' Bacillus piliformis ' (Tyzzer) and Tyzzer’s disease of the laboratory mouse I. Propagation of the organism in embryonated eggs

1966 ◽  
Vol 165 (998) ◽  
pp. 35-60 ◽  
Keyword(s):  
Epithelial Cells ◽  
Laboratory Mouse ◽  
Serial Passage ◽  
Yolk Sac ◽  
Liver Lesions ◽  
Hepatic Cells ◽  
Large Numbers ◽  
Significant Difference ◽  
Hepatic Lesions

A contagious disease of laboratory mice characterized by focal necrotic lesions of the liver was first described by Tyzzer (1917). This author described a pleomorphic sporing organism associated with the disease and called it ' Bacillus piliformis '. He found it intracellularly in hepatic cells bordering the necrotic foci and also in epithelial cells of the large intestine of mice showing liver lesions. Many cells contained large numbers of organisms, and Tyzzer concluded that multiplication was intracellular. B . piliformis has not been cultivated on bacteriological media and hitherto the only sources available for experimental study of Tyzzer’s disease have been infected mouse liver and brain. With such material it is difficult to produce liver lesions in mice by parenteral injection unless the animals are treated with cortisone. This communication describes the isolation and serial passage of two strains of B . piliformis in embryonated eggs. One strain was propagated for 30 serial passages, 26 of which were made in eggs by yolk sac injection and the others in mice. A non-sporing variant obtained from this strain produced hepatic lesions in embryos and mice similar to those produced by the parent sporing strain. No evidence was obtained of extracellular growth of the organism and intracellular growth occurred almost exclusively in epithelial cells of the yolk sac endoderm and hepatic cells of the embryo. Although pleomorphic, B . piliformis has a distinctive morphology which was described in detail by Tyzzer. All the forms described by this author were observed in embryonated eggs. The vegetating form of the organism rapidly lost its infectivity in vitro and also in the egg following death of the embryo. This loss of infectivity proved a limitation to certain in vitro studies and no method of halting it in the fluid state was discovered. However, it proved possible to obtain sufficient survival of the non-sporing strain for inoculation purposes by preserving crude tissue suspensions at – 75 °C. The spores survived heating at 56 °C but were usually inactivated at 65 °C. They survived well at room temperature and the organism was recovered by mouse and egg passage from inoculated eggs kept at this temperature for over a year. Penicillin was found to be highly effective in halting the progress of infection in eggs. In the course of the work there were indications of a cyclic variation in the susceptibility of embryonated eggs to infection, and on one occasion a significant difference was demonstrable between eggs obtained from different pens. The results obtained when yolk sac and chick embryo liver infected with B . piliformis were injected into mice are described in the following paper (Craigie 1966).

' Bacillus piliformis ’ (Tyzzer) and Tyzzer's disease of the laboratory mouse II. Mouse pathogenicity of B . piliformis grown in embryonated eggs

1966 ◽  
Vol 165 (998) ◽  
pp. 61-77 ◽  
Keyword(s):  
Chick Embryo ◽  
Laboratory Mouse ◽  
Preceding Paper ◽  
Serial Passage ◽  
Yolk Sac ◽  
Liver Lesions ◽  
Adult Mice ◽  
Hepatic Cells ◽  
Embryo Liver ◽  
Hepatic Lesions

Hitherto the only sources of ' Bacillus piliformis ’ available for experimental study of Tyzzer’s disease have been infected mouse liver and brain and the sole evidence that this organism is the aetiological agent has been its constant association with the typical liver lesions of the disease. The isolation and serial propagation of B . piliformis in embryonated eggs is described in the preceding paper (Craigie 1966) and this communication reports results obtained when infected yolk sac and chick embryo liver were injected into mice. B . piliformis did not lose its capacity to infect hepatic cells of the mouse and produce typical liver lesions when subjected to serial passage in embryonated eggs. A non-sporing variant proved as pathogenic as the parent sporing strain and produced hepatic lesions in rats and hamsters. In general, suspensions prepared from infected yolk sac and chick embryo liver were sufficiently potent to produce liver infection when injected intraperitoneally and with a number of preparations fatal infection was obtained by intraperitoneal injection of adult mice not treated with cortisone. With active preparations the enhancing effect of cortisone was confirmed. Penicillin was found effective in checking experimental and naturally occurring infection. An increase in the severity of infection was demonstrated in mice im-­planted with a transplantable sarcoma, in which the organism multiplied. B . piliformis was also found to survive in the Krebs 2 ascites tumour preserved at - 75 °C.

