Hemagglutination of human group A erythrocytes by enterotoxigenic Escherichia coli isolated from adults with diarrhea: correlation with colonization factor. Enterotoxigenic Escherichia coli (ETEC) of several different serotypes isolated from adults with diarrhea and known to possess the colonization factor antigen (CFA) were found to cause mannose-resistant hemagglutination (HA) of human group A erythrocytes. CFA-negative E. coli isolated during the same study did not possess the mannose-resistant hemagglutinin, although some non-ETEC, CFA-negative isolates did exhibit mannose-sensitive HA activity. The mannoseresistant hemagglutinin of ETEC was found to possess many characteristics previously associated with CFA, which is a surface-associated fimbriate heatlabile antigen, and the functionally and morphologically similar K88 and K99 antigens of animal-specific ETEC. Mannose-resistant HA and CFA titers were maximal when ETEC cells were grown on an agar medium (CFA agar) composed primarily of 1% Casamino Acids and 0.15% yeast extract, pH 7.4. Neither CFA nor HA were produced at a growth temperature of 18 degrees C; HA was completely inhibited by pretreatment of CFA-positive cells with the anti-CFA serum. The mannose-resistant hemagglutinin was lost spontaneously and simultaneously with CFA when clinical ETEC isolates were passaged on artificial medium in the laboratory, indicating plasmid control of both entities. The mannose-resistant hemagglutinin of ETEC was shown to be thermolabile, i.e., sensitive to heating at 65 degrees C, as was the CFA. Also, there was correlation between possession of CFA, as detected serologically and by demonstration of biological activity (adherence in the infant rabbit small intestine), presence of CFA-type fimbriae, and the ability of various E. coli isolates to cause mannose-resistant HA of human group A erythrocytes. These results indicate that the mannose-resistant HA of ETEC is another manifestation of CFA.


Protection against challenge with Escherichia coli heat-labile enterotoxin by immunization of rats with cholera toxin/toxoid. Rats were used to study dose-response curves to intestinal challenge with cholera toxin or Escherichia coli heat-labile enterotoxin. Prior immunization with cholera toxin/toxoid induced significant and equal protection against both toxins.


Escherichia coli heat-labile enterotoxin: comparison of antitoxin assays and serum antitoxin levels. The mouse adrenal tumor cells (Y-1 strain) and the Chinese hamster ovary cells, two routinely used tissue culture assays for Escherichia coli heat-labile enterotoxin (LT), were used to detect serum antitoxin responses in culture-positive patients from several well-defined sources. There was no correlation between a significant antitoxin response and isolation of LT-producing E. coli in two "domestic" diarrheal outbreaks. Serum samples from a third group of individuals in a rural cholera-endemic area consistently demonstrated significant rises in neutralizing antibody to LT.


Suppression of immunity to Mycobacterium lepraemurium infection. After injection of 10(8) live Mycobacterium lepraemurium (MLM) into the left hind footpad of mice, there is development of local swelling attributable to a granuloma of the cell-mediated immunity type. Concomitant intravenous inoculation of live MLM delays and may even suppress footpad swelling, the effects being proportional to the intravenous dose of organisms. Concomitant footpad infection and intravenous inoculation of 10(9) dead MLM also delays footpad swelling, but over a period of months the feet become excessively swollen. The excessive swelling is due to local enhancement of infection as evidenced by an increase in the number of MLM per footpad. Attempts were made to prevent such immunosuppression by splenectomy or treatment with BCG. Splenectomy was entirely without effect, but 10(7) live BCG administered intravenously 2 to 4 weeks before dead MLM prevented enhancement of infection. The mediator of the immunosuppressive mechanism that results in enhanced infection remains to be elucidated, but it is unlikely to be antibody or immune complexes.


Isolation and properties of complement-resistant strains of Escherichia coli K-12. Several strains that were resistant to the bactericidal action of antibody and complement were isolated from Escherichia coli K-12 W3110/SM by selecting them through the medium containing antiserum and complement. They can be agglutinated by antiserum against the parent strain and showed similar immune adherence reactivity to the parent when sensitized with this antiserum. Few differences were found in the compositions of phospholipids and proteins between both inner and outer membranes of these strains and those of the parent. However, there were fewer short-chain and more long-chain fatty acids in these strains than in the parent. It was also found that unsaturated fatty acide decreased and saturated and cyclopropanoic acids increased in phosphatidylethanolamine and phosphatidylglycerol in both inner and outer membranes of one of these strains when compared with those from the parent. Therefore, the resistance of these strains to the complement-mediated bactericidal action was considered to be due to the rigidity of their membrane structures which might repel the insertion of membrane-attack complement complex C5b-9, although they could fix the earlier complement components up to the step of the formation of C4b,2a,3b complex enzyme.


Mouse virulence of Salmonella typhimurium mutants deficient in two major outer membrane proteins. The role of several outer membrane components as virulence factors is well established. We have now isolated spontaneous mutants and conjugational hybrids of smooth mouse virulent Salmonella typhimurium deficient in two major outer-membrane proteins. The lack of the 34,000- and/or 36,000-dalton proteins was confirmed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Mutants and recombinants were then tested for their mouse virulence by intraperitoneal inoculation and were found to be as virulent as the parent strain.


Nocardia infections in congenitally athymic (nude) mice and in other inbred mouse strains. The mortality rate and histopathological features of Nocardia asteroides and Nocardia brasiliensis infections in congenitally athymic (nude) mice of ICR and C3H/eB origins were quite different from what we found for Swiss white mice and other inbred mouse strains (namely, C57/BL/6J, New Zealand Black, BALB/c, CBA/LAC, and C3H/eB). The immunocompetent littermates of the congenitally athymic mice occupied an intermediate position between their athymic siblings and Swiss white mice in terms of their responses to both these organisms. Macrophage ingestion and destruction of N. brasiliensis, as demonstrated by electron microscopy, was found to occur. The T-lymphocyte appears to be an essential component in normal mouse resistance to infection by both N. asteroides and N. brasiliensis.


Characterization of the attachment of Treponema pallidum (Nichols strain) to cultured mammalian cells and the potential relationship of attachment to pathogenicity. The interaction of Treponema pallidum (Nichols strain) with 19 different cultured mammalian cell types was examined. These types included cells derived from testis, kidney, spleen, lung, epidermis, cervix, urethra, and nerve tissue of human, rabbit, or rat origins. They represented normal and malignant cells, epithelial and fibroblastic morphology, cell lines, and cell strains, Large numbers of organisms attached to the cultured cells; this attachment prolonged the time of retention of active treponemal motility. Attachment was examined in terms of the number of treponemes inoculated, cultured cells present, and actively growing versus stationary cultured cells; the motility of the treponemes; the viability of the cultured cells; and the different cell passages. In sharp contrast to the attachment of T. pallidum, 11 nonpathogenic treponemes failed to attach to cultured cells. Immune syphilitic rabbit serum prevented the attachment of T. pallidum to cultured cells, as indicated by phase contrast microscopy and rabbit inoculations. This blockage of attachment by immune serum occurred without interfering with active motility of the organisms. Results are discussed in terms of the potential relationship of attachment to the pathogenicity of T pallidum.


Toxin from the culture filtrate of Shigella dysenteriae that causes morphological changes in Chinese hamster ovary cells and is distinct from the neurotoxin. A toxin that causes morphological changes in Chinese hamster ovary cells was isolated from a culture filtrate of Shigella dysenteriae 1. It was partially purified by successive column chromatography on diethylaminoethyl-cellulose, Bio-Gel A-5m, and hydroxylapatite. The toxin was separated from neurotoxin by diethylaminoethyl-cellulose column chromatography and was not lethal for mice.


Adherence pharyngeal and skin strains of group A streptococci to human skin and oral epithelial cells. Group A streptococci isolated from skin adhere in greater numbers to human skin epithelial cells than to cells obtained from buccal mucosa whereas streptococci isolated from a throat tend to adhere in greater numbers to buccal epithelial cells than to skin epithelial cells in vitro. M protein-producing strains of group A streptococci did not adhere in significantly greater numbers than M-negative strains. Lipoteichoic acid inhibited binding of streptococci to skin epithelial cells as well as was previously shown for oral epithelial cells. Our results suggest that lipoteichoic acid is more centrally involved than M protein in binding streptococci to skin and mucosal surfaces.


Antigenic cross-reactivity between Mycobacterium bovis (BCG) and Leishmania donovani. Antigenic cross-reactivity was demonstrated between Mycobacterium bovis (BCG) and Leishmania donovani, using delayed hypersensitivity as a criterion.


Surgical pocket therapy.

Surgical pocket therapy. (1) Periodontal pockets can be treated successfully by several modalities of surgery. (2) Periodontal pocket walls do not have to be eliminated surgically in order to stop the progress of periodontal disease. (3) A residual defect which can be probed beyond 3 mm at the site of a previous peridontal pocket, does not signify a progressive lesion as long as it does not appear inflamed or contain, exudate and bleeds when probed. (4) The most important aspect of all pocket therapy is to make the exposed root surface biologically acceptable to the surrounding soft tissues. (5) The gingiva should be so closely adapted to the teeth at the end of the healing following treatment that subgingival plaque does not colonize to the extent of being recognizable clinically. (6) Frequent professional prophylaxis is of paramount importance for success of all pocket therapy.


The causes of heroin addiction. A review of the literature. Part I. The causes of heroin addiction have been studied and debated, and many theories have evolved. This article reviews the extensive literature on the subject and groups the material under the categories of Psychological, Pharmacological, Genetic, Socilogical, Social Protest, The Family, and Stepping Stone theories.


Evaluation of the immune response in mice infected with Mycobacterium marinum. Foot pad infection of mice with Mycobacterium marinum carried out with a view to comparing the immune response on the humoral level of such mice, with that observed previously in mice infected with M. leprae, indicated that there was a similarity in terms of the first appearance and proliferation of immunocytes and the time at which the peak and decline in the antibody-producing cells occurred. The significant difference appeared to be in the immunoglobulin G response, which was absent in the M. leprae infected mice, but occurred simultaneously with the immunoglobulin M response at a high level, both during a primary and after a secondary challenge administered 15 days post-primary infection in the M. marinum infected mice. Further confirmation was obtained through additional studies on the specific immunoglobulin levels and determination of both immunoglobulin M and immunoglobulin G antibodies by hemagglutination. Although the growth temperature requirement of the two organisms and their ability to initiate infection in areas of the body with lower temperatures are similar, it is suggested that the type of infection induced by each one of these species in the mouse may disallow the serious consideration of the M. marinum infection model as a possible alternative experimental model for studying the role of host immunity to M. leprae infections in mice.


