A Real-Time TCP Stream Reassembly Mechanism in High-Speed Network

With the continual growth of the variety and complexity of network crime means, the traditional packet feature matching cannot detect all kinds of intrusion behaviors completely. It is urgent to reassemble network stream to perform packet processing at a semantic level above the network layer. This paper presents an efficient TCP stream reassembly mechanism for real-time processing of high-speed network traffic. By analyzing the characteristics of network stream in high-speed network and TCP connection establishment process, several polices for designing the reassembly mechanism are built. Then, the reassembly implementation is elaborated in accordance with the policies. Finally, the reassembly mechanism is compared with the traditional reassembly mechanism by the network traffic captured in a typical gigabit gateway. Experiment results illustrate that the reassembly mechanism is efficient and can satisfy the real-time property requirement of traffic analysis system in high-speed network.

A patch-clamp study on human sperm Cl^- channel reassembled into giant liposome

Aim: To record the single-channel currents and characterize the electrophysiological properties of the Cl^- channels inhuman sperm membrane. Methods: The membrane proteins extracted from the human sperm were reassembled intoliposome bilayer, and the liposomes were fused into giant liposomes with a diameter more than 10μm by dehydration-rehydration procedure. The giant liposomes were used to study the Cl^- channel activities by patch-clamp technique.Results: By patch clamping the giant liposome in an asymmetric NMDG (N-methyl-D-glucamine)-Cl (bath 100//pipette 200 mmol/L) solution system, three kinds of single-channel events with unit conductances of (74.1 ± 8.3) pS,(117.0±5.7) pS and (144.7±4.5) pS, respectively, were detected. Their activities were voltage-dependent and allwere blocked by SITS (4-acetamido-4'-isothiocyanato-stilbene-2', 2'-disulfonic acid) in a concentration-dependentmanner. By constructing the open and close dwell time distribution histograms and then fitting them with exponentialfunction, two time constants were obtained in both the open and the close states. The burst activity and conductancesubstate of the channels were observed. Conclusion; There exist three kinds of Cl^- channels with different conduc-tance in human sperm membrane at least. (Asian J Androl 2001 Sep; 3: 185 - 191)

RESEARCH ON IP FLOW FORMAT AND SEMANTIC FORMALIZATION

Malformed packets and overlapping fragments are harmful to Intranet end hosts. A formalization engine was introduced to formalize transit packets and reassemble fragments to eliminate the fragment semantic ambiguity. In the formalization engine, malformed packets are formalized by a packet verification engine layer according to protocol standards. In order to eliminate the fragment semantic ambiguity, OS classes of end hosts were collected by an OS detector, each fragment was reassembled according to its OS class. According to the reassembly algorithm of different OS, the pre-forward fields of the cached data were counted with the application of the pre-forward policies and were transmitted to save system resource. Applying the BSD-Linux pre-forward policy, the BSD-right pre-forward policy and the First pre-forward policy, the packet loss rate is dropped and system performance improved. The experiments show that the identification precision can be maintained about 90% in heavy processing load.

Reassembly of the axon initial segment and nodes of Ranvier in regenerated axons of the central nervous system

Myelinated axons of the peripheral and central nervous system（PNS＆CNS）are divided into molecularly distinct excitable domains,including the axon initial segment（AIS）and nodes of Ranvier.The AIS is composed of a dense network of cytoskeletal proteins,cell adhesion molecules,and voltage gated ion channels and is located at the proximal most region of the axon（Koleand Stuart, 2012）.