Progastrin1–80stimulates growth of intestinal epithelial cells in vitro via high-affinity binding sites

2003 ◽  
Vol 284 (2) ◽  
pp. G328-G339 ◽  
Author(s):  
P. Singh ◽  
X. Lu ◽  
S. Cobb ◽  
B. T. Miller ◽  
N. Tarasova ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Binding Sites ◽  
Intestinal Epithelial Cells ◽  
Full Length ◽  
Intestinal Epithelial ◽  
Growth Effects ◽  
High Affinity ◽  
Significant Difference

Proliferation and carcinogenesis of the large intestinal epithelial cells (IEC) cells is significantly increased in transgenic mice that overexpress the precursor progastrin (PG) peptide. It is not known if the in vivo growth effects of PG on IEC cells are mediated directly or indirectly. Full-length recombinant human PG (rhPG1–80) was generated to examine possible direct effects of PG on IEC cells. Surprisingly, rhPG (0.1–1.0 nM) was more effective than the completely processed gastrin 17 (G17) peptide as a growth factor. Even though IEC cells did not express CCK1and CCK2receptors (-R), fluorescently labeled G17 and Gly-extended G17 (G-Gly) were specifically bound to the cells, suggesting the presence of binding proteins other than CCK1-R and CCK2-R on IEC cells. High-affinity ( Kd= 0.5–1.0 nM) binding sites for125I-rhPG were discovered on IEC cells that demonstrated relative binding affinity for gastrin-like peptides in the order PG ≥ COOH-terminally extended G17 ≥ G-Gly > G17 > *CCK-8 (* significant difference; P< 0.05). In conclusion, our studies demonstrate for the first time direct growth effects of the full-length precursor peptide on IEC cells in vitro that are apparently mediated by the high-affinity PG binding sites that were discovered on these cells.

123 GENISTEIN TRANSPORT ACROSS THE BOVINE OVIDUCT EPITHELIUM

2013 ◽  
Vol 25 (1) ◽  
pp. 208 ◽  
Author(s):  
C. Simintiras ◽  
F. L. Courts ◽  
R. G. Sturmey
Keyword(s):  
Epithelial Cells ◽  
Room Temperature ◽  
Cell Monolayer ◽  
Barrier Properties ◽  
Apparent Permeability ◽  
Mammalian Embryo ◽  
Significant Difference ◽  
Apical Compartment ◽  
Oviduct Epithelium

