strains that were trisomic or tetrasomic for chromosomes. I. The disomic segregants of these strains produce tetrasomic spore colonies that can be distinguished by their phenotype. Results indicated the existence of preferential pairing and nonrandom assortment of chromosomes at meisosis I. The frequency of crossing over is apparently normal in at least some regions when nonpreferred pairing occurs.


Requirement for cell dispersion prior to selection of induced azaguanine-resistant colonies of Chinese hamster cells. With V79 Chinese hamster cell cultures treated with a mutagen, the maximum frequency of colonies resistant to 8-azaguanine (AZG) was attained when the cells were dispersed after a suitable expression time before adding the selection medium. V79-4 cells were exposed to 500 muM MMS, 7 muM AFAA, or 10 muM MNNG and allowed to multiply before being reseeded at 4 times 10-4 cells/60 mm dish and selected with 10 mu-g/ml AZG. Maximum frequencies of 4 times 10-5, 4 times 10-4, and 2.4 times 10-3 were obtained about 100, 130, and 200 hrs after exposure to MMS, AFAA, and MNNG, respectively. The maximum frequencies following MMS or MNNG treatments were about 10-fold greater than those obtained when induction and selection of AZG-resistant colonies were performed in the same culture dish. The reseeding of treated cells eliminated the possibility of metabolic cooperation within mosaic colonies of wild-type and mutant cells and achieved expression of the induced changes before intercolony crossfeeding reduced the frequency of resistant colonies. -  AZG-resistant colonies were selected in medium containing dialyzed fetal bovine serum, and the selection medium replacement were necessary for consistent achievement of background frequencies of resistant colonies near 10-6. Reconstruction experiments with AZG-resistant V79 lines showed that the efficiency of recovery of resistant cells in the selection medium was constant over a range of 0-20 colonies observed/dish. A mixed population of V79 and AZG-resistant cells was also correctly analyzed by the procedure used in mutagenesis studies.


Inositol-requiring mutants of Saccharomyces cerevisiae. Fifty-two inositol-requiring mutants of Saccharomyces cerevisiae were isolated following mutagenesis with ethyl methanesulfonate. Complementation and tetrad analysis revealed ten major complementation classes, representing ten independently segregating loci (designated ino1 through ino10) which recombined freely with their respective centromeres. Members of any given complementation class segregated as alleles of a single locus. Thirteen complementation subclasses were identified among thirty-six mutants which behaved as alleles of the ino1 locus. The complementation map for these mutants was circular. - Dramatic cell viability losses indicative of unbalanced growth were observed in liquid cultures of representative mutants under conditions of inositol starvation. Investigation of the timing, kinetics, and extent of cell death revealed that losses in cell viability in the range of 2-4 log orders could be prevented by the addition of inositol to the medium or by disruption of protein synthesis with cycloheximide. Mutants defective in nine of the ten loci identified in this study displayed these unusual characteristics. The results suggest an important physiological role for inositol that may be related to its cellular localization and function in membrane phospholipids. The possibility is discussed that inositol deficiency initiates the process of unbalanced growth leading to cell death through the loss of normal assembly, function, or integrity of biomembranes. - Part of this work has been reported in preliminary form (CULBERTSON and HENRY 1974).


Mating type and sporulation in yeast. I. Mutations which alter mating-type control over sporulation. In Saccharomyces cerevisiae, meiosis and spore formation as well as mating are controlled by mating-type genes. Diploids heterozygous for mating type (aalpha) can sporulate but cannot mate; homozygous aa and alpha-alpha diploids can mate, but cannot sporulate. From an alpha-alpha diploid parental strain, we have isolated mutants which have gained the ability to sporulate. Those mutants which continue to mate as alpha-alpha cells have been designated CSP (control of sporulation). Upon sporulation, CSP mutants yield asci containing 4alpha spores. The mutant gene which allows alpha-alpha cells to sporulate is unlinked to the mating-type locus and also acts to permit sporulation in aa diploid cells. Segregation data from crosses between mutant alpha-alpha and wild-type aa diploids and vice versa indicate (for all but one mutant) that the mutation which allows constitutive sporulation (CSP) is dominant over the wild-type allele. Some of the CSP mutants are temperature-sensitive, sporulating at 32 degrees, but not at 23 degrees. In addition to CSP mutants, our mutagenesis and screening procedure led to the isolation of mutants which sporulate by virtue of a change in the mating-type locus itself, resulting in loss of ability to mate.


Mating type and sporulation in yeast. II. Meiosis, recombination, and radiation sensitivity in an alpha-alpha diploid with altered sporulation control. In wild-type S. cerevisiae, diploid cells must be heterozygous at the mating-type locus in order to sporulate. In the preceding paper, we described a number of mutants (CSP mutants), isolated from nonsporulating aa and alpha-alpha parent strains, in which sporulation appeared to be uncoupled from control by mating type. The characterization of one of these mutants (CSP1) is now extended to other processes controlled by mating type. This mutant is indistinguishable from alpha-alpha cells and unlike aalpha cells for mating factor production and response, zygote formation, intragenic mitotic recombination, and for X-ray sensitivity. The mutant apparently undergoes a full round of DNA synthesis in sporulation medium, but with delayed kinetics. Only 20% of the cells complete sporulation. Among spores in completed asci, the frequency of both intra- and intergenic recombination is the same as it is for spores produced by aalpha cells. However, experiments in which cells were shifted from sporulation medium back to minimal growth medium gave a frequency of meiotic recombination between ade2 or leu2 heteroalleles only 25% to 29% as high for CSP1 alpha-alpha diploid or CSP1 aa disomic cells as for aalpha diploid or disomic cells. Because the latter result, indicating recombination defectiveness, measured recombinant production in the entire cell population, whereas the result indicating normal recombination sampled only completed spores, we infer that all meiotic recombination events occuring in the population of CSP1 alpha-alpha cells are concentrated in those few cells which complete sporulation. This high degree of correlation between meiotic recombination and the completion of meiosis and sporulation suggests that recombination may be required for proper meiotic chromosome segregation in yeast just as it appears to be in maize and in Drosophila.


Mutation of a heterothallic strain to homothallism. Upon mutagenesis, a heterothallic alpha-alpha diploit strain mutated to homothallism. The gene confering homothallism is nuclear, recessive, and unlinked to mating type. This gene is not allelic to the HO gene, which is responsible for previously described instances of homothallism in yeast. We have designated this new gene for homothallism as cmt (change of mating type).


Selection for a large genetic duplication in Salmonella typhimurium. Salmonella typhimurium strains containing a duplication of nearly a third of the genome have been isolated by a simple procedure involving selection for improved utilization of L-malate as sole carbon source. The duplication occurs at a very high spontaneous frequency. Strains containing the duplication can be isolated selectively on malate medium, or by a non-selective procedure involving Hfr conjugation. When strains containing the duplication are maintained on non-selective medium, the duplication is readily lost. Genetic evidence suggests that the duplication is chromosomal and tandem. The fact that the recA gene is included in the duplication has been used to obtain evidence that the recA1 marker is recessive to its wild-type allele. Unlike tandem duplications previously described in I. coli, the duplication described in this report appears to have unique endpoints.


A controlled, randomized trial of highly selective vagotomy versus selective vagotomy and pyloroplasty in the treatment of duodenal ulcer. The results of highly selective vagotomy without drainage and selective vagotomy with pyloroplasty for duodenal ulcer were compared in a randomized, controlled trial of a series of 100 patients. The frequency of dumping, diarrhoea, and epigastric fullness was significantly lower after highly selective (6, 6, and 8 percent) than after selective vagotomy (30, 20, and 28 percent) one year after the operations. Recurrent and persisting duodenal ulcers appearing from one to four years after the operations were significantly more frequent after highly selective (22 percent) than after selective vagotomy (8 percent). No significant relationships were found between recurrent ulceration and gastric acid secretion measurements after the two operations. The Hollander response was early positive in 28 percent and late positive in 30 percent of the patients subjected to highly selective vagotomy, while the corresponding figures after selective vagotomy were 26 and 32 percent. The overall clinical results of the two operations were not different according to the classification of Visick. Excluding the patients with recurrence resulted in significantly better clinical results after highly selective vagotomy.


Pancreatitis after renal transplantation. Pancreatitis is seldom seen as a severe complication of renal transplantation. In a review on 1321 renal transplants, 23 cases with 12 deaths are reported (Johnson and Nabseth, 1970). Single case reports may be added. In our departments pancreatitis has proved to be a fairly frequent complication. It developed in 10 (7 percent) of 147 patients with renal transplantation one week to seven and a half years after transplantation (patients with primary hyperparathyroidism excluded). Three of the eight acute cases had haemorrhagic pancreatitis, in two of them leading to death. Two patients had chronic calcifying pancreatitis. Pancreatitis was complicated in one case by abscess formation and in two by severe haemorrhage into a pseudo-cyst. In two patients the diagnosis was made at necropsy only and death was probably not related to the acute pancreatitis. The exact pathogenesis of pancreatitis after renal transplantation cannot be precisely assessed. Possible contributing factors are treatment with corticosteroids, azathioprin, and L-asparaginase, early hypercalcaemia after transplantation, surgery, infections of bacterial or viral origin, and unknown immunological processes.