Development of Split-Protein Systems: From Binary to Ternary System

Tens of thousands of protein-protein interactions (PPIs) have been found in human cells and many of these macromolecular partnerships could determine the cell growth and death. Thus there is a need to develop the methods to catalogue these macromolecules by detecting their interactions, modifications, and cellular locations. It will be helpful for scientists to compare the difference between a diseased cellular state and its normal state and to find the potential therapy treatment to intervene this status. One technology called split-protein reassembly or protein fragment complementation has been developed in the last two decades. This technology makes use of appropriate fragmentation of some protein reporters and the refolding of these reports could be detected by their function to confirm the interaction of interest. This system has been set up in cell-free systems, E. coli, yeast, mammalian cells, plants and live animals. Herein, I present the development in fluorescence- and bioluminescence-based split-protein biosensors in both binary and ternary systems. In addition, some people developed the split-protein system by combining it with chemical inducer of dimerization strategy (CID). This has been applied for identifying the enzyme inhibitors and regulating the activity of protein kinases and phosphatases. With effort from many laboratories from the world, a variety of split-protein systems have been developed for studying the PPI in vitro and in vivo, monitoring the biological process, and controlling the activity of the enzyme of interest.

Disassembly intermediates of RbsD protein remain oligomeric despite the loss of an intact secondary structure

Many proteins exist as homo-oligomers in living organisms wherein the change of oligomeric status apparently serves as an effective means for modulating their biological activities. We have previously reported that the homo-decameric RbsD from Escherichia coli undergoes stepwise disassembly and non-stepwise reassembly. Here the structural status of the urea-induced RbsD disassembly intermediates was examined, mainly using urea-containing polyacrylamide gel electrophoresis and chemical cross-linking. Such intermediates were found to remain oligomeric while losing their intact secondary structures. Such disassembly intermediates were able to effectively refold when the concentration of the urea denaturant was reduced to a lower level, or to refold/reassemble into the native decamers when urea was completely removed, as detected by non-denaturing polyacrylamide gel electrophoresis. These novel observations strongly suggest that the assembly of oligomeric proteins may occur before the completion of subunit folding.

The microtubule aster formation and its role in nuclear envelope assembly around the sperm chromatin in Xenopus egg extracts

Nuclear envelope is a dynamic structure in the cell cycle. At the beginning of mitosis, nuclear envelope breaks down and its components disperse into the cytoplasm. At the end of mitosis, nuclear envelope reassembles using the dispersed components. Searching for the mechanisms of the nuclear disassembly and reassembly has for a long time been one of the key projects for cell biologists. In this report we show that microtubules take a role in the nuclear envelope assembly around the sperm chromatin in Xenopus egg ex-tracts. Microtubule cytoskeleton has been demonstrated to take roles in the transport of intracellular membranes such as Golgi and ER vesicles. We found that the nuclear envelope assembly needs functional microtubules. At the beginning of the nuclear assembly, microtubules nucleated to form a microtubule aster around the centrosome at the base of the sperm head. Using the microtubule drug colchicine to dis-rupt the microtubule nucleation, nuclear envelope reassem-bly was seriously inhibited. If the microtubules were stabi-lized by taxol, another microtubule drug, the nuclear enve-lope reassembly was also interfered, although a significantly large aster formed around the chromatin. Based on these observations, we propose that microtubules play an impor-tant role in the nuclear envelope reassembly maybe by transporting the nuclear envelope precursors to the chroma-tin surfaces.

Nucleolar Matrix in HeLa Cells

Nucleolus, in which rRNA precursors are synthesized and subunits of ribosomesare assembled, is composed of many copies of rDNA, rRNA and many kinds ofproteins. The processing of rRNA precursors also occurs in nucleolus, which is associ-ated with some nonribosomal proteins. In the 1980s, scientists proposed thatnucleolar skeleton or nucleolar matrix which is mainly constructed with proteins

