Structural studies of proteinase entrapment by human alpha 2-macroglobulin using 3-D electron microscopy

Human a2 -macroglobulin ( a2 M; homotetramer, Mr 720 kDa) is an essential scavenger of proteinases in the serum. Each of its four subunits has a ‘bait region’, with cleavage sequences for almost all endo-proteinases, an unusual thiol ester moiety and a receptor-binding domain (RBD). Bait region cleavage in native a2 M ( a2 M-N) by a proteinase results in rapid thiol ester breakage, with a large-scale structural transformation, in which a2 M uniquely entraps the proteinase in a cage-like structure and exposes receptor-binding domains for rapid endocytosis. Transformed a2 M ( a2 M-TR) contains up to two proteinases, which remain active to small substrates. 3-D electron microscopy is optimally suited to study this unusual structural change at resolutions near (1/30) Å−1. ^ The structural importance of the thiol esters was demonstrated by a genetically-engineered a2 M, with the cysteines involved in thiol ester formation mutated to serines, which appeared structurally homologous to a2 M-TR. This demonstrates that the four highly labile thiol esters alone maintain the a2 M-N structure, while the ‘closed trap’ formed by a2 M-TR is a more stable structural form. ^ Half-transformed a2 M ( a2 M-HT), with cleaved bait regions and thiol esters in only two of its four subunits, provides an important structural link between a2 M-N and a2 M-TR. A comparison with a2 M-N showed the two proteinase-entrapping domains were above and below the plane bisecting the long axis. Both a2 M-N and a2 M-TR consist of two dense, oppositely twisted strands with significant interconnections, indicating that the structural change involves a rotation of these strands. In a2 M-HT these strands were partially untwisted with large central openings, revealing the manner in which the proteinase enters the internal cavity of a2 M. ^ In reconstructions of a2 M-N, a2 M-HT and a2 M-TR labeled with a monoclonal Fab, the Fabs were located on distal ends of each constitutive strand, demonstrating an anti-parallel arrangement of the subunits. Separation between the top and bottom pairs of Fabs was nearly the same on all structures, but the pairs were rotated about the long axis. Taken together, these results indicate that upon proteinase cleavage the two strands in a2 M-N separate. The proteinase enters the structure, while the strands re-twist to encage it. In a2 M-TR, which displays receptor-binding arms, more than two subunits are transformed as strands in the transformed half of a2 M-HT were not separated. ^

Characterization of a Novel Role for the Tousled- like Kinase in Kinetochore Assembly and Function in Caenorhabditis elegans

Chromosome segregation is a critical step during cell division to avoid aneuploidy and promote proper organismal development. Correct sister chromatid positioning and separation during mitosis helps to achieve faithful transmission of genetic material to daughter cells. This prevents improper chromosome partitioning that can potentially result in extrachromosomal fragments, increasing the tumorigenic potential of the cells. The kinetochore is a protenaicious structure responsible for the initiation and orchestration of chromosome movement during mitosis. This highly conserved structure among eukaryotes is required for chromosome attachment to the mitotic spindle and failure to assemble the kinetochore results in aberrant chromosome segregation. Thus elucidating the mechanism of kinetochore assembly is important to have a better understanding of the regulation that controls chromosome segregation. Our previous work identified the C. elegans Tousled-like kinase (TLK-1) as a mitotic kinase and depletion of TLK-1 results in embryonic lethality, characterized by nuclei displaying poor mitotic chromosome alignment, lagging chromosome, and chromosome bridges during anaphase. Additionally, previous studies from our group revealed that TLK-1 is phosphorylated independently by Aurora B at serine 634, and by CHK-1 at threonine T610. The research presented herein reveals that both phosphorylated forms of TLK-1 associate with the kinetochore during mitosis. Moreover, by systematic depletion of kinetochore proteins, I uncovered that pTLK-1 is bona fide kinetochore component that is located at the outer kinetochore layer, influencing the microtubule-binding interface. I also demonstrated that TLK-1 is necessary for the kinetochore localization of the microtubule interacting proteins CLS-2 and LIS-1 and I show that embryos depleted of TLK-1 presented an aberrant twisted kinetochore pattern. Furthermore, I established that the inner kinetochore protein KNL-2 is an in vitro substrate of TLK-1 indicating a possible role of TLK-1 in regulating centromeric assembly. Collectively, these results suggest a novel role for the Tousled-like kinase in regulation of kinetochore assembly and microtubule dynamics and demonstrate the necessity of TLK-1 for proper chromosome segregation in C. elegans.