[Effects of 5-(3-tert-butylamino-2-hydroxy) propoxy-3,4-dihydrocarbostyril hydrochloride(OPC-1085) on coronary circulation and myocardial metabolism]. Effects of a new beta-adrenergic blocking agent, 5-(3-tert-butylamino-2-hydroxy) propoxy-3,4-dihydrocarbostyril hydrochloride (OPC-1085), on the coronary circulation  and myocardial metabolism were investigated in anesthetized open-chest dogs and isolated perfused dog hearts. In anesthetized open-chest dogs, OPC-1085 antagonized the responses to isoproterenol of heart rate, Vmax, mean blood pressure, myocardial oxygen consumption, coronary blood flow and redox potential. The antagonistic potency of OPC-1085 was stronger than propranolol. OPC-1085 3 to 30 mug/kg caused appreciable decreases, but in doses of 100 to 1,000 mug/kg caused increases in heart rate and Vmax. The effect of OPC-1085 on max dp/dt was similar to that on Vmax. However, propranolol 3 to 3,000 mu-g/kg caused only decreases in heart rate, Vmax and max dp/dt. OPC-1085 3,10 mu-g/kg while propranolol 30, 100 mu-g/kg caused a fall in myocardial oxygen consumption and coronary blood flow. In isolated perfused hearts, intracoronary injection of OPC-1085 0.1 mg almost completely suppressed isoproterenol-induced augmentation of heart rate, myocardial contractile force and coronary blood flow and reduction of redox potential. OPC-1085 0.1 mg caused slight increases in heart rate, myocardial contractile force and myocardial oxygen consumption. It is concluded that OPC-1085 is a more potent beta-adrenergic blocking agent than propranolol and possesses a weak negative inotropic effect in doses of 3 to 30 mu-g/kg and a intrinsic sympathomimetic activity in doses of 100 to 1,000 mu-g/kg.


Protein mapping by combined isoelectric focusing and electrophoresis of mouse tissues. A novel approach to testing for induced point mutations in mammals. The protein-mapping method which combines isoelectric focusing in acrylamide gel and gel electrophoresis was previously used mainly for the separation of plant proteins and human serum proteins. We investigated with this technique soluble proteins of mouse tissues (whole embryos, the liver of fetal and adult mice, kidneys) and the proteins of mouse serum. The technique was tested under a number of different conditions to find those best for our purpose; they may represent some general improvements in the method. The protein patterns show high resolution and excellent reproducibility. About 275 spots were found for fetal liver, about 230 for whole embryos (day 14 p.c.) and about 100 for serum. The fact that a high number of protein spots can be evaluated by a single and comparatively simple experiment suggests that this method may be useful as an assay system for induced point mutations. The protein patterns demonstrated are compared and discgs of dominant lethal examinations after acute and subacute application of these three substances.


Experimental Escherichia coli O6 pyelonephritis in rabbits. Effect on O6 antibody quantity and avidity of prior immunization with E. coli O2 bacteria. Haematogenous pyelonephritis was induced in rabbits using Escherichia coli 06:K13:H1 bacteria and the amounts and avidities of antibodies to the 06 antigen were analysed by the ammonium sulphate precipitation technique of Farr. In a group of six animals preimmunized with E. coli 02:K2ab:H1, five developed pyelonephritis and one pyelitis, as determined by histological examination. All aminals showed a considerable antibody response to E. coli 06 antigen during the infection. The animal with pyelitis gave a slightly smaller response than the others. The antibody avidity showed a pronounced variation. In a second group of six rabbits not preimmunized, five animals developed pyelonephritis. The titres of antibodies against E. coli 06 antigen increased during the infection inall of the six animals. However, the increase was significantly smaller than for the animals preimmunized with E. coli 02:K2ab:H1 (P smaller than 0.01). The pattern of the antibody avidities in this group was also heterogenous. The results are consistent with previous findings that exposure to serologically heterologous E. coli bacteria can enhance the development of the homologous antibody titres. This could be of relevance for serological diagnostic work as well as in the determination of the protective capacity of the antibody.


Age-related decline in the antibody response to E. coli lipopolysaccharide in New Zealand Black mice. A thymus-independent immune function in ageing NZB and BALB/c mice was compared by measuring the antibody response to E. coli lipopolysaccharide (LPS). Since it was found that 10-13 month-old NZB mice was particularly sensitive to LPS, this antigen had to be detoxified by alkali treatment. The anti-LPS splenic plaque-forming cell (PFC) response of NZB mice decreases with age and was lower than that of BALB/c mice in all age groups studied. The response of 4- and 10-month-old NZB mice showed an irregular time course and a number of mice showed no response. The present results indicate that, besides an impairment of T-cell functions, an impairment on the B-cell level must also be considered in ageing NZB mice.


Leucoagglutination and cytotoxicity of the serum of infected mice and of extracts of Trichinella spiralis larvae and the capacity of infected mouse sera to prolong skin allografts. The sera of mice infected with the nematode Trichinella spiralis agglutinate and kill homologous lymphoid cells in vitro. The agglutinating activity is present in the sera of infected animals on the 7th day following inoculation with the parasite, rises to a maximum on the 30th day of the infection and then decreases. The leucoagglutinating titre of these sera is related neither to the level of the inoculating dose of parasites nor to the intensity of muscle infection. The agglutinating and leucotoxic activity of the sera of infected mice is probably due to the same factor(s), but target cells require sensitization before cytotoxicity becomes apparent. Skin allografts were prolonged in infected animals or in animals treated with the serum of infected animals. Saline extracts of T. spiralis larvae also have leucoagglutinating and leucotoxic activity.


The effect of lysozyme on the complement-dependent bactericidal action of different antibody classes. Preparations of rabbit, dog and sheep IgA, IgA and IgM were examined for their antibacterial effects using a complement-dependent bactericidal assay. IgM and IgG were efficient bactericidal antibodies in the presence of complement; IgA, however, contained negligible activity. Except for sheep IgG no enhancement of bactericidal activity was observed in the presence of added lysozyme.


Use of the defined antigen substrate spheres system as a model for analysing possible mechanisms of inhibition-blockade of anti-tumour lymphocytotoxicity. The defined antigen substrate spheres (DASS) system was employed for analysing reactions between solid state antibody or antigen and soluble immune complexes. Sepharose beads covalently coupled with ovalbumin were used to represent tumour cells and beads coupled with antibody against ovalbumin were used to represent anti-tumour lymphocytes; the ovalbumin and corresponding antibody simulated tumour-derived antigen and antibody to tumour respectively. Binding of soluble complexes to the beads was measured by fluorimetry and/or radiometry of fluorescein or 125-I-labelled ovalbumin or antibody. Antigen-antibody complexes in antibody excess bound less effectively to the antibody beads than antigen alone, but complexes in slight or moderate antigen excess bound more effectively. Complexes in antibody excess were most effective in the complex before levelling off and then decreased in extreme antibody excess. The model demonstration of augmentation by antibody of antigen binding to solid state antibody might by analogy reflect a mechanism of inhibition of lymphocyte cytotoxicity. Complexes in a wide range of antibody excess should also be effective in blocking lymphocytotoxicity at the target cell level.


In vitro synthesis of lysozyme by human and mouse tissues and leucocytes. Previous studies have shown that lysozyme can be detected in many body fluids, in extracts of tissues, and also in granulocytes, monocytes and macrophages. However, the sites of synthesis of lysozyme have not been defined. In the present report, the synthesis of lysozyme by tissues, and by defined cell populations cultured in vitro has been studied by detecting the incorporation of 14-C-labelled amino acids into lysozyme. This method detects only lysozyme newly synthesized during the incubation of the specimen and therefore shows unequivocally which tissues and cell types are capable of lysozyme synthesis. The validity of the method has been shown by parallel studies using an independent method to detect lysozyme production in vitro. In studies in humans and mice, lysozyme synthesis has been demonstrated in the mucosa of the respiratory and gastrointestinal tracts, and in lymphoid organs. In studies of defined cell populations, monocytes and macrophages (mononuclear phagocytes) have been shown to synthesize lysozyme. Granulocytes from peripheral blood contain lysozyme but do not synthesize it, and lymphocytes neither contain nor synthesize lysozyme. The present findings provide further evidence that lysozyme has an important role in the defence of the host against micro-organisms, and the findings suggest that lysozyme may reach its target by several routes. At an intracellular level it is delivered from lysosomes into the phagocytic granules of granulocytes and macrophages. Local synthesis of the mucous membranes contributes lysozyme to secretions. Synthesis and secretion by mononuclear phagocytes which reach a tissue in response to an inflammatory stimulus contribute lysozyme to the exudate, and the release of lysozyme from breakdown of granulocytes has the same effect.


