Regionalisation of cell fate and morphogenetic movement of the mesoderm during mouse gastrulation

The developmental fate of cells in the epiblast of early-primitive-streak-stage mouse embryos was assessed by studying the pattern of tissue colonisation displayed by lac Z-expressing cells grafted orthotopically to nontransgenic embryos. Results of these fate-mapping experiments revealed that the lateral and posterior epiblast contain cells that will give rise predominantly to mesodermal derivatives. The various mesodermal populations are distributed in overlapping domains in the lateral and posterior epiblast, with the embryonic mesoderm such as heart, lateral, and paraxial mesoderm occupying a more distal position than the extraembryonic mesoderm. Heterotopic grafting of presumptive mesodermal cells results in the grafted cells adopting the fate appropriate to the new site, reflecting a plasticity of cell fate determination before ingression. The first wave of epiblast cells that ingress through the primitive streak are those giving rise to extraembryonic mesoderm. Cells that will form the mesoderm of the yolk sac and the amnion make up a major part of the mesodermal layer of the midprimitive-streak-stage embryo. Cells that are destined for embryonic mesoderm are still found within the epiblast, but some have been recruited to the distal portion of the mesoderm. By the late-primitive-streak-stage, the mesodermal layer contains only the precursors of embryonic mesoderm. This suggests that there has been a progressive displacement of the midstreak mesoderm to extraembryonic sites, which is reminiscent of that occurring in the overlying endodermal tissue. The regionalisation of cell fate in the late-primitive-streak mesoderm bears the same spatial relationship as their ancestors in the epiblast prior to cell ingression. This implies that both the position of the cells in the proximal-distal axis and their proximity to the primitive streak are major determinants for the patterning of the embryonic mesoderm. © 1995 Wiley-Liss, Inc.     

Identification of members of the HSP30 small heat shock protein family and characterization of their developmental regulation in heat-shocked xenopus laevis embryos

In the present study we have characterized the synthesis of members of the HSP30 family during Xenopus laevis development using a polyclonal antipeptide antibody derived from the carboxyl end of HSP30C. Two-dimensional PAGE/immunoblot analysis was unable to detect any heat-inducible small HSPs in cleavage, blastula, gastrula, or neurula stage embryos. However, heat-inducible accumulation of a single protein was first detectable in early tailbud embryos with an additional 5 HSPs at the late tailbud stage and a total of 13 small HSPs at the early tadpole stage. In the Xenopus A6 kidney epithelial cell line, a total of eight heat-inducible small HSPs were detected by this antibody. Comparison of the pattern of protein synthesis in embryos and somatic cells revealed a number of common and unique heat inducible proteins in Xenopus embryos and cultured kidney epithelial cells. To specifically identify the protein product of the HSP30C gene, we made a chimeric gene construct with the Xenopus HSP30C coding sequence under the control of a constitutive promoter. This construct was microinjected into fertilized eggs and resulted in the premature and constitutive synthesis of the HSP30C protein in gastrula stage embryos. Through a series of mixing experiments, we were able to specifically identify the protein encoded by the HSP30C gene in embryos and somatic cells and to conclude that HSP30C synthesis was first heat-inducible at the early tailbud stage of development. The differential pattern of heat-inducible accumulation of members of the HSP30 family during Xenopus development suggests that these proteins may have distinct functions at specific embryonic stages during a stress response.     

T cell recognition of melanoma antigens in association with HLA-A1 on allogeneic melanoma cells

Previous studies have shown that recognition of melanoma by cytotoxic T lymphocytes may be restricted by HLA-A1, A2 and other HLA antigens. The present study examined the cytotoxic specificity and major histocompatibility complex restriction of cloned cytotoxic T lymphocytes (CTL) isolated from a patient with the HLA phenotype A3,31 who had been immunized with a vaccine prepared from HLA-A1,3 melanoma cells. Cytotoxic assays against HLA-typed allogeneic melanoma cells indicated that cloned CTL from the patient were able to kill allogeneic melanoma cells expressing HLA-A1 but not other HLA-A1-positive cells. Studies on a representative clone indicated that proliferation and cytokine (tumour necrosis factor ) production in response to melanoma cells was also associated with HLA-A1 on melanoma cells. Response to the melanoma cells was associated with interleukin-4 (IL-4) rather than IL-2 production. The antigen recognized in the context of HLA-A1 on allogeneic melanoma cells was detected in cytotoxic assays on cells from 9 of 12 HLA-A1⁺ melanoma cell lines and did not appear to be the product of the MAGE-1 or-3 genes. These findings suggest that T cells can recognize melanoma antigens in the context of alloantigens and that allogeneic vaccines containing immunodominant alloantigens may generate CTL that are ineffective against autologous melanoma. The study does not, however, exclude the possibility that CTL with specificity to the latter may be activated by allogeneic vaccines, and further studies are needed to answer this question.     

Mycorrhizal associations in Hong Kong Fagaceae

Polyphenols histochemically detected in fresh uninfected roots of Quercus, Castanopsis and Lithocarpus growing in Hong Kong and shown to be condensed tannins were found mainly as intracellular material in the cells of the root cap, the epidermal layer and the endodermis. The cell walls of the outer cortex and the endodermis also contained suberin. Following invasion by compatible ectomycorrhizal symbionts, condensed tannins disappeared from cells of the root cap and the epidermal layer but hyphae were prevented from colonizing the cortex presumably due to suberin barriers. In vitro experiments indicated that a number of broad-host ectomycorrhizal fungi could utilise various polyphenolic compounds, including tannins found in the root exudates of the host trees, with different degrees of efficiency.     