Are all nonsolid Mycobacterium leprae dead? Does a negative finding in the mouse foot pad indicate that there is actually no growth of M. leprae in the animals? Ever since the discovery of the foot pad technic for growth of M. leprae in mice, investigators have overemphasized the laboratory results in clinical applications. Overenthusiasm has led to some dire results in the leprosy field. Two well-known examples can be cited, which are based on the presumption that: a) all the nonsolidly stained M. leprae are dead, and b) that a negative finding in the mouse foot pad indicates no growth of M. leprae in the animals. The former led clinical investigators to claim a false emergence of drug resistance after one year's treatment with a potent antileprosy drug, B663, which was almost abandoned for later clinical use. The latter led investigators to introduce a low-dose drug treatment, which resulted in a worldwide appearance of DDS resistance in leprosy. This paper outlines the reasoning that not all nonsolid M. leprae are dead, and that not all the organisms in the foot pads are detectable by the present standard foot pad/ M. leprae technic.


Freeze-etching study of human and murine leprosy bacilli. Morphologic features of the electron transparent zone (ETZ) material around human and murine leprosy bacilli were examined by a freeze-etching technic. The ETZ around human leprosy bacilli is composed of spherical droplets of hydrophobic material. These are always liquid at body temperature and they never show crystalline lamellar structure even at the temperature of liquid nitrogen. The ETZ around murine leprosy bacilli is composed of ribbon-like or membranous crystalline structures. This material is solid and crystalline at the body temperature of mice, and this solid material is the chief cause of the random arrangement of murine leprosy bacilli inside the cytoplasm of murine lepra cells. This crystalline structure has also been observed around murine leprosy bacilli grown on cell-free culture media.


Culture and phagocytic characteristics of Schwann cells in vitro. A possible model substrate for cultivation of M. leprae. Tissue cultures of five acoustic and four spinal schwannomas demonstrated good growth and migration of Schwann cells within two to four days. Two types of cells corresponding to Antoni type A tissue and Antoni type B tissue were clearly recognized. Both these cell types showed avid phagocytosis when the cultures were inoculated with mycobacteria, either ICRC bacilli or M. leprae. The phagocytic index was 95% at the end of two hours. The Schwann cells grown in vitro also showed intense acid phosphatase reaction with Gomori's stain, suggesting lysosomal activity. Neither this nor any phagocytosis was evidenced by fetal fibroblasts cultured similarly. Cells from two spinal neurofibromas grew and migrated slowly in vitro and were mainly fibroblasts.


Radiation induced alterations in the endotoxin of S. typhimurium. The lipopolysaccharide (LPS) of S. typhimurium has been shown to be significantly detoxified after in vivo irradiation at 500 krad. Radiation is thus a useful method for converting endotoxin into toxoid. The structural alterations in the detoxified LPS are shown to be mainly in the lipid A molecule, resulting in the loss of beta-hydroxymyristic acid.


Sensitization of Escherichia coli C to gamma-radiation by 5-bromouracil incorporation. Escherichia coli C cells, unifilarly substituted with 5-bromouracil (BrUra) were 2-25 times as sensitive as unsubstituted cells to killing by gamma-irradiation under aerobic conditions. The yield of DNA double-strand breaks in BrUra-substituted cells was increased by a factor only 1-55, suggesting that other lesions also contribute to cell-killing. Alkaline sucrose density gradient analysis of the 3H-thymine labelled DNA strand showed there was less repair of gamma-ray-induced single-strand breaks when BrUra was in the complementary strand. Since there are  more of these unrepaired breaks than can be accounted for by BrUra-induced DNA double-strand breakage, some fraction of the lethal events in BrUra-substituted E. coli cells may be unrepaired DNA single-strand breaks.


Bacterial radiosensitization by 8-methoxypsoralen. 8-Methoxypsoralen has been shown to act as a radiosensitizer of hypoxic bacterial cells with uvrA, recA and uvrB and/or lexA mutations. No effect of the drug on the radiosensitivity of oxic bacteria with these mutations was observed. This drug differs from O2 and electron-affinic radiosensitizers in that its effect is not purely dose-modifying and can exceed the oxygen effect in certain mutants.


Radiosensitization of Serratia marcescens by bipyridinium compounds. Bipyridinium compounds (viologens) have been shown to  radiosensitize hypoxic Serratia marcescens cells by two components. These can be separated on the basis that only the one-electron reduced form of the compounds can penetrate the bacterium cell wall. One component is  associated with sensitization at the membrane and the other with an internal site. The efficiency of sensitization at the membrane-associated site follows the order of increasing one-electron reduction potentials of the compounds. The one-electron reduced forms of the bipyridinium compounds are involved in a mechanism that reduces the initial level of sensitization. No additivity in sensitization is found on combining the bipyridinium compounds with other radiosensitizers, PNAP and Ro 07-0582 at concentrations of each, which  will give sensitization to the level associated with the membrane site. It is concluded that all these  electron-affinic compounds sensitize this site. The protective effect of added glycerol on sensitization by viologens is related to protection at the membrane-associated site.


A mathematical description of causative factors and prevention of elevated intraocular pressure after keratoplasty. In keratoplasty with grafts the same size as the recipient bed, tight sutures and thick recipient corneal periphery distort the angle and may collapse the filtering meshwork. This can cause very high postoperative pressures, which can be avoided by the use of donor grafts larger than the recipient bed. These relationships can be mathematically predicted.


Steroid-induced glaucoma and cataract in renal transplant recipients. Fifteen kidney transplant recipients were studied ophthalmologically for periods of one to five years following the transplantation. All subjects received maintenance immunosuppressive therapy which included azathioprine (IMURAN) and corticosteroids. Two patients developed posterior subcapsular cataracts. Seven patients (47%) developed increased intraocular pressure and one of these had glaucomatous field loss and disk cupping. The increased intraocular pressure responded favorably to topical treatment while the patients were under continuous treatment with systemic steroids and immunosuppressive drugs.


Ultrastructural localization of a new neuronal peptide (VIP). The vasoactive intestinal polypeptide (VIP) represents a neuronal peptide of wide-spread occurrence. By electron microscopic immunocytochemistry VIP has been localized to the granules of the terminals of p-type neurons. This localizations lends support to the postulate that VIP may be a neurotransmitter.


Reduction of Salmonella and fecal contamination of pork during swine slaughter. Experiments were designed to reduce Salmonella and fecal contamination of pork, using several sanitizing methods and agents. Sanitizing the hauling vehicle and holding pens with chlorine or quaternary ammonium compounds proved ineffective in reducing carcass contamination. When the eviscerator wore a plastic glove and sanitized his knife in 82-C water before using it on each carcass, contamination was reduced about 50%. Sanitizing the eviscerator's knife in 500-ppm chlorine solution adjusted to a pH of 6.0, or in 25-ppm iodine solution reduced contamination approximately 75%.


Salmonella-associated conjunctivitis in a cat. Salmonella typhimurium-associated conjunctivitis in an adult female cat was characterized by epiphora and blepharospasm. The cat also harbored intestinal S typhimurium, as did one other cat in the same shipment of 7 cats obtained from a commercial cat dealer. The 3 isolates were resistant to ampicillin, cephaloridine, cephalothin, streptomycin, triple sulfonamides, and tetracycline. All 3 isolates were capable of transferring a part of the antibiotic resistance pattern to an Escherichia coli K-12 recipient. Salmonella were not recovered from the conjunctiva or fecal flora of either cat after 10 days of therapy with chloramphenicol.


Yersiniosis in captive exotic mammals.

Yersiniosis in captive exotic mammals. Within a 2 1/2-month period, Yersinia pseudotuberculosis infection occurred in 3 blesbok (Damaliscus dorcas), 1 dik-dik (Madoqua kirkii), and 1 giant anteater (Myrmecophaga tridactyla) at the National Zoological Park, Washington, DC. Lesions consisted of fibrinonecrotic enteritis and peritonitis, mesenteric lymphadenitis, and embolic pyogranulomatous lesions in the liver, spleen, and lungs. Feed contaminated with the feces of wild rats and pigeons was thought to be the source of infection.


Uptake of 2'-amino-2'-deoxyguanosine by Escherichia coli and its competition by guanosine. The antibacterial activity of a new nucleoside antibiotic, 2'-amino-2'-deoxyguanosine (2AG), is reversed by guanosine and other purine nucleosides. 2AG is apparently taken up by E. coli by a mechanism different from that of guanosine; guanosine inhibits this uptake non-competitively. Insensitive E. coli strains and the resistant mutant obtained from the sensitive strain also took up 2AG.


Mechanism of growth inhibition by 2'-amino-2'-deoxyguanosine in Escherichia coli. The mechanism of inhibition of Escherichia coli by the new nucleoside antibiotic, 2'-amino-2'-deoxyguanosine (2AG), is described. Upon the addition of 2AG, the syntheses of macromolecules continued for 15 minutes. After this lag time, protein synthesis sharply decreased, Rna synthesis slightly decreased, but DNA synthesis was not affected. Tritiated 2AG was readily incorporated into the acid-soluble fraction of cells in the form of the mono-, di- and triphosphates. In the acid-soluble fraction, radioactivity was found only in the RNA fraction. The major part of the radioactivity was found to be guanylate; only 25% existed as the nucleotide of 2AG. In resistant strains of E. coli, there was a lower degree of phosphorylating activity and less incorporation of 2AG into RNA per unit of cell mass increase. These results suggest that 2AG inhibits growth by its incorporation into RNA and the subsequent distrubance of RNA function causing a block in protein synthesis.


Differentiation of rennet from other milk-clotting enzymes by polyacrylamide gel electrophoresis. Polyacrylamide gel electrophoresis was used to differentiate animal rennet and other milk-clotting enzymes. After electrophoresis, the separated components were visualized by staining with aniline blue-black. Two prominent proteins were found in calf and bovine rennet, while only 1 major protein was observed in pepsin and enzymes of microbial origin. These patterns provided a basis for distinguishing animal rennet and the other enzymes as well as a means of identifying each type of enzyme by the characteristic pattern shown.


Effects of a protease inhibitor on biosynthesis of Escherichia coli proteins. A protease inhibitor, tosyl-L-lysine chloromethylketone, altered the composition of newly synthesized protein in Escherichia coli; a significant portion of the proteins synthesized sedimented rapidly, presumably forming aggregates of the abnormal proteins. Protein I of the outer membrane was only slightly synthesized under these conditions.


