Structural studies on yeast RNA polymerases. Existence of common subunits in RNA polymerases A(I) and B(II). The subunits of yeast RNA polymerases A(I) and B(II) were characterized using several techniques. The present studies demonstrate that the A and B enzymes possess three subunits, which are indistinguishable on the basis of molecular weight, isoelectric point, and fingerprint pattern. The three common subunits belong to the small molecular weight components of the enzymes. By polyacrylamide gel electrophoresis with sodium dodecyl sulfate they migrate with apparent molecular weights of 27,000, 23,000, and 14,500, respectively. A two-dimensional subunit mapping technique on polyacrylamide gel was used to separate the subunits according to isoelectric point and molecular weight. The common polypeptides co-migrated on three spots corresponding to isoelectric points of 9.2 (27,000), 4.5 (23,000), and 4.6 (14,500). The fingerprints of the 35S-labeled tryptic peptides of the presumptive common subunits were found to be essentially identical. Finally, the presence of common subunits was supported by the fact that antibodies against pure RNA polymerase A cross-react with and inhibit RNA polymerase B. Except for the common subunits, it is likely that RNA polymerases A and B are primarily made of distinct gene products for the following reasons. A total of 13 polypeptide chains are present in enzyme A, whereas 10 polypeptides are found in enzyme B. The molecular weight, isoelectric point, and sulfur content of the majority of these polypeptide chains are different in the two enzymes. No similarity was found in the 35S-peptide fingerprint from a number of A and B subunits of slightly different molecular weight. Finally, antibodies against the largest subunit from RNA polymerase A do not cross-react with or inhibit RNA polymerase B. The data are discussed in terms of structural organization of eukaryotic RNA polymerases.


Stoichiometry of polypeptide chain elongation. To quantitate the amount of GTP hydrolyzed during polypeptide chain elongation, an in vitro system containing purified endogenous Escherichia coli polysomes has been developed. The polysomes are washed with 1 M NH4Cl to eliminate endogenous GTPase activities and are depleted of subunits and free ribosomes to diminish the uncoupled elongation factor G-dependent GTP hydrolysis. These polysomes, supplemented with elongation factors, aminoacyl-tRNA, and low concentrations of GTP, incorporate amino acids in their nascent peptide chains. After correcting for a background of uncoupled GTP hydrolysis, it has been found that the incorporation of each molecule of amino acid is associated with the hydrolysis of 2 molecules of GTP.


Binding interactions between radiolabeled Escherichia coli elongation factor G and the ribosome. We have prepared homogeneous radiolabeled Escherichia coli Elongation Factor G (EF-G) and examined its interactions with the ribosome. In agreement with earlier indirect observations we found that in the presence of high concentrations of fusidic acid approximately equimolar amounts of [3H]EF-G and [alpha-32P]GDP are stably bound to the ribosome. In the absence of fusidic acid, we observed a previously undescribed nucleotide-independent binding interaction between EF-G and the ribosome. This binding is detectable by rapid elution on small gel columns but is not apparent when reactions are analyzed by sucrose density gradient sedimentation. With the exception of the fact that the nucleotide-independent binding of EF-G to ribosome is apparently unaffected even by high concentrations of fusidic acid, it shares many properties in common with that binding which occurs in the presence of GDP. Nucleotide-independent binding requires magnesium ion (10 to 20 mM ) and does not require a monovalent cation but is strongly inhibited by even moderate concentrations of NH4Cl. This binding requires the presence on the ribosome of Protein L7/L12 and is inhibited by the antibiotic thiostrepton. Although we were unable to examine the binary ribosome.EF-G complex by equilibrium means, the observed stoichiometry under the conditions we employed did not exceed 0.2 mol of EF-G/mole of ribosome. Nonequilibrium measurements revealed that one-half of the EF-G was bound at a ribosome concentration of about 50 muM.


The ribosomal proteins of Saccharomyces cerevisiae. Phosphorylated and exchangeable proteins. Sixty-seven ribosomal proteins of the yeast Saccharomyces cerevisiae have been distinguished by two-dimensional polyacrylamide gel electrophoresis. Five of the ribosomal proteins are phosphorylated in vivo. One of the phosphorylated proteins appears to correspond to the phosphorylated ribosomal protein, S6, of rat liver. Another of the yeast-phosphorylated proteins, which is highly acidic, may be related to the protein L7/L12 of Escherichia coli. The phosphorylation of four of the five ribosomal proteins depends on protein synthesis. Ribosome synthesis, however, is not necessary for the phosphorylation of any of them. Three proteins of the 60 S ribosomal subunit are "exchangeable" in vivo. Two of these are phosphoproteins, one of which is the very acidic protein possibly related to L7/L12 of E. coli. The possible significance of this phosphorylated, exchangeable protein in protein synthesis is discussed.


Further experience with interfascicular grafting of the median, ulnar, and radial nerves. The results of interfascicular nerve-grafting, a procedure developed in 1964, were reported in The Journal in 1972 9. Between 1964 and 1972, defects in forty-three median, fourty-four ulnar, and sixteen radial nerves were grafted using this technique. Final functional results after a minimum follow-up of two years are known for thirty-eight patients with median, thirty-nine patients with ulnar, and thirteen patients with radial-nerve lesions. In the median-nerve cases useful motor recovery (M 3 or higher) was achieved in 82% and protective sensibility was restored in all but one. In all of the patients with ulnar-nerve lesions useful motor recovery (M 2+ or higher) occurred, while in 77% of the patients with radial-nerve lesions an M 4 or M 5 level of function was achieved. The majority of the patients with poor results had some unfavorable circumstance such as advanced age, severity of the original trauma, or long time interval before repair. Comparison of our results with those in other series indicates that interfascicular nerve-grafting gives results which are at least as good as those after epineural nerve suture under ideal conditions and better than those after neurorrhaphy under tension.


Organization of the flagellar apparatus and associate cytoplasmic microtubules in the quadriflagellate alga Polytomella agilis. The organization of microtubular systems in the quadriflagellate unicell Polytomella agilis has been reconstructed by electron microscopy of serial sections, and the overall arrangement confirmed by immunofluorescent staining using antiserum directed against chick brain tubulin. The basal bodies of the four flagella are shown to be linked in two pairs of short fibers. Light microscopy of swimming cells indicates that the flagella beat in two synchronous pairs, with each pair exhibiting a breast-stroke-like motion. Two structurally distinct flagellar rootlets, one consisting of four microtubules in a 3 over 1 pattern and the other of a striated fiber over two microtubules, terminate between adjacent basal bodies. These rootlets diverge from the basal body region and extend toward the cell posterior, passing just beneath the plasma membrane. Near the anterior part of the cell, all eight rootlets serve as attachment sites for large numbers of cytoplasmic microtubules which occur in a single row around the circumference of the cell and closely parallel the cell shape. It is suggested that the flagellar rootless may function in controlling the patterning and the direction of cytoplasmic microtubule assembly. The occurrence of similar rootlet structures in other flagellates is briefly reviewed.


Subcellular localization of cellulases in auxin-treated pea. Two forms of cellulase, buffer soluble (BS) and buffer insoluble (BI), are induced as a result of auxin treatment of dark-grown pea epicotyls. These two cellulases have been purified to homogeneity. Antibodies raised against the purified cellulases were conjugated with ferritin and were used to localize the two cellulases. Tissue sections were fixed in cold paraformaldehyde-glutaraldehyde and incubated for 1 h in the ferritin conjugates. The sections were washed with continuous shaking for 18 h and subsequently postfixed in osmium tetroxide. Tissue incubated in unconjugated ferritin was used as a control. A major part of BI cellulase is localized at the inner surface of the cell wall in close association with microfibrils. BS cellulase is localized mainly within the distended endoplasmic reticulum. Gogli complex and plasma membrane appear to be completely devoid of any cellulase activity. These observations are consistent with cytochemical localization and biochemical data on the distribution of these two cellulases among various cell and membrane fractions.


[Stab wound of the right branch of the portal vein. Suture. Healing]. The authors report a stab wound of the right branch of the portal vein observed at the Lariboisière hospital, and discuss 27 published cases of traumatic lesions of the portal vein or its branches. The treatment of choice of these lesions is mainly suture, which is the best procedure when there is a frank lesion without loss of tissue. Furthermore, we have found that ligature of one branch of the portal vein is possible, even if one does not carry out removal of the liver in the corresponding area. On the other hand, in no case, in a patient without liver disease or portal hypertension, should one carry out sudden total interruption of the portal blood flow.


Gonadotropin secretion in response to low and high doses of LRF in normal and hypogonadal women (functional disparity of the gonadotrophs). A comparison was made of gonadotropin release after an iv bolus of large (150 mug) and small (10 mug) doses of LRF during various phases of the menstrual cycle and in primary hypogonadal women. Gonadotropin release was quantitatively similar to small and large doses of LRF in hypogonadal women with low estradiol levels. In contrast, normal women from the early to late follicular phase of the cycle, the gonadotropin response to a small dose of LRF (10 mug) was not as great as that observed with a large dose of LRF (150 mug) and did not elicit the augmented gonadotropin response of the late follicular phase. During the mid-luteal phase, a significant enhancement of responses to small doses of LRF was observed. The sustained gonadotropin release induced by the large dose of LRF in the late follicular phase was no longer apparent during the mid-luteal phase, although the dose-related discrimination of LH release was maintained. These dose-related disparities in the gonadotropin release to LRF might represent functional expressions of the two-pool dynamics of the gonadotrophs induced by ovarian steroids.


Strength-duration characteristics of estrogen effects on gonadotropin response to gonadotropin-releasing hormone in women. II. Effects of varying concentrations of estradiol. This study was designed to investigate the effect of varying concentrations of estradiol, administered to normal women, upon the gonadotropin response to synthetic gonadotropin-releasing hormone (GnRH or LRF). Beginning at 4 pm on the first day of the menstrual cycle, 19 studies were performed in subjects who received injections of estradiol benzoate (E2B every 12 h for 6 days). Concentrations of E2B administered (mug/kg/12 h) were: 0.3, 0.6, 1.25, 2.5, 3.75, and 5.0. Mean serum estradiol concentrations achieved at these respective concentrations of E2B were 43, 53, 91, 145, 195, and 305 pg/ml. Twelve h after the last E2B injection, an intravenous bolus of 100 mug GnRH was administered. Gonadotropin response to this dose of GnRH after E2B was compared with each subject's response in the early follicular phase of a previous (control) cycle during which no exogenous estradiol was administered.