Nutrients and heavy metal contamination of plants and sediments in Futian mangrove forest

An ecological survey was carried out to determine the levels of nutrients and heavy metals in the sediments and leaf tissues of two dominant mangrove plant species, Kandelia candel and Aegiceras corniculatum, in Futian mangrove forest, Shenzhen, the People's Republic of China. The spatial and seasonal variations of these elements were also investigated. The results show that there was no major difference between two sampling sites 150 m apart. In both sites, the sediment concentrations of total and NH₄ ⁺-N, total and extractable P, total and extractable K, total organic carbon were consistently higher in the landward locations and decreased gradually towards the sea. The sediment sample collected at the seaward edge of the mangrove plant community had the lowest levels of nutrient and organic matter. The vertical variations (from the land to the sea) of sediment heavy metals were less obvious and no particular trend could be identified. Extremely high contents of Cu, Cd, Pb, Cr and Zn were found at certain locations, suggesting the occurrence of some local contamination. The mean total metal concentrations in sediments decreased in the order Mn > Zn > Cu > Cr = Pb > Cd for the sample locations. Most of the heavy metals were not in a bioavailable form as the concentrations of extractable metals were relatively low (< 1% of total metals). Pb, Cr and Cd were not detected in leaf samples. Leaf C, N, P and K contents were similar between the two species and no significant difference was found among locations, although A. corniculatum seemed to have lower Mn concentrations than K. candel. With reference to temporal variations, no significant difference in sediment concentrations of some nutrients and metals was found between the spring and autumn seasons.     

Mapping and characterization of mangrove plant communities in Hong Kong

Ecological surveys were carried out to investigate the distribution and characterization of remaining mangrove stands in Hong Kong. The field studies indicate that 43 mangrove stands, excluding Mai Po Nature Reserve, still remained along the coastline of Hong Kong despite tremendous reclamation and development which occurred in the past 40 years. Most mangrove stands were found in Deep Bay (western part)and Sai Kung District (eastern coasts). The total areas occupied by these mangrove stands were 178 ha,varying from a very small stand (with 1–2 mangrove shrubs) to fairly extensive mangroves in Deep Bay (> 10 ha). It appeared that mangrove stands located in Deep Bay area were larger than those in the eastern coasts. Twenty plant species were identified from these stands, with 13 being exclusive or associate mangrove species. The major constituent species were Kandelia candel, Aegiceras corniculatum, Excoecaria agallocha and Avicennia marina. Rare species such as Heritiera littoralis were only found in a few mangrove stands. Out of the 43remaining mangrove stands, 23 were more worthwhile for conservation and their plant community structures were further investigated by transect and quadrat analyses. The importance values (sum of relative abundance,frequency and dominance) show that K. candel was the most dominant species. Species richness and Simpson's indices together with tree height, tree density and canopy area fluctuated significantly between mangrove stands. These values were used to prioritize the conservation potential of the remaining mangrove stands in Hong Kong. This revised version was published online in August 2006 with corrections to the Cover Date.     

Comparison of the effect of calcium(II) and manganese(II) ions on trypsin autolysis

The effect of Mn²⁺ and Ca²⁺ ions on the rate of trypsin autolysis was studied at pH 7.0 and at 34.4-60.2°C. For comparison, the kinetic constants of esterolytic activity of trypsin in the presence of the metal ion were determined at pH 7.4 and at 36° and 40°C. There was no significant difference in the rate of autolysis between Mn²⁺ and Ca²⁺ in the temperature range 34-47°C, but at 56.8° and 60.2° autolysis was slightly more rapid in the presence of Mn²⁺. The Mn²⁺ or Ca²⁺ ion bound to trypsin is supposed to control the conformation and thereby the stability and the activity of the enzyme. This indirect effect of Mn²⁺ and Ca²⁺ is discussed on a structural basis of the enzyme molecule.     

Oxygen-binding and immunological properties of complexes between dextran and animal haemoglobins.

Complexes of dextran 20 000 with haemoglobins of sheep, rabbit, dog, bovine and human origin were prepared through alkylation of haemoglobin by N-bromoacetylaminoethylamino-dextran. The yields were uniformly high. Complex-formation in each case was accompanied by the disappearance of reactive thiol groups on the haemoglobin, and by an increase in the affinity of the haemoglobin for oxygen. The immunological properties of dog, rabbit and sheep dextran-haemoglobin were investigated in both homologous and heterologous species. The complexes were found to be non-immunogenic in the homologous species. In heterologous species the anti-haemoglobin response induced by each complex was generally of a similar level to that induced by the haemoglobin alone.     

A single mutation affects L-serine deaminase, L-leucyl-, L-phenylalanyl-tRNA protein transferase, and proline oxidase activity in Escherichia coli K-12.

A mutation at a single locus, wyb, results in several phenotypic changes in Escherichia coli K-12. The Wyb- phenotype includes: (i) an increase in L-serine deaminase activity, together with a loss of inducibility by L-leucine; (ii) an absence of L-leucyl-, L-phenylalanyl-tRNA protein transferase activity; (iii) inducibility of proline oxidase by proline; and (iv) a loss of ability to use maltose as a carbon and energy source.