Molecular cloning of the cDNA encoding a murine sialic acid-specific 9-O-acetylesterase and RNA expression in cells of hematopoietic and non-hematopoietic origin.

We describe the isolation of a cDNA encoding a murine sialic acid-specific 9-O-acetylesterase as well as its expression pattern in cells of both hematopoietic and non-hematopoietic origin. This enzyme catalyzes the removal of O-acetyl ester groups from position 9 of the parent sialic acid N-acetylneuraminic acid. The cDNA is 2105 nt in length and encodes a protein of 541 amino acids with a predicted molecular weight of 61 kDa, not including oligosaccharides linked to eight potential N-glycosylation sites. The cDNA encoding the acetylesterase displays a widespread distribution in various cell lines and tissues. Expression studies of B lineage cell lines and primary fetal liver cells revealed a developmentally regulated expression pattern in cells of hematopoietic origin. Given the importance of 9-O-acetylation of sialic acids, the cloning of the cDNA encoding a sialic acid-specific 9-O-acetylesterase will be helpful in understanding further the regulation of this post-translational modification and the biological consequences thereof.     

A graphical method for detecting recombination in phylogenetic data sets

Current phylogenetic tree reconstruction methods assume that there is a single underlying tree topology for all sites along the sequence. The presence of mosaic sequences due to recombination violates this assumption and will cause phylogenetic methods to give misleading results due to the imposition of a single tree topology on all sites. The detection of mosaic sequences caused by recombination is therefore an important first step in phylogenetic analysis. A graphical method for the detection of recombination, based on the least squares method of phylogenetic estimation, is presented here. This method locates putative recombination breakpoints by moving a window along the sequence. The performance of the method is assessed by simulation and by its application to a real data set.      

Variation of physico-chemical parameters along a river transect through the Okavango Delta,Botswana

The Okavango Delta depends on water quantity and quality to sustain its ecosystem services. Whereas many studies have been carried out on its hydrology, few have been done on water quality in the delta. Water pH, electrical conductivity (EC), dissolved oxygen (DO), turbidity, total suspended solids (TSS) and dissolved organic carbon (DOC) were monitored at 10 sites along the Okavango–Boro–Thamalakane–Lake Ngami system almost fortnightly from June 2008 to June 2010. Water quality in the delta was generally good, despite high evapotranspiration rates which would normally produce very saline waters. Electrical conductivity and water temperature increased with distance from Mohembo to Lake Ngami, the former most likely due to evapoconcentration. In contrast, pH, DO, turbidity and TSS decreased with distance from Mohembo to Boro at the lower end of the seasonal floodplain, before increasing again to Lake Ngami. Dissolved oxygen and TSS most likely declined due to biological uptake and particle sedimentation, respectively. Strong and significant relationships were observed between TSS and turbidity and between DOC and EC, indicating that turbidity and EC could be useful proxies for routine estimations of TSS and DOC, respectively, in the delta.     

Holocene emergence in the Cook Islands,South Pacific

There is evidence of Holocene emergence on several of the Cook Islands. On Suwarrow Atoll there are extensive outcrops of emergent, but truncated, reef on the northern atoll rim, radiocarbon-dated 4680–4310 years B. P., overlain by younger cemented boulder conglomerates. On the northeast of the atoll there are fossil algal ridges indicating up to 1 m of emergence; the landwardmost has been dated 4220 years B. P., the intermediate one 3420 years B. P. and the present one 1250 years B. P. On Mitiaro, a makatea island in the Southern Cooks, there are emergent reefal deposits in the centre of the reef flat dated 5140–3620 years B. P. Similar thought poorly preserved deposits occur on Mauke, and an erosional bench and notch occurs on Atiu. Emergence on all islands appears synchronous with that reported on Mangaia, where a relative fall of sea level of at least 1.7 m in the last 3400 years has been reported. The evidence for emergence is broadly similar to that described from French Polynesia, though timing of emergence appears to differ.     

Influence of gibberellic and abscisic acids and the growth retardant,CCC, upon plastid development

Summary Gibberellic acid (GA₃) enhances ultrastructural morphogenesis of plastids in greening cereals whilst abscisic acid (ABA) and CCC have the reverse effect over a shorter period. GA₃ application counteracts the ABA and CCC inhibition of membrane development and, over longer periods of greening, the fastest rate of chloroplast development is shown in the presence of both GA₃ and ABA. Experiments with isolated etioplasts show that the ABA inhibition of development also occurs and can be counteracted with GA₃ treatment but no individual enhancement of plastid morphogenesis by GA₃ was detected.Ribulose 1,5-diphosphate carboxylase (RuDPC) acitvity was also increased with applications of GA₃ and reduced with ABA treatment. The initial levels of RuDPC activity in the presence of CCC were concentration dependant; high in 10^-3 M CCC and low in 10^-6 M CCC but activity returned to normal levels after CCC application was stopped. Experiments with isolated etio-chloroplasts gave similar results but levels of RuDPC activity declined rapidly in both illuminated and un-illuminated incubated suspensions of intact etioplasts.     