Reconstitution and purification of flagellar filaments from Caulobacter crescentus. Filaments from isolated flagella of Caulobacter crescentus have been purified by successive dissociation and reconstitution. After the second and third reconstitutions from subunits in 0.8 M sodium citrate, filament preparations contained only two proteins, flagellin A (26,000 daltons) and flagellin B (28,000 daltons). There was some enrichment for flagellin A during reconstitution by this procedure, since isolated flagella contained flagellin A and flagellin B in a ratio of approximately 3.8:1 and filaments after the third reconstitution contained the two proteins in a ratio of 5.0:1.


Anaerobic electron transport in anaerobic flagellum formation in Escherichia coli. Flagellum formation by ubiquinone- and menaquinone-deficient mutant strains of Escherichia coli K-12 was studied under both aerobic and anaerobic growth conditions. Ubiquinone was found to be obligatory for aerobic flagellum formation but could be replaced by menaquinone for anaerobic flagellum formation. A mutant devoid of both quinones was immotile aerobically as well as anaerobically. Hence, the respective electron transport system is obligatory for flagellum formation in Escherichia coli.


Selection of yeast auxotrophs by thymidylate starvation. A rapid procedure for the recovery of Saccharomyces cerevisiae auxotrophs was developed by exploiting the protection of these mutants from thymineless death when a required metabolite was withheld. The method can be used for thymidine 5'-monophosphate-requiring auxotrophs or wild-type strains blocked in de novo synthesis of thymidylate by folate antagonists.


Formation of HfrH-type donor cells as a result of integrative suppression by R-F recombinant plasmids. Three recombinant plasmids, resulting from recombination between an R plasmid of the FI incompatibility group and the F of HfrH, were introduced in a temperature-sensitive dnaA mutant to isolate Hfr-type-donors. All of the temperature-insensitive clones isolated from two of the three recombinant plasmids had the same origin and transfer pattern as the parental HfrH strain.


Isolation of a lambdadcys transducing bacteriophage and its use in determining the regulation of cysteine messenger ribonucleic acid synthesis in Escherichia coli K-12. A defective specialized lambda transducing phage carrying the cysJ, cysI, cysH, and cysD genes has been isolated from a secondary-site lysogen. Deoxyribonucleic acid-ribonucleic acid (DNA-RNA) hybridization studies utilizing this phage have been carried out to detect cysteine-specific messenger RNA (cys mRNA) synthesized in vivo. A vivo. A 3.5- to 9-fold increase in the rate of synthesis of cys mRNA has been detected in the derepressed wild-type (Cys+) strain grown on glutathione compared with a repressed control grown on cystine. Pleiotropic cysE and cysB mutants grown on glutathione were found to possess rates of synthesis of cys mRNA that were significantly lower than their derepressed isogenic parent. The addition of O-acetyl-L-serine to the cysE strain produced a 5.5-fold increase in the rate of synthesis of cys mRNA. These results indicate that cysteine biosynthesis is controlled at the level of transcription by the inducer O-acetylserine, the cysB protein and cyst(e)ine.


Thymineless death in Escherichia coli dnaB mutants and in a dnaB dnaG double mutant. The interference of dnaB mutations of Escherichia coli with thymineless death is described. All the isogenic Thy- dnaB mutants of E. coli we have tested show a remarkable immunity towards cell death induced by thymine deprivation at the nonpermissive temperature. We have also constructed and tested an isogenic double dnaB dnaG mutant. It loses its viability in the absence of thymine at both permissive and nonpermissive temperatures. The role of the dnaB gene product is discussed.


Genetic mapping of the F plasmid gene that promotes degradation of stable ribonucleic acid in Escherichia coli. F+ Escherichi coli cells that contain an srnA mutant allele degrade their stable ribonucleic acid (RNA) extensively after RNA synthesis is blocked at 42 degrees C. The relevant gene promoting degradation of stable RNA, srnB+, or its promoter was mapped between 1.7 and 2.8 kilobases on the F plasmid by using deleted F' plasmids and chimeric plasmids composed of pSC101 and fragments of F plasmid.


Inhibitory action of a non-metabolizable fatty acid on the growth of Escherichia coli: role of metabolism and outer membrane integrity. The inhibitory action of decanoic acid on both Escherichia coli K-12/154 (normal lipopolysaccharide) and E. coli RC59 (defective lipopolysaccharide) was studied. A correlation was found between the doubling time of E. coli 154 growing in different media and the lethal effect of 0.4% decanoic acid on this bacterium. Decanoic acid (0.4%) exerted a lytic action on glucose-starved and NaN3-inhibited cells of E. coli 154 and RC59. Exponentially growing cultures of both strains were not affected by the addition of 0.4% methyldecanoate, but cells of E. coli RC59 reaching the stationary phase were attacked by that compound. A bactericidal action of 0.4% methyldecanoate on exponential E. coli 154 and RC59 was observed when sodium azide was also present in the media. Concentrations lower than 0.01% methyldecanoate had a lytic effect on spheroplasts from E. coli 154 and RC59. These results indicate that the inhibitory action of a non-metabolizable fatty acid on E. coli depends on the cellular metabolic activity and the outer membrane integrity.


Genetic analysis of components involved in vitamin B12 uptake in Escherichia coli. The products of three genes are involved in cyanocobalamin (B(12)) uptake in Escherichia coli. btuB (formerly bfe), located at min 88 on the Escherichia coli linkage map, codes for a protein component of the outer membrane which serves as receptor for B(12), the E colicins, and bacteriophage BF23. Four phenotypic classes of mutants varying in response to these agents were found to carry mutations that, based on complementation and reversion analyses, reside in the single btuB cistron. In one mutant class, ligand binding to the receptor appeared to be normal, but subsequent B(12) uptake was defective. The level of receptor and rate of uptake were responsive to btuB gene dosage. Previous studies showed that the tonB product was necessary for energy-dependent B(12) uptake but not for its binding. Other than those in tonB, no mutations that conferred insensitivity to group B colicins affected B(12) utilization. The requirement for the btuB and tonB products could be bypassed by elevated levels of B(12) (>1 muM) or by mutations compromising the integrity of the outer membrane as a permeability barrier. Utilization of elevated B(12) concentrations in strains lacking the btuB-tonB uptake system was dependent on the function of the btuC product. This gene was located at 37.7 min on the linkage map, with the order pps-btuC-pheS. Strains altered in btuC but with an intact btuB-tonB system were only slightly impaired in B(12) utilization, being defective in its accumulation. This defect was manifested as inability to retain B(12), such that intracellular label was almost completely lost by exchange or efflux. It is proposed that btuC encodes a transport system for B(12) in the periplasm.


Osmotic imbalance in inositol-starved spheroplasts of Saccharomyces cerevisiae. Physiological states associated with inositol starvation of spheroplasts of Saccharomyces cerevisiae were investigated and compared with conditions preceding death of starved whole cells. In the absence of synthesis of inositol-containing lipids, cell surface expansion terminated after one doubling of whole cells. In spheroplasts, cessation of membrane expansion was apparently followed by rapid development of an osmotic imbalance, causing lysis. Continued synthesis and accumulation of cytoplasmic constituents within the limited cell volume were implicated as a cause of the osmotic imbalance. In whole cells, an increase in internal osmotic pressure also follows termination of membrane and cell wall expansion. The cell wall prevents lysis, allowing a state of increasing cytoplasmic osmotic pressure to persist in the period preceding onset of inositol-less death.


Isolation of a metK mutant with a temperature-sensitive S-adenosylmethionine synthetase. An Escherichia coli metK mutant, designated metK110, was isolated among spontaneous ethionine-resistant organisms selected at 42 degrees C. The S-adenosylmethionine synthetase activity of this mutant was present at lower levels than in the corresponding wild-type strain and was more labile than the wild-type enzyme when heated or dialyzed. A mixture of mutant and wild-type enzyme preparations had an activity equal to the sum of the component activities. These facts strongly suggest that the mutated gene in this strain is the structural gene for this enzyme. Genetic mapping experiments placed the metK110 mutation near or at the site of other known metK mutants (i.e., 63 min), confirming its designation as a metK mutant. A revised gene order has been established for this region, i.e., metC glc speC metK speB serA.


Characterization of fusions between the lac operon and the ilv gene cluster in Escherichia coli: ilvC-lac fusions. By means of the general procedure of Casadaban (J. Mol. Biol. 104: 541-556, 1976), the lac genes carried on a lambda-Mu-1 hybrid phage were inserted into a temperature-inducible Mu-1 prophage that had earlier been inserted into a site near the beginning of the ilvC gene of Escherichia coli strain K-12. Selection of temperature-resistant derivatives of the lysogen resulted in a fusion of the lac genes to a region of deoxyribonucleic acid that is transcribed under the control of the ilvC regulatory elements. A strain bearing the fusion was shown to be inducible for beta-galactosidase by acetohydroxybutyrate, a natural inducer of acetohydroxy acid isomeroreductase. Induction of the lysogen by mitomycin C led to the isolation of a plaque-forming lambda derivative carrying this ilvC-lac fusion.


Genetic analysis of the Escherichia coli K-12 srl region. Specialized transducing lambda derivatives, deletion mapping, and Plkc transductional crosses have been used to analyze the genetic organization and regulation of the srl genes. Transducing phages obtained from a secondary site lambda insertion in srlA are of two types: lambdapsrlC1 and lambdaprecA are substituted in the b2 region of the lambda chromosome (galtype) and carry the srlC gene but not srlD; lambdapsrlD is substituted in the early region of the phage deoxyribonucleic acid (biotype) and carries the srlD gene but not srlC. The lambdapsrlC1 phage, which lysogenizes at attlambda, complements srlC mutants in trans, indicating that this gene codes for a diffusable positive regulatory element. The srl genes have been ordered relative to the cysC, recA, and alaS genes by two- and three-factor P1kc crosses. The order, cysC...srlD-srlA-srlC-recA-alaS, has been obtained. The srlA and srlD genes comprise an operon with srlD operator distal. From the secondary site lysogen, it has been possible to obtain deletion mutants of this region that are sensitive to ultraviolet light and are recombination deficient. Genetic evidence suggests that these deletions extend from srl into the recA gene.