Hypersecretion of LH and FSH by a pituitary adenoma. A 51-year-old man who had a pituitary adenoma that appeared to be hypersecreting LH and FSH is described. Not only were serum LH and FSH concentrations above the normal ranges, but the serum concentrations of testosterone, free testosterone, and dihydrotestosterone were also above normal. Serum LH and FSH concentration increased in response to synthetic thyrotropin-releasing hormone as well as to synthetic gonadotropin-releasing hormone. The elevated hormone concentrations decreased following an initial partial hypophysectomy and decreased further following repeat hypophysectomy.


Effect of cyproterone acetate therapy on gonadotropin response to synthetic luteinizing hormone-releasing hormone (LRH) in girls with idiopathic precocious puberty. 50 mg daily of cyproterone acetate (CA) were orally administered for 8 to 35 months to 7 girls with idiopathic precocious puberty. Plasma levels of FSH and LH, cortisol, testosterone, estradiol, and progesterone were measured in 6 patients before treatment. Compared with control subjects of the same chronological age, significantly higher values were found for gonadotropins, testosterone, and estradiol. After treatment, no significant variation was observed in FSH and LH levels; testosterone was reduced in the majority of the cases without significant decline in mean values; estradiol fell significantly and returned in the prepubertal range. The plasma gonadotropin pattern following exogenously administered luteinizing hormone-releasing hormone (LRH, 100 mug iv) was characterized before treatment by an exaggerated LH response both in terms of maximum level (32.02 +/- 4.35 SE mIU/ml; prepubertal controls: 16.20 +/- 1.45 SE mIU/ml) and maximum increment above baseline values (25.36 +/- 2.84 SE mIU/ml; prepubertal controls: 13.78 +/- 1.71 mIU/ml); plasma FSH response was similar to prepubertal subjects. Treatment with CA caused a significant reduction of mean LH response (P less than .025 in comparison with pre-treatment values for maximum level and maximum increment), whereas effect on FSH response was minimal. In all patients examined, a gonadotropin release from the pituitary after the injection of synthetic LRH was evident also after several months of therapy.


Peripheral blood T and B lymphocytes, in vitro stimulation with phytohemagglutinin, and sensitization with 2,4-dinitrochlorobenzene in Grave's disease. The percentage and absolute numbers of thymos-dependent (T) and thymos-independent (B) peripheral blood lymphocytes, the in vitro response of lymphocytes to phytohemagglutinin (PHA), and the sensitization with 2,4dinitrochlorobenzene (DNCB) was studied in 26 patients with hyperthyroidism due to Graves' disease. The results showed a normal percentage of both T and B cells and increased absoluted numbers of total lymphocytes in patients compared with 30 healthy controls. The in vitro response to PHA was normal in most patients with the exception of 5 of them who showed an impaired response. All patients developed a reaction of delayed hypersensitivity to DNCB. Our study indicated that there is no general abnormality in cell-mediated immunity in Graves' disease.


Effect of clomiphene on basal and LRH-induced gonadotropin secretion in postmenopausal women. LRH tests were performed in 11 postmenopausal women before and after the administration of 200 mg of clomiphene citrate daily by mouth for 7 consecutive days. The basal levels and the maximum increments of serum LH (deltaLH) and the area under the response curve from basal level (deltaarea) after LRH administration, significantly (P less than 0.005) decreased after consecutive administration of this compound. Serum FSH levels were significantly (P less than 0.025) decreased but deltaFSH and deltaarea in LRH test were statistically unchanged. These results suggest that clomiphene citrate in postmenopausal women inhibits the pituitary gonadotropin response to exogenous LRH by its estrogenic effects.


Evaluation of an adenosine 5'-triphosphate assay as a screening method to detect significant bacteriuria. The bioluminescent reaction of adenosine 5'-triphosphate (ATP) with luciferin and luciferase has been used in conjunction with a sensitive photometer (Lab-Line's ATP photometer) to detect significant bacteriuria in urine. This rapid method of screening urine specimens for bacteriuria was evaluated by using 348 urine specimens submitted to the clinical microbiology laboratory at the University of Minnesota Hospitals for routine culture using the calibrated loop-streak plate method. There was 89.4% agreement between the culture method and the ATP assay, with 7.0% false positive and 27.0% false negative results from the ATP assay using 10(5) organisms/ml of urine or greater as positive for significant bacteriuria and less than 10(5) organisms/ml as negative for significant bacteriuria.


Rapid, colorimetric test for the determination of hippurate hydrolysis by group B Streptococcus. A colorimetric test for the determination of hippurate hydrolysis was developed. Brain heart infusion broth made with 1% sodium hippurate served as the test medium. Hydrolysis was determined by the addition of two chemical developers, M (rhodamine B) and A (uranium acetate). A dark pink color indicated hydrolysis; no color change indicated no hydrolysis. The method was efficacious in either rapid or overnight incubation. One hundred twenty-five strains of group B, 44 strains of group A, 15 strains of group C, and 10 strains of group G Streptococcus were tested. By using the Lancefield method as the standard, there was 100% agreement with both the colorimetric and ferric chloride tests for hippurate hydrolysis, and 96% agreement with the CAMP test.


Retention of motility of Treponema pallidum (Nichols virulent strain) in an anaerobic cell culture system and in a cell-free system. Optimum parameters for retention of motility of Treponema pallidum (Nichols virulent strain) were found by anaerobic co-incubation of the treponeme with rat glial cells and anaerobic incubation in spent medium obtained from glial cells originally grown aerobically.


Comparison of amino acid decarboxylase and dihydrolase results by Moeller, rapid, and replicator plate methods. A slightly modified Moeller medium for amino acid decarboxylase and dihydrolase testing by the replicator method correlated 96 to 99% with the reference Moeller broth method. False positive results were not encountered in this study.


Diagnostic value of indirect hemagglutination in the seroepidemiology of Shigella infections. To evaluate the usefulness of the indirect hemagglutination (IHA) test in the epidemiological investigation of shigellosis, single serum specimens were tested from 50 patients with Shigella dysenteriae 1 (Shiga bacillus) infections, 103 asymptomatic contacts of these cases, 267 adult and 100 student control, and serum specimens collected during two outbreaks caused by S. sonnei and one outbreak due to S. flexneri 6. In patients with S. dysenteriae 1, 74% demonstrated titers of greater than or equal to 1:40, with 50% showing titers of greater than or equal to 1:160, whereas in the controls 10.4% had titers of greater than or equal to 1:40 and only 0.3% had titers of greater than or equal to 1:160. IHA titers in serum specimens collected from patients with S. sonnei and S. flexneri 6 were too low to be considered diagnostic for individual patients, but were useful in analysis of group results. Groups of ill individuals yielded titers significantly higher than non-ill groups; however, titers from ill groups were usually less than 1:40. The IHA test for S. dysenteriae 1 antibodies serves as a valuable adjunct to the diagnosis of Shiga bacillus dysentery. In our laboratory, an IHA titer of 1:40 or 1:80 is a "borderline positive." Shiga bacillus dysentery is strongly indicated when IHA titers are greater than or equal to 1:60.


Unusual Enterobacteriaceae: H2S+ Shigella sonnei, one authentic and one false positive due to contamination with the obligate anaerobe Eubacterium lentum. A mixture of Shigella sonnei and Eubacterium lentum produced H2S in triple sugar iron agar; however, neither produced any in pure culture. A second culture of S. sonnei, isolated in Japan, is thought to be the first documented H2S+ Shigella.


Simple procedure for detection of Mycobacterium gordonae in water causing false-positive acid-fast smears. A simple procedure for detecting a few cells of Mycobacterium gordonae in laboratory water that yielded spurious smear results is described.


Evaluation of the hemolysis-in-gel test for the screening of rubella immunity and the demonstration of recent infection. The hemolysis-in-gel method for detection of antibodies to rubella virus gave results which correlated well with results of hemagglutination inhibition and neutralization tests. With a diffusion time of 24 or 48 h, a linear correlation was obtained between the logarithm of antibody concentration and the diameter of the hemolytic zone. Fourfold, and even twofold, differences in serum antibody concentrations were shown to give statistically significant differences in hemolytic zone diameters. It could therefore be concluded that the hemolysis-in-gel test is well suited for the serological diagnosis of primary rubella infection, as well as of reinfection. The sensitivity of the hemolysis-in-gel test was comparable to that of the hemagglutination inhibition test. Pigeon erythrocytes were superior to sheep erythrocytes for use in the test. Studies of the antibody response after natural rubella infection or vaccination showed that the appearance and persistence of antibodies measured by hemolysis in gel is similar to that of hemagglutination inhibition antibodies.


Bone morphology and bone loss in periodontal disease. A clinical study of the distribution of different types of bone defects in chronic periodontitis was carried out on 30 patients. An analysis of 176 defects suggests that their distribution reflects the original morphology of the alveolar bone. The role of function as a determinant of that morphology is examined. While there appears to be a relationship between functional stress and the phylogenesis of the bone, there is no direct relationship between function and ontogenesis. A study of growth remodelling of the mandible in several animals indicates that the pattern of activity is dictated by a drive to achieve a mature form which is the expression of the genetic endowment. Normal function and normal muscle stress are part of the environment in which complete growth can take place, but they do not appear to play a more active role in determining the form of growth; abnormal function does not allow normal growth to take place. The relationship between functional stress and the morphology and activity of the mature bone is not clear. Osteoporosis of age, as measured by the metacarpal index in 101 patients with chronic periodontitis, did not appear to be significantly related to the bone loss score. However, in 54 females patients, aged 35-45 years, a significant correlation was found between the metacarpal index and a "rapidity of bone loss" score.


The effect of systematic plaque control on bone regeneration in infrabony pockets. A clinical trial was undertaken to test the hypothesis that periodontitis can be cured and that bone regeneration occurs in infrabony pockets in patients maintained on an optimal standard of oral hygiene. The material comprised 24 patients with advanced periodontal disease. After an initial examination, the patients were randomly distributed into one test group and one control group. All the patients were given instruction and practice in a proper oral hygiene technique, and then subjected to periodontal surgery using the modified Widman flap procedure. Following treatment, during a 2-year period the patients of the test group were recalled once every second week for professional tooth cleaning. The control patients were recalled once every 12 months for prophylaxis. The results showed that all osseous defects of the patients of the test group were refilled with bone. The control patients, on the other hand, could not maintain a high standard of oral hygiene, and exhibited a progressive deterioration of the periodontal tissues during the postsurgical observation time.