The oviduct plays a vital role in regulating the environment surrounding the gametes and early mammalian embryo. However, the permeability of the oviduct to circulating dietary-derived compounds remains relatively unknown. The present study has investigated the barrier properties of the oviduct epithelium in vitro to the movement of genistein, a soya isoflavone and analogue of 17β-estradiol that naturally occurs in the diet and has been reported to exert teratogenic effects. Bovine oviduct epithelial cells (BOECs) were isolated from abattoir-derived reproductive tracts at the mid-luetal phase. The purity of the cell isolate was confirmed using flow cytometry to determine the proportion of cells that expressed CK18 (epithelia), and Vimentin (fibroblasts). Cells were seeded at a density of 1 × 106 mL–1 on polyethylene terephthalate transwell porous supports (Corning) and maintained between two media-filled chambers for 10 to 12 days, until they formed a polarised confluent monolayer, as confirmed by transepithelial resistance (TEER) greater than 700 Ωcm2. To assess the rate of transport, genistein was added to the basal compartment at physiologically relevant levels (100 µM) and the apical compartment was sampled at regular time points for 120 mins. The concentration of genistein in the apical and basal media was measured by HPLC. Furthermore, we compared the rate of genistein transport at physiological (39°C) and room temperature to indicate whether transport was temperature dependent. Rates of transport are expressed as mean apparent permeability coefficient and were compared between groups by a Student’s t-test. Genistein crossed the bovine oviduct epithelium at a linear rate that was higher than spontaneous diffusion across a blank membrane support (12.7 × 10–3 cm–2 µM–1 v. 7.32 × 10–3 cm–2 µM–1, n = 4; P = 0.0075). The rate of genistein transport by epithelial cells was unchanged when cells were assayed at room temperature (12.7 × 10–3 cm–2 µM–1 v. 13.11 × 10–3 cm–2 µM–1, n = 3; P = 0.76), respectively. No significant difference in the directionality of transport was found. Furthermore, TEER was maintained at approximately 700 Ωcm2, indicating that the cells remained confluent for the duration of the experiment. These data suggest that the bovine oviduct epithelial cell monolayer facilitates genistein movement from the basal to the apical compartment in vitro. Furthermore, the observation that the rate of transport is unchanged by temperature suggests a passive, trans-cellular, physicochemical mechanism, rather than an active biological process. Regardless of the mechanism, the oviduct epithelium is permeable to genistein, and may even facilitate its transport into the lumen, suggesting that gametes and early embryos could be exposed to this compound in vivo. This is relevant given the previously reported finding indicating that this naturally occurring dietary isoflavone has detrimental effects on early development (Newbold et al. 2001 Cancer Res. 61, 4325–4328). Furthermore, the results demonstrate the potential use of this epithelial model in characterising the transport or barrier properties of the oviduct epithelium towards a range of circulating xenobiotics.

Capacity to adhere to and invade human epithelial cells, as related to the presence of virulence genes in, motility of, and biofilm formation ofCampylobacter jejunistrains isolated from chicken and cattle

2019 ◽  
Vol 65 (2) ◽  
pp. 126-134 ◽  
Author(s):  
Mauricio Farfán ◽  
Natalia Lártiga ◽  
María Belén Benavides ◽  
Raúl Alegría-Morán ◽  
Leonardo Sáenz ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Dairy Cattle ◽  
Broiler Chickens ◽  
Virulence Genes ◽  
Animal Reservoir ◽  
Wilcoxon Rank Sum Test ◽  
Significant Difference ◽  
T84 Cell Line ◽  
The Relationship

Campylobacter jejuni is a zoonotic pathogen transmitted through the “farm to fork” route. Outbreaks are generally associated with the consumption of chicken meat; however, dairy cows, birds, wild and domestic food animals, and pets are other important sources. Currently, there are not enough data comparing the virulence of strains isolated from these reservoirs. In this study, we compared C. jejuni strains isolated from broiler chickens and dairy cattle by determining their ability to adhere to and invade in vitro human colonic epithelial cells in the T84 cell line with their motility, formation of biofilms, and presence of eight virulence genes. A Wilcoxon Rank Sum test was performed to establish the relationship between presence of the studied genes and cellular invasion and adhesion, as well as differences between the animal species of origin of the isolate. A Spearman correlation was performed to assess the relationship between invasion and motility, along with invasion and biofilm generation. The virB11 gene was positively associated with the adherence capacity of the strains (mean difference = 0.21, p = 0.006), and strains isolated from chickens showed a significant difference for adherence compared with strains isolated from cattle (p = 0.0001). Our results indicate that strains of C. jejuni have a difference in their adherence capacity depending on the animal reservoir from which they came, with chicken isolates displaying higher virulence than dairy cattle isolates.

scholarly journals Epithelium of mouse yolk sac placenta lacks H-2 complex alloantigens.