Escherichia coli K-12 mutants hyperproducing chromosomal beta-lactamase by gene repetitions. Escherichia coli K-12 ampicillin-resistant mutants hyperproducing chromosomal beta-lactamase arose spontaneously from strains carrying ampA1 ampC(+). Such mutants were found even in a recA background. Two Amp(r)-100 strains were analyzed genetically. The Amp(r)-100 resistance level of both strains could be transduced by direct selection for ampicillin resistance. Several classes of ampicillin-resistant transductants were found that differed from one another in the beta-lactamase activity and the ampicillin resistance mediated by an ampA1 ampC(+)-carrying strain. The data suggested that beta-lactamase hyperproduction was due to repetitions of the chromosomal amp genes. The size of the repeated region was calculated from cotransduction estimates, using the formula of Wu (Genetics 54:405-410, 1966), and was found to be about 1 min in one strain and 1.5 min in the other. Second-step Amp(r)-400 mutants were isolated from an Amp(r)-100 strain. The resistance of these mutants was apparently also due to repetitions, each mediating a resistance to about 10 mug/ml. Mutants of wild-type strains that were moderately resistant to ampicillin also gave rise to intermediate-resistance classes, suggesting repetitions of the wild-type amp alleles. F' factors hyperproducing chromosomal beta-lactamase by gene repetitions were constructed. They mediated levels of ampicillin resistance comparable to that of naturally occurring resistance plasmids. The expression of beta-lactamase hyperproduction was not affected by the presence of ampA and ampC alleles in trans and did not act in trans on the other alleles.


Characterization of hybrid plasmids carrying individual ribosomal ribonucleic acid transcription units of Escherichia coli. We have screened the strains with ColE1 hybrid plasmids constructed by Clarke and Carbon (Cell 9:91-99, 1976) for the presence of ribosomal ribonucleic acid (rRNA) genes on the plasmids and identified 16 strains whose plasmids carry rRNA genes. The structures of these 16 plasmids were compared by heteroduplex analysis, and the plasmids were classified into six groups on the basis of their chromosomal origins. Homology with known transducing-phage deoxyribonucleic acids and genetic mapping have assigned locations on the Escherichia coli chromosome to three of the six groups. These are rrnB near rif at 88 min, rrnC near ilvE at 83 min, and rrnD near aroE at 71 min. A fourth group is probably rrnA at 85 min (T. Ikemura and M. Nomura, Cell, 11:779-793, 1977). We conclude that the minimum number of rRNA transcription units per haploid chromosomes is seven, that is, the six groups identified in this work plus a known operon (rrnE near metA at 89 min) that we failed to find among the hybrid plasmids. This heteroduplex analysis also suggests  that there are only two kinds of rRNA operons with respect to their spacer region; three of the six rRNA operon groups studied here have one kind, whereas the remaining three have the other kind.


Synthesis of mot and che gene products of Escherichia coli programmed by hybrid ColE1 plasmids in minicells. Hybrid Escherichia coli ColE1 plasmids carrying the genes for motility (mot) and chemotaxis (che) were transferred to a minicell-producing strain. The mot and che genes on the hybrid plasmid directed protein synthesis in minicells. Polypeptides synthesized in minicells were identical to the products of the motA, motB, cheA, cheW, cheM, cheX, cheB, cheY, and cheZ genes previously identified by using hybrid lambda and ultraviolet-irradiated host cells (Silverman and Simon, J. Bacteriol. 130:1317-1325, 1977), thus confirming these gene product assignments. The products of some che genes (cheA and cheM) appeared as more than one band on polyacrylamide gel electrophoresis, but analysis of partial peptide digests of these polypeptides suggested that the multiple forms were coded for by a single gene. Measurement of the physical length of the hybrid plasmids allowed an estimate of the amount of coding capacity of the cloned deoxyribonucleic acid, which was devoted to the synthesis of the mot and che gene products. These estimates were also consistent with the hypothesis that the multiple polypeptides corresponding to cheA and cheM were the products of single genes.


Purification and characterization of active component and active fragment of colicin E3. 1. Two components of colicin E3, namely proteins A and B, were prepared by means of an improved method. 2. Protein A thus obtained was more than a thousand times as active as native colicin E3 when they were assayed in terms of activity for ribosome inactivation. 3. Protein A was reconstituted to colicin E3 simply by mixing with protein B. 4. Trypsin digestion of colicin E3 yielded two fragments, T1 and T2, probably by cleaving one specific bond of the A moiety of colicin E3. 5. T2 was a complex of T2A and B proteins. T2A showed an activity equivalent to that of protein A when assayed in the in vitro system, and its activity was neutralized by protein B. Thus T2A was assigned as an active fragment of protein A. 6. T2A has a characteristic amino acid composition rich in the basic amino acid, lysine. 7. The structure and function of the colicin E3 molecule is discussed based on the results obtained with its components as well as with fragments of the components.


Interaction of S-sulfonated human IgG with human complement and its components. S-sulfonated human IgG (S-sIgG) was prepared by treating IgG with sodium sulfite and sodium tetrathionate. The treatment resulted in the selective cleavage of interchain disulfide bonds of the IgG to give S-sulfonate groups. Complement fixing activities of aggregated S-sIgG and the immune complex formed with the S-sIgG antibody were very weak. S-sIgG at a high dose reduced the activity of the first complement component (C1) in normal human serum without any reduction of other complement components activites, but S-alkylated IgG at the same dose did not. Loss of C1 activity was not caused by either S-sulfonated myeloma proteins (IgA and IgE) or urea-treated S-sIgG, in which both inter- and intra-chain disulfide bonds were cleaved. These results suggest that the selective reduction of C1 by S-sIgG is due to a conformational change of the immunoglobulin.


Changes in phospholipids of Saccharomyces cerevisiae associated with inositol-less death. Two inositol-requiring strains of Saccharomyces cerevisiae were examined for changes in levels of phospholipids occurring after inositol deprivation. Lack of inositol results in loss of cell viability (inositol-less death) and in very large increases in two phospholipid precursors, phosphatidic acid and CDP-diacylglycerol; the accumulation of other glycerophospholipids continues for a considerable time at normal rates. Phosphatidylinositol accumulation does not occur in the absence of inositol; however, the further metabolism of this lipid continues, with 80 to 90% of this lipid disappearing. This disappearance is matched by increases in the phosphoinositol containing sphingolipids and extracellular glycerophosphoinositol. These changes are not observed when growth is blocked by cycloheximide or by omission of lysine from a lysine auxotroph, most lipids continuing to accumulate long after growth stops. There appears to be no close coordination in the synthesis of the major yeast phospholipids or between protein synthesis and phospholipid synthesis. However, despite very large changes in the composition of yeast phospholipids that can be achieved by altering culture conditions, it appears that the average charge per phospholipid molecule remains fairly constant.


Mechanism of T5-induced DNA polymerase. I. Replication of short primer templates. Bacteriophage T5-induced DNA polymerase shows an initial phase of rapid synthesis, followed by a slower steady rate for much longer periods, with short DNA primer-templates (400 to 600 nucleotides long), in vitro. On extrapolating the line of steady rate back to 0 min, an intercept is obtained on the ordinate. With large DNA primer-templates, such as denatured T5 DNA (average chain length approximately 50,000 bases), the rate of synthesis remains constant and is equal to the initial rate obtained with short primer-templates. The zero time intercept was proportional to the amount of enzyme used and independent of temperature. Polymer challenge experiments indicate that the initial phase of rapid synthesis can be attributed to the processive mode of synthesis by T5 DNA polymerase. After synthesizing a stretch of DNA processively for about 200 nucleotide residues, the enzyme apparently forms a "dead-end complex" with the primer-templates used and must dissociate from the primer-template in order to resume synthesis. The average size of the product made processively, during various phase of synthesis, remains invariant and is in good agreement with the size of the zero time intercept per enzyme molecule.


Purification and some novel properties of Escherichia coli RNase II. RNase II of Escherichia coli (EC 3.1.4.23) has been purified to apparent homogeneity. The K+-activated diesterase activity against poly(U), which defines RNase II, cochromatographs with activity against T4 mRNA or pulse-labeled E. coli RNA successively on DEAE-cellulose, hydroxyapatite or phosphocellulose, and Sephadex G-150 columns. Activities with both substrates are selectively reduced to less than 2% of the wild type level in a newly isolated mutant strain, S296, or after thermal inactivation in a mutant strain with temperature-sensitive RNase II. RNase II releases 5'-XMP without a lag as its only detectable alcohol-soluble produce from all substrates and has an apparent molecular weight of 80,000 to 90,000 in both nondissociating and sodium dodecyl sulfate-polyacrylamide gels. The pure enzyme shows the standard K+ activation against poly(A), poly(U), or poly(C), but only a slight preference for K+ over Na+ ions with T4 mRNA or pulse labeled E. coli RNA as substrate. Uniformly labeled E. coli rRNA or tRNA is degraded little if at all.


Translation of the L-species dsRNA genome of the killer-associated virus-like particles of Saccharomyces cerevisiae. Virus-like particles containing the L (P1)-species of double-stranded RNA (dsRNA) were isolated from Saccharomyces cerevisiae, and the translational activity of the virus-like particle-derived dsRNA was analyzed in the wheat germ cell-free system. Denaturation of the dsRNA immediately prior to in vitro translation resulted in the synthesis of one major and at least three minor polypeptides, whereas undenatured dsRNA, as expected, did not stimulate [35S]methionine incorporation into polypeptides, but actually slightly inhibited endogenous activity. The major in vitro translation product of the denatured L-dsRNA was shown to be identical with the major L-dsRNA containing virus-like particle capsid polypeptide on the basis of three criteria: co-electrophoresis on sodium dodecyl sulfate polyacrylamide gels, immunoprecipitation, and tryptic peptide analysis. We have therefore established that the L-dsRNA genome encodes the major virus-like particle capsid polypeptide. This result adds considerable support to the hypothesis that the L-dsRNA genome acts as a helper genome to the smaller (1.6 x 10(6) dalton) M-dsRNA genome in killer strains of yeast by providing the M-dsRNA containing virus-like particles with their major coat protein.


Antemortem diagnosis of massive air embolism in the newborn. A case is presented of massive air embolism complicating respiratory distress syndrome of the newborn. This uncommon complication of respiratory therapy has occurred only with high inflation pressures and has been uniformly fatal in this and all reported cases. Antemortem radiographs are necessary for accurate diagnosis.


Pulmonary vascular air embolism in hyaline membrane disease. Pulmonary vascular air embolism is a rare, fatal complication of positive-pressure ventilation in hyaline membrane disease. Sudden deterioration of an infant on respiratory therapy should alert the clinician and radiologist to the possibility of this entity. A case history with typical radiographic findings is presented.