The use of chlorhexidine mouthwash compared with a periodontal dressing following the gingivectomy procedure. The clinical results achieved were compared when a group of 21 patients requiring gingivectomies in comparable bilateral segments received a dressing or a chlorhexidine mouthwash during the first post-operative week. Initial pre-operative conditions were comparable. In each of the two treatments highly significant reductions in pocket depths were achieved; the sizes of the reductions were of clinical significance (greater than 1.5 mm). The observed difference (0.16 mm) between the two treatments in favour of the mouthwash, although significant at the conventional 5% level, was clinically unimportant. More patients preferred the dressing as a post-operative treatment, and the clinical implications of the patients' preferences are discussed.


Odontohypophosphatasia: report of two cases. Reports of a mild form of hypophosphatasia have been presented in which the oral findings were the only clinical and radiographic manifestations of the disease. It is important to support the clinical interpretations with laboratory findings in order to make a differential diagnosis from such diseases as Papillon-LeFevre Syndrome and cyclic diseases and noncyclic neutropenia. The term odontohypophosphatasia is suggested in those cases where the dental findings are the predominant manifestations of the disease.


Relation of musical wind instruments to malocclusion. Eighty-four seventh graders of both sexes, not undergoing orthodontic treatment, participated in a double-blind study to determine the effect on the anterior teeth of playing a musical wind instrument. The instrument players were just beginning their studies. Seventh graders not playing a musical wind instrument served as the control group. Impressions and plaster casts were made of each mouth at the beginning and conclusion of the study. The plaster casts were marked with the same numerical code assigned to each student. Overjet and overbite measurements of the anterior teeth were taken from the plaster casts and recorded. Findings indicate that, on an individual basis, the effect on the anterior teeth of playing a musical wind instrument is unpredictable. Only on a group basis can a class of musical wind instruments be theorized to have a certain effect on the anterior teeth. The playing of a musical wind instrument should not be used as a substitute for orthodontic treatment.


[Plasma concentrations of LH and of sex steroids during the normal menstrual cycle and during contraceptive treatment]. The authors have studied the long-term effects of combined oestrogen-progestogen on the secretion of gonadotrophins and on ovarian function. Estimation of the radio-immune levels of the plasma concentrations of the pituitary luteinizing hormone (LH), of oestradiol (E2), of progesterone (P) and of 20 alpha hydroxyprogesterone (20 alpha OHP) and of testosterone (T) served as a base for this study. A test cycle and 8 cycles in which a combination of 50 micrograms of ethinyl-oestradiol and 0.5 mg of norgestrel were administered were studied in 4 normal volunteer women. A pituitary stimulation test using 50 micrograms of gonadotrophin "releasing factor" (LH-RH) was given during the 7th cycle. Ovarian stimulation using human menopausal gonadotrophins (HMG) was given during the 8th cycle of treatment in 3 of the women and in 3 other subjects. Pituitary secretion of LH and ovarian secretion of E2 and P are partially inhibited. Pituitary response to the injection of LH-RH stays normal but 5 out 6 patients had no response to stimulation by gonadotrophins. These results allow us to conclude that the lowering of production of gonadotrophins during treatment with combination oestrogen and progestagens is responsible for inhibition of ovarian activity, and that there is a delay before the latter respond to stimulation by either endogenous or exogenous gonadotrophins.


Laser stroboscopic photography. Technique for cell orientation studies in flow. Squamous cells in fluid flow are photographed using a laser stroboscopic technique. The technique provides either dark field or phase contrast multiple images of a cell on a single frame of film at resolutions permitted by the thickness of the cell stream. Cell orientation, velocity and acceleration are obtained from these photographs for evaluation of flow system performance.


Flow microfluorometric quantitation of the blastogenic response of lymphocytes. A method for quantitating the specific stimulation of peripheral lymphocytes has been developed using the techniques of flow microfluorometry. Peripheral bovine lymphocytes were collected and specifically stained for deoxyribonucleic acid (DNA) content using a low-salt propidium iodide procedure. Flow microfluorometry was used to determine, on the basis of DNA content, the percentage of cells in a population that was stimulated. Extremely uniform staining of the lymphocytes (coefficient of variation of less than 2%) provides a high resolution between proliferating and nonproliferating cells. The method provides a rapid, highly repoducible technique for determing the fraction of lymphocytes stimulated in response to tuberculin antigens based on an increase in cellular DNA content. Specific and nonspecific stimulation by a defined antigen can be measured and resolved.


A new method for the cytochemical demonstration of peroxidase for light, fluorescence and electron microscopy. A new technique for the cytochemical demonstration of peroxidase is presented. Monomeric homovanillic acid is converted by H2O2 and peroxidase into its dimeric form, which is then precipitated as a complex salt of lead and rhodamine 6G or rhodamine B. The reaction product can be visualized by conversion to lead sulphide or viewed directly under the fluorescence microscope, since it emits a red fluorescence when excited with green light. The reaction is rapid and results in good localization at the cytologic level of peroxidase activity in granulocytes. The technique can be applied for the ultrastructural localization of enzymatic activity, but in its present form it does not match the localization sharpness of the diaminobenzidine method. This fluorescent cytochemical technique will also detect horseradish peroxidase activity and may provide a usefull probe in peroxidase immunohistochemistry. The principle of complexing metal salts with fluorescent dyes may find a more general application in enzyme cytochemistry.


Humoral immunity in aged mice. I. Age-related decline in the secondary response to DNP of spleen cells propagated in diffusion chambers. The secondary response of young-adult and old BC3F1 mice to DNP was assessed by boosting spleen cells from primed mice with DNP-KLH, propagating them in diffusion chambers implanted into irradiated recipient mice, and by counting the DNP-specific PFC. There was no difference between young-adult and old cells in the antigen dose response and in the kinetics of the immune response. There were marked differences in the magnitude of the peak response. The response of 2-year-old mice was significantly smaller than the response of young-adult mice. Two and one half-year-old mice had the smallest response. In the old mice, there was an inverse correlation between the number of nucleated cells in the spleen and the log of the number of PFC.


The Goodale white-spotted mice: a historical report. After tracing the history of Dr. Goodale's selection study of white-spotted mice, it appears that the white spotting selected for was controlled by a complex of genetic factors. Spot occurrence was controlled by a single gene pair, causing small white areas on the forehead. The size of head spot was determined by a few modifier gene loci, among which some nonadditive gene action may have taken place. The B locus appeared to be associated with head-spot size, either through linkage or pleiotropic effects. There may also have been some maternal effects on head-spot size. Selection for increased head-spot size was successful, the heritability of the trait being between 0.1 and 0.2. The increase in spot size was fairly slow; a total score of about 13 units was reached over 16 years of selection. Accumulation of factors favoring increased head-spot size led eventually to the appearance of body spots, and selection began to concentrate on these. Body-spot size was controlled by a larger number of gene pairs; gene action appeared to be more nearly additive, and there appeared to be no maternal effect or association with B locus alleles. Heritability of body-spot size was 0.3 to 0.5. Total score increased from 13 to 56 units over the 11 years of selection.


Humoral immunity in aged mice. II. Increased suppressor T cell activity in immunologically deficient old mice. Mixed cultures of DNP-primed spleen cells from old and young BC3F1 mice in diffusion chambers yielded significantly fewer secondary PFC than expected from the numbers of PFC found when young and old spleen cells were cultured alone. There was a negative correlation between the log of the number of PFC found in cultures of old spleen cells alone and the degree of immunosuppression exerted by the cells. The immunosuppressive effect was abolished by pretreatment of the old spleen cells with anti-theta serum and complement. Thus, the effect was attributed to the presence of large numbers of suppressor T cells in the spleen of primed old mice. No suppression was found in mixed cultures of spleen cells from old unimmunized mice and young primed mice.


Role of monosaccharides in the interaction of two lymphocyte mediators with their target cells. The role of monosaccharides on the interaction of two lymphocyte mediators related to the human cell migration inhibition systems with their respective target cells has been investigated. Human MIF activity on blood monocytes was significantly reduced by several 5-methylpentose sugars indcluding L-fucose, L-rhamnose and 6 deoxy-D-glucose. Certain structurally unrelated monosaccharides such as D-galactose, methyl alpha-D-mannoside and N-acetyl-D-glucosamine had no effect on this system. Incubation of human monocytes with alpha-L-fucosidase, a glycosidase which cleaves terminal L-fucose moieties, reversibly prevented their response to MIF. These results indicate that alpha-L-fucosyl residues form an integral part of the receptor for MIF on human monocytes, and furthermore, this interaction may be somewhat nonspecific in that the 5-methylpentose configuration perse is recognized. In contrast, human LIF activity on PMN leukocytes was significantly reduced by N-acetyl-D-galactosamine, but not by 5-methylpentoses, D-glucose, N-acetyl-D-galactosamine, methyl alpha-Dmannoside or D-galactose. The latter finding suggests that a subterminal sugar may form part of the receptor to LIF on human PMN leukocytes.


Carrier determined tolerance with various subclasses of murine myeloma IgG. Groups of BALB/c mice were treated with various conjugates of 2,4 dinitrophenyl (DNP) and BALB/c myeloma proteins belonging to the four subclasses of IgG (IgG1, IgG2, IgG2b, IgG3). Immediately therafter, they were challenged with DNP-keyhole limpet hemocyanin in complete Freund's adjuvant and antibody to the hapten was measured by direct and indirect hemolytic plaque assay. The results show that all subclasses of IgG are effective as tolerance-inducing carriers. However, the ability of induce tolerance is dependent upon the concentration of hapten bound to each myeloma protein. Tolerogenic conjugates suppress both direct and indirect plaque-forming cells in all types of antibodies measured (IgG1, IgG2a, IgG2b, IgGa), whereas, non-tolerogenic conjugate failed to suppress them. The intact molecule of IgG but not its fragments (Fab, F(ab)2', Fc) appear necessary as tolerance-inducing carriers. It is suggested that the ability to induce tolerance is related to the capacity of the tolerogenic conjugates to cause receptor blockade.