Diatoms, total organic carbon, and total nitrogen of sediment core Dethlingen, Germany

To provide insights into the long-term evolution of aquatic ecosystems without human interference, we here evaluate a decadal- to centennial-scale-resolution diatom record spanning about 12 ka of the Holsteinian interglacial (Marine Isotope Stage 11c). Using a partially varved sediment core from the Dethlingen palaeolake (northern Germany), which has previously been studied for palynological and microfacies signals, we document the co-evolution of the aquatic and surrounding terrestrial environment. The diatom record is dominated by the genera Stephanodiscus, Aulacoseira, Ulnaria and Fragilaria. Based on the diatom assemblages and physical sediment properties, the evolution of the Dethlingen palaeolake can be subdivided into three major phases. During the oldest phase (lasting ~1900 varve years), the lake was ~10-15 m deep and characterized by anoxic bottom-water conditions and a high nutrient content. The following ~5600 years exhibited water depths >20 m, maximum diatom and Pediastrum productivity, and a peak in allochtonous nutrient input. During this phase, water-column mixing became more vigorous, resulting in a breakdown of anoxia. The youngest lake phase (~4000-5000 years) was characterized by decreasing water depth, turbulent water conditions and decreased nutrient loading. Based on our palaeolimnological data, we conclude that the evolution of the Dethlingen palaeolake during the Holsteinian interglacial responded closely to (i) changes within the catchment area (as documented by vegetation and sedimentation) related to the transition from closed forests growing on nutrient-rich soils (mesocratic forest phase) to open forests developing on poor soils (oligocratic forest phase), and (ii) short-term climate variability as reflected in centennial-scale climate perturbations.

Vegetation in the coastal area of East Greenland

This paper deals with the syntaxonomy and ecology of debris, scree and alluvium vegetation of the Ammassalik district, Southeast Greenland, on more or less moist soil. The Oxyria digyna- and Chamaenerion latifoliumvegetation types are classified as Saxifrago-Oxyrietum digynae (Böcher 1933 ap. Nordh. 1943) Gjaerevoll 1950 respectively Chamaenerietum latifolii Böcher 1933 in the class Thlaspietea rotundifolii Br.-BI. ap. Br.-BI. et al. 1947. The chionophytic Saxifrago-Oxyrietum digynae and the Chamaenerietum latifolii occurring on river-banks are classified in the alliance Saxifrago stellaris-Oxyrion digynae Gjaerevoll 1950. This alliance belongs to the order Androsacetalia alpinae Br.-BI. ap. Br.-BI. & Jenny 1926, Thlaspietea rotundifolii Br.-BI. ap. Br.-BI. et al. 1947. The following syntaxa are described as new: Saxifrago-Oxyrietum digynae stellarietosum humifusae and typicum with two variants and one variant of the subassociation inops De Molenaar 1976, and the Chamaenerietum latifolii typicum with two variants and salicetosum herbaceae with three variants.

Agatón : La Belleza de Narciso

El encomio del trágico Agatón es un intermedio musical, una canción que detiene la avanzada conceptual del Banquete, pero con gravísimo poder proléptico. Se recupera todo lo dicho hasta allí, pero tergiversado por su enfoque de marcada impronta gorgiana. Cada comensal conlleva la concepción compuesta Amor-Belleza, donde todo amor es amor de cierta belleza. Distinguiendo el éros presentado por Agatón (erótica narcisista) intentaremos mostrar qué tipo de belleza nos presenta este comensal, y cuál es la poesía que le corresponde. Entonces se dará un agón paratextual, una referencia al gran combate que Platón sostuvo toda su vida: el conflicto entre filosofía y poesía. Mostraremos que Agatón, a la vez Narciso, poeta mimético, inspirado, poseso y cosmético, es la imagen del poeta que el filósofo ateniense detesta, que, como paradigma construido para este diálogo, tiene todos los defectos criticados a lo largo de la obra platónica y sirve para tematizar este enfrentamiento

Profiles of diatoms, pollen, XRD, XRF and organic composition of two sediment cores (PG2022 and PG2023) from Lake Kyutyunda in Yakutia, NE Siberia, Russia

Although the climate development over the Holocene in the Northern Hemisphere is well known, palaeolimnological climate reconstructions reveal spatiotemporal variability in northern Eurasia. Here we present a multi-proxy study from north-eastern Siberia combining sediment geochemistry, and diatom and pollen data from lake-sediment cores covering the last 38,000 cal. years. Our results show major changes in pyrite content and fragilarioid diatom species distributions, indicating prolonged seasonal lake-ice cover between ~13,500 and ~8,900 cal. years BP and possibly during the 8,200 cal. years BP cold event. A pollen-based climate reconstruction generated a mean July temperature of 17.8°C during the Holocene Thermal Maximum (HTM) between ~8,900 and ~4,500 cal. years BP. Naviculoid diatoms appear in the late Holocene indicating a shortening of the seasonal ice cover that continues today. Our results reveal a strong correlation between the applied terrestrial and aquatic indicators and natural seasonal climate dynamics in the Holocene. Planktonic diatoms show a strong response to changes in the lake ecosystem due to recent climate warming in the Anthropocene. We assess other palaeolimnological studies to infer the spatiotemporal pattern of the HTM and affirm that the timing of its onset, a difference of up to 3,000 years from north to south, can be well explained by climatic teleconnections. The westerlies brought cold air to this part of Siberia until the Laurentide ice-sheet vanished 7,000 years ago. The apparent delayed ending of the HTM in the central Siberian record can be ascribed to the exceedance of ecological thresholds trailing behind increases in winter temperatures and decreases in contrast in insolation between seasons during the mid to late Holocene as well as lacking differentiation between summer and winter trends in paleolimnological reconstructions.