Changes in ionic movements across rabbit polymorphonuclear leukocyte membranes during lysosomal enzyme release. Possible ionic basis for lysosomal enzyme release. Changes in the movements of Na+, K+, and Ca+2 across rabbit neutrophils under conditions of lysosomal enzyme release have been studied. We have found that in the presence of cytochalasin B, the chemotactic factor formyl methionyl leucyl phenylalanine (FMLP) induces within 30 s large enhancements in the influxes of both 22Na+ and 45Ca+2 and an increase in the cellular pool of exchangeable calcium. The magnitude of the changes induced by cytochalasin B and FMLP exceeds that induced by FMLP or cytochalasin B alone, and cannot be explained on the basis of an additive effect of the two agents. However, these compounds either separately or together produce much smaller enhancements in 45Ca efflux. The divalent cation ionophore A23187 also produces a rapid and large increase in the influxes of both 22Na and 45Ca+2 in the presence and absence of cytochalasin B. We have also found an excellent correlation between calcium influx and lysosomal enzyme release. 42K influx is not significantly affected by any of these compounds. On the other hand, a large and rapid increase of 42K efflux is observed under conditions which give rise to lysosomal enzyme release. A flow diagram of the events that are thought to accompany the stimulation of polymorphonuclear leukocytes (PMNs) by chemotactic or degranulating stimuli is presented.


Structural changes in the epidermal cells of Rhodnius during tracheole capture. Epidermal cells deprived of their oxygen supply by tracheal section give off cytoplasmic processes which become attached to air-filled tracheoles in neighbouring areas and draw these into the oxygen-deficient zone. Many of these cytoplasmic strands exceed 100 micrometer in length but may be no more than 50 nm in diameter; they contain mitochondria, ribosomes, microtubules and microfilaments. In basis structure they resemble the tendon cells; and also the tapering conical epidermal cells along the intersegmental borders of the abdomen, the terminal strands of which are inserted into the basement memebrane behind and in front of the segmental boundaries. The cytoplasmic walls of those tracheoles most exposed to tension during the process of tracheole capture become thickened and packed with microtubules. In all these structures the microtubules are believed to be concerned in resistance to tension. Contraction is presumably effected by microfilaments, but no new evidence is given. The possible role of the epidermal strands in the transport of energy-rich metabolites is discussed.


The induction of cytoplasmic petite mutants of Saccharomyces cerevisiae by hydrostatic pressure. This study demonstrates that hydrostatic pressure is a potent inductive agent of the petite mutation in cultures of Saccharomyces cerevisiae. The inductive capacity of this mutagen is dependent on the magnitude and the duration of the pressure treatment. Furthermore, the extent of petite induction varies with the growth stage of the culture. Induction occurs in pressure-treated (1-4 X 1-(4) lbf in.-2 or 9-66 X 10(4) kN m-2 for 4 h) log growth cultures but not in stationary or lag phase cultures. Petite induction and cell survival are also dependent on the particular strain of yeast which is pressure-treated. Tetrad analysis and complementation assays demonstrate that pressure-induced petite cells are cytoplasmic in nature. Moreover, induced petite cells show a wide range of suppressivity (2--99%) with a large proportion of the petite cells being highly suppressive.


Immunofluorescent localization of urease in the cotyledons of jack bean, Canavalia ensiformis. Urease has been localized in sections of cotyledons from germinating seeds of jack bean, using FITC-labelled immunoglobulin prepared from urease antiserum raised in rabbits. The complication of lectin binding to the immunoglobulins was resolved by treatment of the sections with specific glycosides. Urease is localized in 2 sites: within the cytoplasm of storage parenchyma cells in spherical granules up to 3 micrometer in diameter, and within the intercellular spaces in spherical granules. Although similar in size, the latter are distinguished from the cytoplasmic granules by the presence of beta-lectin and appear to function as an extracellular lytic compartment or lysosome.


[New technic of retro-peritoneal renal transplantation in the left lumbar position in the child. Report of two cases (author's transl)]. In the child, renal transplantation is almost always carried out by placing the kidney within the peritoneum for a simple reason: the donors being adults, the kidneys are too large to be placed in extra-peritoneal position. However, all authors agree that placing the kidney in an extra-peritoneal position offers undoubted advantages.


Enzyme-linked immunosorbent assay for detection of Escherichia coli heat-labile enterotoxin. The development of an enzyme-linked immunosorbent assay (ELISA) for the detection of heat-labile Escherichia coli enterotoxin is described. The assay, which is based on the immunological similarity between Vibrio cholerae toxin and heat-labile E. coli enterotoxin, is similar in design to a radioimmunoassay but utilizes enzyme-labeled rather than radioactive isotope-labeled reagents. The ELISA system is as sensitive as both radioimmunoassay and the y-1 adrenal cell assay for the detection of heat-labile E. coli enterotoxin but requires neither radioactive reagents nor tissue culture techniques. The ELISA is easy to perform and is adaptable for use in small laboratories.


Evaluation of a caffeic acid-ferric citrate test for rapid identification of Cryptococcus neoformans. An evaluation of a rapid caffeic acid-ferric citrate paper disk test for the identification of Cryptococcus neoformans, using 474 isolates of yeasts and yeastlike organisms, showed that 96.6, 97.7, and 98.3% of 176 isolates of C. neoformans produced brown to dark-brown pigment on disks incubated for 6 h at room temperature, 30 degrees C, and 37 degrees C, respectively. All C. neoformans produced brown to dark-brown pigment within 24 h. However, nonspecific pigmentation was encountered at all temperatures of incubation with one isolate of Trichosporon cutaneum and, at room temperature only, with one isolate of C. luteolus after 6 h. Other genera of yeasts produced similar pigmentation after 24 h at all temperatures. The age of the cultures tested or the types of media used before testing did not significantly affect the ability of C. neoformans to produce pigmentation at 37 degrees C. A positive test may prove useful for presumptive identification of C. neoformans, but a negative reaction should not be used to rule out an identification of this yeast. Since a number of false-negative and false-positive tests occur, it is necessary to confirm, by other biochemical tests, the identification of all organisms suspected of being C. neoformans, to reduct the serious risk of missing or misidentifying this important pathogen.


Acetylene reduction (dinitrogen fixation) by clinical isolates of Klebsiella pneumoniae. Freshly isolated clinical strains of Klebsiella were tested for the ability to fix dinitrogen by the acetylene reduction assay. Ability to detect this trait was markedly affected by cultural conditions. When the test was run at 37 degrees C in the presence of yeast extract (50 mg/liter), only 1.6% of the organisms were diazotrophs, whereas this temperature without yeast extract yielded 12.9% positive cultures. The optimum condition found was 28 degrees C without yeast extract (21.9% positive); therefore, search for diazotrophy in clinical strains should not be conducted at the usual incubation temperature. There was a high incidence of indole-positive strains among diazotrophs. No such correlation was noted with any other biochemical trait or antibiotic susceptibility tested. The significance of this correlation is not apparent.


Yersinia enterocolitica: biochemical, serological, and gas-liquid chromatographic characterization of rhamnose-, raffinose-, melibiose-, and citrate-utilizing strains. Thirteen atypical Yersinia enterocolitica isolates, all fermenting rhamnose, raffinose, and melibiose and utilizing sodium citrate within 24 to 48 h at 22 degrees C (Y.e.rh+), were examined biochemically-serologically, and by gas-liquid chromatography. These data, as well as cultural, biochemical, and antibiotic susceptibility data gathered from two previous studies involving (i) these same atypical Y.e.rh+ isolates, (ii) Y. enterocolitica serotypes O:1 through O:15 (rhamnose, raffinose, and citrate negative [Y.e.rh-]), (iii) Y. enterocolitica serotype O:16 (rhamnose positive but raffinose and citrate negative), and (iv) Yersinia pseudotuberculosis serogroups I through V were statistically compared. Both preand postabsorption agglutination studies demonstrated the serological distinctiveness of Y.e.rh+ from Y.e.rh- and Y. pseudotuberculosis. At the same time, three immunological groups among the 13 Y.e.rh+ strains were seen; 8 corresponded to Y. enterocolitica serotype O:17; 1 to Y. enterocolitica serotype O:16; and the remaining four were nontypable in antisera against known Y. enterocolitica antigen types. Each of the three Yersinia groups tested chromatographically produced acetic and lactic acids. Both Y.e.rh- and Y.e.rh+ formed propionic acid, but only Y.e.rh+ produced detectable amounts of succinic acid. Based on 49 variables, statistical analysis of the three Yersinia groups studied placed each of the Y.e.rh+ strains in a homogeneous group separate from both Y.e.rh- and Y. pseudotuberculosis. These data, coupled with deoxyribonucleic acid homology studies of Brenner and co-workers (D. J. Brenner, A. G. Steigerwalt, D. F. Falcao, R. E. Weaver, and G. R. Fanning, Int. J. Syst. Bacteriol. 26:180-194, 1976), support the distinctiveness of Y.e.rh+ from typical Y. enterocolitica and Y. pseudotuberculosis.


Rapid detection of bacterial growth in blood samples by a continuous-monitoring electrical impedance apparatus. A growth detection method utilizing an automated apparatus capable of rapidly detecting bacterial growth by measuring changes of electrical impedance in bacteriological medium was utilized with "mock" blood cultures containing various gram-negative and gram-positive bacteria. Measurement of changes of electrical impedance was found to ba as accurate and comparable for time of growth detection as the radiometric method for detection of the same bacteria using mock blood cultures. In a limited clinical trial the use of the electrical impedance apparatus detected in 1 positive specimen from 40 clinical blood specimens as rapidly as by radiometric measurement. Both methods were considerably faster for detecting bacterial growth as compared with conventional culture methods. The selected species of gram-positive and -negative organisms tested were all detected by the electrical impedance method, including aerobes and anerobes. However, addition of 5% CO2 to the incubation atmosphere enhanced detection of gram-positive organisms.


Antibody response to Serratia marcescens isolated from patients with malignant diseases. Sixty-four patients with malignant diseases from whom Serratia marcescens was isolated from various sources were studied regarding their antibody responses to somatic O antigens of this microorganism. Antibodies were titrated by the passive hemagglutination test. An antibody response was considered present when either a fourfold or greater rise in antibody titers between two consecutive serum specimens was demonstrated, or when elevated titers (greater than or equal to 40 for serogroup O14 and greater than or equal to 160 for all others) were present in the first available specimen. Overall, 31% of subjects mounted an immune response, but there were differences depending upon the infection site. Seventy-one percent of patients with S. marcescens bacteremia responded immunologically; whereas the percentage for patients with Serratia present in the respiratory tract was only 22%, in the urinary tract, 31%, and in wounds, 26%. Documentation of an immune response to the patient's own infecting strain of Serratia aids in the differentiation between infection and contamination and possibly also between clinical disease and colonization. In addition, immunoglobulin samples collected in different decades were examined to determine whether the background level of antibodies to S. marcescens had changed in the general population over the years. No difference in antibody titers to 13 O antigens was observed in immunoglobulin preparations from 1951, 1962, 1971, and 1975.