1980 ◽  
Vol 152 (4) ◽  
pp. 945-955 ◽  
Author(s):  
E L Parr ◽  
R V Blanden ◽  
R S Tulsi
Keyword(s):  
Epithelial Cells ◽  
Cytotoxic Effect ◽  
Apical Membrane ◽  
Vascular Endothelial Cells ◽  
Yolk Sac ◽  
Mhc Antigens ◽  
Mhc Molecules ◽  
Apical Membranes ◽  
Yolk Sac Placenta

The only fetal cell membrane exposed to the mother in the mouse yolk sac placenta is the apical membrane of the endodermal epithelial cells. In yolk sac preparations in vitro, this apical membrane was exposed to reagents or cells in the incubation medium. By using several techniques we were not able to detect fetal major histocompatibility complex (MHC) antigens in this membrane. Immunoferritin labeling with and without prefixation and after neurominidase and trypsin digestion indicated that the apical membrane could contain no more than approximately 1% of the H-2 complex antigens that were present on peritoneal macrophages. Incubation of yolk sac preparations in anti-H-2 complex antiserum and complement had no cytotoxic effect on the endodermal epithelium, nor did incubation in an excess of alloreactive lymphocytes. Dissociated preparations of prefixed yolk sac contained endodermal epithelial cells and vascular endothelial cells whose entire surface membranes were exposed to the medium. H-2-complex antigens were not detected by immunoferritin labeling in either the apical or the laterobasal membrane of the yolk sac endoderm, but they were present in low density on the vascular endothelium. Also, incubation of unfixed, dissociated cells in anti-H-2-complex serum and complement had no detectable cytotoxic effect on endodermal epithelial cells. These observations indicate that H-2 antigens are sparse or absent in both the apical and laterobasal membranes of endodermal epithelial cells. The deficiency of MHC antigens in the apical membrane may account for the failure of sensitized females to reject the yolk sac, whereas the composition of the laterobasal membrane is probably less important to maternal-fetal relations. The present observations are consistent with labeling studies of adult-lining epithelial cells, which indicate that self-marker MHC molecules are absent from the apical membranes oriented toward the outside world and variably expressed in the laterobasal self-side membranes. It is suggested that the corresponding exclusion of fetal self-marker molecules from the apical membranes of some kinds of placental epithelia would deprive the mother of target sites for an alloimmune reaction at the maternal-fetal interface.

301 EFFECT OF CO-CULTURE WITH DIFFERENT STAGE EMBRYOS ON DEVELOPMENT OF ALGINATE-ENCAPSULATED SMALL NUMBER BOVINE EMBRYOS

2007 ◽  
Vol 19 (1) ◽  
pp. 266
Author(s):  
S. Kobayashi ◽  
M. Sakatani ◽  
Y. Inaba ◽  
S. Kobayashi ◽  
K. Imai ◽  
...  
Keyword(s):  
Blastocyst Stage ◽  
Developmental Competence ◽  
Bovine Embryos ◽  
Alginate Gel ◽  
Large Numbers ◽  
Significant Difference ◽  
Calcium Alginate Gel ◽  
Recorded Data ◽  
The Individual

