First record of organic-walled microfossils from the Tonian Shiwangzhuang Formation of the Tumen Group in western Shandong,North China

Organic-walled microfossils offer important information on the biospheric evolution in pre-Cryogenian and provide biostratigraphic implications for many Proterozoic fossiliferous sequences that are poorly age constrained for the lack of reliable radiometric date. Recently, macroscopic carbonaceous compression fossils have been reported for the first time from the Tonian Shiwangzhuang Formation of the Tumen Group in western Shandong, North China. However, organic-walled microfossils have never been discovered from this formation up till now. To improve our knowledge about Proterozoic biodiversity in North China, we conducted a micropaleontological survey on the argillaceous limestone samples of the Shiwangzhuang Formation, which also contain macroscopic carbonaceous compression fossils, from the Baishicun section in Anqiu, western Shandong, North China. Our investigation shows that the Shiwangzhuang microfossil assemblage is dominated by smooth-walled sphaeromorphic acritarchs and cyanobacterium-like filaments and relatively low abundance of other acritarchs, including 16 taxa, such as Polysphaeroides filliformis, Ostiana microcystis, Simia annulare, ?Jacutianema sp., Arctacellularia tetragonala, Pellicularia tenera, Polythrichoides lineatus, and Navifusa actinomorpha. The Shiwangzhuang organic-walled microfossil assemblage, although consisting of long-ranging and not age diagnostic taxa, is consistent with a Tonian age suggested by macroscopic carbonaceous compression fossils, including the Chuaria-Tawuia and Sinosabellidites-Protoarenicola-Pararenicola assemblages, revealed from the same fossiliferous horizon of the Shiwangzhuang Formation and by organic-walled microfossil assemblage, including the late Mesoproterozoic to Tonian index fossil Trachyhystrichosphaera aimika, from the underlying Tongjiazhuang Formation. However, it is also worth noting that a Cryogenian or Ediacaran age cannot be completely excluded based just on the Shiwangzhuang microfossils because of their limited biostratigraphic utility.     

Ubc9 acetylation modulates distinct SUMO target modification and hypoxia response

While numerous small ubiquitin‐like modifier (SUMO) conjugated substrates have been identified, very little is known about the cellular signalling mechanisms that differentially regulate substrate sumoylation. Here, we show that acetylation of SUMO E2 conjugase Ubc9 selectively downregulates the sumoylation of substrates with negatively charged amino acid‐dependent sumoylation motif (NDSM) consisting of clustered acidic residues located downstream from the core ψ‐K‐X‐E/D consensus motif, such as CBP and Elk‐1, but not substrates with core ψ‐K‐X‐E/D motif alone or SUMO‐interacting motif. Ubc9 is acetylated at residue K65 and K65 acetylation attenuates Ubc9 binding to NDSM substrates, causing a reduction in NDSM substrate sumoylation. Furthermore, Ubc9 K65 acetylation can be downregulated by hypoxia via SIRT1, and is correlated with hypoxia‐elicited modulation of sumoylation and target gene expression of CBP and Elk‐1 and cell survival. Our data suggest that Ubc9 acetylation/deacetylation serves as a dynamic switch for NDSM substrate sumoylation and we report a previously undescribed SIRT1/Ubc9 regulatory axis in the modulation of protein sumoylation and the hypoxia response.     

WormFarm: a quantitative control and measurement device toward automated Caenorhabditis elegans aging analysis

Caenorhabditis elegans is a leading model organism for studying the basic mechanisms of aging. Progress has been limited, however, by the lack of an automated system for quantitative analysis of longevity and mean lifespan. To address this barrier, we developed ‘WormFarm’, an integrated microfluidic device for culturing nematodes. Cohorts of 30–50 animals are maintained throughout their lifespan in each of eight separate chambers on a single WormFarm polydimethylsiloxane chip. Design features allow for automated removal of progeny and efficient control of environmental conditions. In addition, we have developed computational algorithms for automated analysis of video footage to quantitate survival and other phenotypes, such as body size and motility. As proof‐of‐principle, we show here that WormFarm successfully recapitulates survival data obtained from a standard plate‐based assay for both RNAi‐mediated and dietary‐induced changes in lifespan. Further, using a fluorescent reporter in conjunction with WormFarm, we report an age‐associated decrease in fluorescent intensity of GFP in transgenic worms expressing GFP tagged with a mitochondrial import signal under the control of the myo‐3 promoter. This marker may therefore serve as a useful biomarker of biological age and aging rate.     

Novel phage display-derived mycolic acid-specific antibodies with potential for tuberculosis diagnosis

Tuberculosis is a major cause of mortality and morbidity due to infectious disease. However, current clinical diagnostic methodologies such as PCR, sputum culture, or smear microscopy are not ideal. Antibody-based assays are a suitable alternative but require specific antibodies against a suitable biomarker. Mycolic acid, which has been found in patient sputum samples and comprises a large portion of the mycobacterial cell wall, is an ideal target. However, generating anti-lipid antibodies using traditional hybridoma methodologies is challenging and has limited the exploitation of this lipid as a diagnostic marker. We describe here the isolation and characterization of four anti-mycolic acid antibodies from a nonimmune antibody phage display library that can detect mycolic acids down to a limit of 4.5ng. All antibodies were specific for the methoxy subclass of mycolic acid with weak binding for α mycolic acid and did not show any binding to closely related lipids or other Mycobacterium tuberculosis (Mtb) derived lipids. We also determined the clinical utility of these antibodies based on their limit of detection for mycobacteria colony forming units (CFU). In combination with an optimized alkaline hydrolysis method for rapid lipid extraction, these antibodies can detect 10⁵ CFU of Mycobacterium bovis BCG, a close relative of Mtb and therefore represent a novel approach for the development of diagnostic assays for lipid biomarkers.     