Inhibition of T-cell differentiation by an antibody to a soluble thymic factor. An antiserum to chicken soluble thymic factor (STF) was prepared in rabbits and absorbed with chicken thymocytes until it reacted only with thymic reticuloepithelial cells. When injected into 6-day-old embryonated eggs, this antiserum caused a severe depletion of small lymphocytes in the cortical region of the thymus which was filled with large undifferentiated cells. In addition, reticuloepithelial cells displayed a marked disruption of cytoplasmic elements. The percentage of thymic lymphocytes bearing the T antigen was greatly reduced. It is believed that a factor responsible for differentiation of stem cells into T cells has been inhibited.


T cell-dependent mediator and B-cell clones. Supernatants were collected from keyhole limpet haemocyanin (KLH) primed spleen cells which had been incubated in tissue culture medium with their priming antigen KLH. These non-specific factor (NSF) containing supernatants were then tested in a microculture system for their ability to facilitate an anti-SRBC response of nu/nu or AT times BM spleen cells. It was concluded that:(a) NSF was able to engage a large number of the available pool of sheep erythrocyte (SRBC) specific B cells into an antibody response (b) this response was characterized by the development of clones expressing plaque-forming cells (PFC), the number of PFC produced within a clone being dependent upon the amount of NSF available in that culture; (c) NSF probably acted directly on B cells, and not via other accessory cell types.


Binding of aggregated human immunoglobulin to murine thymocytes and T cells through receptors for the Fc region. Normal thymus lymphocytes and T cells of mice have the ability to bind heat-aggregated IgG of human origin (aggHIgG), as shown by indirect immunofluorescence. At 4 degrees, the cells bind aggHIgG with an irregular speckled appearance; at 37 degrees, the aggregates are incorporated into the surface membrane, inducing rearrangement of the receptors and capping. At the point of maximum binding capacity, thymocytes show a fairly homogeneous fluorescence pattern, whereas T cells show a heterogeneous appearance. Aggregated pure human Fc fragments, but not Fab fragments, retained not only the binding capacity of the complete molecule but also the ability to induce cap formation on the surface of thymusceds, at 37 degrees.


Immune response in aged and young subjects, following administration of large doses of tuberculin. Different parameters of the immunologic response have been studied in young and aged subjects before, during and after administration of increasing doses of tuberculin purified protein derivative (PPD). In both age groups, a considerable increase in skin reaction was observed with the progression of antigen administration. This is particularly striking among old subjects, who before the treatment had either no reaction to tuberculin or a very weak one. Increased production of circulating antibodies to tuberculin was also consistently observed. The treatment resulted, especially in the old patients, in an increase in the lymphocyte sensitivity to phytohemagglutinin. The production of migration inhibitory factors was studied with lymphocytes from 6 treated subjects, in all cases this production was of the same degree as the one obtained from lymphocytes of PPD-positive untreated subjects. Lymphocyte cultures done during the treatment showed an absence of response in the presence of tuberculin, and in young patients only a slight increase in the spontaneous synthesis of desoxyribonucleic acid in the absence of tuberculin. These results demonstrate that in old people the depression of cellular immunity seems to be small. Adequate and prolonged antigenic stimuli are able to reactivate to approximately normal levels many features of this altered function.


Purified antibodies to collagen: an immunofluorescence study of their reaction with tissue collagen. Antibodies to soluble calf or rat skin collagen were purified by appropriate immunoadsorption and separated into three subfractions directed either to antigenic sites on unfolded alpha-chains (denatured collagen), to nonhelical sites, or to helical sites exposed on the triple helical molecule. In indirect immunofluorescence tests each of the antibody solutions reacted with collagen of skin and kidney tissue, although the latter two antibody solutions appeared to be more active. Distinct activity was also observed with antibodies to the N-terminal antigenic determinant of rat skin collagen alpha2-chain, a structure usually involved in cross-linking. Indirect immunofluorescence tests with anti-collagen sera on sections of skin resulted in the staining of the whole dermis, while anti-procollagen sera revealed binding only to the uppermost subepithelial layer of the dermis (stratum papillare). On kidney sections only the interstitial connective tissue reacted with purified anti-collagen or anti-procollagen sera. Both skin and glomerular basement membranes remained unstained with either kind of purified antibodies. However, antisera not subjected to immunoadsorption do react with the glomerular basement membrane. Antibodies to noncollagenous contaminants are considered to be responsible for this finding which emphasizes the necessity to use purified antibodies exclusively for this type of immunofluorescence analysis.


Changes for Bicêtre and its elderly residents: the paradox of progress. Recent changes at Bicêtre, the historic French institution, exemplify an old paradox in the care of the elderly: improvements which benefit part of a society can mean more misfortunes for others.  The effect of relocation of its elderly residents to make way for renovations poses important questions of priorities in societal values.


Spread of Trichophyton mentagrophytes var. gran. infection to man. The occurrence of T. mentagrophytes var. gran. infection of small mammals in their natural habitat was studied. The most common host of the exoanthropic mammals was Clethrionomys glareolus (bank vole); the next most common hosts were hemisynanthropes Sorex araneus (common shrew) and eusynathropes Mus musculus (common mouse). Occurrence of infection increased during the winter months. Because small mammals gather in farm outbuilding during cold weather, man is more likely to be exposed to infection from them during this time. Infection from T. mentagrophytes var. gan. is more likely to occur among agricultural workers than among others. The type of work influences the location of skin infection.


Clinical evaluation of desoximetasone in treatment of dermatoses and psoriasis. A comparative clinical evaluation of desoximetasone, a corticosterone derivative by a double blind study showed that desoximetasone has potent topical activity as effective as betamethasone valerate and probably more effective than triamcinolone acetonide.


Studies of the mechanisms of radiosensitization of bacterial and mammalian cells by Diamide. Diamide sensitizes bacterial and mammalian cells to radiation by at least two mechanisms. Sensitization of V79-GL1 Chinese Hamster cells is due mainly to a reduction of a reduction of the survival-curve shoulder, is observed both in oxygen and in hypoxia,and is additive to the sensitization of hypoxic cells by some nitroimidazoles. In contrast, sensitization of the radioresistant organism, Micrococcus sodonensis, which has apronounced shoulder, is entirely dose-modifying. In a rapid-mix study using Serratia marcescens, two mechanisms of sensitization have been identified. After 4 to 40 msec preirradiation contact with Daimide, the enhancement ratio is constant at 1.6; after 10min contact, it is 2.5. With V79-GL1 cells, no sensitization occurs in the short-time-range; likewise non is seen when Diamide is added to bacteria or mammalian cells 4 msec after irradiation. It is concluded that Diamide sensitizes by at least two mechanisms, one of which resembles that of the electron-affinic type.


Postoperative monitoring following critical trauma. This review has focused on several aspects of monitoring post-injury patients. A few of the more commonly used monitoring devices have been discussed to emphasize that their use carries some risk and, as such, proper indications, based on the benefit to be achieved, must be available before they are used. Guidelines for the safe use of these devices have been outlined. The postoperative injury patient is at risk for developing certain complications as a result of the magnitude of injury. This review has dealt with respiratory insufficiency, renal insufficiency, and stress ulceration. Guidelines have been presented to aid in the prevention, early diagnosis, and management of these problems.


Bone induction in implants of decalcified bone and dentine. The fate of decalcified bone and dentine implanted in muscle and beneath the kidney capsule has been studied in young rats. Quantitatively speaking there was a great deal of variation, but in general the implants became surrounded and invaded by young vascular connective tissue; then tunnels were eroded and cavities enlarged by multi-nucleated giant cells; then the matrix around erosion chambers became recalcified; and finally new bone was induced on the eroded recalcified surfaces. Erosion was much more extensive, and bone was much more readily induced in the intramuscular than in the subcapsular implants. It is concluded that the presence of an eroded, recalcified surface is a pre-requisite for bone induction under these conditions.


Tropomyosin antibody: the specific localization of tropomyosin in nonmuscle cells. An antibody against purified chicken skeletal muscle tropomyosin is used in indirect immunofluorescence to visualize the localization of tropomyosin in a variety of nonmuscle cells. The antibody produces a fluorescent pattern which is very similar to that obtained with an actin-specific antibody. This pattern is composed of fluorescent fibers which are shown to be coincident with the fibers seen with phase-contrast optics. High resolution epifluorescent microscopy reveals that fibers stained with the actin antibody show a continuous fluorescence, while fibers reacted with the tropomyosin antibody show a periodic fluorescence. Measurements indicate that the lengths of the fluorescent segments are variable with an average of 1.2 mum while the spacing between segments is approximately 0.4 mum.


