N-acetyl-D-galactosaminyl-β-(1→4)-D-galactose: A terminal non-reducing structure in human blood group Sda-active Tamm-Horsfall urinary glycoprotein

Human Sd^a-active Tamm-Horsfall urinary glycoprotein labelled with galactose oxidase and tritiated sodium borohydride was found to contain both galactose and N-acetylgalactosamine as [³H]-labelled terminal non-reducing sugars. Fragmentation of the macromolecule achieved by hydrazinolysis and acid hydrolysis was followed by fractionation of the degradation products by gel filtration, ion exchange and paper chromatography. A major product was a disaccharide which contained unlabelled galactose and [³H]-labelled N-acetylgalactosamine. Sugar analysis, sodium borohydride reduction, methylation analysis and enzymic degradation enabled the structure N-acetyl-D-galactosaminyl-β-(1→4)-D-galactose to be assigned to the disaccharide.     

A genetic reconstruction of the invasion of the calanoid copepod <Emphasis Type="Italic">Pseudodiaptomus inopinus</Emphasis> across the North American Pacific Coast

The rate of aquatic invasions by planktonic organisms has increased considerably in recent decades. In order to effectively direct funding and resources to control the spread of such invasions, a methodological framework for identifying high-risk transport vectors, as well as ruling out vectors of lesser concern will be necessary. A number of estuarine ecosystems on the North American Pacific Northwest coast have experienced a series of high impact planktonic invasions that have slowly unfolded across the region in recent decades, most notably, that of the planktonic copepod crustacean Pseudodiaptomus inopinus. Although introduction of P. inopinus to the United States almost certainly occurred through the discharge of ballast water from commercial vessels originating in Asia (the species’ native range), the mechanisms and patterns of subsequent spread remain unknown. In order to elucidate the migration events shaping this invasion, we sampled the genomes of copepods from seven invasive and two native populations using restriction-site associated DNA sequencing. This genetic data was evaluated against spatially-explicit genetic simulation models to evaluate competing scenarios of invasion spread. Our results indicate that invasive populations of P. inopinus exhibit a geographically unstructured genetic composition, likely arising from infrequent and large migration events. This pattern of genetic patchiness was unexpected given the linear geographic structure of the sampled populations, and strongly contrasts with the clear invasion corridors observed in many aquatic systems.     

Enhanced somatic single embryo formation by plasmolyzing pretreatment from cultured ginseng cotyledons

Cotyledon explants of Panax ginseng zygotic embryos directly produced somatic embryos on Murashige and Skoog medium without growth regulators. Somatic embryos were formed only near the proximal excised region of cotyledons. Multiple and/or single embryos were formed and the frequency of these formations differed according to the degree of maturity of the zygotic embryos used as the explant source. When cotyledon explants pre-plasmolysed (1.0 M sucrose for 24 h), the frequency of single embryo formation was enhanced regardless of cotyledon maturity. In addition, the distribution pattern of somatic embryos changed markedly because the embryos were formed over the whole surface of the cotyledons. Histological observation revealed that plasmolyzing pretreatment broke the plasmodesmatal connection between cells and when the embryogenic cell divisions commenced, plasmodesmatal strands were hardly observed except for newly formed cell walls. This indicates that the enhanced single embryo formation over the entire surfaces of cotyledon explants might be the result of an interruption of cell–cell interaction by plasmolyzing pretreatment.     

Platycopia compacta n. sp., the Second Species of Platycopioida (Crustacea: Copepoda) in the Indo-Pacific Region, with Remarks on Development, Feeding, Swimming, and Zoogeography

Platycopia compacta new species (Copepoda: Platycopioida) is described from the hyperbenthic zone in the Tokara Islands, southern Japan. This is the second species of the order in the Indo-Pacific region, and the eighth species of the genus. The present new species is distinguishable from the closely related P. inornata from the Bahamas by (1) the formation of allobasis in the antenna; (2) two unequal setae on the basal exite of the maxillule; (3) the intermaxillipedal process with only 3 pairs of prominences at tip; (4) the fusion of the coxa and basis in leg 3; and (5) the fusion of the second and third endopod segments in female legs 3-5. Copepodid stages III, V, and VI are compared with those of P. orientalis collected from Okinawa, and differ in the developmental patterns of the antennule and legs. The presence of an intermaxillipedal process and gut content analysis revealed that Platycopia is carnivorous. The armature of legs 2-5 of the hyperbenthic genus Platycopia is stouter than that of cavernicolous platycopioids, which indicates that Platycopia may penetrate into loose sediment, whereas the latter group might be more adapted to pelagic life. The horizontal distribution of the genus Platycopia apparently exhibits a full Tethyan track.     