Local and systemic cell-mediated immunity against transmissible  gastroenteritis, an intestinal viral infection of swine. The occurrence of local and systemic cell-mediated immunity (CMI) in swine against transmissible gastroenteritis (TGE) virus, a coronavirus that infects the epithelial cells of the small intestine of swine, has been investigated. Leukocyte suspensions prepared from intestinal lamina propria (LP) and spleen were each incubated with and without inactivated TGE virus. The supernatants from these cultures were tested for the presence of macrophage migration inhibition factor (MIF) by the indirect MIF test using guinea pig peritoneal exudate cells as the source of macrophages. All swine serologically negative for TGE antibodies (i.e., unexposed to TGE virus) were also negative for MIF production by lymphocytes from both sources. In 10 of 10 animals given TGE virus subcutaneously, MIF production by splenic lymphocytes was significantly greater (p less than or equal to 0.01) than by LP lymphocytes, and in 9 of 10 of these animals MIF production by LP lymphocytes was negligible (percentage of inhibition of migration less than 3%).


Artifactual plaque formation in vitro and in vivo to passive transfer of specific antibody. The localized hemolysis in gel (LHG) assay for antibody-secreting cells was used to evaluate antibody production in heterologous erythrocyte-immunized cultures of human peripheral blood lymphocytes supplemented with 10% human plasma. Antigen-specific plaques were detected in such cultures and the number of plaques varied with different lymphocyte donors and were inhibited by heat killing the cultures before assay. However, plaque number varied directly with the number of immunizing erythrocytes added to the cultures and inversely with the number of lymphocytes added. Maximum plaque production occurred within 10 min of initiation of the cultures and occurred in the absence of any lymphoid cells. Absorption of the plasma used as culture supplement with the immunizing erythrocyte resulted in complete abrogation of subsequent specific plaque formation. Mice previously passively immunized with high titered anti-erythrocyte antibody had large numbers of plaques detected 30 min after i.v. immunization with the appropriate erythrocyte. These plaques detected by LHG are the result of carry-over of aggregates of antibody-coated erythrocytes and subsequent release of this antibody in the LHG assay and are not the result of active antibody synthesis. This "pseudoplaque" production may lead to the false interpretation of plaque formation as indicating active synthesis of antibody in vivo and in vitro where such experiments are carried out in immune animals or with antibody containing serum culture supplements.


The effect of serum inhibitor on the antigenic and mitogenic responses to E. coli bacteria. The effect of serum inhibitory factors on the induction of the in vitro immune response and mitogenic response to Escherichia coli antigens in murine spleen cell cultures was investigated. Both normal and congenitally athymic mice served as spleen cell and serum donors. Cultured CBA/J spleen cells produced anti-bacterial antibody in the presence of mouse serum, but the mitogenic response to the bacterial antigen was suppressed by the addition of mouse serum. The addition of mouse serum from either CBA/J or nude mice to nude spleen cell cultures led to suppression of both the antigenic and mitogenic responses to bacterial antigen. The polyclonal antibody response by CBA/J spleen cell cultures to SRBC induced by bacteria was significantly enhanced by the addition of either CBA/J or nude mouse serum. Conversely, the nude spleen cell polyclonal response to SRBC was significantly inhibited by high concentrations of nude or CBA/J mouse serum. These data indicate that inhibition of the specific E. coli and nonspecific polyclonal antibody responses by mouse serum occurs only in cultures of spleen cells taken from mice lacking a population of T lymphocytes.


Klebsiella L-forms: effect of growth as L-form on virulence of reverted Klebsiella pneumoniae. A strain of Klebsiella pneumoniae virulent for mice was serially passaged in vitro as a penicillin-induced L-form. Periodically, the L-form was reverted to the bacterial form by removal of penicillin, and the revertant was tested for virulence. Virulence was gradually reduced, and after 109 passages the revertant was totally avirulent. Virulence was not restored by 40 passages of the revertant through mouse peritoneum. The revertant grew less vigorously and did not infect mice when inoculated subcutaneously; in contrast, the parental form was found in all tissues examined. After phagocytosis in vitro or in vivo, the revertant survived but did not increase in numbers, whereas the parental form did increase in numbers. The revertant had a much smaller capsule than did the parental form. Immunization with the live revertant resulted in type-specific protection against infection with the parental form.


Murine pulmonary alveolar macrophages: rates of bacterial ingestion, inactivation, and destruction. The component processes of phagocytosis (ingestion, inactivation, and destruction of bacteria) were studied in mice by histological and microbiological techniques after aerosol infection with Staphylococcus aureus. Rates of bacterial ingestion and inactivation were (respectively): 0 hr, 37.7% and 0; 1 hr, 64.5% and 45.8%; 2 hr, 75.9% and 67.9%; 4 hr, 82.4% and 84.1%; and 8 hr, 90.7% and 94.8%. Bacterial destruction began 2-4 hr after aerosol infection and affected 80% of the bacteria by 8 hr. Comparison of these processes indicated that bacterial ingestion occurred before inactivation and was 76% complete at 2 hr. Inactivation resulted in death of 84% of the bacteria at 4 hr, and 80% of the bacteria were at least partially destroyed by 8 hr. The mechanism of X-irradiation-induced depression of pulmonary bacterial inactivation was studied in syngeneic mice protected from lethal effects of X-irradiation by chest and pelvic shields or by transplantation of 2 times 10(6) bone marrow cells. Impairments in bacterial inactivation resulted from diminished ingestion of bacteria by macrophages.


Interactions of choleragenoid and GM1 ganglioside with enterotoxins of Vibrio cholerae and Escherichia coli in cultured adrenal cells. The heat-labile enterotoxins of Vibrio cholerae and Escherichia coli induce morphologic changes and steroidogenesis in clonal lines of adrenal tumor cells in tissue culture; these effects are preventable by prior incubation of either toxin with GM1 ganglioside (galactosyl-N-acetylgalactosaminyl [sialosyl] lactosyl ceramide; GGnSLC) but are not preventable by prior incubation of adrenal cells with choleragenoid. Choleragenoid, however, is capable of interfering with the ability of GM1 ganglioside to neutralize the effects of either toxin. These results suggest that the GM1 ganglioside may not be the true receptor for the toxins on adrenal cells, but that it is acting as a pseudoreceptor. The ability of a subunit (A) of the cholera enterotoxin molecule to induce in adrenal cells morphologic changes and steroidogenesis similar to those effects inducible by the whole toxin indicates the possibility that separate receptor sites on these adrenal cells may exist for the binding and active fragments of the molecule.


Isolation and properties of heat-labile enterotoxin(s) from enterotoxigenic Escherichia coli. Various techniques have been applied to the detection of skin reactivity associated with heat-labile Escherichia coli enterotoxin in fermenter-grown cultures of enterotoxigenic strains in syncase medium and in trypticase soy broth. Isolated products that were homogeneous, as determined by disc electrophoresis and sodium dodecyl sulfate gel electrophoresis, differed immunologically and in physicochemical characteristics depending on the strain and medium used, even though the products had similar specific activities in skin tests and in Chinese hamster ovary cells. In support of observations on porcine E.coli enterotoxin, and in contrast to the enterotoxin of Vibrio cholerae, the products were single polypeptide chains with molecular weights ranging from approximately 35,000 ot over 100,000 daltons. Proteolytic cleavage during culture and purification might account for some of the variations observed. The isolated products were almost 10(6)-fold less active than purified choleragen in causing morphologic alterations of Chinese hamster ovary cells, approximately 1,000-fold less active in skin tests, and at least 100-fold less active in rabbit ileal loops. In addition, only 1/100 as much active protein was produced by the strains employed as by V. cholerae. It is possible that accessory or host-derived factors are required. The most effective large-scale procedure for isolation from trypticase soy broth culture supernatants was a sequence of concentration by ultrafiltration, judicious (NH4)2SO4 precipitation, gel filtration on Sephadex G-150, and gel filtration on Agarose A5m, from which the E. coli products (like choleragen) are retarded in their elution. Toxin was isolated from an enterotoxigenic strain that caused diarrhea (total volume, 60-liters) for four days in a patient who had visited Mexico.


Purification of heat-labile enterotoxin from Escherichia coli O78:H11 by affinity chromatography with antserum to Vibrio cholerae toxin. Concentrated culture filtrate of Escherichia coli O78:H11, strain H10407, was applied to an affinity column prepared with IgG antibodies to the toxin of Vibrio cholerae. Elution of the retained material with 3 M KCNS yielded a nonenterotoxic protein that precipitated with antiserum to V. cholerae toxin and had three major protein components on sodium dodecyl sulfate gels. After treatment with 2-mercaptoethanol, two protein components were observed. Elution of the affinity column with 5 M guanidine yielded an enterotoxic protein that precipitated with antiserum to V. cholerae toxin. After treatment with 2-mercaptoethanol, only one protein component, with mobility identical to that of the slower component of the eluate (treated with 3 M KCNS and 2-mercaptoethanol), was observed.


Activation of Heat-labile Escherichia coli enterotoxin by trypsin. Trypsin-treated, cell-free filtrates derived from enterotoxigenic Escherichia coli, strain H197 (O78:H11), exhibited a fourfold or greater increase in heat-labile vascular permeability factor activity and a 10-fold or greater increase in the ability to stimulate secretion of growth hormone by cultured rat pituitary cells. In contrast, trypsin-treated filtrates were not different from untreated filtrates in their ability to elicit a secretory response in ligated rabbit intestinal loops. However, incubation of culture filtrate in ligated intestinal loops, or with rabbit intestinal fluid (in vitro), resulted in at least a twofold increase in permeability factor that did not occur in the presence of trypsin inhibitor or with heat-inactivated intestinal fluid. Moreover, trypsin inhibitor could reduce the secretory response to culture filtrate. These findings suggest that activation of heat-labile E. coli enterotoxin by host enzymes may play a role in the development of a full pathogenic effect.


Estimation of bile acid excretion in man: comparison of isotopic turnover and fecal excretion methods. Bile acid excretion was studied in 9 human subjects simultaneously by the Lindstedt (Lindstedt, S. 1957. Acta Physiol. Scand. 40:1-9) isotopic turnovermethod and by fecal chemical analysis during a balance study. The identities of the fecal bile acids were confirmed by combined gas-liquid chromatography/mass spectrometry. Under the steady state conditions of the patient studies, bile acid excretion values obtained by fecal analysis were lower (by 18.1 to 44.2%) than the values obtained by the isotopic turnover method. This difference persisted even in those patients given [14C]chenodeoxycholic acid instead of [3H]chenodeoxycholic acid. The fecal excretion values were similar when calculated using either beta-sitosterol or chromium sesquioxide as fecal flow markers. The fecal excretion values during the earlier part of the isotopic study were higher than those during the latter part of the study. The lower values of bile acid excretion obtained from fecal analysis could not be explained by the loss of bile acids as sulfate conjugates or by losses due to bile acid degradation in the intestine. This study suggests that bile acid turnover is consistently higher than bile acid excretion under experimental conditions. It is recommended that the data obtained from the isotopic turnover method should not be compared with fecal excretion data.


