ENHANCEMENT OF SALMONELLOSIS AND EMERGENCE OF SECONDARY INFECTION IN MICE EXPOSED TO COLD.

Miraglia, Gennaro J. (Bryn Mawr College, Bryn Mawr, Pa.) and L. Joe Berry. Enhancement of salmonellosis and emergence of secondary infection in mice exposed to cold. J. Bacteriol. 84:1173-1180. 1962.-The ld(50) dose for mice of Salmonella typhimurium, strain RIA, is 4.1 x 10(5) for animals individually housed without bedding and maintained at 25 C. It is 3.8 x 10(3) for animals similarly housed but kept at 5 C. An intravenous injection of 0.1 ml of Proferrin 2 hr prior to infection with RIA lowers the ld(50) to 4.9 x 10(3) and to 4.0 x 10(1) for mice kept, respectively, at 25 and at 5 C. Low environmental temperature and "blockade" of the reticuloendothelial system (RES) lower the resistance of mice to about the same degree, but low temperature and RES impairment together lower resistance as if each were acting independently. No effect of cold could be detected in mice infected with the highly virulent SR-11 strain of S. typhimurium, since all animals died after infection with only a few cells. Mice that were natural carriers of salmonellae, as judged by fecal discharge, were highly resistant to challenge and responded to cold in a manner similar to normal mice infected with RIA. Strain RIA could be isolated from the tissues of infected animals with greater frequency and persisted longer in mice maintained at 5 C than those at 25 C. Staphylococci were isolated from livers of animals that survived salmonella infection for 14 days at 5 C, and the incidence of staphylococci was proportional to the number of salmonellae injected. At 25 C, only a small percentage of mice had staphylococci in tissues and these occurred independent of the infectious dose of salmonellae. These observations were made on normal mice infected with RIA and on carrier mice infected with SR-11.     

Some metabolic aspects of tolerance to bacterial endotoxin

Berry, L. Joe (Bryn Mawr College, Bryn Mawr, Pa.), and Dorothy S. Smythe. Some metabolic aspects of tolerance to bacterial endotoxin. J. Bacteriol. 90:970-977. 1965.-The tolerance to bacterial endotoxins which is produced in mice given a series of daily injections of heat-killed Salmonella typhimurium failed to occur when actinomycin D was administered with the heat-killed cells. Neither ethionine nor 2-thiouracil, when given with endotoxin, altered the development of tolerance. An injection of endotoxin, actinomycin D, or ethionine lowered the activity of the liver enzyme tryptophan pyrrolase more significantly at either 4 or 17 hr postinjection in normal mice than in tolerant mice. Similarly, an injection of either saccharated iron oxide or Thorotrast lowered liver tryptophan pyrrolase activity more extensively in normal than in tolerant animals. Activation of the reticuloendothelial system (RES) of tolerant mice, as determined by an accelerated rate of carbon clearance from the blood, was observed, but this was prevented by the appropriate dose of actinomycin D. Similar results were obtained when saccharated iron oxide, rather than endotoxin, was used to activate the RES, but these animals were not resistant to endotoxin and their tryptophan pyrrolase was normally diminished after an injection of endotoxin. Thus, RES activation may occur without tolerance developing. A more nearly normal level of enzyme activity appears to be characteristic of the tolerant state.     

Inhibition of inducible liver enzymes by endotoxin and actinomycin D

Berry, L. Joe (Bryn Mawr College, Bryn Mawr, Pa.), Dorothy S. Smythe, and Louise S. Colwell. Inhibition of inducible liver enzymes by endotoxin and actinomycin D. J. Bacteriol. 92:107-115. 1966.-Bacterial endotoxin at the ld(50) level lowers liver tryptophan pyrrolase in mice, it prevents for 16 to 20 hr the induction of the enzyme by a concurrent injection of cortisone, it lowers significantly but does not prevent substrate induction, and it reduces the enzymatic activity promptly and significantly when administered during the course of hormonal induction. The ld(50) amount of actinomycin D has a similar effect on tryptophan pyrrolase, except that its inhibition of induction by cortisone persists for a longer period of time. Endotoxin in the intact mouse induces tyrosine-alpha-ketoglutarate transaminase almost as well as cortisone, but not in the adrenalectomized animal, a fact that suggests induction of this enzyme is due to release of endogenous adrenal hormones. Actinomycin D, on the other hand, has an effect on this transaminase similar to that on tryptophan pyrrolase. The site of action of endotoxin and actinomycin D would appear to be similar for one of the two enzymes studied and different for the other, a relationship that requires a specificity difficult to imagine for a material as complex as endotoxin.     

