Surface functionalization of inorganic nano-crystals with fibronectin and E-cadherin chimera synergistically accelerates trans-gene delivery into embryonic stem cells

Stem cells holding great promises in regenerative medicine have the potential to be differentiated to a specific cell type through genetic manipulation. However, conventional ways of gene transfer to such progenitor cells suffer from a number of disadvantages particularly involving safety and efficacy issues. Here, we report on the development of a bio-functionalized inorganic nano-carrier of DNA by embedding fibronectin and E-cadherin chimera on the carrier, leading to its high affinity interactions with embryonic stem cell surface and accelerated trans-gene delivery for subsequent expression. While only apatite nano-particles were very inefficient in transfecting embryonic stem cells, fibronectin-anchored particles and to a more significant extent, fibronectin and E-cadherin-Fc-associated particles dramatically enhanced trans-gene delivery with a value notably higher than that of commercially available lipofection system. The involvement of both cell surface integrin and E-cadherin in mediating intracellular localization of the hybrid carrier was verified by blocking integrin binding site with excess free fibronectin and up-regulating both integrin and E-cadherin through PKC activation. Thus, the new establishment of a bio-functional hybrid gene-carrier would promote and facilitate development of stem cell-based therapy in regenerative medicine.     

Novel basic protein, PfN23, functions as key macromolecule during nacre formation

The fine microstructure of nacre (mother of pearl) illustrates the beauty of nature. Proteins found in nacre were believed to be "natural hands" that control nacre formation. In the classical view of nacre formation, nucleation of the main minerals, calcium carbonate, is induced on and by the acidic proteins in nacre. However, the basic proteins were not expected to be components of nacre. Here, we reported that a novel basic protein, PfN23, was a key accelerator in the control over crystal growth in nacre. The expression profile, in situ immunostaining, and in vitro immunodetection assays showed that PfN23 was localized within calcium carbonate crystals in the nacre. Knocking down the expression of PfN23 in adults via double-stranded RNA injection led to a disordered nacre surface in adults. Blocking the translation of PfN23 in embryos using morpholino oligomers led to the arrest of larval development. The in vitro crystallization assay showed that PfN23 increases the rate of calcium carbonate deposition and induced the formation of aragonite crystals with characteristics close to nacre. In addition, we constructed the peptides and truncations of different regions of this protein and found that the positively charged C-terminal region was a key region for the function of PfN23 Taken together, the basic protein PfN23 may be a key accelerator in the control of crystal growth in nacre. This provides a valuable balance to the classic view that acidic proteins control calcium carbonate deposition in nacre.     

洪湖水体藻类藻相特征及其对生境的响应

藻类是水生态系统的重要成分,它的群落结构、细胞密度变化与水环境相适应,随水环境的变化而改变,因此藻相变化是评价水体质量的一项重要指标。2009—2010年在洪湖水面不同方位布点采样,对水体的浮游藻类藻相(群落结构、密度)和水质状况(水位(water level,H)、水温(water temperature,tw)、透明度(transparency,SD)、总氮(total nitrogen,TN)、总磷(totalphosphorus,TP)、高锰酸盐指数(permanganate index,CODMn)和叶绿素含量(chlorophyll a,Chl-a)等)进行了逐月的调查、监测,采用湖库营养状态指数法对湖泊富营养化评价,并运用多元逐步回归方法建立水体藻类与生境因子的回归方程。结果表明:洪湖水体总体上已达到轻度富营养化状态;藻类以蓝藻门、硅藻门和绿藻门为主,共鉴定出7门65属,藻细胞密度为1.14×106—3.24×107个/L,藻相季节变化特征明显。藻类组成以蓝藻门密度最高(52.93%),硅藻门(25.96%)和绿藻门(16.83%)次之,隐藻门(1.98%)、金藻门(1.64%)、裸藻门(0.42%)和甲藻门(0.25%)相对较低。藻类藻相及优势种在不同季节有所差异,冬春季节(12—5月)以硅藻门的直链藻、小环藻和针杆藻为优势种,夏秋季节(7—11月)以蓝藻门的鱼腥藻和微囊藻为优势种;湖泊藻类细胞密度与tw、CODMn呈正显著相关关系,与H、TN、TP、SD相关关系不显著;藻细胞生长逐步回归方程为y=3.7815+0.0794tw+0.5670CODMn-1.3467SD,藻细胞生长主要受水温和有机污染物等的影响,氮磷不是藻类生长的控制因子;在湖泊不同区域,环境条件存在差异,导致藻类生长过程中起主导作用的环境因子存在一定的差异,蓝藻生长主要受tw、CODMn和SD控制,绿藻生长主要受H、tw和CODMn影响,而硅藻生长决定于水体CODMn。     