A new miniature hydrostatic pressure chamber for microscopy. Strain-free optical glass windows facilitate phase-contrast and polarized-light microscopy of living cells. Optional fixture permits simultaneous control of pressure and temperature. This paper describes the development of a miniature, temperature-controlled, stainless steel pressure chamber which uses strain-free optical glass for windows. It is directly adaptable to standard phase-contrast and polarized-light microscopes and requires a minimum amount of equipment to generate and measure pressure. Birefringence retardation (BR) og 0.1 nm up to 3,000 psi, 0.4 nm up to 5,000 psi and 1.0 nm up to 10,000 psi can be detected over a 0.75-mm central field with two strain-free Leitz 20 times UM objectives, one used as a condenser. In phase-contrast studies a Nikon DML 40 times phase objective and Zeiss model IS long working-distance phase condenser were used, with little deterioration of image quality or contrast at pressures as high as 12,000 psi. The actual design process required a synthesis of various criteria which may be categorized under four main areas of consideration: (a) specimen physiology; (b) constraints imposed by available optical equipment and standard microscope systems; (c) mechanical strength and methods for generating pressure; and (d) optical requirements of the chamber windows. Procedures for using the chambers, as well as methods for shifting and controlling the temperature within the chamber, are included.


Production of a comprehensive research directory from multiple secondary sources. Printed, automated, and manually maintained sources of information about cancer research and control activities were identified. Data were consolidated into an automated file to characterize the organizations active in such work and to identify individual projects and scientists. A computer-formatted directory was produced, with access via geographic location, personal name, organizational name, and keyword.


Chromatographic hydrophobic parameters in correlation analysis of structure-activity relationships. The use of RM and delta RM parameters as indices of hydrophobicity for inclusion in quantitative structure activity relationships has been studied. The relationship between these parameters and other free-energy related parameters is illustrated theoretically and experimentally. It is suggested that the chromatographically obtained parameters could find a wider applicability in structure-activity relationships, and that their use would result in improved correlation of data.


Chromatographic analysis of fungicides.

Chromatographic analysis of fungicides. The separation and analysis of a wide range of fungicides, by gas, liquid, column, paper, and thin-layer chromatography have been reviewed. Major attention has been given to methods for the identification and quantitation of individual and multiresidues of fungicides in environmental and agricultural samples.


Myotonia dystrophica: Studies on gonadal function using luteinizing hormone-releasing hormone (LRH). Eight men and four women with myotonia dystrophica were investigated. The gonadotropin responses to 100 mug luteinizing hormone-releasing hormone (LRH) were studied, and serum testosterone assayed. In the men with testicular atrophy a high basal follicle stimulating hormone was found with an exaggerated response to LRH. Although the mean basal luteinizing hormone was not significantly elevated, there was an excessive response to LRH, which was possibly conditioned by the lower mean testoerone levels in this group. This study provides biochemical indices of both the primary nature of the testicular damage and the disproportionately greater involvement of the seminiferous tubules. The pituitary-gonadal axis was normal in the women.


Human pituitary gonadotropin index. I. Standardized LRH test criteria for evaluation of functional amenorrhea. LRH test were carried out by giving 339 amenorrheic women and 74 normally menstruating volunteers an intravenous injection of 25 mug LRH (Hoe 471). Plasma LH and FSH were measured by RIA in two laboratories (Tuebingen and Ulm) using two standard reference preparations: LER 907 and 2nd IRP-HMG. The average conversion factors between the two standard preparations were calculated at 5.0 for LH and 25.0 for FSH. Furthermore, the estradiol-17beta levels were measured in 139 out of the 339 patients immediately before and 60 minutes after LRH injection. Taking the episodic and cyclic plasma gonadotropin fluctuations into consideration a shorthand system classifying the gonadotropin baseline (BI-BIV) and LH responses to 25 mug LRH (R0-R2) has been established and is referred to as Human Pituitary Gonadotropin Index (HPGI). It is possible to achieve reproducible gonadotropin results in two different laboratories using two different standard reference preparations. Two separate, randomly selected groups of amenorrheic women were found to have the same percent distribution of the HPGI. A correlation coefficient of r equal 0.67 between basal and LRH stimulated plasma LH levels does not sufficiently characterize the individual LH response behavior. A significant increase of plasma LH and FSH within the test period (60') reveals that the iv administration of 25 mug LRH represents an adequate dose for the LRH test in women. The HPGI which characterizes the functional state of gonadostat, may become a useful diagnostic index for evaluating women with anovulatory disease before, during, and after therapy.


Experimental gingivitis in young and elderly individuals. The development of experimental gingivitis was studied in young elderly humans during a 21-d period of oral hygiene abstention. The state of the gingiva was assessed by the Gingival Index and by measurements of the amount of gingival exudate on filter paper strips placed at the entrance of the gingival sulcus of the lower lateral incisors and cuspids. Soft deposits were assessed by the Plaque Index and by differential counts of microorganisms in gram stained smears od ento-gingival plaque. At the end of the plaque growth period, the patients were given a thorough dental prophylaxis. Gingival condition and plaque were assessed at regular intervals during a subsequent period of controlled oral hygiene. The development of gingivitis during the oral hygiene abstention period was more rapid and more severe in old than in young individuals. Plaque accumulation was greater in the older persons. A definite difference in plaque consistency was alos observed. However, microscopic counts of various types of microorganisms did not reveal any differences throughout the period of plaque accumulation. When active oral hygiene was reinstituted, the state of the gingiva rapidly returned to pre-experimental levels in both groups. The findings of this study indicate that with age ther is an altered host response to the microorganisms of the plaque.


Inhibitory effect of benzethonium and zinc chloride mouthrinses on human dental plaque and gingivitis. In a double-blind study the plaque- and gingivitis-inhibiting capacities of three aqueous mouthrinse solutions of benzethonium chloride, zinc chloride, and benzethonium chloride combined with zinc chloride were compared to those of a placebo mouthrinse. Forth-three adults with clean teeth and healthy gingiva were randomly assigned to the treatment and control groups. All participants received an initial prophylaxis. Assessments of plaque and gingivitis were made before and after a 10-day test period. During this time the subjects suspended oral hygiene procedures and rinsed daily with 20 ml of their respective rinse solutions. According to the measurements made before and after the test period, the mouthrinse containing benzethonium chloride produced 42 to 42.9% less plaque than any of the other three formulations. The differences were statistically significant ( P smaller than 0.05). Gingivitis scores of the four groups did not differ significantly. These findings indicate that benzethonium chloride is a moderately effective antiplaque agent.


Effect of professional tooth cleaning on healing after periodontal surgery. The investigation was undertaken to find out whether favourable conditions for healing after periodontal surgery would develop in patients whose oral hygiene was professionally maintained at a high standard. The study was performed on 20 patients with advanced periodontal disease. Following an initial examination, comprising plaque index and gingival index scoring, measurement of pocket depths and loss of attachment, the patients were randomly distributed between a test and a control group. The patients first received professional cleaning of the teeth once every 2 weeks. The patients of the control group were recalled for scaling of the teeth once every 6 months. All patients were reexamined after 6, 12 and 24 months. It was found that the control patients were unable to maintain a high standard of oral hygiene with the result that the treatment of the periodontal disease failed. The patients of the test group maintained a high standard of oral hygiene, and the treatment of the periodontal disease was, therefore, successful.


A comparative clinical study of three periodontal surgical techniques. A comparative 4-month trial was performed after initial preparation. The procedures used were curettage, replaced flap and apically repositioned flap procedures with osseous recontouring respectively in a split-mouth technique in 17 male and 23 female patients, 22 to 65 years old. Changes were assessed with plaque and gingival indices, and measurements of pocket depths, attachment levels and tissue contours. All procedures reduced pocket depths. Apically repositioned flaps were the most successful, reducing overall mean pockets from 3.5 to 1.9 mm. Changes in attachment levels post-operatively showed only a small degree of variation among the three surgical procedures, and would not affect the choice of the apically repositioned flap as the most effective method for pocket reduction. A marked improvement in tissue contour was obtained in those areas treated by apically repositioned flap procedures and osseous recontouring. Despite failure to improve tissue contour, replaced flap procedures showed an equal degree of success in maintaining plaque control when compared to apically repositioned flaps, as evidenced by static plaque indices. Replaced flap procedures were also the only group to show improved gingival health over the 4-month post-operative period with lower gingival indexes. It would seem that good healing, followed by a high standard of oral hygiene, may overcome the presence of gingival and marginal osseous deformities.