A note on the food of the Norway lemning

Norway lemming droppings remaining from the high 1963 population in Jämtland (mid-Sweden) were examined microscopically for an evaluation of food species present. The findings from this were compared with the occurrence of these species growing on the mountains. It is tentatively concluded that lemmings are exercising a degree of selectivity in their choice of food species, but the reasons for this choice still remain obscure.     

Lectin histochemistry of the mast cell: A light microscopical study

Summary The purpose of this study was to determine if human mast cell granules contain non repeating oligosaccharide sequences. The binding of lectins to human mast cell granules was studied using a panel of 11 lectins variously selective for bothN- andO-linked oligosaccharide sequences. The tissues were principally derived from cutaneous neurofibromata and benign and malignant breast diseases, that is, readily available human material with a known high content of mast cells. Lectin-binding sites in the formalin-fixed paraffin-embedded or resin-embedded material were visualized by means of biotinylated lectins and an avidin—peroxidase technique for light microscopy. The results indicate that human mast cell granules contain abundantN-linked sequences, but few or noO-linked residues. These sequences appear to be mostly in the form of non-bisected highly branched or smaller biantennate sequences, although variable positive binding with erythrophyto-haemagglutinin was observed, indicating some degree of bisection.     

Sodium butyrate suppresses the transforming activity of an activated N-ras oncogene in human colon carcinoma cells

The transforming activity of DNA from a newly established undifferentiated human colon carcinoma cell line (MIP-101) was tested in the NIH-3T3 transfection assay. Southern blot analysis of the transfectant DNA revealed the presence of a human N-ras oncogene. Treatment of MIP-101 cells with the maturational agent sodium butyrate induced a more normal phenotype, including diminished growth rate, elimination of anchorage independent growth, and decreased tumorigenicity (R. Niles, S. Wilhelm, P. Thomas, and N. Zamcheck (1988) J. Cancer Invest. 6, 39). Here we report that there is a significant reduction in the transforming efficiency of the DNA from butyrate-treated MIP-101 cells. A nonspecific reduction in total DNA uptake as an explanation for these findings was eliminated by showing that there was similar uptake and expression of the thymidine kinase gene from the DNA of butyrate-treated and control MIP cells. Butyrate treatment had no detectable effect on the overall structure, methylation, and level of expression of the human N-ras gene from MIP-101 cells. An NIH-3T3 transformant ability after treatment with sodium butyrate. Although butyrate suppressed several transformed properties similar to MIP-101 cells, DNA from control and treated cultures had an identical level of transforming activity. The results suggest that the environment of the MIP cells may contain additional elements not present in the NIH-3T3 transformants which are required to observe the effect of butyrate on reduction of transforming activity.     

CCR5- and CXCR4-Utilizing Strains of Human Immunodeficiency Virus Type 1 Exhibit Differential Tropism and Pathogenesis In Vivo

CCR5-utilizing (R5) and CXCR4-utilizing (X4) strains of human immunodeficiency virus type 1 (HIV-1) have been studied intensively in vitro, but the pathologic correlates of such differential tropism in vivo remain incompletely defined. In this study, X4 and R5 strains of HIV-1 were compared for tropism and pathogenesis in SCID-hu Thy/Liv mice, an in vivo model of human thymopoiesis. The X4 strain NL4-3 replicates quickly and extensively in thymocytes in the cortex and medulla, causing significant depletion. In contrast, the R5 strain Ba-L initially infects stromal cells including macrophages in the thymic medulla, without any obvious pathologic consequence. After a period of 3 to 4 weeks, Ba-L infection slowly spreads through the thymocyte populations, occasionally culminating in thymocyte depletion after week 6 of infection. During the entire time of infection, Ba-L did not mutate into variants capable of utilizing CXCR4. Therefore, X4 strains are highly cytopathic after infection of the human thymus. In contrast, infection with R5 strains of HIV-1 can result in a two-phase process in vivo, involving apparently nonpathogenic replication in medullary stromal cells followed by cytopathic replication in thymocytes.