Quantum Attack-Resistent Certificateless Multi-Receiver Signcryption Scheme

The existing certificateless signcryption schemes were designed mainly based on the traditional public key cryptography, in which the security relies on the hard problems, such as factor decomposition and discrete logarithm. However, these problems will be easily solved by the quantum computing. So the existing certificateless signcryption schemes are vulnerable to the quantum attack. Multivariate public key cryptography (MPKC), which can resist the quantum attack, is one of the alternative solutions to guarantee the security of communications in the post-quantum age. Motivated by these concerns, we proposed a new construction of the certificateless multi-receiver signcryption scheme (CLMSC) based on MPKC. The new scheme inherits the security of MPKC, which can withstand the quantum attack. Multivariate quadratic polynomial operations, which have lower computation complexity than bilinear pairing operations, are employed in signcrypting a message for a certain number of receivers in our scheme. Security analysis shows that our scheme is a secure MPKC-based scheme. We proved its security under the hardness of the Multivariate Quadratic (MQ) problem and its unforgeability under the Isomorphism of Polynomials (IP) assumption in the random oracle model. The analysis results show that our scheme also has the security properties of non-repudiation, perfect forward secrecy, perfect backward secrecy and public verifiability. Compared with the existing schemes in terms of computation complexity and ciphertext length, our scheme is more efficient, which makes it suitable for terminals with low computation capacity like smart cards.     

量子点荧光技术标记肿瘤细胞中HSP的应用研究

目的探讨量子点荧光技术对人肾癌细胞株(ACHN)中不同HSP进行标记的可行性应用。方法利用量子点的荧光特性,免疫细胞化学方法检测体外培养的ACHN细胞中量子点特异性标记的HSP70、HSP90、HSPgp96的表达情况。结果共聚焦荧光显微镜下可见ACHN细胞中HSP70、HSP90、HSPgp96均有明显表达,呈现均匀分布的橙红色强荧光,且量子点在持续激发30分钟后无荧光淬灭发生。结论量子点荧光标记技术能够对不同HSP进行标记,且与传统的标记方法相比具有显著优点,可作为一种新型的检测技术应用于科研及临床标记检测中。     

珍稀濒危物种堇叶紫金牛高效快繁体系的建立

筛选堇叶紫金牛(Ardisia violacea)野生优株,以其当年新发带休眠腋芽茎段为外植体,通过启动培养、丛生芽诱导增殖、壮苗培养、生根培养和炼苗移栽等过程建立其组培快繁技术体系。研究结果表明,最佳启动培养基为MS+0.80 mg·L~(–1)KT+0.10 mg·L~(–1) NAA+0.10 mg·L~(–1) IBA,腋芽萌发率达92.60%;最佳丛生芽诱导增殖培养基为MS1+0.50 mg·L~(–1) TDZ+0.10mg·L~(–1) NAA,平均增殖系数达8.60;最佳壮苗培养基为MS+1.00 mg·L~(–1) KT+0.50 mg·L~(–1) NAA;最佳生根培养基为1/2MS+2.00 mg·L~(–1) IBA+1.00 mg·L~(–1) NAA+1.00 mg·L~(–1) AC,平均生根率达98.70%;采用松鳞和泥炭(2:1,v/v)作为炼苗基质,炼苗成活率可达85.30%。实验成功建立了堇叶紫金牛高效组培快繁技术体系,经验证该体系能够满足规模化生产的需求。     

High-resolution solution structure of the 18 kDa substrate-binding domain of the mammalian chaperone protein Hsc70.

The three-dimensional structure for the substrate-binding domain of the mammalian chaperone protein Hsc70 of the 70 kDa heat shock class (HSP70) is presented. This domain includes residues 383-540 (18 kDa) and is necessary for the binding of the chaperone with substrate proteins and peptides. The high-resolution NMR solution structure is based on 4150 experimental distance constraints leading to an average root-mean-square precision of 0.38 A for the backbone atoms and 0.76 A for all atoms in the beta-sandwich sub-domain. The protein is observed to bind residue Leu539 in its hydrophobic substrate-binding groove by intramolecular interaction. The position of a helical latch differs dramatically from what is observed in the crystal and solution structures of the homologous prokaryotic chaperone DnaK. In the Hsc70 structure, the helix lies in a hydrophobic groove and is anchored by a buried salt-bridge. Residues involved in this salt-bridge appear to be important for the allosteric functioning of the protein. A mechanism for interdomain allosteric modulation of substrate-binding is proposed. It involves large-scale movements of the helical domain, redefining the location of the hinge area that enables such motions.     

Genomic instability and aging-like phenotype in the absence of mammalian SIRT6

The Sir2 histone deacetylase functions as a chromatin silencer to regulate recombination, genomic stability, and aging in budding yeast. Seven mammalian Sir2 homologs have been identified (SIRT1-SIRT7), and it has been speculated that some may have similar functions to Sir2. Here, we demonstrate that SIRT6 is a nuclear, chromatin-associated protein that promotes resistance to DNA damage and suppresses genomic instability in mouse cells, in association with a role in base excision repair (BER). SIRT6-deficient mice are small and at 2-3 weeks of age develop abnormalities that include profound lymphopenia, loss of subcutaneous fat, lordokyphosis, and severe metabolic defects, eventually dying at about 4 weeks. We conclude that one function of SIRT6 is to promote normal DNA repair, and that SIRT6 loss leads to abnormalities in mice that overlap with aging-associated degenerative processes.