Sterilization of infected bone by lyophilization and rehydration with antibiotic solutions. In vitro experiments were carried out with artificially infected rib-fragments in order to find out, whether freeze-drying and subsequent sterilization with ethylene oxide and antibiotic solutions will eliminate bacteria inside the bone. Ethylene oxide will kill bacterial surface contaminants, but not bacteria inside the bone; rehydration with appropriate antibiotic solutions will eliminate bacteria inside the graft. Some clinical implications are mentioned.


Probucol, a new cholesterol lowering drug. The purpose of this investigation was to determine the ability of Dh-581 (Procubol) (See Figure 1) to reduce the level of serum cholesterol for extended periods of time and to maintain the reduction without loss of effectiveness. The effects of DH-581 on triglycerides and the safety and tolerance of daily doses were also assessed.


Plasma and red cell cholinesterase activity in uremic patterns (effects of hemodialysis and renal transplantation). Plasma butyrylcholinesterase (BuChE) and red cell acetylcholinesterase (AChE) activity was measured in patients with endstage kidney disease undergoing hemodialysis or renal transplantation. All patients had significantly lower values of BuChE activity than normals. A single hemodialysis did not influence the level of BuChE activity but caused a moderate, though statistically significant increase of red cell AChE activity. Chronic hemodialysis significantly increased BuChE but did not affect AChE activity. After successful renal transplantation a sharp fall of BuChE activity occurred, a completely unexpected finding. This decrease coincided with the administration of large doses of glucocorticoids. Once the dose of these drugs was decreased the activity of plasma BuChE returned toward preoperative values. No similar changes in red cell AChE were seen. The possible effect of the steroid drugs on protein synthesis in the liver is discussed.


Interaction between oestrogen and gonadotrophin- releasing hormone on the release and synthesis of luteinizing hormone and follicle-stimulating hormone from incubated pituitaries. The release and synthesis of LH and FSH were studied in adenohypophyses from ovariectomized rats incubated for a period of 4 h in flasks containing 1 ml Eagle's medium. One hemipituitary was used as the experimental gland and the other half served as a control. Glands from ovariectomized untreated animals showed a spontaneous release of LH and FSH and the amount of hormones released (per mg gland) by both the hemipituitaries was not significantly different. Also the content of the hormones at the end of the incubation period was similar in both halves. Gonadotrophin-releasing hormone (Gn-RH) added to the incubation medium stimulated the release of LH and FSH. A dose--response relationship was obtained between doses of 0-51 and 8-00 ng/ml medium. Although lower doses were required to increase the release of LH, the amount of FSH released was higher when expressed as a percentage of gland content. Pituitary glands from ovariectomized rats treated with 5 mug oestradiol benzoate 24 h before being killed showed an increase in sensitivity to Gn-RH, but the response decreased when oestrogen was injected 2 h before death. Also the addition of oestradiol-17beta to the incubation medium inhibited LH and FSH release induced by Gn-RH. Gonadotrophin-releasing hormone increased the spontaneous synthesis of LH and FSH observed in the incubated pituitaries. This effect of Gn-RH was stimulated by the injection of oestrogen into the donor animals whereas administration of oestrogen into the medium enhanced the synthesis of LH and partially inhibited that of FSH. These results provide evidence for a dual effect of oestrogen on the release of LH and FSH induced by Gn-RH. They also show that synthesis of gonadotrophic hormones was favoured by oestrogen or by increased gonadotrophin release.


Immunoreactive luteinizing hormone releasing factor in rat pituitary stalk blood: effects of electrical stimulation of the medial preoptic area. Blood was collected from the cut pituitary stalk of male and female rats before and during the application of an electrical stimulus to the medial preoptic area. The plasma was assayed for immunoreactive LH releasing factor (RF) by a double antibody radioimmunoassay using a specific antiserum raised in rabbits against the free acid derivative of the decapeptide LH-RF conjugated to bovine serum albumin. The decapeptide (used as a standard) and pituitary stalk plasma cross-reacted in a similar manner with the antiserum. Stimulation of the preoptic area increased significantly the amount of LH-RF in pituitary stalk plasma in both male and female rats. The increase in LH-RF was linearly related to the strength of the stimulating current, and the amount of LH-RF liberated diminished on cessation of the stimulus. The concentration of LH-RF in pituitary stalk plasma from female rats was significantly greater than that in jugular venous plasma. The magnitudes of the mean increments of LH-RF in pituitary stalk plasma (stimulation minus pre-stimulation values) at various times of the oestrous cycle in female rats suggest that between 18.00 h of dioestrus and 13.00 h of pro-oestrus there is an increase in sensitivity of the LH-RF secretory mechanism to electrical stimulation. However, the increments decreased in magnitude between 13.00 and 18.00 h of pro-oestrus, indicating that the marked increase in responsiveness of the hypothalamo-hypophysial system to electrical stimulation which occurs during this period is due mainly to a change in sensitivity of the pituitary gonadotrophs to LH-RF. The LH-RF in pituitary stalk plasma collected before application of the stimulus was higher at some of the times examined during pro-oestrus than at other times of the oestrous cycle. A higher level of the secretion of the factor may be important for the full development of the priming effect of LH-RF and, consequently, the marked increase in responsiveness of the pituitary gland which occurs during the afternoon of pro-oestrus.


Absence of an immunologically distinct follicle-stimulating hormone releasing factor in rat stalk median eminence extracts. Rat stalk median eminence (SME) extract was incubated with various quantities of an antiserum raised against synthetic luteinizing hormone (LH-RH) and the resulting material was used at two or more dose levels to stimulate release of LH and FSH from anterior pituitary tissue, obtained from male rats, incubated in vitro. The resulting dose-response lines were use to obtain the LH- and FSH-releasing potency of the material, expressed as a percentage of SME extract pre-incubated with normal rabbit serum. Treatment with various doses of antiserum reduced LH-releasing and FSH-releasing potencies to a similar extent. Stalk median eminence extract and synthetic LH-RH were incubated with antiserum and directly compared in the same experiment in vitro. The changes in LH and FSH release caused by pre-treatment with antibody were similar for SME extract and synthetic LH-RH. In a final experiment, SME extract was passed through a column of Sepharose 2B particles to which was coupled anti-LH-RH antibody. The resulting material, when mixed with synthetic LH-RH and used to stimulate rat pituitary tissue in vitro caused a similar increase in the LH- and FSH-releasing potencies of the synthetic decapaptide. It is concluded that rat SME extract contains a single releasing factor for LH and FSH immunologically indistinguishable from synthetic LH-RH.


Inhibition by oestradiol of the pituitary response to luteinizing hormone releasing hormone in the dog. Intravenous injections of synthetic LH-RH stimulated LH secretion in male dogs. Pretreatment with 50 mug oestradiol-17beta, 60 or 165 min but not 15 min before LH-RH, inhibited LH secretion. Injection of 0.1-10 mug oestradiol 165 min before administration of LH-RH, Blocked LH secretion. Basal plasma LH levels before LH-RH administration were lower in dogs given oestradiol than in controls.


Epithelial cell binding of group A streptococci by lipoteichoic acid on fimbriae denuded of M protein. Group A streptococci were treated with various enzymatic and chemical agents in an attempt to dissociate the type-specific M protein from intact surface "fimbriae." Mild peptic digestion at pH 5.8, which was previously shown to extract serologically active M antigen from intact streptococci had little visible effect on the fimbriae even though virtually all of the M protein was removed as demonstrated by (a) increased susceptibility to phagocytosis, (b) lack of opsonic effect of homologous M antibody on the treated streptococci, and (c) loss of HCl-extractable M protein. These fimbriated streptococci which lacked M protein adhered to human oral mucosal cells equally as well as untreated, fimbriated organisms which retained their M protein. Removal of both fimbriae and M protein by digesting organisms with HCL at pH 2.0 at 94 degrees C. or with trypsin abolished their ability to bind mucosal cells. Electron microscopy of streptococci bound to epithelial cells demonstrated fimbriae radiating from the surface of the organisms to the membrane of the epithelial cells. It is apparent, therefore, that the determinants of streptococcal fimbriae involved in resistance to phagocytosis can be dissociated from those involved in epithelial cell binding. These results are consistent with our previous studies which suggested that fatty acids ester linked with glycerol teichoic acid rather than M protein of streptococci binds the organisms to epithelial cells.


Idiotypic analysis of lymphocytes in vitro. I. Specificity and heterogeneity of B and T lymphocytes reactive with anti-idiotypic antibody. Guinea pig anti-idiotypic antibodies (anti-Id) of the IgG1 class, directed to an A/J antibody to Group A streptococcal carbohydrate (A-CHO), or directed to a BALB/c myeloma protein that binds the same antigen, stimulate B-precursor cells as well as T-helper cells when injected into mice of the appropriate strain. The strain-specific induction of both precursor and helper activity was detected by in vitro secondary responses of primed spleen cells to A-CHO or to 2,4,6-trinitrophenyl (TNP) upon challenge with Group A streptococcal vaccine (Strep.A) or with TNP-Strep.A, respectively. B- and T-cell populations primed with anti-Id were uniform with respect to the binding of antigen and of anti-Id. This was in contrast to cells primed with Strep.A, which were heterogenous. Taken together, B and T cells that possess the same antigen-binding specificity share idiotypic determinants, reveal the same idiotypic polymorphism, and may display similar degrees of heterogeneity with respect to the binding of antigen and anti-Id. Since the anti-Id used in this study detect Id determinants associated with the heavy chain of the variable region of mouse antibodies, the data suggest that this region of the immunoglobulin molecule is shared between T- and B-cell antigen receptors.


Octopamine: presence in firefly lantern suggests a transmitter role. The lantern of the adult firefly is innervated by a nerve supply with endings similar to other known monoaminergic nerves. However, catecholamines could not be detected in the nerve endings and noradrenaline and adrenaline were relatively weak agonists in eliciting a flash of light when applied to the lantern in vitro. Firefly lanterns were assayed for octopamine using a sensitive, specific enzymatic assay in which octopamine is enzymatically converted to 3H-synephrine and quantitated by liquid scintillation counting. The presence of substantial amounts (125 ng/g tissue) of octopamine was demonstrated and confirmed by thin layer chromatography. On the basis of this and earlier work, it is suggested that the nerves to the firefly lantern are octopaminergic.