Scanning electron microscopy of the subarachnoid space in the dog. III. Cranial levels. Young dogs were anesthetized by intrathoracic injection of sodium pentobarbital and perfused with buffered aldehydes. Lining tissue samples from the cranial subarachnoid space were prepared for scanning electron microscopy (SEM) by postfixing in buffered OSO4. Samples were then dehydrated, dried in a Critical Point Drying System, and coated with carbon and palladium-gold. Specimens were viewed in a Cambridge S4 scanning electron microscope. After thorough scanning, selected samples were routinely prepared for transmission electron microscopy (TEM) and viewed in a Philips EM-200 transmission electron microscope. This study depicts the surface morphology of the meningeal linings of the cranial subarachnoid space. The cranial pia mater possesses natural gaps or fenestrations between cells. SEM reveals a more complex morphology of arachnoid trabeculae than previously interpreted from light and transmission electron microscopy. Many free cells are observed on the meningeal linings of the subarachnoid space. The present study establishes that these free cells are macrophages by means of definitive TEM correlates. Microvillous-like processes extending between macrophages and the pial surface are present. The frequency and the nature of these thin processes suggest the possibility of a plasmalemma-mediated system of communication.


Results of a one-year trial of tolmetin in patients with rheumatoid arthritis. The safety and effectiveness of tolmetin was assessed in ten patients with active rheumatoid arthritis for 12 months. All patients met the requirements of the American Rheumatism Association's diagnostic criteria for definite rheumatoid arthritis. None of the patients had received gold, antimalarials, or glucocorticoids. All had been stabilized, more or less, with aspirin. Monthly measurements were made of grip strength, duration of morning stiffness, time to the onset of fatigue, Westergren sedimentation rates, the number of swollen joints, number of hot joints, the time to walk 50 feet, and the circumference of proximal interphalangeal (PIP) joints (measured in millimeters with a plastic-loop arthrocircameter). The initial dosage of tolmetin in all patients was 600 mg a day, given orally in three divided doses. Upon completion of the study, the average daily oral dosage was 1400 mg tolmetin. No other antiinflammatory drugs were allowed. The results indicate that tolmetin affords symptomatic relief of rheumatoid arthritis; its use results in global improvement in 80 per cent of the patients, and a net improvement in one or more of the parameters used to assess disease activity in 100 per cent of the patients; it seems to be sufficiently safe to administer to patients over a long period of time; there was no evidence of severe toxicity of any kind; gastrointestinal side effects were uncommon, and epigastric distress, in particular, was not observed.


Cements for permanent luting: a summarizing review. Three cement systems are favored for permanent luting of cast restorations. These include zinc phosphate, reinforced zinc oxide-eugenol, and polycarboxylate cements. Although others have been used in the past and new luting media are anticipated for the future, the status of currently used cement for luting is based mainly on the results of laboratory research and on clinical experience. Zinc phosphate cement, with an impressive 100-year history, currently holds the advantage. Future comprehensive clinical studies, correlated with results of physical and biological testing, may eventually direct the clinican's preference to another, newer material. It is hoped that properties and strength values that are clinically significant also will be identified, so that laboratory tests can be more predictive of clinical success.


Abnormalities in cell-mediated immunity in patients with Cryptococcus neoformans infection. Cryptococcin, streptokinase-streptodornase (SK-SD), mumps, and purified protein derivative (PPD) were used for skin testing and, with whole killed Cryptococcus neoformans, were used in migration inhibition and lymphocyte transformation assays of control subjects and patients with past or present disseminated C. neoformans infection. Cryptococcin was found to differentiate control subjects grouped by known Cryptococcus exposure. Cryptococcin and C. neoformans were effective in stimulating leukocyte migration inhibition and lymphocyte transformation in the cryptococcin skin test-positive control subjects. Fifteen apparently normal patients who had been cured of cryptococcosis were found, as a group, to have impaired responsiveness to skin testing with cryptococcin and mumps, minimal leukocyte migration inhibition when stimulated with cryptococcin or C. neoformans, but normal group responses to cryptococcin in  Cryptococcus-induced lymphocyte transformation. Six patients with known predisposing conditions to disseminated infection (sarcoid, lymphoma, leukemia, steroid therapy) had markedly diminished responses to most skin tests and in vitro assays. It is suggested that the apparently normal individual who develops disseminated cryptococcal infection has subtle defects in cellular immunity that may have antedated and predisposed to infection.


Primary interaction between antibody and components of Alternaria. II. Antibodies in sera from normal, allergic, and immunoglobulin-deficient children. Antibodies in sera from normal, allergic, and immunoglobulin-deficient children were studied for binding to radiolabeled components of Alternaria tenuis. Significant binding levels were found in 103 of 105 sera from normal children. The levels were age-dependent, rising from a low point in the 7- to 12-month age group to adult levels by the age of 8 years. Levels of binding to two antigens, a culture filtrate derivative (125I-CLF) and a mycelial derivative (125-I-HS), were similar. Sera from asthmatic children with strong immediate skin test reactions to Alternaria extracts bound significantly higher levels of 125-I-CLF than did sera from allergic children with negative skin tests or from control children. Binding levels in sera from children with hypogammaglobulinemia were significantly less than binding levels in sera from normal children in any age group. Sera from children with selective IgA  deficiency bound 125-I-CLF at normal levels. The almost universal occurrence of anti-Alternaria antibodies in children was partly explained by the finding of partial cross-reactivity and/or shared antigens among several fungal species, including A. tenuis, Stemphylium sp., Curvularia sp., and Aspergillus fumigatus. The biological significance of these antibodies is not clear, but the procedures described lend themselves to further investigations.


Absence of hepatitis B antigens from feces and sewage as a result of enzymatic destruction. Hepatitis B surface antigen (HBs Ag) was not detected in sewage by means of a sensitive affinity chromatography method combined with radioimmunoassay. The antigen was also absent from the feces and urine of 23 patients with HBs Ag antigenemia; this observation indicates that HBs Ag is rarely discharged into sewage. The absence of HBs Ag from feces is ascribed to antagonists of an enzymatic nature or to carboxypeptidase A, which destroys the antigen. Antagonists with similar effects were produced by three species of Pseudomonas but were not produced by various other bacteria. HBs Ag was also destroyed by two subtilisin enzymes. When hepatitis B sera were incubated with these enzymes or with the antagonists, small spherical particles, tubules, and the coats of Dane particles disappeared first, and Dane cores disappeared later. Although sewage or activated sludge did not affect the stability of HBs Ag, the results indicate that even Dane cores are not excreted in feces and that sewage plays a negligible role in the spread of HBs Ag, Dane cores, and viral hepatitis type B.


Light and electron microscopic study of Chlamydia trachomatis infection of the uterine cervix. Cervical biopsy specimens from two patients with infections caused by Chlamydia trachomatis (TRIC agent) were studied by means of light and electron microscopy. Intraepithelial vesicles containing C. trachomatis were present in the cervical mucosa. These intramucosal vesicles were readily recognized in paraffin sections by means of light microscopy. The developmental stages of C. trachomatis, which have previously been seen in tissue cell culture, chicken embryo, and conjunctival infections, were also apparent in cervical infection.


Superiority of MacConkey's agar over salmonella-shigella agar for isolation of Shigella dysenteriae type 1. The efficiency of MacConkey's agar in the isolation of various types of Shigella was compared with that of salmonella-shigella (SS) agar during an extensive 18-month outbreak of disease caused by Shigella. In all, 1,580 isolates of Shigella were obtained from 12,307 rectal swab and fecal samples of patients with diarrhea and their contacts by direct plating onto MacConkey's and SS agars. Shigella dysenteriae type 1 and Shigella flexneri constituted 55 percent and 33 percent of all isolates, respectively, with a smaller number of Shigella boydii and Shigella sonnei. MacConkey's agar was superior to SS agar in the detection of S. dysenteriae type 1; 83 percent of the isolates were detectable on MacConkey's agar, compared with 40 percent on SS agar. In contrast, 84 percent of S. flexneri isolates were detectable on SS agar, compared with 51 percent on MacConkey's agar alone. These findings confirm that, for the culture of fecal specimens thought to contain S. dysenteriae type 1, one of the media used should be noninhibitory. Shigella was isolated at a consistently and significantly higher rate from fecal samples than from rectal swabs.