Previous studies show that embryos cultured in large numbers have better developmental competence than those in small numbers in mice, sheep, and cattle. We have reported that co-culture of bovine embryos encapsulated in calcium-alginate gel (microcapsule) improves the development of embryos cultured in small numbers (Kobayashi et al. 2006 Reprod. Fertil. Devel. 18, 248). This method is beneficial for culture of small numbers of embryos such as OPU-derived embryos by recognizing the individual donor cows with abattoir-derived unidentified IVF embryos. In the previous study, we used the same stage embryos for co-culture of encapsulated embryos. However, in the case of unavailability of the same stage embryos, encapsulated embryos may be co-cultured with different stage embryos. Effect of different stage embryos on co-culture of encapsulated embryos is not clear. In the present study, we investigated the effect of co-culture of different stage embryos on development of encapsulated small number embryos. In vitro-matured and fertilized zygotes from abattoir derived ovaries were used for the experiment. Small numbers of zygotes were encapsulated by alginate-gel microcapsule to distinguish from co-cultured embryos. Encapsulation was carried out by putting the 1% sodium alginate solution containing zygotes slowly into 0.1% calcium chloride solution (microcapsule). The embryos used for co-culture were produced by IVF 1-3 days before preparation of encapsulated zygotes (Day 1, Day 2, and Day 3). Five encapsulated zygotes were cultured with 15 embryos for co-culture in one droplet (100 �L) made by CR1aa + 5% CS, at 38.5�C, CO2 in air. Encapsulated zygotes co-cultured with the same stage of zygotes were assigned as a control (Day 0). The rates of cleavage on Day 2 and development to blastocyst stage on Day 9 were recorded. Data were analyzed by Student&apos;s t-test. No significant difference was observed in the rate of cleavage in all experimental groups compared with control (Day 1: 72.5&percnt; (n &equals; 80) vs. control: 75.7&percnt; (n &equals; 70); Day 2: 76.3&percnt; (n &equals; 80) vs. control: 82.5&percnt; (n &equals; 80); and Day 3: 78.7&percnt; (n &equals; 75) vs. control: 70.8&percnt; (n &equals; 65). There was not a significant difference in the rate of development to the blastocyst stage in all experimental groups compared with control (Day 1: 42.5&percnt; vs. control: 44.3&percnt;; Day 2: 43.8&percnt; vs. control: 38.8&percnt;; Day 3: 44.0&percnt; vs. control: 35.4&percnt;). These results indicate that co-culture of different stages of embryos can normally support the development of small numbers of encapsulated embryos. These methods are useful to improve the development of small numbers of embryos derived from OPU-IVF embryos without synchronization of the developmental stage of co-cultured embryos.

High Mobility Group Box-1 Protein and Interleukin 33 Expression in Allergic Rhinitis

ORL ◽  
2022 ◽  
pp. 1-9
Author(s):  
Nongping Zhong ◽  
Qing Luo ◽  
Xiaoyan Huang ◽  
Jieqing Yu ◽  
Jing Ye ◽  
...  
Keyword(s):  
Allergic Rhinitis ◽  
Epithelial Cells ◽  
Nasal Mucosa ◽  
High Mobility ◽  
Normal Group ◽  
High Mobility Group ◽  
Hmgb1 Protein ◽  
Significant Difference

<b><i>Background:</i></b> Allergic rhinitis (AR) is characterized by an inflammatory reaction. High mobility group box 1 (HMGB1) protein and interleukin (IL)-33 are damage-associated molecular pattern molecules and have many characteristics similar to pro-inflammatory cytokines. However, the role of IL-33 and HMGB1 in AR remains unclear. The aim of this study is to explore the role of HMGB1 and IL-33 in AR. <b><i>Methods:</i></b> Twenty patients with AR (AR group) and 10 normal controls (normal group) were enrolled in this study. HMGB1 and IL-33 expression were analyzed by immunohistochemistry in epithelial cells of the inferior turbinate mucosa samples. Then, the human nasal mucosa epithelial cells (HNECs) were cultured in vitro, and the house dust mite allergen (Derp1) was used to stimulate the cells. Quantitative real-time PCR and ELISA assay were performed to detect HMGB1 and IL-33 expression in HNECs. <b><i>Results:</i></b> The expression of HMGB1 and IL-33 in the nasal mucosa was higher in the AR group than in the normal group, with a statistically significant difference (<i>p</i> &#x3c; 0.05). In HNECs of AR, the expression of both HMGB1 and IL-33 in stimulated groups was higher than that in non-stimulated groups. The differences were statistically significant (<i>p</i> &#x3c; 0.05). In addition, they increased gradually with the prolonging time and the concentration of the added Derp1. <b><i>Conclusions:</i></b> The expression of HMGB1 and IL-33 were both increased in AR. HMGB1 and IL-33 may have a close relationship in AR.