Misinformation from sputum cultures without microscopic examination. Only 13 of 38 hospital laboratories surveyed include a Gram stain routinely in microbiological sputum examination. In a prospective three-hospital study, 60% of over 1,200 "sputum" specimens consisted predominantly of saliva, as judged by cell composition. Compared with the results of cultures in which microorganisms presumptively identified on sputum smears were specifically sought ("directed cultures"), cultures of the same specimens processed in the routine manner missed pneumococci 61%, haemophili 23%, and yeasts 44% of the time. The findings were similar in all three hospitals despite differences in administration, staffing, primary culture media, and workload. Unless microscopic examination is routinely included, half of all microbiological information rendered on sputum specimens is meaningless and subject to dangerous misapplication. Furthermore, culture must be guided by microscopic findings, or respiratory pathogens will frequently be missed. Finally, when routine culture and smear disagree, the culture cannot be assumed to be correct. Microscopic examination should be mandatory in sputum microbiology, both for specimen evaluation and as a guide to what to look for in culture.


Evaluation of the Wampole Streptonase B test. The Streptonase B test was compared with the Center for Disease Control streptococcal anti-deoxyribonuclease B test. A total of 96% of anti-deoxyribonuclease B titers obtained with the Streptonase B test were within +/- 1 dilution step of those obtained with the Center for Disease Control anti-deoxyribonuclease B test.


Bacto-TB hydrolysis reagent (Tween 80) for identification of Serratia. Among 21 species of Enterobacteriaceae, only Serratia hydrolyzed the commercially available Bacto-TB hydrolysis reagent (Tween 80). The procedure allowed accurate differentiation of Serratia within 24 h.


Direct manipulation of physiological arousal in induced anxiety therapy - biofeedback approach. This study investigated the role of physiological arousal in the affect induction phase of Induced Anxiety therapy by using biofeedback to facilitate arousal. Twenty-one college students who were suffering from free-floating anxiety were assigned randomly to one of three groups: (1) a no-treatment control group simply completed the measures before and after therapy; (2) a conventional Induced Anxiety group went through five standard Induced Anxiety sessions; and (3) biofeedback Induced Anxiety group went through a similar procedure except that biofeedback was used in the affect induction phase to facilitate heart rate increase. It was found that the biofeedback procedure did result in a greater heart rate increase during the affect induction phase arousal than did the conventional procedure (.01 level of significance), but did not facilitate subjective emotional arousal. Biofeedback Induced Anxiety resulted in a greater reduction of trait anxiety as measured by the Multiple Affect Adjective Check List than did the no-treatment group or the conventonal Induced Anxiety group. The conventional Induced Anxiety group did not differ significantly from the no-treatment control group.


The effect of the Transcendental Meditation technique on anxiety level. Two weeks of twice-daily practice of the transcendental Meditation (TM) technique was compared with 2 weeks of twice-daily practice of passive relaxation as a means of reduction of anxiety, as measured by the Trait scale of the State-Trait Anxiety Inventory. Thirty-three graduate and undergraduate students were assigned randomly to a relaxation group and a TM group. After a 2-week experimental interval, the relaxation and meditation Ss, as well as between conditions of the relaxation-mediation group; TM was significantly more effective in reducing anxiety level. Thus, the anxiety-reducing effect of the practice of TM cannot be attributed merely to sitting quietly twice daily, although additional research must determine the extent to which S expectations for change contributed to this effect.


Developmental assessment of cognitive style in mentally retarded psychotics. Evaluated 55 mentally retarded psychotic adults with the Color-Form Preference Test, a developmentally based index of cognitive style, and other measures of intellectual, social, and psychiatric functioning. Results indicated that different grades of retardation were associated with distinct cognitive styles, which showed a systematic progression that corresponded to the normal developmental sequence. In contrast, no significant relationships were observed between cognitive style and psychotic manifestations. The findings suggested that fresh insights into mental retardation and psychosis are made possible by qualitative, developmental assessment. The implications for clinical application, theory formulation, and research were discussed.


The effect of acid etching on enamel, dentin, and the inner surface of the resin restoration: a scanning electron microscopic investigation. Buccal cavities were prepared in vivo on 22 contralateral tooth pairs. The cavities and the surrounding enamel were either etched or cleaned with a surface-active solution (the control). The cavities were filled with Concise Enamel Bond and composite or left unrestored. The teeth were extracted immediately after treatment. Treated surfaces or the inner surface of the restorations were studied in the scanning electron microscope (SEM). An "in-pair" evaluation of the results of 320 photographs was done. The etched axial wall had a smooth surface with widened and patent dentinal tubule apertures. The corresponding inner resin surfaces were irregular, sporadically showing tags of varying length corresponding to the dentinal tubules and alternating with pits of varying size. Etching for 15 seconds or two minutes gave no apparent difference in appearance of enamel, dentin, and inner surface of the restoration.


Guidelines for antiinflammatory drug research. Guidelines have been developed by the Bureau of Drugs, FDA, for antinflammatory drug development. They address the problem of efficacy and safety testing but leave the definition of qualified investigators and appropriate subjects unstated. The natural history of rheumatoid arthritis, the most-studied chronic inflammatory disorder, is not sufficiently appreciated, and "controlled" studies are too narrowly structured to give adequate information. By contrast, other countries speed the review process by emphasizing early toxicity studies and letting the market place determine relative efficacy while long-range safety data are carefully monitored.


Pharmacokinetics and clinical effects of amantadine in drug-induced extrapyramidal symptoms. Plasma amantadine concentrations were assessed in a series of hospitalized schizophrenic patients receiving this drug during a double-blind trial of amantadine and benztropine in the treatment of neuroleptic-induced extrapyramidal symptoms (EPS). Mean (+/- S.E.) plasma amantadine concentrations were 0.54 +/- 0.08 microgram/ml on day 7 and 0.43 +/- 0.08 microgram/ml on day 14. Overall improvement of EPS was not correlated with plasma level, but improvement in the target EPS of rigidity was correlated with plasma amantadine concentration on day 7 (r = 0.75) and day 14 (r = 0.68). There was no evidence that the overall improvement in schizophrenic symptomatology was influenced by plasma amantadine concentrations.


Adhesive bonding of various materials to hard tooth tissues: XIII Synthesis of a polyfunctional surface-active amine accelerator. Surface-active amine polymerization accelerators can be prepared by the reaction of polyepoxy resins with the sodium salt of N-phenylglycine and N-methyl-p-toluidine. These materials are expected to promote adhesion through complexation with surface calcium (or other metal ions), utilizing several chelating groups per molecule, and by functioning as polymerization accelerators for dental resins; they can also function as catalysts for the anionic polymerization of cyanoarylate monomers.


Improved adhesion of acrylic restorative materials to dental enamel by precoating with monomers containing phosphonate groups. The adhesion of acrylic composite restorative materials to etched dental enamel is increased by a factor of two or more when the enamel is coated with a thin layer of vinyl-benzyl phosponic acid (VBPA) prior to forming the enamel-restorative bond. The present results support the suggestion that the enhanced adhesion is attributable to bonding between calcium ions in the enamel surface and phosphonate groups in the VBPA.


A subjective and objective method assessing the efficacy of hypnotic medications in insomniacs. The response of chronic insomniacs to 100 mg pentobarbital, 300 mg methyprylon, 500 mg glutethimide, and placebo was assessed using our previously described subjective and objective techniques. The purpose of the study was to examine (1) the presence or absence of the subjects' reported insomnia; (2) the subjects' ability to discriminate between active hypnotic drugs and placebo; and (3) whether any preference existed among active medications. Statistically significant findings included a high degree of correlation (P less than 0.001) between subjective and objective data and greater response to active medications as compared to placebo shown on all parameters except objective onset of sleep. In no case was there significant difference between the two nights of placebo. Although methyprylon was most frequently superior to placebo, there was no significant patient preference for any of the active medications.


Long-term evaluation of a new aerosol bronchodilator, carbuterol, and comparison with isoproterenol. A new aerosol bronchodilator, carbuterol, was compared with isoproterenol during a three-month double-blind trial in 15 matched pairs of patients with relatively stable and partially reversible obstructive airways disease. Pulmonary function tests were performed at baseline and at 30 and 120 minutes after bronchodilator administration on days 1, 15, 28, 56, and 84. Isoproterenol and carbuterol produced improvement in all parameters at 30 and 120 minutes. The per cent changes produced by isoproterenol were superior at 30 minutes, while carbuterol was as effective as isoproterenol at 120 minutes. Atrial and ventricular premature contractions and palpitations were more frequent in the isoproterenol group. Evaluation of the patients' symptomatic responses showed carbuterol to be superior to isoproterenol. We conclude that carbuterol is a useful bronchodilator when aerosolized.


The effect of Procion Blue on certain metabolic activities of Streptococcus mutans. Metabolic activities of S mutans were selectively affected by Procion Blue, known to cause covalent bonds and stable cross links in relation to carbohydrate and protein. In this study, despite the reduction in extra polysaccharide, the level of activity of glucosyltransferase was not significantly changed from the control level and colonial morphology was not transformed from smooth to rough. The results of the investigation imply at least interference with formation of extra cellular polysaccharide, possibly in the bacterial cell wall.


Measurement and synthesis of insoluble and soluble dextran by Streptococcus mutans. Total and insoluble dextransucrase activities of 10 strains of oral streptococci were measured by a modified filter disk assay. Strains that were nonadherent to hard surfaces had only low levels of insoluble dextransucrase activity. A physical rather than metabolic mechanism is suggested to explain the decreased insoluble and increased soluble activities observed when dextran T-10 is added to the media.


Adherence controlling elements in ceramic-metal systems. I. Precious alloys. Elemental analysis of two precious alloyceramic systems revealed the accumulation of Sn atoms within the interaction zone of couples subjected to a range of metal oxidation time; and varying total firing times. The source of Sn atoms which accumulated within the interaction zone was the opaque porcelain layer which contained an initially higher Sn concentration than the alloy. It is hypothesized that SnO2 provides continuity of electronic structure between alloy and ceramic structures and is a primary factor in controlling ceramic-metal adherence.


Adherence controlling elements in ceramic-metal systems. II. Nonprecious alloys. Nickel-chromium alloy-ceramic couples used in this study exhibited Cr, Ni-Cr, and Ti-Cr interactions with bonding agent-ceramic complexes. Bonding agents may broaden or suppress the width of interaction zones. Metal oxides such as NiO, Cr2O3, or more complex forms such as TiO-Cr2O3 and NiCr2O4 may play a prominent role in controlling ceramic-metal adherence.