Thomas Hunt Morgan as an Embryologist: The View from Bryn Mawr

Thomas Hunt Morgan taught at Bryn Mawr College from 1891 until1904. During his years there he concentrated his research interestson embryology; he included regeneration as an integral partof development. This article maintains that Morgan did not abandonhis interest in embryology when he became a geneticist at Columbia,but it deals mainly with his teaching and research while atBryn Mawr. He worked on the development of a great diversityof organisms, plant and animal, he used widely differing experimentalmethods to investigate them, and he concerned himself with manydifferent general and special problems. He strove to investigateproblems that were directly soluble by experimental intervention,and was highly critical, in the best possible way, of the experimentsand interpretations made by his contemporaries, who regardedhim as a leader. He exerted his influence on developmental biologynot only through his research, but also through a number offine textbooks, and by his teaching. During his Bryn Mawr yearshe encouraged his students, undergraduate and graduate, to carryout independent research. He sometimes published with them asco-author, but dozens of articles by his students were publishedwithout carrying Morgan's name as co-author.     

Genetic transformation of the blue-green bacterium,Anacystis nidulans

Evidence is presented which confirms the existence of genetic transformation in the blue-green bacterium, Anacystis nidulans. This process has been demonstrated for three mutations: streptomycin resistance, a phenylalanine requirement and an ornithine requirement. The optimal conditions under which transformation occurs are also investigated, and the potential of this system for genetic mapping is discussed.This work was submitted by K. G. O. in partial fulfillment of the requirements for the degree of Doctor of Philosophy in the Department of Biology, Bryn Mawr College.     

Photokinesis in the teleost fish Danio malabaricus

Summary Photokinesis as shown by the rapidly moving fish Danio malabaricus was examined in two experiments.In Experiment I, which involved sequences of short periods of light and darkness, the Ss for the most part showed a positive relationship between light intensity and level of activity. This reaction included a well differentiated response to abrupt intensity change, the effect of the transition from darkness to light being marked. Level of activity in both light and dark periods tended to fall; after a certain point, however, it rose again in the light but continued to fall in the darkness.In Experiment II, successive increases in the total area of lighted space available were matched by increases in the total length of time spent there by the Ss. The overall reaction, however, was initially photonegative and became more so as the experiment progressed. The range of most frequent periods of time spent in the lighted area increased gradually, while the length of entries into the shaded area gradually decreased, in the course of the experiment. But for a lighted area the Ss' length of entry was shorter than for the corresponding dark area, the choice of period was more precise, there were fewer longer periods, and the range was less. In the last condition, the Ss divided their time approximately equally between the lighted 7/8 and the shaded 1/8 of the area available. The number of times the Ss crossed from one area to the other almost doubled during the experiment.The author wishes to express his gratitude to Prof. G. Thinès for his help and encouragement. He was supported by a research grant from N. A. T. O., administered through the Science Research Council, London. The findings reported formed part of a study submitted as a doctoral dissertation at the Université de Louvain. The raw data is tabulated in this study and may be consulted at the Institut de Psychologie et des Sciences Pédagogiques. Address after 1 October: Department of Psychology, Bryn Mawr College, Bryn Mawr, Pennsylvania, U.S.A.     

Lilian Vaughan Morgan (1870-1952): Her Life and Work

Lilian Vaughan Morgan was an important geneticist of the firstpart of the 20th century. She discovered both the attached-Xand closed-X chromosomes as part of her extensive research onthe X chromosome of Drosophila. She received A.B and M.S. degreesin Biology from Bryn Mawr College and became an independentinvestigator in the 1890s at the Marine Biological Laboratory,Woods Hole, Massachusetts. Although her research at the turnof the century was in embryology, she made the successful transitionto genetics. In 1904 she married T. H. Morgan, raised four children,managed the Morgan household, and when the children were grownresumed full time research. She always worked independently,and in her lifetime published 16 single author papers.     

Inbreeding,eugenics, and Helen Dean King (1869–1955)

Helen Dean King’s scientific work focused on inbreeding using experimental data collected from standardized laboratory rats to elucidate problems in human heredity. The meticulous care with which she carried on her inbreeding experiments assured that her results were dependable and her theoretical explanations credible. By using her nearly homozygous rats as desired commodities, she also was granted access to venues and people otherwise unavailable to her as a woman. King’s scientific career was made possible through her life experiences. She earned a doctorate from Bryn Mawr College under Thomas Hunt Morgan and spent a productive career at the Wistar Institute of Anatomy and Biology in Philadelphia where she had access to the experimental subjects which made her career possible. In this paper I examine King’s work on inbreeding, her participation in the debates over eugenics, her position at the Wistar Institute, her status as a woman working with mostly male scientists, and her involvement with popular science.     