Myeloma light chains are ligands for cubilin (gp280)

1998 ◽  
Vol 275 (2) ◽  
pp. F246-F254 ◽  
Author(s):  
Vecihi Batuman ◽  
Pierre J. Verroust ◽  
Gabriel L. Navar ◽  
James H. Kaysen ◽  
Fatime O. Goda ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Light Chain ◽  
Brush Border ◽  
High Capacity ◽  
Inhibitory Effect ◽  
Yolk Sac ◽  
Light Chains ◽  
Endosomal Trafficking ◽  
Brush Border Membranes

Although myeloma light chains are known to undergo receptor-mediated endocytosis in the kidney, the molecular identity of the receptor has not been characterized. We examined the interaction between cubilin (gp280) and four species of light chains isolated from the urine of patients with multiple myeloma. Four lines of evidence identify cubilin, a giant glycoprotein receptor, which is restricted in distribution to endocytic scavenger pathways and which has potent effects on endosomal trafficking, as a potentially physiologically relevant binding site for light chains: 1) light chains coeluted during immunoaffinity purification of cubilin; 2) polyclonal antisera to cubilin but not control sera, displaced human light chain binding from rat renal brush-border membranes; 3) cubilin bound to multiple species of light chains during surface plasmon resonance; 4) anti-cubilin antiserum interfered with light chain endocytosis by visceral yolk sac epithelial cells. However, both binding of light chains to brush-border membranes and endocytosis of light chains by yolk sac epithelial cells were only partially inhibited by anticubilin antibodies, suggesting presence of additional or alternate binding sites for light chains. Excess light chain had a potent inhibitory effect on endosomal fusion in vitro. Binding showed dose and time-dependent saturability with low-affinity, high-capacity equilibrium binding parameters. These data demonstrate that cubilin plays a role in the endocytosis and trafficking of light chains in renal proximal tubule cells.

scholarly journals Prevalence of ExoY Activity in Pseudomonas aeruginosa Reference Panel Strains and Impact on Cytotoxicity in Epithelial Cells

2021 ◽  
Vol 12 ◽  
Author(s):  
Hazel Silistre ◽  
Dorothée Raoux-Barbot ◽  
Federica Mancinelli ◽  
Flora Sangouard ◽  
Alice Dupin ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Epithelial Cells ◽  
Bacterial Species ◽  
Lung Epithelial Cells ◽  
Reference Panel ◽  
Host Cells ◽  
Cytotoxic Effects ◽  
Infection Models ◽  
Significant Difference

ExoY is among the effectors that are injected by the type III secretion system (T3SS) of Pseudomonas aeruginosa into host cells. Inside eukaryotic cells, ExoY interacts with F-actin, which stimulates its potent nucleotidyl cyclase activity to produce cyclic nucleotide monophosphates (cNMPs). ExoY has broad substrate specificity with GTP as a preferential substrate in vitro. How ExoY contributes to the virulence of P. aeruginosa remains largely unknown. Here, we examined the prevalence of active ExoY among strains from the international P. aeruginosa reference panel, a collection of strains that includes environmental and clinical isolates, commonly used laboratory strains, and sequential clonal isolates from cystic fibrosis (CF) patients and thus represents the large diversity of this bacterial species. The ability to secrete active ExoY was determined by measuring the F-actin stimulated guanylate cyclase (GC) activity in bacterial culture supernatants. We found an overall ExoY activity prevalence of about 60% among the 40 examined strains with no significant difference between CF and non-CF isolates. In parallel, we used cellular infection models of human lung epithelial cells to compare the cytotoxic effects of isogenic reference strains expressing active ExoY or lacking the exoY gene. We found that P. aeruginosa strains lacking ExoY were in fact more cytotoxic to the epithelial cells than those secreting active ExoY. This suggests that under certain conditions, ExoY might partly alleviate the cytotoxic effects of other virulence factors of P. aeruginosa.

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