Influence of carbonate on the binding of carboplatin to DNA

The reaction of aged carboplatin (reaction of carboplatin in 24 mM NaHCO(3) for 45 h, 37 degrees, pH 8.6) with pBR322 DNA at 0 < r < 2.8, where r = [drug]/[DNA-bp], in 24 mM HEPES buffer, pH 7.4, for 24 h, followed by agarose gel electrophoresis showed DNA mobility changes consistent with unwinding closed circular DNA. However, identical experiments conducted in a two-buffer system, 24 mM HEPES plus 24 mM carbonate, showed no DNA mobility changes, indicating that carbonate blocks formation of the 1,2 intrastrand cross-link on DNA. Studies with aged carboplatin and with cisplatin carried out with ca. 4.0 < r < 10.0 in the two-buffer system show that some DNA binding and unwinding occurs for both drugs. Since carbonate inhibits the binding of aged carboplatin and cisplatin to DNA, carbonate present in the body likely modulates the reactivity of these drugs with a variety of biological targets including DNA.     

Bioerosional structures and pseudoborings from Late Jurassic and Late Cretaceous-Paleocene shallow-water carbonates (Northern Calcareous Alps, Austria and SE France) with special reference to cryptobiotic foraminifera

Examples of bioerosional processes (boring patterns) are described from shallow-water limestones of the Late Jurassic Plassen Carbonate Platform (PCP) and the Late Cretaceous to Paleocene Gosau Group of the Northern Calcareous Alps, Austria. Some micro-/macro-borings can be related to distinct ichnotaxa, others are classified in open nomenclature. In the Alpine Late Jurassic, bioerosional structures recorded from clasts in mass-flows allow palaeogeographical conclusions concerning the source areas. In particular, these are borings of the Trypanites-ichnofacies detected from clasts (Barmstein limestones) of the PCP or special type of bored ooids of unknown source areas or restricted autochthonous occurrences. In the Lower Gosau Subgroup, Gastrochaenolites macroborings occur in mobile carbonate clast substrates of shore zone deposits (“Untersberg Marmor”). Different types of borings are recorded from rudist shells and coral skeleton, some of which are referable to the ichnotaxon Entobia produced by endolithic sponges. In the present study, special attention is paid to the occurrences of the cryptobiotic foraminifera Troglotella incrustans Wernli and Fookes in the Late Jurassic and Tauchella endolithica Cherchi and Schroeder in the Late Cretaceous. The latter is so far only known to be from the Early Cenomanian of France and is reported here for the first time from the Late Turonian-Early Coniacian stratigraphic interval where it was found in turbulent carbonate deposits within borings penetrating bivalve shells or coralline algae. The records of cryptobiotic foraminifera from the Northern Calcareous Alps are supplemented by a single finding from the Middle Cenomanian of SE France. A palaeoenvironmental interpretation of the occurrences of the cryptobiotic foraminifera is provided.     

Facies development of a Late Ordovician mixed carbonate-siliciclastic ramp proximal to the developing Taconic orogen: Lourdes Formation, Newfoundland, Canada

The Upper Ordovician (late Whiterockian to Mohawkian) Lourdes Formation represents a narrow (tens of kilometers), short-lived [∼5–7 million years (my)], open-ocean (high-energy) mixed siliciclastic-carbonate ramp that onlapped allochthonous strata along the orogen side of the local Taconic foreland basin. Platform development followed a 6–8 my hiatus during which weathering had concentrated chemically mature siliciclastics that were admixed with initial carbonate sediments. A cross-platform facies gradient contains paleokarst and peritidal carbonates and sandstones, shallow-ramp carbonate bioherms and skeletal shoals, and deeper ramp calcareous shales. Transgressive systems tracts are marked by ramp-wide sheets and shoals of skeletal grainstone and low accumulation rates, and highstand systems tracts are marked by significant admixture and interbedding of siliciclastics with cross-ramp carbonate facies. Platform demise coincides with increased siliciclastic input, which is likely tectonically influenced. The Lourdes platform is equivalent to epicontinental foreland ramps along eastern Laurentia, but its narrower width precluded formation of oceanographically restricted platform-interior facies.     

Applications of nummulitids and other larger benthic foraminifera in depositional environment and sequence stratigraphy: an example from the Eocene deposits in Zagros Basin,SW Iran

The Tale-Zang Formation in Zagros Mountains (south-west Iran) is a Lower to Middle Eocene carbonate sequence. Carbonate sequences of the Tale-Zang Formation consist mainly of large benthic foraminifera (e.g. Nummulites and Alveolina), along with other skeletal and non-skeletal components. Water depth during deposition of the formation was determined based on the variation and types of benthic foraminifera, and other components in different facies. Microfacies analysis led to the recognition of ten microfacies that are related to four facies belts such as tidal flat, lagoon, shoal and open marine. An absence of turbidite deposits, reefal facies, gradual facies changes and widespread tidal flat deposits indicate that the Tale-Zang Formation was deposited in a carbonate ramp environment. Due to the great diversity and abundance of larger benthic foraminifera, this carbonate ramp is referred to as a “foraminifera-dominated carbonate ramp system”. Based on the field observations, microfacies analysis and sequence stratigraphic studies, three third-order sequences in the Langar type section and one third-order sequence in the Kialo section were identified. These depositional sequences have been separated by both type-1 and type-2 sequence boundaries. The transgressive systems tracts of sequences show a gradual upward increase in perforate foraminifera, whereas the highstand systems tracts of sequences contain predominantly imperforate foraminifera.     