Candida glabrata and Candida albicans; dissimilar tissue tropism and infectivity in a gnotobiotic model of mucosal candidiasis

Germ-free transgenic epsilon 26 (Tgepsilon26) mice, deficient in both natural killer (NK)- and T-cells, were inoculated (orally) with each of two Candida glabrata (BG2 or BG1003) or Candida albicans (CAF2-1 or SC5314) strains. Candida glabrata- or C. albicans-colonized mice exhibited similar numbers of viable Candida in the alimentary tract. Neither C. glabrata nor C. albicans caused systemic candidiasis of endogenous (alimentary tract) origin. Candida albicans invaded oroesophageal (tongue, palate, esophagus) and keratinized gastric tissues, evoked hyperkeratosis and a prominent, chronic, granulocyte-dominated, inflammatory response in all infected tissues, stimulated the production of splenic granulocytes and was lethal for the mice within 3-5 weeks after oral colonization. The two C. glabrata strains colonized the alimentary tract and penetrated into the keratinized (cardia-antrum) gastric tissues, but in contrast to C. albicans, were unable to infect oroesophageal tissues. Furthermore, C. glabrata strains were not lethal for the Tgepsilon26 mice, and did not evoke an inflammatory response in colonized gastric tissues or stimulate the production of splenic granulocytes. This 'stealth-like' behavior could explain the ability of C. glabrata to persist in infected tissues and survive as a commensal in the alimentary tract.     

Differential radioprotection of tissues in C3H/J mice by a combination of 5-hydroxy L-tryptophan with 2-aminoethylisothiuronium bromide hydrobromide.

Differential radioprotection between normal tissues and carcinoma was observed in C3H/J mice treated with a combination of 5-hydroxy L-tryptophan (5-HTP, 100 mg/kg) and 2-aminoethylisothiuronium bromide hydrobromide (AET, 20 mg/kg). Protection to normal tissues was judged by LD50(30) and by radiation induced damage to bone marrow(BM) using clonogenic ability of blood forming stem cells (10 day CFUs) as the criteria. Pretreatment with 5-HTP + AET combination 30 min before whole body gamma radiation (WBGR) enhanced the recoveries of the number of blood forming stem cells in BM of irradiated mice after 0, 7th and 10th day of irradiation. LD50(30) for C3H/J mice was 7.3 Gy and the dose modifying factor (DMF) of 5-HTP + AET combination was 1.76. On the contrary, pretreatment with this combination did not protect the mammary carcinoma transplanted in C3H/J mice, when exposed to 80 Gy soft X-rays.     

Chlamydial Induction of Hydrosalpinx in 11 Strains of Mice Reveals Multiple Host Mechanisms for Preventing Upper Genital Tract Pathology

The female lower genital tract is constantly exposed to microbial infection, some of which can ascend to and cause pathology such as hydrosalpinx in the upper genital tract, which can affect fertility. To understand host mechanisms for preventing upper genital tract pathology, we screened 11 inbred strains of mice for hydrosalpinx induction by C. muridarum. When examined on days 60 to 80 after intravaginal infection, the 11 strains fell into 3 groups based on their hydrosalpinx severity: CBA/J and SJL/J mice were highly susceptible with a hydrosalpinx score of 5 or greater; Balb/c, C57BL/6J, C57BL/10J, C3H/HeJ and C3H/HeN were susceptible with a score between 1 and <5; NOD/ShiLtJ, DBA/1J, DBA/2J and A/J were resistant with a score of <1. The diverse range of mouse susceptibility to hydrosalpinx induction may reflect the varied clinical outcomes of C. trachomatis-infected women. When the 11 strains were infected via an intrauterine inoculation to bypass the requirement for ascension, higher incidence and more severe hydrosalpinges were induced in most mice, indicating that the interaction between chlamydial ascension and host control of ascension is critical for determining susceptibility to hydrosalpinx development in many mice. However, a few mouse strains resisted significant exacerbation of hydrosalpinx by intrauterine infection, indicating that these mice have evolved ascension-independent mechanisms for preventing upper genital tract pathology. Together, the above observations have demonstrated that different strains of mice can prevent upper genital tract pathology by using different mechanisms.     