The strengthening of aluminous porcelain with bonded platinum foils. Results are presented to demonstrate the effect of bonding thin platinum foils to aluminous dental porcelain. The increased strength is explained in terms of a reduction in surface and subsurface pores in the porcelain brought about by improved wetting.


A study of "broken appointment" patients in a children's hospital dental clinic. Part I of this study investigated possible variables determining "High Risk No Show" behavior on the part of patients in a dental clinic. Only "source of payment" seemed important; namely, that Medicaid patients were statistically more likely to be "High Risk No Show" patients (P less than .001). Race was associated but was found to be confounded with Medicaid. In an attempt to investigate the reasons for the Medicaid group being "High Risk No Show" patients, Part II of the study compared three distinct groups of patients at the Children's Hospital Medical Center Clinic. These included the self-paying group and the Medicaid group of the original sample of 90 in addition to the Union plan group (patients receiving free third-party payment care as an employee benefit). It was shown that both groups with third-party coverage were associated with "High Risk No Show" behavior. The type of "No Show" behavior, however, was different for the two groups. Union plan patients tended to cancel appointments or at least notify in advance of inability to keep an appointment. Medicaid patients, conversely, were just as likely to fail appointments and not file advance notice of inability to keep an appointment.


Nutritional regulations and legislation--past developments, future implications. Laws to prohibit adulteration of foods were enacted before 1900, but the first steps in the development of nutritional regulations were taken only after passage of the Food, Drug and Cosmetic Act of 1938. This act authorized the Food and Drug Administration (FDA) to establish Standards of Identity for foods, including standards for enrichment with vitamins and minerals to meet Minimum Daily Requirements (MDRs). The legal definition of nutrients, however, has remained unclear. Vitamins are foods when used as supplements but drugs when prescribed as therapeutic agents. To protect consumers, in 1962 the FDA proposed Standards of Identity for Special Dietary Foods, e.g., vitamin supplements. Opposition to the proposal has been vigorous and was the subject of the longest hearings on record. Implementation of standards has been contested in the courts and curtailed by Congressional action. A bill passed in 1975 prohibits the FDA from establishing Standards of Identity for special dietary foods for adults. During the legal and political controversy, the validity of Recommended Dietary Allowances as nutritional standards was challenged but subsequently confirmed. Revised regulations for vitamin and mineral preparations, taking into account the Congressional and court actions, will become effective January 1, 1978, sixteen years after first being proposed.


Cyclandelate in the treatment of senile mental changes: a double-blind evaluation. The treatment of cerebrovascular insufficiency and its many symptoms in the ever-increasing numbers of the aged, is of major concern to physicians engaged in such care. Despite past skepticism as to the degree of efficacy of cerebral vasodilators, there is renewed interest in this form of therapy. Our investigation was designed to assess the effectiveness of cyclandelate, under strict double-blind conditions, in 58 geriatric patients. The cyclandelate and placebo groups (32 and 26 patients respectively) received either 1,600 mg/day of cyclandelate in fractional doses, or identical-appearing placebo capsules--over a period of 12 weeks. During the initial examination and every four weeks thereafter, patients were assessed for possible changes in vital signs and for evidence of adverse reactions. In addition, the Sandoz Clinical Assessment-Geriatric (SCAG) and the Nurses Observation Scale for Inpatient Evaluation (NOSIE) were completed, with particular attention to symptom clusters. A final global assessment was made in which the physician rated patients according to their overall clinical condition. The results of our study and analysis indicate that cyclandelate is a safe and effective agent for treating certain symptoms of senility in properly selected patients, provided the therapy is carried on for at least eight weeks and, if indicated, for a longer period. Clinical evidence suggests that the prudent use of this drug may definitely delay deterioration.


Repair of tissue defects resulting form removal of cutaneous neoplasms. The repair of large defects resulting from ablation of a primary malignancy of the skin requires judicious decision. Failing reasonable surety of extirpation, permitting spontaneous healing, if feasible, is the best cours. Grafting or primary closure that does not require extensive undermining are usually second choices. Flap repair is least desirable, except in instances where immediate function is necessary.


Experience with a "continuous" laser in the treatment of suitable cutaneous conditions: preliminary report. A clinical trial with a "continuous" CO2 laser in the treatment of cutaneous lesions was instituted 2 1/2 years ago. The laser was used as a "light" knife for excision and for gross destruction of some lesions directly. Details of technique and a preliminary report assessing clinical results and general effects are given. Some results thus far have proved satisfactory, and we feel that further investigation of the laser as a surgical modality is warranted.


Repair of flexor tendons by intratendinous tendon suture. In a three year period, severed flexor tendons in the fingers and palm of 50 patients were repaired using an intratendinous suture method. The majority were secondary repairs, and 21 of 50 were in children less than 15 years of age. Results were satisfactory, with 79% being rated as good or excellent. The technique apparently causes little disturbance in the circulation, and if adhesions develop, they are limited and good results can be expected from tenolysis.


Primary flexor tendon repair followed by immediate controlled mobilization. A study was made of the results of immediate repair and controlled mobilization in 156 severed flexor tendons in 68 patients occuring over an 18-month period. Eight patients with 16 tendon injuries could not be followed. Results were obtained from examinations done 6 weeks to 18 months (mean, 5.3 months) after repair. Thirty-one of the 60 patients were less than 20 years old, and 44 of the 60 were less than 30 years old. Seventy-nine (56%) of the injuries occurred in the area known as "no man's land"; 28 patients with repair of tendons in this area were rated by our standards as "excellent" or "good"--75% of patients as compared to 84.4% for the results of repair in other areas. In one fourth of the cases of severance of both tendons, because of local conditions in the wound, the superficialis was excised, but in all others it was repaired.


Delayed flexor tendon repair in no man's land. Thirty seven digital flexor tendon injuries in 31 patients were treated by closure of the skin and delayed repair from 24 hours to 21 days later. All skin wounds healed without serious complication, and there were no infections. On examination at a minium of 4 months after repair, 36% had total active motion (TAM) of 220 degrees, 32% from 200 degrees to 220 degrees, 6% from 180 degrees to 200 degrees and 26% with less than 180 degrees. Under proper conditions, repair of flexor tendons can be carried out with the expectation of results comparable to more complex reconstruction procedures.


G.H. Mead's social behaviorism.

G.H. Mead's social behaviorism. This paper seeks to clarify those conceptual foundations of G.H. Mead's social behaviorism which are assumed, but not made explicit, in that writer's well-known volume Mind, Self and Society. These foundations are shown to be an outgrowth of Mead's early commitment to the organic conception of conduct underlying the psychological functionalism of the Chicago School. Further light is shed upon Mead's position by pointing out the fundamental differences between his model of conduct and that characteristic of the behaviorist tradition in American psychology.


Psychology's first paradigm.

Psychology's first paradigm. By the end of the nineteenth century, psychology had become a mature scientific discipline unified around a common paradigm. Although the underlying unity of the field was recognized at the time, it has generally been obscured in later historical writings. In this article, the first "mentalist" paradigm of scientific psychology is examined. The mentalists shared a common definition of the field, two accepted modes of observation, and a common conception of the relationship between modes of observation and data thus observed. Rival schools advanced competing articulations of this shared paradigmatic structure.


A pictorial concordance to "important psychologists, 1600-1967". Pictorial resources are cited for Annin, Boring, and Watson's published list of historically important psychologists from 1600-1967.


The nonwhite as child: G. Stanley Hall on the education of nonwhite peoples. Although he was a significant educational reformer during the progressive era, a founder of various journals in psychology and pedagogy, a profile writer, and the individual who brought Freud and Jung to the United States, G. Stanley Hall's ideas on the education of nonwhites were, for his period, quite conventional. Unlike those of some of his contemporaries, Hall's racial ideas were not overtly vicious. He argued that nonwhites were the children of the human race and that their education, like that of children of particular ages, should be based on an understanding of their indigenous culture and inherent capabilities. This argument, although reformist in tone, supported a policy of nonwhite subservience.


The role of mental illness in the European witch hunts of the sixteenth and seventeenth centuries: an assessment. Historians of psychiatry have propagated the view that the witch hunts of sixteenth and seventeenth-century Europe were primarily a persecution of the mentally ill and that demonological concepts of possession and witchcraft impeded psychiatric progress for centuries. The author reviews the evidence marshaled by these historians and examines additional historical material bearing on the psychopathological view. He concludes that the role of mental disorder in the witch hunts has been overinflated by authors with an interest in promulgating the medical model of abnormal behavior. Furthermore, the psychopathological paradigm is based on an outmoded philosophy of science, which results in historical distortion and paradoxes, and on restriction and selectivity in the choice of evidence.


The beginnings of Gestalt psychology in the United States. Robert M. Ogden's unrecognized contributions to the beginnings of Gestalt psychology in the United States are briefly discussed. It is proposed that he not only introduced Gestalt psychology to this country through his books, but was also instrumental in bringing Gestalt psychologists to America.


Weber, Durkheim, and the comparative method. This essay compares and contrasts the means by which Durkheim and Weber dealt with methodological issues peculiar to the comparative study of societies, what Smelser has called "the problem of sociocultural variability and complexity." More specifically, it examines how Weber and Durkheim chose appropriate comparative units for their empirical studies. The approaches that Weber and Durkheim brought to theproblem of cross-cultural comparison have critical implications for more current procedures used in the comparative study of contemporary and historical societies.


The real F.M. Urban.

The real F.M. Urban. F.M. Urban made important contributions in psychometry and is best known for his introduction of a correction in the "Müller-Urban weights." Born in Austria, he came to the University of Pennsylvania where he made most of his contributions in psychology. He was trapped in Europe by World War I and never returned to the United States. Myriad hardships forced him to abandon his original project of a study of the entire field of probability and its application to other sciences. Whenever possible, however, he pursued his interests in psychometry and psychophysics and strove to remain active in psychology.


The concept of childhood in the middle ages. The question is raised of whether the Middle Ages held a view of the nature of childhood as distinct from adulthood. Evidence is offered from legal documents, medical writings, and church and monastic chronicles to support the viewpoint that the special nature of the child was perceived (although with great ambivalence) as being fragile, vulnerable, and naive, while also possessing a potential for closeness to God and the supernatural world.


A technique for the immunohistochemical localization of prostaglandin E. Prostaglandin E (PGE) has been localized via the unlabeled antibody technique in freeze-dried and ethanol-fixed cryostat sections. Discrete perivascular and stromal localization was present in the uterus prepared by the method presented, but not in classically fixed specimens. Absorption of the anti-PGE by addition of free PGE was ineffective; whereas, removal of PGE-reactive antibodies from the anti-serum was effectively accomplished with an Affigel-101-PGE immunoadsorbant column.