Cyclic sedimentation across a Middle Ordovician carbonate ramp (Duwibong Formation), Korea

Summary The Middle Ordovician Duwibong Formation (about 100 m thick), Korea, comprises various lithotypes deposited across a carbonate ramp. Their stacking patterns constitute several kinds of meter-scale, shallowing-upward carbonate cycles. Lithofacies associations are grouped into four depositional facies: deep- to mid-ramp, shoal-complex, lagoonal, and tidal-flat facies. These facies are composed of distinctive depositional cycles: deep subtidal, shallow subtidal, restricted marine, and peritidal cycles, respectively. The subtidal cycles are capped by subtidal lithofacies and indicate incomplete shallowing to the peritidal zone. The restricted marine and peritidal cycles are capped by tidal flat lithofacies and show evidence of subaerial exposure. These cycles were formed by higher frequency sea-level fluctuations with durations of 120 ky (fifth order), which were superimposed on the longer term sea-level events, and by sediment redistribution by storm-induced currents and waves. The stratigraphic succession of the Duwibong Formation represents a general regressive trend. The vertical facies change records the transition from a deep- to mid-ramp to shoal, to lagoon, into a peritidal zone. The depositional system of the Duwibong Formation was influenced by frequent storms, especially on the deep ramp to mid-ramp seaward of ooid shoals. The storm deposits comprise about 20% of the Duwibong sequence.     

Stratigraphy, geometry, and facies of a Middle Devonian ramp-to-basin transect (Eastern Anti-Atlas, SE Morocco)

Summary In the castern Anti-Atlas (SE Morocco), a small sedimentary basin (Mader Basin) evolved during the late Palacozoic. The Middle Devonian deposits consist of shales and limestones with a thickness up to 700 m in the depocentre. Sedimentary structures and sole marks of Middle Devonian limestones indicate transport from the northwest and the south towards the basin centre, located in the central Mader area. Lithostratigraphic, biostratigraphic, and dynamic stratigraphic approaches were applied to correlate stratigraphic sections. Five correlatable large-scale base-level hemicycles were recognized in the Middle Devonian succession. Thickness trends of Middle Devonian deposits, regional correlations, and facies-trends reveal the geometry of a carbonate ramp. The carbonate ramp was slightly inclined (<1°) to the NE. A sedimentary wedge, consisting of limestones and limestone/marl alternations, was deposited during the Eifelian and marks the transition from the ramp to the adjacent basin. Middle Devonian water depths are estimated as close to and within the storm wave-base at the southern area of the ramp and far below storm wave-base in the northeastern part of the ramp. Shallowest conditions (inner-ramp environment), close or within the fairweather wave-base, existed during the early Givetian as documented by the abundance of recfal fauna (stromatoporoids, corals) and calcimicrobes (lumps, micritic envelopes) in the eastern and southeastern area of the ramp.     

Facies variability in Lower Liassic carbonate successions of the Western Dinarides (Croatia)

Summary In the Western Dinarides the Lower Liassic carbonates are underlain by Upper Triassic “Hauptdolomit”, whereas the first appearance of the foraminiferOrbitopsella praecursor (Gümbel) marks the beginning of the Middle Liassic. Their composition, observed at several localities in Western Croatia, shows a correlation of sedimentation events, which took place during Early Liassic on the Adriatic-Dinaridic carbonate platform. Facies variability is interpreted as result of autocyclic sedimentary processes on which the carbonate platform reacted by periodical oscillations of sea-bottom near the fair-weather wavebase. As a consequence, the Lower Liassic carbonate successions in the Dinarides is characterized by stacking of two main types of coarsening-upward parasequences: (1) the basal part of the Lower Liassic succession is represented by parasequences composed of mudstones or pelletal-bioclastic wackestones as their lower members, and peloidal-bioclastic wackestone/packstones to grain-stones as their upper members; and (2) the upper part of the Lower Liassic succession with parasequences consisting of mudstones or pelletal-bioclastic wackestones overlain by ooid grainstones. Judging from the composition of parasequences and thickness relations of their members, the first type is interpreted to comprise late transgressive system tract (ITST) and/or early highstand system tract (eHST), while the second type corresponds to a late highstand system tract (1HST) and/or early lowstand system tract (eLST) of a third-order sequence.