Early ontogeny of vascular cambium

Observations were made on structural changes of procambium and the subsequent appearance of cambium in the developing shoot. The procambium in early stages shows radial seriation of cells as a result of repeated tangential divisions. In tangential view the procambium has initially a rather homogeneous structure and later becomes organized into two distinct systems, one composed of long cells and the other of short cells. The latter cells are arranged tangentially in axial files and transversely in radial files. They show repeated transverse divisions. Some of the short cells elongate to intrude between neighboring cells. Therefore, long cells may be derived both from cells of homogeneous structure in the first stage and from elongation of some of the short cells in axial files. Long cells have mostly tapering end walls and elongate actively. In the subsequent stages, the frequency of transverse divisions of short cells in axial files decreases and these cells expand radially. Short cells in axial files are separated from one another vertically by the elongation of neighboring long cells which break up the axial files as seen in tangential view. The vascular meristem in this stage is believed to initiate the cambium. Eventually, short cells remain mostly single, or form files of two or three cells in height in tangential view. The observations are discussed in relation to the structure of the vascular meristem in other plants.     

Quantitative patterns of Hsps in tubular adenoma compared with normal and tumor tissues reveal the value of Hsp10 and Hsp60 in early diagnosis of large bowel cancer

Large bowel carcinogenesis involves accumulation of genetic alterations leading to transformation of normal mucosa into dysplasia and, lastly, adenocarcinoma. It is pertinent to elucidate the molecular changes occurring in the pre-neoplastic lesions to facilitate early diagnosis and treatment. Heat shock proteins (Hsps), many of which are molecular chaperones, are implicated in carcinogenesis, and their variations with tumor progression encourage their study as biomarkers. There are many reports on Hsps and cancer but none to our knowledge on their systematic quantification in pre-neoplastic lesions of the large bowel. We performed immunohistochemical determinations of Hsp10, Hsp60, Hsp70, and Hsp90 in biopsies of large bowel tubular adenomas with moderate grade of dysplasia and compared to normal mucosa and adenocarcinoma with a moderate grade of differentiation (G2). A significant elevation of Hsp10 and Hsp60 only, i.e., in the absence of elevation of Hsp70 or Hsp90, in both epithelium and lamina propria was found in tubular adenoma by comparison with normal mucosa. In contrast, adenocarcinoma was characterized by the highest levels of Hsp10 and Hsp60 in epithelium and lamina propria, accompanied by the highest levels of Hsp70 only in epithelium and of Hsp90 only in lamina propria, by comparison with normal and tubular adenoma counterparts. Hsp10 and Hsp60 are promising biomarkers for early diagnosis of tubular adenoma and for its differentiation from more advanced malignant lesions. Hsp10 and Hsp60 may be implicated in carcinogenesis from its very early steps and, thus, are potentially convenient targets for therapy.     

Improving solubility of NR2B amino-terminal domain of N-methyl-d-aspartate receptor expressed in Escherichia coli

The amino-terminal domains (ATDs) of N-methyl-d-aspartate (NMDA) receptors contain binding sites for modulators and may serve as potential drug targets in neurological diseases. Here, three fusion tags (6xHis-, GST-, and MBP-) were fused to the ATD of NMDA receptor NR2B subunit (ATD2B) and expressed in Escherichia coli. Each tag's ability to confer enhanced solubility to ATD2B was assessed. Soluble ATD2B was successfully obtained as a MBP fusion protein. Dynamic light scattering revealed the protein (1mg/ml) exists as monodispersed species at 25 degrees C. Functional studies using circular dichroism showed that the soluble MBP-ATD2B bound ifenprodil in a dose-dependent manner. The dissociation constants obtained for ifenprodil were similar in the absence (64nM) and presence (116nM) of saturating concentration of maltose. Moreover, the yield of soluble MBP-ATD2B is 18 times higher than the refolded 6xHis-ATD2B. We have reported a systematic comparison of three different affinity tagging strategies and identified a rapid and efficient method to obtain large amount of ATD2B recombinant protein for biochemical and structural studies.