Thyroid Epithelial Morphogenesis in Vitro: A Role for Bumetanide-Sensitive Cl Secretion during Follicular Lumen Development

The structural and functional unit of the thyroid gland is the follicle, consisting of a closed lumen surrounded by a single layer of polarized epithelial cells. In this paper we have attempted to characterize the process of lumenal development when primary cultures of porcine thyroid cells reorganized to form follicles. Cells incubated with the loop diuretic, bumetanide, an inhibitor of NaK2Cl cotransport, aggregated but failed to form normal follicles. Laser scanning confocal microscopy combined with immunohistochemical markers of thyroid cell-surface proteins demonstrated that in the presence of bumetanide cells polarized and assembled ZO-1-containing tight junctions separating their apical and basolateral membrane domains. Cultures formed small lumena but their subsequent growth was inhibited by bumetanide. Electrophysiological studies confirmed that bumetanide-sensitive Cl^- transport was the major contributor to the transepithelial electrical potential difference across the follicular wall after 48 h incubation. Other potential mechanisms did not contribute significantly to follicular lumenal growth. In particular, bumetanide did not affect cell proliferation and, in contrast to tissue follicles, thyroglobulin could not be detected within the lumena of cultured follicles. We conclude that thyroid follicular reorganization involves two distinct and separate phases of lumenal development: initial lumen formation which probably reflects the assembly of a specialized apical membrane domain; and subsequent lumenal growth which is mediated by the inward transport of Cl^- by polarized epithelial cells.     

Evolutionary rates for tuf genes in endosymbionts of aphids

The gene encoding elongation factor Tu (tuf) in aphid endosymbionts (genus Buchnera) evolves at rates of 1.3 x 10(-10) to 2.5 x 10(-10) nonsynonymous substitutions and 3.9 x 10(-9) to 8.0 x 10(-9) synonymous substitutions per position per year. These rates, which are at present among the most reliable substitution rates for protein-coding genes of bacteria, have been obtained by calibrating the nodes in the phylogenetic tree produced from the Buchnera EF-Tu sequences using divergence times for the corresponding ancestral aphid hosts. We also present data suggesting that the rates of nonsynonymous substitutions are significantly higher in the endosymbiont lineages than in the closely related free-living bacteria Escherichia coli and Salmonella typhimurium. Synonymous substitution rates for Buchnera approximate estimated mutation rates for E. coli and S. typhimurium, as expected if synonymous changes act as neutral mutations in Buchnera. We relate the observed differences in substitution frequencies to the absence of selective codon preferences in Buchnera and to the influence of Muller's ratchet on small asexual populations.      

DNA Polymorphism Revealed by Arbitrary Primers Polymerase Chain Reaction Among Blastocystis Strains Isolated from Humans, a Chicken, and a Reptile

DNA polymorphisms of different strains of Blastocystis isolated from humans, a chicken, and a reptile were examined by an arbitrary primer PCR method. Two strains of Blastocystis hominis isolated from humans in the USA and Japan yielded nearly identical PCR products. However, one strain of B. hominis (isolated from a human in Singapore) yielded quite different PCR products. Blastocystis sp. isolated from a chicken yielded PCR products similar to those of the former two strains, while Blastocystis lapemi, isolated from a reptile, shared no bands with any of the other isolates. These results indicate the possibility that our isolate from the chicken is a zoonotic strain, and that there is intraspecific variation of Blastocystis hominis.     

Seedling Establishment of a Canopy Tree Species in Malaysian Tropical Rain Forests

Abstract Unlike many other canopy tree species in tropical rain forests, Dryobalanops aromatica (Dipterocar-paceae, local name: Kapur) establishes monospecific dominant (monodominant) forests in Peninsular Malaysia. In natural conditions, monodominance of Kapur does not necessarily mean low species diversity of the Kapur forests. While the emergent canopy layer is occupied by Kapur, many other plant species, which are common to lowland dipterocarp primary forests in the same region, are found in lower canopy layer and understory.
To understand the ecological implications of the monodominance of Kapur, we monitored post dispersal survival and seedling establishment in a pure stand of Kapur in a plantation in Kepong, near Kuala Lumpur. Immediately after seed fall, seeds and cotyledon-stage seedlings suffered high predation by vertebrates such as rodents. The predation pressure was higher in a more general fruiting year (1991) than in a sporadic fruiting year (1992). In contrast to the high mortality of seeds and newly emerged seedlings, seedlings surviving to the six-leaf stage showed low mortality, which allowed the establishment of a sapling bank.
The occurrence of saplings of Kapur with a wide range of size classes in natural forests indicates that this species is more shade tolerant than other dipterocarp species such as the Shorea group and that it could well respond to enhanced light conditions caused by canopy opening. These characteristics may partly contribute to maintaining monodominance of Kapur.     

Characterization of the adenosinetriphosphatase and calsequestrin isolated from sarcoplasmic reticulum of normal and dystrophic chickens.

The Ca2+ +Mg2+-dependent adenosinetriphosphatase (EC 3.6.1.3) and calsequestrin have been isolated from the sarcoplasmic reticulum of normal and dystrophic chicken muscle. The adenosinetriphosphatases, isolated from the two lines of chickens were identical in molecular weight, enzyme activity and in Ca2+ +Mg2+-dependence. Calsequestrins isolated from the two lines bound identical amounts of calcium. There were no differences in the Ca2+ transport functions of the sarcoplasmic reticulum membrane, isolated from the two lines of chickens. These results indicate that morphological differences in dystrophic chicken sarcoplasmic reticulum, described by Sabbadini et al (Sabbadini, R., Scales, D. Inesi, G. FEBS Lett. 54, 8 (1975), cannot be ascribed to qualitative differences in the adenosinetriphosphatase or calsequestrin.     

Serological evidence on the distinctness of the monocotyledonous families Flagellariaceae, Hanguanaceae and Joinvilleaceae

Serological comparisons of the seed proteins of Flagellaria indica L., Joinvillea bomeensis Becc. and Hanguana malayanum Merr. were made in order to provide evidence for the systematic evaluation of these taxa. Lack of serological cross-reactivity suggests the placement of these genera in three separate monogeneric families, Flagellariaceae, Joinvilleaceae and Hanguanaceae. Additional serological analyses are needed to evaluate their systematic position in the monocotyledons.