Modified Spinning Top Homogeneous Spray Apparatus for Use in Experimental Respiratory Disease Studies

The May spinning top generator was adapted to a modified Henderson tube for producing large aerosol particles (>4 mum) to obtain almost exclusive upper respiratory tract deposition of infectious aerosols in exposed mice. The system was installed in a biological safety cabinet to permit experimentation with pathogens. A novel mechanism utilizing parts from a machinists micrometer and the mechanical stage from a light microscope was developed for the spinning top generator as a means for precisely positioning the liquid feed needle. Aerosol light-scatter properties were continuously analyzed to provide relative measures of particle size distribution and aerosol concentration. When mice were exposed to influenza virus aerosols in which none of the virus was contained in particles with aerodynamic diameters <4 mum, essentially all of the virus was deposited in the upper respiratory tract tissues.     

Induction of penicillinase by staphylococci in vitro and in vivo

Eyckmans, Luc (Cornell University Medical College, New York, N.Y.), and Edward W. Hook. Induction of penicillinase by staphylococci in vitro and in vivo. J. Bacteriol. 91:997-1003. 1966.-Staphylococci in mice with peritoneal infection showed no significant increase in penicillinase activity 6 hr after administration of penicillin G. In contrast, induction of penicillinase was readily demonstrated in leukopenic animals under similar conditions. Induction of penicillinase by staphylococci in vitro was inhibited by including leukocytes and immune serum in the mixtures. The role of leukocytes in inhibiting induction of penicillinase by staphylococci in response to penicillin was investigated.     

Metabolomic analysis of liver and skeletal muscle tissues in C57BL/6J and DBA/2J mice exposed to 2,3,7,8-tetrachlorodibenzo-p-dioxin

2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) has been demonstrated to have the adverse effects on human health. In this study, we applied a metabolomic approach in conjunction with unsupervised and supervised machine learning methods to investigate the toxic effects of TCDD. By using liquid chromatography/quadrupole time-of-flight mass spectrometry, non-targeted metabolomic analysis revealed the metabolic signatures of the toxicity in aryl hydrocarbon receptor (AhR)-high affinity C57BL/6J (C6) mice as well as low affinity strain-DBA/2J (D2) mice. Lysophospholipids and long chain fatty acids were strikingly elevated in the C6 mice exposed to TCDD in both liver and skeletal muscle tissues. Meanwhile, the level of palmitoylcarnitine, which is one of the important indicators in fatty acid β-oxidation, increased significantly. Moreover, several nucleosides and amino acids decreased markedly. On the other hand, much less differentiating metabolites were highlighted in another strain-D2 mouse model. Taking liver and skeletal muscle tissues together, the levels of inosine, valine and glutamine decreased significantly. One lysophospholipid and two fatty acids were found to be enhanced. The principal components analysis and support vector machine clustering results also exhibited discriminations in the liver and skeletal muscle tissues of the mice. The obtained results indicated that TCDD could disrupt several metabolic pathways, including fatty acid biosynthesis and amino acid metabolism in both C6 and D2 mice. The increased rate of fatty acid beta-oxidation, however, was only observed in the liver and skeletal muscle tissues of C6 mice. The perturbation of the tricarboxylic acid (TCA) cycle was testified in two strains but the change was much slighter in D2 mice. It was of particular interest to note that the succinate level was enhanced in the liver tissues of both strains, and particularly, the change was up to 11.49-fold in the liver of C6 mice treated with TCDD. Collectively, the discrimination of D2 mice was not as distinct as that of C6 mice when exposed to the same dosage. Furthermore, D2 was confirmed to be less-sensitive rather than resistant to a high dose of TCDD.     

Biochemical characterization of endogenously formed eosinophilic crystals in the lungs of mice

Crystals seldom form spontaneously within tissues of mammals, except in the urinary tract or in association with eosinophil-rich diseases in humans (Charcot-Leyden crystals). Endogenously formed eosinophilic crystals have been reported in respiratory tract and other tissues of several strains of mice, but the biochemical characterization of these crystals has not been reported. In this study, eosinophilic crystal formation was examined in homozygous C57BL/6J viable motheaten mice, lung-specific surfactant apoprotein C promoter/soluble human tumor necrosis factor p75 receptor type II fusion protein transgenic mice (C57BL/6NTac x Sv/129), and CD40L-deficient mice with spontaneous Pneumocystis carinii infection. In viable motheaten but not wild type mice, rapidly developing crystals represented a major feature of the fatal lung injury induced by macrophage dysregulation. Conversely, eosinophilic crystals did not form until 4-8 months of age in transgenic and CD40L-deficient mice and were present in 10-30% of age-matched wild type controls. Mass spectrometry analysis of proteins from bronchoalveolar lavage fluid identified the crystals as Ym1, sometimes referred to as T-lymphocyte-derived eosinophil chemotactic factor. The Ym1 sequence was homologous to chitinase, and enzymatic assays indicated a 3-5-fold increase in chitinase activity compared with control mice. Intracellular and extracellular crystals associated with epithelial damage suggested that the crystals may contribute to lung inflammation through mechanical damage and enzymatic degradation.     