Vitamin B6-dependent Streptococcus mitior (mitis) isolated from patients with systemic infections. Strains of nutritional-variant viridans Streptococcus were isolated from two patients with bacterial endocarditis and from one patient with a pancreatic abscess. All three strains grew as satellite colonies of other bacteria but did not grow as pure cultures in media lacking sufficient thiol compounds. For one-half maximal growth these organisms required the addition of one of the active forms of the B6 vitamin, pyridoxal with HCl (0.16-1.45 mug/ml) or pyridoxamine dihydrochloride (0.67-2.0 mug/ml), or addition of l-cysteine (0.235-0.425 mg/ml) to routine bacteriological media. The active forms of vitamin B6 are essential coenzymes in the synthesis of l-cysteine and of other thiol compounds. Use of media supplemented with 0.001 percent pyridoxal with HCL led to identification of the strains as Streptococcus mitior (mitis). Incorporation of pyridoxal with HCl, pyridoxamine dihydrochloride, or l-cysteine into media is recommended for the isolation and identification of vitamin B6-dependent viridans streptococci.


Combined type-specific antisera in the identification of Mycoplasma hominis. In the usual serologic test for Mycoplasma hominis, disks containing antibody to a prototype strain are applied to an agar plate that has been inoculated with the microorganism to be tested, and inhibition of growth is studied. The recent demonstration that M. hominis is not serologically homogeneous but consists of at least seven serotypes suggested the need for reexamination of the specificity of the disk test. The inhibition of growth of M. hominis on solid medium by disks containing seven antisera to the seven prototype strains was studied. Antisera to strains 1, 2, and 3 significantly inhibited all seven prototype strains, but the other four antisera did not inhibit the growth of all heterologous strains. The individual antisera 1, 2, and 3 did not inhibit all of a group of 19 recent genital isolates of M. hominis. However, when the three antisera were combined, they inhibited all 19 isolates as much as or more than any of the three alone. The growth of the prototype strain 5 (PG 21) and of some recent clinical isolates was not completely prevented by the homologous or reference antisera, but the growth of all was partially inhibited.


Serum dopamine-beta-hydroxylase in familial dysautonomia. The mean value of serum dopamine-beta-hydroxylase (D beta H) in patients with familial dysautonomia, 1 to 5 years of age, does not differ significantly from control children of the same age (24.0 plus or minus 21.06 S.D. as compared to 34.0 plus or minus 33.12). Among patients 6 years of age and over, the mean value was slightly but significantly lower than in control subjects (62.7 plus or minus 49.61, as compared to control values of 86.1 plus or minus 54.31 p less than 0.025). However, the determination of serum D beta H does not contribute to the diagnosis of familial dysautonomia because well over half the children have levels within 1 S.D. of the mean levels of the control subjects. There is no correlation with clinical symptomatology. The disease process may tend to depress the level of serum D beta H but the effect is neither consistent nor decisive.


Prevention by glucocorticoids of disseminated intravascular coagulation induced by endotoxin: mechanisms. The therapeutic efficiency of two glucocorticoids (hydrocortisone and dexamethasone) on endotoxin-induced intravascular coagulation was investigated in the rat. Coagulation and platelet aggregation studies were performed and plaminogen was assayed. Our results indicate that pretreatment of the animals with large doses of these steroids within a few hours prior to endotoxin totally prevents the consumption in Hageman factor, measurable contact product activity, platelets, fibrinogen, plasminogen, and the loss in platelet aggregability and serotonin. In addition to this, the hypercoagulable state  consecutive to endotoxin, characterized here by shortenings in the partial thromboplastin and recalcification times and by an increase in the availability of platelet procoagulant activity, was also totally prevented by the steroid pretreatment. On the other hand, it is shown that these glucocorticoids do not interfere in the normal rat with platelet aggregation (tested with thrombin,adenosine diphosphate, and collagen), but with the availability of platelet procoagulant activity. This last phenomenon, in addition to that of an interference in vivo with the mechanism of activation of Hageman factor, are believed to be responsible for prevention by glucocorticoids of endotoxin-induced disseminated intravascular coagulation.


Abnormal renal acidification in alcoholic liver disease. After ingestion of 150 mEq. of calcium chloride (CaCl-2), urinary acidification was studied for 6 hours in 22 normokalemic patients with alcoholic liver disease (L) of varying severity, and in 7 control (C) subjects during 10 studies. The degree of the induced systemic acidosis was similar in all groups. Nine L patients were unable to normally lower urine pH below 5.25 (L-I) and these were compared with the 13 L patients achieving lower pH (L-II) and with control subjects. This defect was consistently reproduced. Titratable acid excretion was less in L-I than in the other groups. The percentage contribution of ammonium to maximal net acid excretion was significantly higher in L-I and L-II than in C. No L-I patient had spontaneous metabolic acidosis, nor was there evidence of encephalopathy or of proximal tubular dysfunction. Sodium excretion was significantly lower in L-I than in either L-II or C. Sodium sulfate and sodium phosphate infused after acid-loading rapidly reduced urine pH into the appropriately acidic range in L-I patients with alcoholic liver disease by means of a simple, safe, and short acid-loading test. Although the mechanism of this renal tubular acidfying defect remains unknown a low distal delivery of sodium by limiting the transtubular potential difference may have been partially responsible.


Mutations in Salmonella typhimurium conferring resistance to Felix O phage without loss of smooth character. Several mutants obtained from smooth Salmonella typhimurium strains by selection for resistance to Felix O (FO) phage [whose receptor site includes the N-acetylglucosamine branch of the lipopolysaccharide (LPS) core] were smooth in cultural properties, antigenic character and phage sensitivity pattern (except for their FO resistance). However, the affected genes of several such 'FOR' (FO-resistant) mutants were shown by transduction of map in the short cysE-pyrE segment, which includes nearly all known rfa genes responsible for synthesis of LPS core. All of seven FOR mutants differed from their parents, and resembled rfa mutants with defects in the deeper part of the LPS core, by increased sensitivity to various antibiotics. One FOR mutant was non-virulent (LD50 greater than 10-7, compared with smaller than 100 for its parent); LT7 derivatives given this FOR gene by co-transduction with cysE+ were likewise non-virulent. It is inferred that FOR mutations affect the assembly of the inner part of the LPS core, perhaps causing incomplete blocks in glycosyl transferase reactions.


Transmissible substrate-utilizing ability in enterobacteria. Three of 152 strains of Escherichia coli transmitted their ability to utilize sucrose (Sac+) to other strains by conjugation. The transfer factor of one of them and of a Sac+ Salmonella thompson strain was thermosensitive. The raffinose-utilizing ability of 27 of 163 E. coli strains was also transmissible. Transmissible raffinose-utilizing ability was a feature of porcine enterpathogenic strains possessing the K88 antigen. The determinants controlling raffinose utilization (Raf) and K88 antigen production were commonly transmitted together from these strains; so also was the determinant controlling enterotoxin production, but to a lesser extent. It was not possible to transfer lactose-utilizing ability from 320 strains of E. coli, salicin-utilizing ability from 12 strains of E. coli or dulcitol-utilizing ability from 99 strains of E. coli and 88 strains of salmonellae. Sucrose- and raffinose-utilizing ability were transmitted separately to several Salmonella sp., including Salm. typhi, to Shigella flexneri and Sh. sonnei and to a variety of strains of E. coli. A strain of Salm. typhimurium in which Sac had been established and a strain of E. coli in which Raf had been established survived less well in the alimentary tract of chickens than their Sac minus or Raf minus parent strains.


The effect of culture age, chloramphenicol and B6 inhibitors on intra- and extracellular keto and amino acids of Escherichia coli B. Keto acids and free amino acids were assayed in the cells and the medium of Escherichia coli B growing in the presence of chloramphenicol, cycloserines, aminooxyacetate, and limiting nitrogen source. Under these growth-limiting conditions the cells accumulated ketoglutarate and 'ketovaline' but no other keto acids. In all experiments only ketoglutarate, pyruvate, and 'ketovaline' were found in the medium. Amino acids are released into the medium in the early phases of growth and the composition of the extracellular amino acids is similar to that of the amino acid pool. The concentrations of free amino acids were 10-3-10-4 times higher in the cell than in the medium. The internal pool composition is fixed under all growth-limiting conditions. In the presence of the drugs the cells release amino acids into the medium.