Genetic transfer of Salmonella O antigens to Escherichia coli O8. His+ hybrids from a cross between a Salmonella typhimurium donor and an Escherichia coli O8 recipient expressed E. coli O8 specificity and in addition Salmonella O4,12-specificity. This indicated that the recipients had received the his-linked donor rfb cluster determining the synthesis of S. typhimurium O-specific repeat units and that the rfb genes of both mating partners are functional in these hybrids. Chemical analyses showed that the hybrids contained an E. coli O8 lipopolysaccharide (O antigen) and a S. typhimurium specific lipopolysaccharide with only one O-specific repeat unit (SR antigen). O8-negative mutants selected from the O8-positive hybrids retained the Salmonella O-specificity and represent semi-rough (SR) forms, because the rfc gene(s) determining the polymerization of repeat units has not been transferred. Attempts to introduce the S. typhimurium rfc locus into E. coli O8 remained unsuccessful. Crosses between a S. typhi donor and E. coli O8 gave rise to smooth (S) and SR His+ recombinants exhibiting only S. typhi O-specificity. The smooth recombinants are assumed to have obtained the his-linked rfb cluster and in addition the rfc gene(s) of the donor. The exchange of the rfb region of such smooth recombinants by that of a S. typhimurium donor led to smooth hybrids with O4,(5), 12-specificity. The phenotypically smooth recombinants exhibited concomitantly S- and SR-lipopolysaccharides of S. typhi and S. typhimurium O-specificity, respectively.


Further observations on the association of the colicine V plasmid of Escherichia coli with pathogenicity and with survival in the alimentary tract. A high proportion of invasive strains of Escherichia coli produced colicine V. This property was easily eliminated from 20 of 21 of these strains by 'curing' agents, especially sodium lauryl sulphate, indicating that the genes determining it were located on a plasmid (ColV) which was transmitted from ten of the strains by conjugation. Inoculated intramuscularly, the ColV- forms of all 17 strains tested were less pathogenic for chickens than the corresponding ColV+ forms. The pathogenicity of the ColV- forms of four strains was increased, usually to that of the ColV+ form from which they were derived, by implanting other ColV determinants in them. Much higher concentrations of organisms were found in the blood and liver of chickens infected with ColV+ forms than in chickens infected with ColV- forms. Inoculated intraperitoneally, the ColV+ form of one of the strains, BI88, was more pathogenic for mice than the ColV- form; much higher concentrations of organisms were found in the peritoneal fluid and blood of ColV+-inoculated mice than of ColV--inoculated mice. Inoculated orally and intravenously, ColV+ forms were more pathogenic for colostrum-deprived calves than the corresponding ColV- forms. After mixtures of ColV+ and ColV- organisms of the same strain in ratios of 1:10,1:100 or 1:1000 were given orally, they were found in a similar ratio in the contents of the alimentary tract 1 to 2 days later, when the calves were near to death. Many more ColV+ than ColV- organisms were found in the mesenteric lymph nodes, the deeper tissues and the blood; in the urinary and gall bladders, locations remote from the defence mechanisms of the body, the numbers of ColV- organisms sometimes exceeded those of ColV+ organisms. Colicine V, although demonstrated in the blood at death, did not appear adversely to influence the concentration of ColV- organisms in these calves. Several days after mixtures of ColV+ and ColV- organisms of strain BI88 were taken orally by two human beings, the ColV+ organisms became much more numerous in their faeces than the ColV- ones. Similar results were obtained when colVr organisms of BI88 were included in the inoculum or when a strain whose ColV- form was completely colicine V-resistant was studied. A ColE+ form of BI88 ColV- was no more pathogenic for chickens, mice or colostrum-deprived calves than was a ColE- form of this strain, and it did not persist in the faeces of the two human beings for longer than the ColE- form.


Transduction of leucine auxotrophs of Proteus mirabilis to prototrophy or antibiotic resistance by P. mirabilis high frequency transducing bacteriophages. High frequency transducing (HFT) phages 5006MHFTk and 5006MHFTak for kanamycin or ampicillin plus kanamycin resistance, derived from Proteus mirabilis strains PM5006(R394) and PM5006(R394) respectively, transduced (at low multiplicities of infection, m.o.i.) antibiotic resistance and prototrophy to PM5006 leu-I at high frequency. Simultaneous transduction of these markers occurred at very much lower frequencies. The latter result was correlated with the proportion of multiply-infected bacteria which, due to the great transducing potential of the phage, could register as transductants. Each HFT lysate was thus heterogenous with regard to high frequency transducing phage. Apart from the additional antibiotic resistance marker carried by one phage, no other difference between the two lysates was detected. High segregation frequencies of antibiotic-resistant or prototrophic transductants indicated transduction by lysogenization. Although antibiotic-sensitive segregants of antibiotic-resistant prototrophic transductants occurred at high frequency, no auxotrophic segregants of these transductants were found. This suggests transduction by a double cross-over event in the leucine region. Most transductants, even at low m.o.i., were lysogenically converted to homologous phage non-adsorption as a result of interaction between the transducing phage genome and the resident cryptic prophage. They could, however, be retransduced by appropriate phage lysates; thus, lysogenic conversion to non-adsorption was not absolute. Some prototrophic transductants were non-lysogenic although their segregants liberated low-titre phage. The latter anomaly, and the fact that the leucine marker and antibiotic resistance were not cotransduced, are explained by the mode of integration of the phage into the host chromosome in relation to the resident cryptic prophage and the leucine region.


Attempted transfer of antibiotic resistance between Bacteroides and Escherichia coli. Transfer of antibiotic resistance between Bacteroides organisms and E. coli in mixed culture under optimal anaerobic conditions was attempted. Donor strains used were E. coli with R factors of a number of compatibility groups and Bacteroides with unusually high antibiotic resistance. Recipient strains included E. coli strains with characteristics favouring conjugal transfer. Although control experiments verified that conjugal transfer of derepressed R factors could occur at a high frequency between E. coli in an anaerobic environment, transfer of antibiotic resistance between E. coli and Bacteroides was never demonstrated.


Stereotactic extralemniscal myelotomy.

Stereotactic extralemniscal myelotomy. A procedure is reported in which radiofrequency lesions were stereotactically placed in the central cord region at the cervicomedullary junction to interrupt selectively the extralemniscal system. Physiological recognition of the target site was based upon the homuncular organization of the dorsal funiculi, which is analysed. Only subjective (segmentally unrelated) analgesia was achieved, usually without demonstrable sensory loss. Stimuli were not painful, while ability to localize it and the discrimination between sharp and blunt components were preserved. A rational approach is offered to certain central pain phenomena which resulted in abolition of hyperpathia and disappearance of, or marked reduction in, deep background pain. Both upper and lower body pain were easily dealt with. Pathways for automatic respiration or for micturition were not encroached upon. Therefore it was electively used for cases with diminished respiratory reserve and for cases with midline and/or bilateral pain.


Tumor volume, luxury perfusion, and regional blood volume changes in man visualized by subtraction computerized tomography. Computer and photographic methods for producing subtractions of computerized axial tomographic (CAT) scans have been developed. By subtracting point for point a normal scan from one taken after intravenous infusion of contrast material, a picture of the contrast in the cerebral vessels is created. By this method, tumor size and degree of vascularity may be assessed. Furthermore, abnormalities in perfusion and changes in blood volume due to mass effects and edema may be detected. Subtracting scans should add to the diagnostic potential of CAT and provide a noninvasive way to study vascular changes in cerebral disease.


Inclusion of physiologic data into computerized nuclear medicine dynamic studies. A general technique has been developed for the multiplexing of time-dependent analog signals into the individual frames of a scintillation camera dynamic function study. A pulse train, frequency-modulated by the physiologic signal, is injected into a test input of a preamplifier servicing an outer phototube of the camera head. These tail pulses mimic photoevents occurring at a point outside of the camera's field of view, chosen to occupy a data cell in an unused corner of a computer-stored image. By selecting a region of interest encompassing this pulser peak, the resulting time-activity curve displays the desired physiologic variable in temporal synchronism with the radiotracer distribution.


Oral sarcoidosis: review of the literature. Patients with symptoms consistent with sarcoidosis who are seen by a dentist are not common. Generally, the disease is documented in a patient before the expression of oral symptoms. The most common oral or para-oral symptom appears to be an asymptomatic swelling--usually of the parotid glands or cervical nodes. Often these lesions are first brought to the attention of our medical colleagues. However, in four instances reported in the literature, systemic sarcoidosis was discovered because of the initial oral symptoms. There are often changes in the minor salivary glands under clinically normal-appearing mucosa in patients with systemic sarcoidosis. We have reviewed the concepts of etiology, criteria for diagnosis, and rationale of treatment, along with a review of the dental literature of sarcoidosis. An illustrative case report of a sarcoid lesion on the buccal mucosa is presented.


The localisation of hepatitis B antigen and immunoglobulin G in liver tissue: an immunofluorescence, light and electron microscopic study. Liver tissue specimens for 43 HBAg seropositive patients and 22 HBAg seronegative individuals were investigated for the presence of HBAg with the immunofluorescence technique. Histological diagnoses were established with light microscopy on routinely stained frozen liver sections. The liver specimens of 42 out of the 43 HBAg-positive patients showed positive fluorescence. Three distinct fluorescence patterns could be observed: diffuse, focal and solitary. The diffuse pattern appeared to occur particularly in chronic aggressive or persistent hepatitis, the focal pattern was seen only in the group of patients without obvious alterations and the solitary pattern especially in acute hepatic necrosis.


Experimental contribution to the pathogenesis of spina bifida. Reopening of the central canal in 3-day-old chick embryos results in the development of a typical myeloschisis within 5 min. Focal continuity between the ectoderm and neural tissue at the site of experimentally induced spina bifida (SB) may be achieved in 2 hr and increases in consecutive hours. In 24 hr a secondary neuro-ectodermal junction at the site of SB is fully formed and can be misinterpreted as evidence of a failure of the neural tube to close. Therefore the continuity between ectoderm and neural tube to close. The detachment of the notochord from the open portion of the spinal cord is a secondary phenomenon.


Pathogenesis of trypan-blue-induced spina bifida. Congenital malformations induced in rats by the application of trypan blue were studied at variable times of their development of congenital malformations is a continuous process of structural and functional adaptation of the development embryo to focal tissue degeneration and necrosis (cell death). The primary degeneration is a transitory phenomenon, because the dead cells are phagocytosed by surviving cells. However, the damage triggers off the process of abnormal development.


