Tuning of the porin expression under anaerobic growth conditions by his-to-Asp cross-phosphorelay through both the EnvZ-osmosensor and ArcB-anaerosensor in Escherichia coli

BACKGROUND: Widespread bacterial signal transduction circuits are generally referred to as 'two-component systems' or 'histidine (His)-to-aspartate (Asp) phosphorelays.' In Escherichia coli, as many as 30 distinct His-to-Asp phosphorelay signalling pathways operate in response to a wide variety of environmental stimuli, such as medium osmolarity and anaerobiosis. In this regard, it is of interest whether or not some of them together constitute a network of signalling pathways through a physiologically relevant mechanism (often referred to as 'cross-regulation'). We have addressed this issue, with special reference to the osmo-responsive EnvZ and anaero-responsive ArcB phosphorelay signalling pathways in E. coli. RESULTS: Under standard aerobic growth conditions, it is well known that the osmoregulatory profile of the outer membrane porins (OmpC and OmpF) is mainly regulated by the EnvZ-OmpR phosphorelay system in response to medium osmolarity. In this study, it was found that, under anaerobic growth conditions, E. coli cells exhibit a markedly altered expression profile of OmpC and OmpF This profile was significantly different from that observed for the cells grown aerobically. Results from extensive genetic studies showed that, under such anaerobic growth conditions, the arcB gene encoding the anaero-sensory His-kinase appears to be an auxiliary genetic determinant that regulates the expression profile of porins. We then provided several lines of in vivo and in vitro evidence, which taken together, supported the following conclusions. CONCLUSIONS: Under anaerobic growth conditions, porin expression is tuned not only by the authentic osmo-resposive EnvZ sensor, but also by the anaero-responsive ArcB sensor, in an OmpR-dependent manner. It is suggested that such ArcB-mediated cross-regulation plays a physiological role by integrating anaerobic respiratory signals into the porin regulation in E. coli anaerobiosis. The proposed model is a clear example of the interplay of two distinct His-to-Asp phosphorelay signalling pathways.     

Mouse Colon Carcinoma Cells Established for High Incidence of Experimental Hepatic Metastasis Exhibit Accelerated and Anchorage-Independent Growth

Highly metastatic variants of mouse colon 38 colon carcinoma cells were established by repeated selection in vivo for liver metastasis and designated as SL4 cells. The SL4 cells formed colonies in the liver of 100% of syngenic mice when injected intrasplenically, while the incidence of liver metastasis was 27% of mice injected with parental cells. The weight of livers, which is an indicator of experimental hepatic metastasis formation, was significantly higher after intrasplenic injection and subsequent splenoctomy with SL4 cells than colon 38 cells. The incidence of hepatic metastasis after intracecal injection of SL4 cells was significantly higher than that of colon 38 cells. The SL4 cells were tested in vitro for their properties. Differences were not detected in the motility and invasive behavior between colon 38 cells and SL4 cells. SL4 cells showed a higher proliferation rate than colon 38 cells under adherent conditions. SL4 cells maintained a capacity to proliferate under non-adherent conditions whereas parental cells did not. SL4 cells should be a useful tool to study the mechanism of hepatic metastasis of colon carcinoma cells and to develop methods to prevent hepatic metastasis.     

Gene-inducing program of human dendritic cells in response to BCG cell-wall skeleton (CWS), which reflects adjuvancy required for tumor immunotherapy

Adjuvants induce the expression of a number of genes in dendritic cells (DCs), which facilitate effective antigen-presentation and cytokine/chemokine liberation. It has been accepted that the toll-like receptor (TLR) family governs the adjuvant activity in DCs. An adjuvant with a long history is mycobacteria in an oil-in-water emulsion, namely Freund's complete adjuvant. Since the active center for the adjuvancy in mycobacteria is the cell-wall skeleton (CWS), we used the bacillus Calmette-Guerin cell-wall skeleton (BCG-CWS) to test DC maturation by GeneChip analysis. We identified the genes supporting an efficient DC response and output. Approximately 2000 genes were up-regulated by BCG-CWS stimulation. BCG-CWS-, peptidoglycan (PGN)- and lipopolysaccharide (LPS)-stimulation generally up-regulated some gene clusters including genes for inflammatory cytokines (TNF, IL1alpha, IL1beta, IL6, IL12 p40, IL23 p19, etc.), chemokines (CCL20, IL8, etc.), cell adhesion molecules (ICAM-1, etc.), apoptosis-related proteins (GADD45B, BCL2A1, etc.), metabolic enzymes (PTGS2, SOD2, etc.) and miscellaneous proteins (EHD1, TNFAIP6, etc.). LPS-stimulation, but not BCG-CWS- or PGN-stimulation, up-regulated the interferon-inducible antiviral proteins, including IFIT1, IFIT2, IFIT4, CXCL10, ISG15, OASL, IFITM1 and MX1. We also found that the BCG-CWS- or PGN-stimulation up-regulated CXCL5, MMP1, etc. We discussed their properties in association with TLRs and recently discovered TLR adapters.     