Factors affecting the changes in amphotericin sensitivity of Candida albicans during growth. The sensitivity of Candida albicans, grown in batch culture at 37 degrees C, to amphotericin methyl ester (AME), judged by the concentration of AME required to induce a standard rate of leakage of K+ from suspensions of organisms, decreased with the time of growth. Organisms in exponential growth were sensitive to 0-I to 0-2 mug AME/ml while organisms in the stationary phase were resistant to 4 to 60 mug AME/ml, depending on the initial concentration of glucose in the medium and the length of time for which incubation had been continued. When the initial concentration of glucose was low (0-I%, w/v), the AME resistance rose during the early stationary phase and then, after 40 h incubation at 37 degrees C, decrease again. Sphaeroplasts were prepared from organisms at different phases of growth and did not show these changes in AME sensitivity, but remained highly sensitive for growth up to 40 h. Sphaeroplasts were prepared by treating suspensions of organisms with mercaptoethanol and then digesting with Streptomyces enzyme preparation. Addition of the material extracted by the digestion to suspensions of exponential-phase organisms or sphaeroplasts increased their AME resistance. Fractionation of the digest showed that the antagonistic material was contained in the neutral lipid fraction. Pure lipids fell into the following order of decreasing antagonism to AME when added together with the antibiotic to suspensions of exponential-phase organisms: sterol esters (ergosterol esters greater than cholesterol esters; unsaturated fatty acid esters greater than saturated fatty acid esters), sterols, triglycerides, unsaturated fatty acids, saturated fatty acids. The amount of antagonistic material released from stationary organisms was not markedly greater than that from exponential-phase organisms and analysis of the lipid content of wall preparations showed that the content of total lipid, neutral lipid and triglyceride of 40 h organisms was not more than 75, 25 and 30%, respectively, greater than that of exponential-phase organisms. The AME resistance of stationary-phase organisms decreased rapidly if suspensions were incubated with glucose or mercaptoethanol. The decrease in the presence of glucose was prevented by metabolic inhibitors, especially SH binding agents. Treatment of organisms with either iodoacetamide or N-ethylmaleimide gave a rapid increase in AME resistance, amounting in some cases to 5- to 15-fold. The effect of iodoacetamide decreased as the organisms passed into the stationary phase and their intrinsic resistance increased. Evidence is presented which suggests that the degree of reduction of SH groups in the cell surface is an important factor in determining AME resistance.


Reversible inactivation of the isocitrate dehydrogenase of Escherichia coli ML308 during growth on acetate. During aerobic growth of Escherichia coli ML308 on acetate as sole carbon source, the apparent synthesis of isocitrate dehydrogenase was repressed relative to cultures on other carbon sources, such as glucose, which do not employ the glyoxylate bypass as an anaplerotic sequence. When cells were removed from an acetate medium, or when compounds were added which made the operation of the glyoxylate bypass unnecessary, the activity of isocitrate dehydrogenase rapidly increased 3- to 4-fold but fell again on restoration to an acetate medium. Changes in activity were rapid and, furthermore, could be demonstrated in the absence of protein synthesis. It is thus improbable that the mechanism involved degradation or de novo synthesis of the enzyme protein. Oxaloacetate and glyoxylate showed concerted inhibition of isocitrate dehydrogenase which could be relieved by dialysis. Because extracts of low enzyme activity, derived from acetate-metabolizing cells, could not be stimulated by dialysis or by addition of a wide range of metabolites, it is unlikely that low molecular weight, freely dissociable effectors were responsible for stimulation or inhibition of activity. Control of isocitrate dehydrogenase permitted the efficient utilization of acetate as sole source of carbon and energy but perserved the capacity of the cell to respond rapidly to an improvement in nutritional conditions. A limited survey showed that the mechanism is common but not universal among strains of E. coli and occurs in at least one strain each of Klebsiella aerogenes, Salmonella typhimurium and Serratia marcescens.


Antidermatophytic activity of 2-thiotetrahydro-1,3,5-thiadiazines and isothiocyanates. Benzyl, furfuryl, tetrahydronaphthyl, alpha-picolyl, and alpha-homopicolyl derivatives of 2-thiotetrahydro-1,3,5-thiadiazines were studied and found to be more active than beta-picolyl, and pyridyl analogs on dermatophytes, including the more resistant Microsporum canis.


Antimicrobial evaluation of diastereoisomeric 2-dimethylaminomethyl-6-phenylcyclohexanols and related esters. 2-Dimethylaminomethyl-6-phenylcyclohexanone was reduced to give the isomeric 2-dimethylaminomethyl-6-phenylcyclohexanols, which were then converted to a number of novel esters. These derivatives were screened against a wide range of microorganisms. The alcohol possessing an axial hydroxy group had high activity against Candida albicans, in contradistinction to the isomer having an equatorial hydroxy function, which was approximately 30 times less active. The esters from the isomeric alcohols demonstrated a low level of antimicrobial activity with the exception of the 10-undecenoyl ester of 2-dimethylaminomethyl-6-phenylcyclohexanol (possessing the equatorial hydroxy group), which showed significant activity against three pathogenic fungi.


Effect of intracerebroventricular bradykinin and related peptides on rabbit operant behavior. The dose-effect relationships of intraventricularly injected bradykinin, Gly-Arg-Met-Lys-bradykinin (GAML-bradykinin), synthetic substance P and angiotensin II on lever-lifting behavior of rabbits in a variable-interval (VI) 72-second schedule of sweetened water presentation were determined. All peptides used caused dose-dependent decreases in overall rates of VI responding during the experimental session in the following order of potency: angiotensin II greater than bradykinin = substance P greater than GAML-bradykinin. The angiotensin II dose-effect curve was less steep than those of the other peptides. The administration of nearly equimolar doses of the bradykinin potentiating peptides, BPP5a and BPP9a, slightly decreased overall VI response rates and caused a 10- to 20-fold potentiation of the rate-decreasing effect of bradykinin on VI responding. Both angiotensin II and bradykinin caused pauses in responding of dose-dependent duration at the beginning of the experimental session that were followed by normal VI responding. The effect of GAML-bradykinin on VI performance was similar to that of bradykinin and angiotensin II but had a delay of onset of 3 to 6 minutes. In contrast, substance P caused actual decreases in response output and pauses of variable duration interspersed between periods of regular VI responding. At the doses used, both bradykinin-potentiating peptides caused uniform decreases in VI responding throughout the experimental session. Gross behavioral changes caused by the peptides were also observed. After the intraventricular injection of bradykinin or GAML-bradykinin, rabbits showed decreased motility, ptosis, miosis and lowered ears; after angiotensin II, animals remained motionless but with wide open eyes, fully raised ears and no miosis. In turn, substance P caused restlessness and increased locomotion. These results together with reported evidence on other powerful central actions of bradykinin, angiotensin and substance P and on the existence of components of their releasing and destroying enzymatic systems in the brain suggest that linear peptides may play a role in the functioning of the central nervous system.


An enzymatic derivative double isotope assay for measuring tissue levels of taurine. The development of a new assay for taurine is described. The procedure utilizes the formation of taurocholic acid by rat liver microsomes and is dependent upon the dilution of the specific activity of radioactive taurine by the amount of taurine present in perchloric acid homogenates of tissues. Known amounts of taurine (21/2-300 nmol) are added to incubation vessels to establish a standard curve so that unknown quantities of taurine present in tissue homogenates can be calculated by a graphical comparison. The possible interference by other compounds present in tissue extracts and their removal by passage through columns containing both cation and anion exchange resins is discussed. Conditions are established for the quantitative recovery of taurine after passage through the exchange resins. Measurement of the taurine levels of several rat tissues is presented along with duplicate assays performed on the amino acid analyzer for comparison purposes.


Regional sensitivity of the rat brain to the inhibitory effects of morphine on wet shake behavior. The aim of this investigation was to determine the brain regions which were most sensitive to the inhibitory effects of morphine on the shaking response of pentobarbital-anesthetized rats to ice water. The median inhibitory dose (ID50) of morphine sulfate administered intraventricularly was found to be 0.35 mug/rat. When morphine was bilaterally injected into different regions of the brain, the ID50 values ranged from 0.04 to 17.9 mug/rat. The lowest ID50 values (0.04-0.20 mug) were found in the periaqueductal gray, the medial preoptic area and the locus ceruleus. The ID50 values ranged from 0.65 to 1.6 mug for areas around the nucleus accumbens, the fasciculus retroflexus, the medical thalamus and the septal area; from 5.6 to 7.3 mug for various hypothalamic nuclei; and from 11.0 to 17.9 mug for the basal ganglia, reticular formation substantia nigra and the reticular nucleus of the thalamus. The brain areas with the lowest ID50 values are known to have thermoregulatory functions. The similarity of the shaking response to shivering is discussed. It is concluded that the central inhibitory effects of morphine on shaking are subserved by discrete neuroanatomical substrates located in medial subcortical structures.


Synthesis and evaluation of 6-arylactamido-2,4-diaminoquinazolines and related compounds as folic acid antagonists. A series of 2,4-diaminoquinazolines bearing an aryl function attached to the 6 position through an acetamido or related linkage was synthesized. Each compound was evaluated as an inhibitor of rat liver dihydrofolate reductase as well as for suppressive antimalarial effects against Plasmodium berghei in mice. Significant in vivo activity was found to reside primarily with 5-chloro-6-arylacetamido derivatives. Most of these compounds were also tested for prophylactic activity against sporozoite-induced Plasmodium gallinaceum in chicks. Thirteen compounds, each of which possesses a 5-Cl or 5-CH3 group, displayed curative activity in this test system. Since several of these showed markedly greater potency against the avian infection, selective inhibitory action upon preerythrocytic forms of the malaria parasite is thus implied.