Immunocytochemistry of the olfactory marker protein. The olfactory marker protein has been localized, by means of immunohistochemical techniques in the primary olfactory neurons of mice. The olfactory marker protein is not present in the staminal cells of the olfactory neuroepithelium, and the protein may be regarded as indicative of the functional stage of the neurons. Our data indicate that the olfactory marker protein is present in the synaptic terminals of the olfactory neurons at the level of the olfactory bulb glomeruli. The postsynaptic profiles of both mitral and periglomerular cells are negative.


Two-wavelength cytophotometry: the choice of the wavelengths from a practical point of view. Methods in use for the choice of the two-wavelengths and the effect of a faulty choice of the two wavelengths in the two-wavelength cytophotometric method of Patau and Ornstein are studied by means of a computer simulation and in practice upon pararosaniline-Schiff stained nuclei. A simplification of the measuring strategy is proposed without affecting the accuracy.


Formation of the chorion (zona pellucida) in the teleost, Oryzias latipes. II. Polysaccharide cytochemistry of early oogenesis. It has been proposed that the mechanism of formation of the chorion of Oryzias latipes involves the transfer of precursor material from the Golgi bodies of the oocyte to the chorion by means of a population of dense cored vesicles. Thin sectioned ovarian material was investigated by means of several techniques available for the resolution of periodate reactive material at the ultrastructural level. The techniques employed were modifications of the thiocarbohydrazide, thiosemicarbazide, alkaline bismuth subnitrate and silver methenamine reactions. These all demonstrated a positive reaction in the material of the chorion and that of the dense cored vesicles. This finding is in accord with the hypothesis that the dense cored vesicles are transporting material from the Golgi to the growing chorion.


Survival of Salmonella eastbourne and Salmonella typhimurium in milk chocolate prepared with artificially contaminated milk powder. Milk chocolate mass containing salmonellas was prepared by mixing artificially contaminated milk powder with the other ingredients at a temperature of about 40 degrees C. From this mass bars were made. Two series were prepared, with S. eastbourne and S. typhimurium respectively. The number of surviving salmonellas was counted after various periods of storage, up to 19 months. S. eastbourne was reduced in numbers during 19 months from an initial count of ca. 3 x 10(4) to ca. 3 x 10(2) per 100 g of chocolate. S. typhimurium died off more rapidly, and was not detectable in about 55 g after 15 months, in spite of an initial count of ca. 10(5) per 100 g.In these experiments the salmonellas in the milk powder had had to survive the spraying procedure and the adverse conditions in the dried powder. This may be the reason why S. eastbourne showed a distinctly better survival on storage than the same serotype showed in previous experiments in which the organism was added as a broth culture to the chocolate mix. With S. typhimurium, however, such a difference was hardly detectable.Possible explanations of these results are discussed.


Effects of antibiotics on the survival of Salmonella in the American cockroach. The faecal survival of Salmonella typhimurium orally administered to restrained American cockroaches maintained on an antibiotic diet was investigated. Significant reductions in median numbers of total aerobic faecal microorganisms were noted in insects fed antibiotics daily, and when Salmonella was introduced to this modified micro-environment, it persisted for 44 days in all specimens. Multiplication of the pathogen was also observed in these insects, with numbers often exceeding 10(3) times that of the initial input. This differs significantly from our previous results showing that the pathogen is unable to multiply in conventional cockroaches. Attempts to restore the normal flora by feeding a faecal suspension from untreated cockroaches resulted in a decrease in numbers of Salmonella excreted, but did not result in their elimination. Carcasses of infected cockroaches retained viable Salmonella for at least 60 days post mortem, or 104 days after the infective meal.


Microbiological evaluation of South Australian rock lobster meat. Samples of frozen precooked rock lobster meat from five South Australian fish-processing plants situated in the West Coast and south-east regions were tested over a period of six months during the 1974/5 lobster  fishing season. The most probable number (MPN) of E. coli and coliforms, Staphylococcus aureus and Salmonella, as well as total plate count (TPC) were determined in 480 samples. Monthly geometric mean TPC ranged from 1600/g to 25,000/g. The highest geometric mean of the MPN of coliforms and E. coli were 4.9/g and 1.8/g respectively. The highest geometric mean number of staphylococci was 18.6/g. Salmonella was not detected in the 480 units tested. Only 0.4% of the samples had TPC exceeding 100,000/g. Coliforms and E. coli were not present in 76.1% and 92.7% respectively of the samples tested. Staphylococcus aureus was not detected in 67.7% of the samples. The numbers of organisms in 82% of the samples fall within the microbiological standards proposed by the National Health and Medical Research Council of Australia for frozen precooked foods. The results of this study demonstrate the microbial quality of precooked lobster meat attainable when good manufacturing practices are used.


Serotype-related enterotoxigenicity in Escherichia coli O6.H16 and O148.H28. The ability of certain Escherichia coli strains to produce enterotoxin is determined by transmissible plasmids. It is therefore possible that any E. coli strain might be able to acquire such a plasmid and that the correlation between enterotoxigenicity and serotype might be random. However, recent studies show that the enterotoxigenic strains so far describe belong to a restricted range of serotypes. Enterotoxigenic strains of E. coli O6.H16 and E. coli O148.H28 have been associated with outbreaks of diarrhoea in several countries, therefor strains of E. coli belonging to these serotypes were selected for further study. Twenty-three strains of E. coli O6.H16 and 14 strains of E. coli O148.H28 were examined; 20 strains of E. coli O6.H16 and all 14 strains of E. coli O148.H28 were enterotoxigenic but strains of E. coli O6 wit flagellar antigens other than H16 and strains of E. coli O148 wit flagellar antigens other than H28 were not enterotoxigenic. The examination of single colony subcultures derived from the E. coli O6.H16 strains showed that in some strains loss of enterotoxigenicity had occurred in a proportional of colonies.


Clonal distribution of resistance plasmid-carrying Salmonella typhimurium, mainly in the Middle East. Strains of Salmonella typhimurium of predominantly Middle Eastern origin, but distributed from England to India, were found to carry at least three types of resistance plasmid. The most important was initially identified as an F(I) plasmid by compatibility tests, but differs from the F factor on the one hand and the F(I) factors R162 and ColV on the other. The three groups of F(I) plasmids can be distinguished by their compatibility reactions with the MP10 plasmid of S. typhimurium (Smith, Humphreys, Grindley, Grindley & Anderson, 1973) and group H(1) factors: the F factor is unilaterally incompatible with group H(1) (Smith, Grindley, Humphreys & Anderson, 1973; Anderson, 1975b); the F(I) factors are compatible with MP10 and group H(1); and F(I)me factors are incompatible with MP10 but compatible with H(1). The majority of S. typhimurium cultures belonged to phage type 208; most of those that did not, belonged to types related to 208. Only a minority of their F(I)me plasmids were autotransferring. The remainder were mobilizable by F-like plasmids, and by group H(1) and H(2) factors, but not by the fi(-) I(1) factor Delta, or by plasmids of the I(2), B, P, W, N and com 7 groups. The compatibility reactions of the autotransferring F(I)me plasmids were identical with those of the non-transferring members of the group, and both were large, single-copy plasmids.The S. typhimurium strains of this series carried A or AK, and SSu resistance determinants: small, probably multicopy, non-transferring plasmids similar to those originally described in phage type 29 of S. typhimurium (Anderson & Lewis, 1965b).These S. typhimurium cultures probably represent a clone of wide geographical distribution. The accurate epidemiological study of such clonal outbreaks requires, in addition to phage typing, precise identification of the plasmids harboured by the epidemic strains, and may have to be carried to the molecular level.F(I)me plasmids were identified in other drug-resistant salmonellas, notably in a strain of S. wien which caused large outbreaks of mainly paediatric infection in Algeria, and also spread to Britain. An F(I)me plasmid was found in S. typhi phage type 44 from Algeria, in which the phage-restricting properties of the plasmid are responsible for the specificity of the type.


Recurrent aphthous stomatitis: clinical characteristics and evidence for an immunopathogenesis. Recurrent aphthous stomatitis is one of the most common diseases affecting the oral mucosa, and one of its variants is the most painful affliction of the oral mucosa. The lesions of recurrent aphthous stomatitis can be manifested as part of a broad spectrum of clinical disease ranging from the common minor aphthous ulcers to Behçet's syndrome. Differential diagnosis, although not often difficult, must include many conditions capable of producing erosive and ulcerative oral mucosal lesions. The salient features of recurrent aphthous stomatitis suggest that it is a heterogeneous entity. Recent immunologic investigations have focused attention on a possible immunopathogenesis and the evidence for this is reviewed.


Immunohistochemical studies of the human cutaneous basement membrane-anchoring fibril complex. Antiserum to the human cutaneous basement membrane-anchoring fibril complex specifically binds to the basement membrane zone of normal human skin as demonstrated by indirect immunofluorescence and, in addition, cross-reacts with normal human glomerulus. This antiserum does not prevent binding of pemphigoid antibody to its specific antigen; thus the antigen of bullous pemphigoid is unlikely to be a component of the cutaneous basement membrane (basal lamina).


Microwave irradiation: a new rapid technique for separating epidermal tissue from mouse skin preserving RNA, DNA, protein and phorbol ester-induced ornithine decarboxylase levels. Exposure of mouse skin to microwave radiation for 10 sec allows the epidermis to be separated from the dermis by gentle scraping of the skin with a scapel. At the end of the irrradiation period the subcutaneous temperature was 55 degrees C. Histologically, the microwave irradiated skin resembled skin exposed to an established heat separation procedure (i.e., immersion in 55 degrees C water followed by immersion in 0-4 degrees C water). The recoveries of DNA, RNA, and protein on a per area basis were the same for the microwave and conventional heat separation procedures, and the TPA-induced ornithine decarboxylase levels were comparable.


Circulating IgA anti-basement membrane zone antibodies in dermatitis herpetiformis. Criculating anti-basement membrane zone antibodies of the IgA class were detected in the sera of 2 or 6 dermatitis herpetiformis (DH) patients who had linear in-vivo-bound IgA deposits and in 1 of 42 DH patients who had granular vivo-bound IgA deposits. In the former 2 patients the circulating antibodies were localized ultrastructurally to the identical site where the in vivo-bound antibodies were localized and were bound by antigens in either the lamina lucida or the subbasal lamina anchoring fibril area of the basement membrane zone of normal human skin.