Mechanical ventilation of infants of less than 1,501 gm birth weight: Health, growth, and neurologic sequelae. A two-year follow-up study of 73 low-birth-weight ( less than 1,501 gm) infants treated with positive pressure ventilation as neonates revealed the following: 24% incidence of lower respiratory tract infections during the first year; weight and height at two years averaging between tenth and twenty-fifth percentiles; major neurologic defects diagnosed in 14 boys (39%) and seven girls (18%) with one-year Bayley scores of less than 80. Major neurologic sequelae were closely associated with a neonatal history of seizures and intracranial hemorrhage and were more common in boys, survivors weighing more than 1,000 gm and following high-risk pregnancies.


The use of a self-polymerizing resin with enamel etchant for temporary stabilization. The indications and characteristics of ideal temporary splints have been discussed. A technique has been presented using a self-polymerizing resin with an enamel etchant for extracoronal splinting of anterior teeth. When properly fabricated, the restoration has most of the properties of an ideal temporary splint: economy, simplicity of fabrication, lack of irritation, compactness, durability and esthetics.


A comparative study of ultrasonic generators and hand instruments. A 6-month study project utilized four human subjects and a total of 63 biopsies which were obtained from gingiva treated by hand scaling, Alpha-Sonic scaling and Cavitron. These were compared with controls (untreated specimens). Microsections of soft tissues taken immediately and after 1 and 2 weeks were evaluated. The clinical acceptability of Alpha-Sonic as a working tool for periodontal scaling is to be noted.


Clinical evaluation of freeze-dried bone allografts in periodontal osseous defects. Freeze-dried cortical bone allografts of a fine particle size were implanted into wide three-wall, two-wall, one-wall, combination, and furcation defects. Of the 97 defects treated, 23 manifested complete bone regeneration; 39 showed greater than 50%; and 24, less than 50% osseous repair. Twelve defects failed to demonstrate any bony regeneration, of which nine were furcation involvements. From the preliminary data available, there is strong evidence which indicates that freeze-dried bone allografts have definite potential as grafting  material in certain periodontal osseous defects. However, final determination must await the outcome of a larger number of cases and also histologic evidence.


Structure of surface coatings on teeth. A review. Classification by origin and structure of the many different coatings which occur on the surfaces of teeth and artificial surfaces exposed to the oral environment necessitate a revision of the terminology used to describe these coatings. Numerous gaps still exist in our understanding of the origin, morphological structure and chemical composition of many of these structures, notably the ones in the "acquired" category. The proposed classification of surface coatings, although not definitive, may provide a more accurate categorization of these structures in the light of contemporary knowledge.


Aggression in children in an urban clinic. Seeking to identify causal factors and practical means of controlling aggressive behavior, the clinical records of 35 aggressive and 35 non-aggressive boys were examined for evidence of parental and peer models, frustating life experiences, and capacity for fantasy on projective tests. Records of 15 girls were also studied. The most striking difference between groups appeared in the high frequency of aggressive behavior in parents and peers of the aggressive children and the relative absence of such behavioral models for the nonaggressive groups. Support for fantasy and frustation theories was also found. The reduction of aggressive behavior would necessitate going beyond individual treatment and enlisting the cooperation of aggressive families.


Discriminating adolescent male delinquents through the use of kinetic family drawings. Studied the discriminatory potential of the Kinetic Family Drawing technique (KFD). The drawings of 20 male adolescent delinquents were compared to a group of 20 normal male adolescents in order to discover whether any hypothesized differences existed. Significant chi square differences were found on only three of the variables: (a) body omission, (b) lack of family, and (c) akinesis. The delinquent group tended to manifest all three of these traits in their KFDs. The value of the KFD in terms of normative discrimination and/or prediction was found to be somewhat inconclusive in light of the low number of distinguishing traits and the small size of the sample.


Effects of glucose on 45Ca2+ uptake by pancreatic islets as studied with the lanthanum method. 1. Fluxes of 45Ca2+ were studied in pancreatic islets from non-inbred ob/ob-mice. Because La3+ blocked the transmembrane fluxes of 45Ca2+ in islet cells, incubations aimed at measuring glucose-induced changes of the intracellular Ca2+ were ended by washing the islets with 2 mM-La3+ for 60 min. 2. Uptake of 45Ca2+ progressed for 2 hr; the intracellular concentration of exchangable Ca2+ was about 7 m-mole/kg dry wt., as estimated from the isotope distribution at apparent equilibrium in islets exposed to 3 mM D-glucose. Raising the D-glucose concentration to 20 mM enhanced the 45 Ca2+ uptake whether or not the islets had first been equilibrated with the isotope. The stimulatory effect of D-glucose was observed in Tris buffer containing no anions but Cl- as well as in polyanionic bicarbonate buffer. The effect could not be reproduced with equimolar L-glucose. 3. The rate of 45Ca2+ release was the same whether the islets had been pre-loaded in the presence of 3 or 20 mM D-glucose. Thus the 45Ca2+ that had been taken up in response to 20 mM D-glucose appeared to be released much more slowly than the bulk of intracellular 45Ca2+. The release of 45Ca2+ was not significantly influenced by D-glucose during the release period. Incubation for 30 min was require for half of the radioactivity to be released. 4. The rates of insulin secretion were about the same in uni-anionic Tris buffer as in polyanionic bicarbonate buffer. A marked insulin secretory response to 20 mM D-glucose was observed in either buffer. 5. It is concluded that 20 mM D-glucose causes a net uptake of Ca2+ from the extracellular fluid into the interior of the beta-cells. This uptake is probably not regulated at the level of the plasma membrane but more likely reflects an increased affinity of some intracellular phase or compartment for the ion. Because the observed uptake and release of intracellular 45Ca2+ are slow processes in comparison with the rapid effects of extracellular Ca2+ on insulin secretion, insulin secretion may also depend on a more superficial and La3+-displacable Ca2+ pool.


Muscle-formed complete mandibular dentures. A method has been presented whereby a patient's musculature is used to indicate the position of the teeth and to develop the shape and thickness of the denture base. Of 30 patients tested, 28 experienced improved stability of their lower dentures.


The effects of etching on the margins of cast gold restorations. 1. Any etching, either chemical or electrical, sufficient to remove an adequate bulk of metal from the tissue surface of a casting will have a measurable and, hence, adverse effect on the margins of the restoration. 2. During any etching process, all margins and contact and occluding surfaces should be protected. 3. For aqua regia etching, Type 2 inlay wax appeared to provide sufficient protection. For electrochemical milling, fingernail polish appeared to provide sufficient protection. 4. Ceramco gold exhibited more resistance to the action of both electrochemical milling and aqua regia etching than did the "C" bridge alloy. 5. Aqua regia etching required more time than electrochemical milling on the two types of gold tested.


Design and technique variables affecting fracture resistance of metal-ceramic restorations. An attempt was made in this study to make a clinically meaningful measure of fracture resistance in metal-ceramic restorations. Forty-four solid metal-ceramic crown forms were fabricated and subjected to compressive load testings. Variables included the presence or absence of a metal coating agent, the type of metal preparation (using stones of different abrasives), and three designs of the underlying metal. The following conclusions were arrived at: (1) The design of the underlying metal structure had a significant relation to the ultimate fracture strength. (2) A design with a definite acuteness of the underlying metal structure failed at significantly lower ultimate fracture strengths. (3) A metal conditioning agent did not decrease fracture resistance if applied properly. (4) Fracture strength was severely decreased when (a) improper thickness of the coating agent was used and (b) porcelain was fused to an unoxidized metal surface. (5) Bond strength, although a contributing factor, may not be as important as metal design and proper manipulation of materials during fabrication of the restoration.


A technique for autogenous root transplantation. A technique of surgically creating a socket and transplanting an autogenous root to provide a stable abutment for a fixed partial prosthesis was described. Radiographic evidence and the demonstration of physiologic mobility of the transplanted root at a three and a half year follow-up of one patient give an indication of the development of a periodontal ligament. One transplant has successfully served as an abutment for seven years.


Reproducing the vertical-lateral defect space in obturator construction. A method of incorporating vertical and horizontal tissues in an obturator impression using an open palate impression tray is presented. The laboratory technique of obturator construction from this impression allows for a definitive (heat-cured) or interim (cold-cured) prosthesis for an edentulous or dentulous patient.


Prosthodontic aspects of palatal elevation and palatopharyngeal stimulation. The prosthodontic aspects of palatal elevation and palatopharyngeal stimulation were discussed. Following are important factors that must be considered. (1) Elevation of the soft palate should be gradual to avoid placing pressure upon the teeth retaining the prosthesis and to reduce mucosal irritation. (2) Prosthetic stimulation should be initiated as soon as palatal paralysis is noted to prevent palatal disuse atrophy. (3) The palatal lift prosthesis may be used as a temporary or definitive treatment for palatal incompetency. When adequate elevation of the soft palate has been achieved, the prosthesis may be discarded. Otherwise, the patient would wear the prosthesis as a permanent supportive device. (4) The construction of the combined palatal lift/pharyngeal section prosthesis includes the gradual palatal elevation and molding of the pharyngeal section to reduce the gag reflexes and to increase palatopharyngeal muscle adaptation to the prosthesis. After the initial placement, adjustment to the pharyngeal section is easier for the patient. (5) Speech and myofunctional therapy should be instituted in conjunction with prosthetic treatment. (6) The palatal lift and combination prostheses are more effective for patients with less severe neurologic impairment and speech articulatory problems. (7) The palatal lift prosthesis is more effective for those patients with palatal incompetency who have no involvement of the other oropharyngeal muscles. The combination type of prosthesis is more effective for patients with palatopharyngeal insufficiency without marked speech articulatory disorders. There are still several questions which require further investigation. These include: (1) What is the relationship between the palatal stimulation and the degree of neuromuscular function and recovery? (2) What is the relationship between the palatal stimulation and the degree of palatal disuse atrophy? (3) What is the relationship between pharyngeal stimulation and muscle contraction? (4) What is the degree of stability of palatopharyngeal function and muscle contraction after stimulation?


Evaluating refractory cast wax-ups for removable partial dentures. A review of the waxed refractory cast and blocked-out master cast by the dentist is an effective method for controlling the quality of framework castings. Major errors in wax-ups have been shown to occur frequently, but they are corrected easily if detected prior to casting. This critique can be incorporated readily into the private practice of dentistry.