Synthesis and biological activity of LH-RH analogs modified at the carboxyl terminus. Des(Pro9,Gly10)-LH-RH ethylamide, des(Pro9,Gly10)-LH-RH butylamide, desGly10-LH-RH 2-aminoethylamide, and desGly10-LH-RH hydrazide were synthesized by a solid-phase method involving cleavage of protected peptide intermediates from their resin support by reaction with ethylamine, butylamine, ethylenediamine, and hydrazine, respectively. In the assay utilizing steroid pretreated, ovariectomized rats, the peptides were found to have the following LH-releasing activities when compared with natural LH-RH: ethylamide, 0.2%; butylamide, 0,1%; 2-aminoethylamide, 2.4%; hydrazide, 12%. DesGly10-LH-RH hydrazide was used as a precursor in the synthesis of desGly10-LH-RH allylamide and desGly10-LH-RH propargylamide by conversion to the azide and reaction with allylamine and propargylamine, respectively. LH and FSH levels were measured over a 4-hr period after subcutaneous injection of these two peptides into immature male rats in order to detect any prolongation of activity. The allylamide analog was quite active, releasing 1.7 times more LH and 1.3 times more FSH than the same dose of LH-RH. The propargylamide analog was considerably less active, exhibiting 50% LH-releasing activity and 64% FSH-releasing activity. Neither peptide appeared to be longer acting than LH-RH.


Synthesis and antitrichomonal activity of certain pyrazolo (1,5-a) pyrimidines. Several bridgehead nitrogen heterocycles were synthesized to be screened as antimicrobial agents, modeled after nalidixic acid. The activity of these new compounds, all derivatives of 3-nitro-4,6-disubstituted pyrazolo (1,5-a)pyrimidin-7-ones (3,7,8, and 9), however, was found to be highly specific for Trichomonas foetus and completely lacking in activity against bacteria, fungi, and parasites other than Trichomonas. Of the nine componds synthesized, including the intermediate 4,6-disubstituted pyrazolo (1,5-a) pyrimidin-7-ones (2-6) and the 6-substituted or unsubstituted pyrazolo (1,5-a) pyrimidin-7-ones (1 and 4), only 6-carbethoxy-4-ethyl-3-nitropyrazolo(1,5-a)pyrimidin-7-one (7) was found to be a potent antitrichomal agent, being comparable or perhaps better than metronidazole. From a tentative structure-activity relationship study, it was apparent that the combination of the 3-mitro, 4-ethyl, and 6-carbethoxy groups imparted specific activity, wheras other substitutions imparted little or no antitrichomonal activity.


Preparation of a new immunosuppressant, 4-5-bis(aminomethyl)acridine. 4,5-Bis(aminomethyl)acridine, useful as an immunosuppressant, was prepared in 31% overall yield by the reaction of acridine with N-(hydroxymethyl)phthalimide and subsequent decomposition with excess 6 N HCl. The drug was found to produce a suppression of the humoral antibody response comparable to several known immunosuppressive agents.


Analysis of expert diagnosis of a computer simulation of congenital heart disease. Analysis of data from five expert cardiologists indicates substantial agreement on the first step to be taken in response to an emergency case of computer-simulated congenital heart disease. There was little or no agreement among these same subjects on the specific steps undertaken following their initial response. The problem was solved in 11, 10, 8, and 6 steps by four of the cardiologists. The problem-solving sequence employed by one expert resulted in the patient's death. It is concluded that while the behavior of experts in the hospital and clinic is the primary means of evaluating successful student performance, computer simulations of patient cases offer the opportunity to use expert data in the calibration of student error.


Stabilization and clarification of sections mounted on formvar-coated slot grids by deliberate irradiation in the electron beam. Epon-Araldite sections supported on Formvar-coated slot grids can be stabilized by exposing the area of interest in the column of the electron microscope to irradiation of 0-5C/cm-2. With this procedure one can obtain electron micrographs of high quality, without drift or distortion, from any part of the open slot area. After treatment the support film is not only stable and transparent but robust enough to permit repeated removal and replacement of the specimen in the microscope column. We illustrate the method with a series of electron micrographs with initial magnifications ranging from x300 to x500,000 to show the quality of image that can be achieved. The effect of the irradiation on tissue architecture is checked by resectioning the irradiated sections. We recommend this method of local irradiation of sections for investigations using low or intermediate magnifications for the wide field it permits, for the durability it confers on the specimen and for its convenience.


High resolution scanning electron microscopy at the subcellular level. Recently developed scanning electron microscopes provide sufficient resolution to allow useful observation of subcellular biological objects. Preparation methods for such objects need not be limited to the traditional coating and mounting procedures. Many methods developed for transmission electron microscopy are immediately adaptable to scanning electron microscopy. We show that a number of techniques are available to the microscopist which yield adequate contrast and high resolution. As examples we show skeletal muscle myofibrils dispersed to reveal thick filaments, uncoated on a thin carbon film; a tropomyosin tactoid, negatively stained with uranyl acetate; oncornavirus, conventionally coated; and T4 bacteriophage on an aluminium substrate.


A technique for processing mucous coated marine invertebrate spermatozoa for scanning electron microscopy. A technique for preparing heavily mucous coated marine invertebrate spermatozoa for scanning electron microscopy (SEM) is described. This technique involves washing in 1500 NF units/ml hyaluronidase in millipored sea water to remove mucus, followed by fixation in glutaraldehyde and osmium tetroxide. Following primary fixation, spermatozoa are enclosed in Nuclepore membrane bags positioned within Teflon specimen capsules allowing them to be processed and critical point dried without excessive mechanical damage or loss.


Comparative survival of lethally irradiated inbred male mice inoculated with marrow from virgin or multiparous female donors. The response of marrow from virgin female donors to the antigens controlled by the Y chromosome of syngeneic male recipients varied with different strains of mice and was independent of their H-2 genotype. Marrow from syngeneic multiparous donors might produce a greater response to male antigens than that observed with marrow from virgin females. The sensitization of the mother to her male progeny indicated that these antigens were expressed before parturition. The reduced responsiveness of mothers to some alloantigens and the accompanying increased responsiveness to male antigens suggested a difference in the time during embryonic development when these antigens were expressed.


Response of congenitally athymic (nude) mice to infection with Mycobacterium bovis (strain BCG). Mice homozygous for the mutation "nude" (nu/nu) are athymic and lack thymus-dependent lymphocytes. Heterozygote (nu/+) controls, which are phenotypically and immunologically normal, and (nu/nu) mice were infected with 1.0 times 10-6 colony-forming units of Phipps strain BCG. In the spleens of nu/+ mice, there was a progressive increase in number of BCG up to the second week, followed by a gradual decline. However, in the nu/nu mice, BCG growth was gradual and continuous until termination of the experiment at 5 weeks. In the lung, significant differences were not noted until after the second week, at which time the nude mice showed a rapid increase (of more than 2 log10) in the number of BCG. However, the number of BCG was not significantly greater in the livers of either group. Changes in the normal histology of the lung included a massive influx of monocytes during the first 2 weeks which peaked at day 21. In the lungs of the nu/+ mice by day 28, there was considerable granuloma formation consisting of monocytes and small lymphocytes. However, in the lungs of nu/nu animals, the granulomas were made up primarily of monocytes with a lack of small lymphocytes. Acid-fast stains confirmed the presence of large numbers of organisms in the macrophages of nu/nu mice, with gradual destruction of these phagocytic cells.


Transport in bacteriophage P22-infected Salmonella typhimurium. There was rapid efflux of L-leucine, L-phenylalanine, and alpha-methyl-D-glucoside after infection of Salmonella typhimurium with the clear plaque mutant C1 of phage P22. The efflux was similar to that observed with cyanide or arsenate treatment except that there was partial recovery in the case of phage infection and almost complete recovery under the condition of lysogeny. There was no efflux after infection with the temperature-sensitive mutant ts16C1 at nonpermissive temperature. Superinfection of superinfection exclusion negative lysogen (sie A minus sie B minus) with C1 led to efflux, whereas the efflux was much less on superinfection of sie A+ Sie B+ lysogen. These results indicate that an effective injection process is enough to cause depression in the cellular transport processes.


Effect of DNA delay mutations of bacteriophage T4 on genetic recombination. Studies have been made of the effect of the DNA delay mutations of bacteriophage T4 on growth and genetic recombination in a number of Escherichia coli hosts. DNA delay mutations in genes 39, 52, 58 (61), and 60 result in abnormally high recombination frequencies. These high recombination frequencies are discussed in the context of other observations.