Ultrastructure,in vitro and in vivo,of staphylococci exposed to antibiotics

The ultrastructure of staphylococci from the peritoneal fluid of mice treated with oxacillin and from sputum of a patient treated with ampicillin was comparable to the structure of staphylococci grown on filter membranes exposed to oxacillin, but was different from the same organisms grown in broth with oxacillin. The selection of a solid phase support growth medium, such as a filter membrane for in vitro studies of drug induced morphology, appears necessary if such studies are to reflect bacterial ultrastructure in vivo.     

A predominant role for antibody in acquired immunity to chlamydial genital tract reinfection

Acquired immunity to murine Chlamydia trachomatis genital tract reinfection has long been assumed to be solely dependent on cell-mediated immunity. However, in this study, we identify a previously unrecognized protective role for Ab. Immunity develops in Ab-deficient mice following the resolution of primary chlamydial genital infection. Subsequent depletion of CD4+ T cells, but not CD8+ T cells, in those immune Ab-deficient mice before secondary infectious challenge, resulted in an infection that did not resolve. Passive immunization with immune (convalescent) serum conferred a marked level of protective immunity to reinfection, which was characterized by a striking decrease in bacterial shedding, from >100,000 inclusion forming units to fewer than 10 inclusion forming units, and a shortened duration of infection. Furthermore, mAbs to the chlamydial major outer membrane protein and LPS conferred significant levels of immunity to reinfection and reduced chlamydial shedding by >100-fold. Anti-heat shock protein 60 mAb had no protective effect. In contrast to the marked protective efficacy of immune serum on reinfection, the course of primary infection was essentially unaltered by the passive transfer of immune serum. Our results convincingly demonstrate that Abs contribute importantly to immunity to chlamydial genital tract reinfection, and that Ab-mediated protection is highly dependent on CD4+ T cell-mediated adaptive changes that occur in the local genital tract tissues during primary infection. These results impact our understanding of immunity to chlamydial genital infection and may provide important insight into vaccine development.     

In vivo localization of Staphylococcus aureus in nasal tissues of healthy and influenza A virus-infected ferrets

An in vivo ferret model was used to study the association of Staphylococcus aureus with specific tissues of the nasal cavity in both control and influenza A virus-infected animals. Ferrets were inoculated intranasally with various doses of influenza A3/Hong Kong/1/68 virus. On Days 2, 5, 9 and 14, four or five virus-inoculated and two uninoculated controls were challenged intranasally with a 1-ml volume of radiolabeled S. aureus (3 mg dry wt), a clinical isolate of low passage history. Ferrets were allowed to clear the staphylococci in vivo for 60 to 90 min before sacrifice. The animals were anesthetized, exsanguinated, and decapitated, and the lower jaw was removed. The nasal fossae were exposed by dissection and turbinates from the left nasal fossa were used for virus isolation. The median septum and tissues from the right nasal fossa, which included vestibule and anterior and posterior turbinates, were harvested and processed for radioassay. The percentage of recoverable staphylococci from virus-infected ferrets (Days 2 and 5) was greater than or equal to 10-fold higher compared with controls and animals infected with suboptimal doses of virus; greater than or equal to 76% of the recoverable staphylococci, whether from controls or virus-infected animals, was associated with the anterior turbinates. Histologic examination of the anterior turbinates from virus-infected ferrets, particularly on Days 2 and 5 postexposure to virus, showed that the staphylococci were adhering to desquamating respiratory epithelial cells. In contrast, the anterior turbinates from control ferrets uninoculated with virus and posterior turbinates from both control and virus-infected animals showed no evidence of bacteria adhering to host cells; instead, the staphylococci were found in association with the mucus gel layer of respiratory mucosa. Examination of vestibular tissue showed staphylococci in association with cells of the stratum granulosum in both virus-infected and control animals. Results of this study suggest that the early events of S. aureus interaction with different sites of ferret nasal tissues are effected by different mechanisms, and that the interaction is significantly enhanced by virus-infection.