Agatón : La Belleza de Narciso

El encomio del trágico Agatón es un intermedio musical, una canción que detiene la avanzada conceptual del Banquete, pero con gravísimo poder proléptico. Se recupera todo lo dicho hasta allí, pero tergiversado por su enfoque de marcada impronta gorgiana. Cada comensal conlleva la concepción compuesta Amor-Belleza, donde todo amor es amor de cierta belleza. Distinguiendo el éros presentado por Agatón (erótica narcisista) intentaremos mostrar qué tipo de belleza nos presenta este comensal, y cuál es la poesía que le corresponde. Entonces se dará un agón paratextual, una referencia al gran combate que Platón sostuvo toda su vida: el conflicto entre filosofía y poesía. Mostraremos que Agatón, a la vez Narciso, poeta mimético, inspirado, poseso y cosmético, es la imagen del poeta que el filósofo ateniense detesta, que, como paradigma construido para este diálogo, tiene todos los defectos criticados a lo largo de la obra platónica y sirve para tematizar este enfrentamiento

Phytoplankton in the Nhatrang Bay, South China Sea

Species composition, phytoplankton abundance, and relative yield of variable fluorescence (F_v/F_m) were determined in the mesotrophic Nhatrang Bay in October-November 2004. Species diversity (250 taxonomic units) and heterogeneity of the phytoplankton structure were high. With respect to number of species and their abundance, diatoms prevailed. In selected parts of the bay, dinoflagellates dominated. Average biomass in the water column under 1 m**2 (Bt) varied from 2.3 to 64.4 mg C/m**3 (av. 31.0 mg C/m**3). Bt values were the lowest at stations nearest to the river mouth. Seaward, Bt increased. Bt values increased with depth at some stations and decreased at others. In surface layers biomass was lower than that in the underlying waters. F_v/F_m values ranged from 0.10 to 0.64 (av. 0.49). The lowest F_v/F_m values were observed in the area close to the seaport. Over greater part of the bay, F_v/F_m values were higher than 0.47. Such values are indicative of relatively high potential of photosynthetic activity of phytoplankton. Abundance and species diversity were higher than those in the dry season (March-April).

Agatón : La Belleza de Narciso

El encomio del trágico Agatón es un intermedio musical, una canción que detiene la avanzada conceptual del Banquete, pero con gravísimo poder proléptico. Se recupera todo lo dicho hasta allí, pero tergiversado por su enfoque de marcada impronta gorgiana. Cada comensal conlleva la concepción compuesta Amor-Belleza, donde todo amor es amor de cierta belleza. Distinguiendo el éros presentado por Agatón (erótica narcisista) intentaremos mostrar qué tipo de belleza nos presenta este comensal, y cuál es la poesía que le corresponde. Entonces se dará un agón paratextual, una referencia al gran combate que Platón sostuvo toda su vida: el conflicto entre filosofía y poesía. Mostraremos que Agatón, a la vez Narciso, poeta mimético, inspirado, poseso y cosmético, es la imagen del poeta que el filósofo ateniense detesta, que, como paradigma construido para este diálogo, tiene todos los defectos criticados a lo largo de la obra platónica y sirve para tematizar este enfrentamiento

List of plant species found on Victoria Island

While engaged in geoecological field work on Victoria Island, 277 new plants could be recorded for the vicinities of Holman, Cambridge Bay, Wellington Bay, Mt. Pelly, Richardson Islands, Hadley Bay, and Minto lnlet; 8 of them were new for Victoria Island, 6 for the western Canadian arctic archipelago.

Distribution of diatoms in surface sediments and some limnological characteristics of studied lakes in the Siberian arctic

The investigation of the species composition and ecology of diatoms of modern bottom sediments in water bodies of arctic polygonal tundra in three subregions of North Yakutiya has been carried out. As a result, 161 taxons of diatoms were determined; the determinant role of the depth, conductivity, pH of the water, and geographic latitude in their distribution was confirmed, and two complexes of species with respect to the leading abiotic factors were distinguished. The diatoms of the first complex prefer shallow water bodies of high latitudes with neutral and slightly alkaline water and relatively high conductivity. The second complex is confined to the water bodies of lower latitudes with small conductivity, as well as neutral and slightly acidic water.