Paneth Cells in Intestinal Metaplasia of the Stomach and Duodenum of Man

The Paneth cells in intestinal metaplasia of stomach and the duodenum in human subjects were studied ultrastructurally, and the fine structures of these cells were compared. Paneth cells showed the ultrastructure of serozymogenic cells and secreted their secretory granules by merocrine process. The rod or tubular dense bodies were observed in the apical region of some Paneth cells. The structures may have some relation to the secretion of the secretory granules. The secretory granules with less dense layer in the periphery, which had never been described in the Paneth cell of man, were also observed. Morphologically intermediate cells between Paneth cell and goblet cell were found. Some of the Paneth cells might be phagocytized by undifferentiated crypt cells. The Paneth cells in intestinal metaplasia were fundamentally the same as those in duodenum at least in morphology. Difference between them was that Paneth cells with many phagolysosomes in the lower cytoplasm were observed more frequently in the duodenum than in intestinal metaplasia of the stomach. The physiological functions of the Paneth cell have been discussed. ACTA PATH. JAP. 27:677–695, 1977.     

Thyroid dysfunction and histologic correlation using thyroid needle biopsy specimens

In this article, we describe the usefulness of thyroid needle biopsy in the differential diagnosis of thyroid disorders revealing unusual thyroid function. Firstly, we describe the relationship between thyroid function and its histology in 601 cases of chronic thyroiditis. In the histologic group A, the majority of the cases showed a latent or overt hypothyroidism and in histologic group B, hyperthyroid, euthyroid and latent hypothyroid cases were found in nearly equal frequency, respectively. In histologic group C, most cases were in euthyroid and in histologic group D, most cases showed a hyperthyroidism. In the silent thyroiditis and postpartum thyroiditis, known to show a characteristic clinical and laboratory finding, the histologic features of thyroid gland were as follows: the observed characteristic histologic changes in both diseases were an extensive follicular destruction associated with chronic diffuse thyroiditis. These destructive changes disappeared in association with the clinical and laboratory recovery. In the patients with iodine-excess hypothyroidism, the thyroid glands also revealed characteristic histologic changes. A marked hyperplastic change of follicular cells and a lack of colloid material in the follicular lumen was the predominant histologic feature. Based on our personal experiences, thyroid needle biopsy should be recommended as a useful tool for differentiation of causes of hyper- or hypothyroidism.     

PRIMARY LEIOMYOSARCOMA OF BONE

A 62-year-old female with primary leiomyosarcoma of the left femur is reported with a review of 21 cases reported in the literature. The resected specimen showed that the tumor extended from the femoral head to the diaphysis for 13cm in length. The tumor showed mainly intramedullary proliferation, but extraosseous growth was also noted at the great trochanter. Microscopic examination revealed well differentiated leiomyosarcoma characterized by interlacing bundles of fusiform cells with eosinophilic cytoplasm and rod-shaped hyperchromatic nuclei. PAP stain of actin on the tumor cells was positive. On electron microscopy, microfilament of 6–8 nm in diameter, dense bodies, plnocytotic vesicles, marginal attachment plate, and basal lamina were noted. The patient died with pulmonary metastasis, 1 year and 7 months after the operation. An autopsy showed metastases in the right pelvic cavity and bilateral lungs, and confirmed the primary site to be the left femur.     

Transcapillary fluid movement in human calf after drinking hypertonic saline

1. Transcapillary absorption of interstitial fluid was demonstrated with a pressure plethysmograph applied to the human calf after the ingestion of 200 ml. hypertonic (5.1%) saline. Capillary absorption began within 15 min after ingestion and lasted for about 2 hr. The maximum rate of absorption (0.019 ml./min. 100 ml. tissue) was attained 30-75 min after ingestion.2. The total amount of fluid absorbed into capillary blood vessels in the calf was 1.11 ml./100 ml. tissue. The amount of fluid thus absorbed in the whole body was estimated to be 677 ml.3. The capillary filtration coefficient (CFC) of the calf was also measured by the pressure plethysmograph. This was 0.0038 ml./min. mm Hg. 100 ml. tissue.4. The peak value of capillary absorption pressure was 5.2 mm Hg.5. The total osmotic pressure of the plasma rose by 12.6 m-osmole/kg H(2)O after ingestion. This rise was accompanied by transcapillary fluid absorption.6. The plasma protein concentration and packed cell volume were almost unchanged by ingestion, indicating that the plasma volume was unaltered.7. It was estimated that the net shift of fluid between intracellular and interstitial compartments during the period of transcapillary fluid absorption was very small.8. It is concluded that the volume of fluid moving from plasma into intestinal lumen is the same as that flowing from interstitial fluid into plasma, and that the transcapillary absorption is caused by a difference in osmotic pressure between the plasma and the interstitial fluid.