Aspects of shell formation in eggs of the tuatara,Sphenodon punctatus

The flexible shell from eggs of the tuatara (Sphenodon punctatus) is comprised of both calcareous and fibrous components. The calcareous material is organized into columns that extend deep into the fibrous shell membrane. Many of the fibers of the membrane are enclosed within the crystalline matrix of the columns. Columns widen and flatten slightly at the outer surface of the eggshell to form cap-like structures composed of a compact crystalline matrix containing no fibers. The outer surface of eggs laid prior to completion of shell formation consists of a series of nodes obscured by a densely fibrous matrix. Similar nodes also are found at the inner surface of partially shelled eggs. The nodes represent the outer and inner aspects of columns that had not completed formation prior to oviposition. Our interpretation is that a layer (or layers) of the shell membrane forms first, with nucleation of columns occurring shortly thereafter. Columns grow into the membrane a short distance and enclose fibers of the membrane, but the primary direction of column growth is toward what will become the outer aspect of the shell. Calcareous columns and the shell membrane form more or less in concert until crystal growth outstrips that of the membrane and a cap-like apex of compact crystalline material is formed. The end result is an eggshell in which the shell membrane and calcareous material form a single unit for much of the thickness of the shell.     

Patterns of mobilization of calcium,magnesium, and phosphorus by embryonic yellow-headed blackbirds(Xanthocephalus xanthocephalus)

Summary Embryonic blackbirds(Xanthocephalus xanthocephalus) obtain most of their calcium from the eggshell (85 90%), but all of their phosphorus comes from reserves in the yolk (80–85%) and albumen (15–20%). Approximately equal amounts of magnesium are supplied by the eggshell, the yolk, and the albumen. Yolk is depleted of magnesium and phosphorus during embryogenesis, but excess calcium absorbed from the eggeshell is stored in the yolk. Consequently reserves of calcium in the yolk actually increase 8-fold during embryonic development. Our results reveal that altricial birds manifest patterns of mobilization and deposition of calcium and other elements similar to those described for precocial species. Evolution of altriciality from precocity evidently did not entail major changes in how embryonic birds meet the challenge of obtaining the calcium, magnesium, and phosphorus required for development.     

The influence of axial structures on chick somite formation

The influence of the axial structures on somite formation was investigated by culturing, on a nutritive agar substrate, segmental plates from chick embryos having 8 to 20 pairs of somites. In the first set of experiments, segmental plate was explanted together with adjacent notochord and approximately the lateral halves of the neural tube and node region. These explants formed 18 to 20 somites within 30 hr. In a second series of experiments, the notochord and neural tube were included as before, but further regression movements in the explants were prevented by removing the node region. These explants formed only 11.9 ± 1.1 somites. Finally, explants of segmental plate that included no neural tube, notochord, or node region were made. These explants had formed 10.7 ± 1.1 somites 14 to 17 hr later. When such explants were cultured for periods longer than 17 hr, there was a marked tendency for the more posterior somites to disperse and for all of the somites to develop a peculiar “hollow” morphology. It was concluded from these results that during the period of development when chick embryos possess 8 to 20 pairs of somites, the segmental plate mesoderm (1) represents about 12 prospective somites, (2) may segment into its full complement of somites without further contact with the axial structures, but (3) requires continued intimate contact with the axial structures for normal somite morphologic differentiation and stability.     

Diminished transmission of drug resistant HIV-1 variants with reduced replication capacity in a human transmission model

Background

Different patterns of drug resistance are observed in treated and therapy naïve HIV-1 infected populations. Especially the NRTI-related M184I/V variants, which are among the most frequently encountered mutations in treated patients, are underrepresented in the antiretroviral naïve population. M184I/V mutations are known to have a profound effect on viral replication and tend to revert over time in the new host. However it is debated whether a diminished transmission efficacy of HIV variants with a reduced replication capacity can also contribute to the observed discrepancy in genotypic patterns.As dendritic cells (DCs) play a pivotal role in HIV-1 transmission, we used a model containing primary human Langerhans cells (LCs) and DCs to compare the transmission efficacy M184 variants (HIV-M184V/I/T) to HIV wild type (HIV-WT). As control, we used HIV harboring the NNRTI mutation K103N (HIV-K103N) which has a minor effect on replication and is found at a similar prevalence in treated and untreated individuals.

Results

In comparison to HIV-WT, the HIV-M184 variants were less efficiently transmitted to CCR5⁺ Jurkat T cells by both LCs and DCs. The transmission rate of HIV-K103N was slightly reduced to HIV-WT in LCs and even higher than HIV-WT in DCs. Replication experiments in CCR5⁺ Jurkat T cells revealed no apparent differences in replication capacity between the mutant viruses and HIV-WT. However, viral replication in LCs and DCs was in concordance with the transmission results; replication by the HIV-M184 variants was lower than replication by HIV-WT, and the level of replication of HIV-K103N was intermediate for LCs and higher than HIV-WT for DCs.

Conclusions

Our data demonstrate that drug resistant M184-variants display a reduced replication capacity in LCs and DCs which directly impairs their transmission efficacy. As such, diminished transmission efficacy may contribute to the lower prevalence of drug resistant variants in therapy naive individuals.