A systematic approach to the control of esthetic form. A systematic, orderly approach to the problem of establishing harmonious phonetics, esthetics, and function in fixed restorations has been described. The system requires an initial investment of time in performing an adequate diagnostic waxing, but recoups that time in many clinical and laboratory procedures. The method has proved a valuable asset in fixed prosthodontic care. The technique can be expanded and combined with other techniques with a little imagination and artistic bent.


The effect of varnish on dentinal bonding strength of five dental cements. 1. The following dental cements are listed in order of decreasing mean bonding strenghts on varnish-free dentin: polycarboxylate, zinc phosphate, EBA, Fynal, and CBA. 2. The application of either Copalite or Varnal decreased the dentinal bonding strength of polycarboxylate and zinc phosphate cements. There was no change for CBA cement. 3. The application of Copalite and Varnal increased the dentinal bonding strength for Fynal to the level of zinc phosphate cement on varnish-free dentin. 4. The dentinal bonding strength of EBA cement increased with the application of Varnal but not with Copalite.


Accuracy of six elastic impression materials used for complete-arch fixed partial dentures. 1. The accuracy of four types of impression materials used to make a complete-arch fixed partial denture was evaluated by visual comparison and indirect measurement methods. 2. None of the tested materials allows safe finishing of a complete-arch fixed partial denture on a cast poured from one single master impression. 3. All of the tested materials can be used for impressions for a complete-arch fixed partial denture provided it is not finished on one single cast. Errors can be avoided by making a new impression with the fitted castings in place. Assembly and soldering should be done on the second cast. 4. In making the master fixed partial denture for this study, inaccurate soldering was a problem that was overcome with the use of epoxy glue. Hence, soldering seems to be a major source of inaccuracy for every fixed partial denture.


Effect of continuous infusion of gonadotrophin-releasing hormone in ewes at different times of the year. Gn-RH (3 mug/hr for 8 hr) evoked a surge of LH in ewes during the anoestrous season which was of similar peak height to that found at oestrus but of shorter duration. When Gn-RH was given on 3 consecutive days, the response was considerably reduced on the 2nd and 3rd day. Follicles grew as a result of Gn-RH infusion but peripheral plasma oestrogen concentration did not increase. During the anoestrous season 9/18 ewes ovulated but only 1/6 did so at mid-anoestrus. Mature follicles or a CL were found in 15/18 ewes that had increased peripheral plasma progesterone concentrations for 1-5 days (11 ewes) or 9-14 days (4 ewes). The concentrations of progesterone found were always lower than those observed during the normal cycle. In the breeding season 10/11 ewes ovulated but there was no evidence that the induced CL were maintained.


The interrelation of LH-RH, ovarian steroids and the adenohypophysis. In this presentation experimental evidence is reviewed and new information is presented concerning the question of cyclic fluctuations in pituitary responsiveness to endogenous and exogenous hypothalamic-releasing hormones as well as the direct role of steroids in modulating pituitary sensitivity. Experiments are described which show that four-day and five-day cyclic rats respond differently to LH-RH, presumably because of differences in secretion patterns of ovarian steroids. The direct role of these steroids in this process was investigated both from the morphological (light and electron microscopy) and physiological (radioimmunoassay measurement of LH) viewpoint in hypophysectomized rats bearing pituitary homografts. It was found that steroids may directly alter both the morphology and the secretory function of pituitary gonadotrophs. Estradiol was found to depress the amount of LH that was secreted while progesterone was seen to influence primarily the time of LH release following LH-RH injection. Thus, it appears that direct feedback effects of ovarian steroids at the level of the pituitary are important regulators of the sensitivity of pituitary gonadotrophs to the LH- releaseing action of hypothalamic LH-RH.


The acute effects of Dramamine on uterine contractability during labor. The acute effects of intravenously administered Dramamine or placebo on uterine activity were studied in 32 primigravida patients during the active phase of spontaneous labor. Uterine activity was monitored by invasive quantitative techniques. The results showed no change in uterine activity in the placebo group, but there was a significant increase in activity in the Dramamaine-treated women. The increased activity was primarily the result of an increase in the frequency of contractions.


African swine fever: an epizootiological review with special reference to the South African situation. The most important characteristics and the distribution of the viruses of African swine fever and hog cholera are reviewed. Both viruses were probably present simultaneously in South Africa in the first two decades of the century. While hog cholera was eradicated by 1918, African swine fever persists to the present day because it has a different epizootiology. The role played by wild pigs and the argasid tick (Ornithodoros moubata porcinus) in the epizootiology of African swine fever is discussed and an account of the outbreaks of the disease in South Africa from 1926 to 1974 is given. It appears that the disease in the Transvaal has had a cyclic occurrence.


Antimalarials. 8. Synthesis of amino ethers as candidate antimalarials. Based upon the antimalarial activities demonstrated by compounds I and II a series of amino ethers represented by structures III-VI was synthesized. These structures incorporated several modifications of compound II. The compounds prepared displayed no activity in either the Rane P. berghei mouse screen or the Rane P. gallinaceum sporozoite-induced chick test.


Antimalarials. 11. 2-Vinylogs of substituted 2-aryl-4-quinoline amino alcohols. 3-(P-Chlorobenzylidene)-5, 7-dimethyl-2, 3-dihydro-1H-cyclopenta[b]quinoline-9-(di-n-butylaminomethyl)methanol and 2-[beta-(p-chlorostyryl)]-6, 8-dimethylquinoline-4-(di-n-butylaminomethyl)methanol were synthesized from 6, -8-dimethyl-4-hydroxycarbostyril by 3, 3-dichlorination, dimethoxylation th the 3-ketal, basic hydrolysis to the glyoxal acetal, Pfitzinger condensation with cylopentanone or acetone to the 2, 3-trimethylene or 2-methylquinoline, condensation with p-ClPhCHO at the 2-methylene or 2-methyl group, hydrolysis to the 4-quinaldehyde, methylenation to the epoxide, and condensation with Bu2NH. Both were curative against P. berghei in mice. The first was the more effective: active at 10 mg/kg, completely curative at 40 mg/kg, and only mildly phototoxic in animals.


Synthesis and microbiological activities of beta-(1-chloro-2-naphthyl) alanine and beta-(1-bromo-2-naphthyl) alanine. beta-(1-Chloro-2-naphthyl)alanine and beta-(1-bromo-2-naphthyl) alanine were synthesized by ammonolysis of the corresponding alpha, 1-dihalo-2-naphthalenepropanoic acids derived from 1-nitro-2-naphthylamine by diazotization and condensation with acrylic acid in the presence of cuprous halides. The two analogs as well as the previously reported beta-(2-naphthyl)alanine and beta-(1-naphthyl)alanine were studied as growth inhibitors of Escherichia coli 9723, Leuconostoc dextranicum 8086, and Lactobacillus plantarum 8014. In general, the chloro and bromo analogs were more effective than the unsubstituted naphthylalanines as growth inhibitors of the three microorganisms studied.


Dyskeratosis congenita: clinical features and genetic aspects. Report of a family and review of the literature. A large family with dyskeratosis congenita is reported. There were nine affected males, the findings in five of who are reported. We review 46 cases selected from the literature. The cardinal findings of this inherited multisystem disorder are delineated from these 51 cases. The complications of the disease, including opportunistic infection, are described. The parallel is made between dyskeratosis congenita and Fanconi's anaemia. The X-linked transmission of dyskeratosis congenita is confirmed by the family pedigree in this report. From the analysis of the families reported in the literature, there appears to be genetic heterogeneity in this disease. This study in our family indicates absence of close linkage between the Xga locus and the X-linked recessive form of dyskeratosis congenita.


Genetics of the complement system.

Genetics of the complement system. The complement system, unlike the coagulation system, was largely characterized by in-vitro techniques which did not make use of genetically deficient plasmas. The existence of the genetically deficient plasmas. The existence of the genetically deficient subjects therefore has served largely to increase our knowledge of the in-vivo role of complement. At the present time its clearest role is in the resistance to infection; obviously in the case of C3 deficiency and bacterial infection and possibly more subtly in the case of deficiency of the early active complement components and low virulence organisms. There is so far no evidence that genetic complement deficiency interferes with antibody formation or with the generation of tolerance as has been suggested in the pas (Azar et al, 1968; Dukor and Hartmann, 1973).


Communicating drug information to physicians. In this paper the authors present a review of the multitudinous studies of researchers who have attempted to identify the sources of drug information utilized by physicians. Consideration is given to both professional sources (for example, journal articles, colleagues, meetings, and pharmacists) and commercial sources (for example, detailmen, journal advertisements, mail advertisements, and samples). Also included is a discussion of the relationship between physicians' sources of drug information and their choice of drugs and of the relationship between the sources of information and the severity of the conditions being treated. The paper concludes with a brief review of some of the new sources of drug information being considered for utilization by the Food and Drug Administration.


Spinal nerves and their bearing on salamander phylogeny. Examination of the vertebral columns of representatives of all families of salamanders revealed that, in contrast to the condition found in most other vertebrates, salamander spinal nerves of often pass through foramina in the vertebrae. Two kinds of spinal nerve foramina were found: those in the anterior halves of vertebrae, and those in the posterior halves. In addition, many salamanders retain intervertebral nerves. However, within each family or, in a few cases, subfamily there is a characteristic pattern of spinal nerve-vertebral relationships. The first spinal nerve of all salamanders exits through a foramen in the anterior half of the atlas. All more posterior nerves are intervertebral in the families Cryptobranchidae, Hynobiidae and Proteidae. The posterior caudal nerves exit through the posterior halves of the caudal vertebrae in the family Amphiumidae, while in the subfamilies Dicamptodontinae and Rhyacotritoninae all post-sacral nerves exit through the posterior halves of the vertebrae. All but the first three nerves exit through posterior foramina in the family Plethodontidae and the subfamily Ambystomatinae, while all but the first two nerves pass through posterior foramina in the families Salamandridae and Sirenidae. Several fossil salamanders were also examined. These showed that the amphiumid and dicamptodontine-rhyacotritonine nerve patterns had evolved by the Late Cretaceous, and the sirenid pattern had probably evolved by that time. Other Cretaceous genera associated with the Ambystomatoidea still possessed the primitive intervertebral pattern. Using spinal nerve patterns and several other previously described morphological characters, a new hypothesis of the phylogeny of recent and fossil salamanders is presented and compared to earlier proposed phylogenies of the group. A new classification of salamander families is presented.