Nuclear reassembly in vitro is independent of nucleosome/chromatin assembly

It was shown that nuclear reassembly was induced by small pieces of DNA fragments in cell free extracts of Xenopus. In an attempt to learn the relationship between the nuclear reassembly and nucleosome/chromatin assembly, limited amounts of CM Cellulose are used to eliminate the capacity of the egg extract S 150 to assemble chromatin, while the forming of nucleosomes is checked with DNA supercoiling by plasmid DNA pBR322 incubated in the extract, and further analysed by micrococcal nuclease digestion. This depleted extract is then used to induce nuclear reassembly around demembraned sperms with membrane vesicles. It is found that CM Cellulose depletes histones H2A and H2B efficiently and blocks the assembly of nucleosomes, the demembraned sperms are yet reconstituted into nuclei in the treated S 150, although the chromatin in reassembled nuclei does not produce protected DNA fragments when digested with micrococcal nuclease. It suggests that in the cell free system of Xenopus, DNA can be formed into nuclei without assembly of nucleosomes or chromatin.

Biological activity assays of cell-free reassembled nuclei——Injecting cell-free reassembled nuclei into unfertilized eggs can induce the eggs to cleave and reconstitute asters,and the injected nuclei undergo cell cycle changes

Nucleus may reassemble spontaneously in cell-free mixture of HeLa metaphase chromosomes,Xenopus egg extracts and ATP-regenerating system,and the nucleus shows some biological activities.It isfound that,after being injected into unfertilized mature eggs,the cell-free reassembled nuclei can cause theeggs to cleave and reconstitute asters in their cytoplasm,and the injected nuclei undergo changes in response tocell cycle regulators stored in the eggs,and that reinjecting cytostatic factors(CSF)into the eggs can stabilizethe eggs in mitotic phase,cause the nuclei disassembly and chromatin condensation to chromosomes.

Chromatin and Nucleosome Organizations and DNA Replication of Nucleus Reassembled in vitro Using Purified Exogenous DNA and Xenopus Egg Extracts

It has been demonstrated in the last ten years that the nuclear reassembly may occur in the cell-free systems from frog egg extracts added with exogenous naked DNA. However, there remains an open question : is the cell-free reassembled nucleus structurally similar to the nucleus in the intact cell ? That is, does the cell-free reassembled nucleus contain nucleosomes and chromatin? For this issue, we have designed experiments for identifying the internal structures of the cell-free reassembled nucleus. These experiments show that the nucleus reassembled in vitro also contains chromatin which is composed of typical 10 nm nucleosome fibers of 'beads-on-a-string', 30 nm filaments and the next higher-order structures. The digestion experiment with the enzyme micrococcal nuclease has demonstrated that the DNA in the nucleosome of the reconstituted chromatin is about 200 base pairs (bp) in length, of which 165 bp may be in the nucleosome particle, and 35 bp may be in the linker between two particles. Prolonging digestion of the 165-bp particle DNA fragment will yield a 146-bp fragment, which may be wounded in the nucleosome core. Experiments also provide evidence that the chromatin reconstitution in the cell-free reassembled nucleus is a progressive process, and that the nucleus can replicate its DNA. Based on these observations, we suppose that the chromatin of the cell-free reassembled nucleus may be structurally and functionally similar to the chromatin of the intact cells.

Silver Ion-Induced Formation of Unprecedented Thorium Nonamer Clusters via Lacuna-Construction Strategy

Herein,we report the synthesis and structures of two novel mixed-metal clusters denoted as Th_(9)Ag_(6)and Th_(9)Ag_(12).Both clusters feature unprecedented Th_(9)cores.The cores are tricapped trigonal prism moieties that are novel among actinides.Attempted alternative synthesis routes indicate that the Th_(9)clusters are accessible only through slow introduction of Ag_(+)into a solution containing a Th6 cluster modified with 2-picolinic acid.Alternative rapid addition of Ag_(+)leads to dissociation of the Th6 cluster with formation of a high-purity(ThAg)_(∞)two-dimensional layered structure material.A mechanism for cluster dissociation and reassembly to yield Th_(9)from Th6 is proposed that is consistent with spectroscopic observations and computational results.Because of Ag⋯Ag andπ–πinteractions,the Th_(9)Ag_(12)cluster exhibits high stability in air,at elevated temperature,underγ-irradiation,and in common solvents.