Monoclonal antibodies to rat renal tissue: an approach to the immunohistological analysis of the nephron-collecting duct system, and ultra-structural localization of antigens

Summary To obtain more accurate information on the nephron-collecting duct system, monoclonal antibodies against renal tissue were prepared. BALB/c mice were immunized every two weeks with rat renal tissue, either cortex or medulla. Spleen cells were collected and fused with myeloma cells sensitive to hypoxanthine-aminopterin-thymidine medium. Hybrids were selected for production of antibodies by indirect immunofluorescence and cloned by the limiting dilution method. Tissue reactivity of the antibodies obtained was defined by immunofluorescence. The intracellular localization of antigenic determinants was ascertained by immunoelectron microscopy. The antibodies were classified into four major groups: (1) antibodies against proximal tubules; (2) antibodies against distal tubules and the loop of Henle; (3) antibodies against collecting duct system; and (4) antibodies against glomeruli. Using immunoelectron microscopy, various intracellular antigenic determinants were recognized, such as brush border, apical canaliculi, vacuolar apparatus, luminal and basolateral plasma membranes. The results obtained indicated that electron microscopy is indispensable for the immunohistological study of the nephroncollecting duct system. The observations help to understand morphological and functional diversity of the nephron-collecting duct system.     

Colicine Typing of Shigella sonnei

The colicine typing method of Shigella sonnei is described with experimental evidence supporting it as well as the manner of selection of eight indicator strains. The disparity of principle between this method and that of Abbott and Shannon's method is (1) selection of the indicators only from wild strains existing in this country, (2) employment of heart infusion broth for colicine production, (3) performance of the typing within 48 hours, and (4) determination of types and subtypes of test-strains by combining their colicinogenic activity against the indicators and their sensitivity to colicines produced by the indicators. A modification of the method is advocated which requires three days to extract colicine by cultivation and one day for sensitivity tests and which uses peptone as the sole nutriment in media. The efficiency of the technique of Abbott and Shannon, McGeachie and McCormick, and the authors' two methods was compared using the selected indicators. Only the technique of McGeachie and McCormick showed some discrepancies.     

The effects of water pollution on the phylogenetic community structure of aquatic plants in the East Tiaoxi River,China

1. Water pollution is one of the most serious aquatic environmental problems worldwide. In China, recent agricultural and industrial development has resulted in rapid changes in aquatic ecosystems. Here, we reveal the effects of water pollution on the phylogenetic community structure of aquatic macrophytes in the Tiaoxi River, China.
2. We placed a rectangular plot at 47 sites within the Tiaoxi River from the mouth of the river to 88.5 km upstream, in which we recorded species abundance and measured 22 physico-chemical variables. Bayesian phylogeny using the rbcL and matK gene sequences was employed to quantify phylogenetic α- and β-diversity, and test the phylogenetic signal in four growth forms: emergent, floating-leaved, free-floating, and submerged.
3. Within communities, water contamination and phytoplankton abundance decreased species richness and phylogenetic diversity, which resulted in phylogenetic clustering; species within communities were more closely related to each other than expected. Between communities, differences in geographical distance and phytoplankton abundance resulted in phylogenetic dissimilarity among plots. Aquatic macrophytes showed phylogenetic signals in which related species responded more similarly to disturbance.
4. Thus, the observed patterns could be explained by environmental filtering and suggested that water pollution by human activity has added more filters to the existing environmental filters that drive the species assembly of macrophyte communities.

     

Sufficient intake of high amylose cornstarch maintains high colonic hydrogen production for 24 h in rats

Colonic hydrogen (H₂) can suppress oxidative stress and damage in the body. We examined the minimum requirement of high amylose cornstarch (HAS) to maintain high colonic H₂ production for 24 h. Ileorectostomized and sham-operated rats were fed a control diet supplemented with or without 20% HAS for 7 days. Colonic starch utilization was determined. Next, rats were fed the control diet with or without 10% or 20% HAS for 14 or 28 days, respectively. Breath and flatus H₂ excretion for 24 h was measured. 1.04 g of resistant fraction in HAS was utilized for 24 h by colonic bacteria. High H₂ excretion was not maintained for 24 h in rats fed the 10% HAS diet, from which only 0.89 g of resistant starch was estimated to be delivered. High colonic H₂ production for 24 h would be maintained by delivering more HAS to the large intestine than is utilized.     

Root endophyte symbiosis in vitro between the ectomycorrhizal basidiomycete Tricholoma matsutake and the arbuscular mycorrhizal plant Prunus speciosa

We previously reported that Tricholoma matsutake and Tricholoma fulvocastaneum, ectomycorrhizal basidiomycetes that associate with Pinaceae and Fagaceae, respectively, in the Northern Hemisphere, could interact in vitro as a root endophyte of somatic plants of Cedrela odorata (Meliaceae), which naturally harbors arbuscular mycorrhizal fungi in South America, to form a characteristic rhizospheric colony or “shiro”. We questioned whether this phenomenon could have occurred because of plant–microbe interactions between geographically separated species that never encounter one another in nature. In the present study, we document that these fungi formed root endophyte interactions and shiro within 140 days of inoculation with somatic plants of Prunus speciosa (=Cerasus speciosa, Rosaceae), a wild cherry tree that naturally harbors arbuscular mycorrhizal fungi in Japan. Compared with C. odorata, infected P. speciosa plants had less mycelial sheath surrounding the exodermis, and the older the roots, especially main roots, the more hyphae penetrated. In addition, a large number of juvenile roots were not associated with hyphae. We concluded that such root endophyte interactions were not events isolated to the interactions between exotic plants and microbes but could occur generally in vitro. Our pure culture system with a somatic plant allowed these fungi to express symbiosis-related phenotypes that varied with the plant host; these traits are innately programmed but suppressed in nature and could be useful in genetic analyses of plant–fungal symbiosis.