Doxycycline-inducible expression of SPARC/osteonectin/BM40 in MDA-MB-231 human breast cancer cells results in growth inhibition

SPARC (secreted protein acidic and rich in cysteine)/BM40/Osteonectin is a matricellular protein with multiple effects on cell behaviour. In vitro, its major known functions are anti-adhesive and anti-proliferative, and it is associated with tissue remodelling and cancer in vivo. SPARC is overexpressed in many cancers, including breast cancer, and the effects of SPARC seem to be cell type-specific. To study the effects of SPARC on breast cancer, we transfected SPARC into the MDA-MB-231 BAG, human breast cancer cell line using the Tet-On inducible system. By western analysis, we found low background levels in the MDA-MB-231 BAG and clone X parental cells, and prominent induction of SPARC protein expression after doxycycline treatment in SPARC transfected clones X5, X21, X24 and X75. Induction of SPARC expression did not affect cell morphology or adhesiveness to collagens type I and IV, but it slowed the rate of proliferation in adherent cultures. Cell cycle analysis showed that SPARC slowed the progression to S phase. Doxycycline induction of SPARC also slowed the rate of monolayer wound closure in the cultured wound healing assay. Thymidine inhibition of proliferation abrogated this effect, confirming that it was due to anti-proliferation rather than inhibition of migration. Consistent with this, we were unable to detect any differences in migration and Matrigel outgrowth analysis of doxycycline-stimulated cells. We conclude that SPARC is inhibitory to human breast cancer cell proliferation, and does not stimulate migration, in contrast to its stimulatory effects reported for melanoma (proliferation and migration) and glioma (migration) cells. Similar growth repression by SPARC has been reported for ovarian cancer cells, and this may be a common feature among carcinoma.

Use of the wound-inducible NtQPT2 promoter from Nicotiana tabacum for production of a plant-made vaccine

The wound-inducible quinolinate phosphoribosyl transferase promoter from Nicotiana tabacum (NtQPT2) was assessed for its capacity to produce B-subunit of the heat-labile toxin (LTB) from enterotoxigenic Escherichia coli in transgenic plant tissues. Comparisons were made with the widely used and constitutive Cauliflower Mosaic Virus 35S (CaMV35S) promoter. The NtQPT2 promoter produced somewhat lower average concentrations of LTB protein per unit weight of hairy root tissue but allowed better growth thereby producing similar or higher overall average yields of LTB per culture batch. Transgenic tobacco plants containing the NtQPT2-LTB construct contained LTB protein in roots but not leaves. Moreover, wounding NtQPT2-LTB transgenic plants, by removal of apices, resulted in an approximate 500% increase in LTB levels in roots when analysed several days later. CaMV35S-LTB transgenic plants contained LTB protein in leaves and roots but wounding made no difference to their LTB content.

Intramuscular heat shock protein 72 and heme oxygenase-1 mRNA are reduced in patients with type 2 diabetes: evidence that insulin resistance is associated with a disturbed antioxidant defense mechanism

To examine whether genes associated with cellular defense against oxidative stress are associated with insulin sensitivity, patients with type 2 diabetes (n = 7) and age-matched (n = 5) and young (n = 9) control subjects underwent a euglycemic-hyperinsulinemic clamp for 120 min. Muscle samples were obtained before and after the clamp and analyzed for heat shock protein (HSP)72 and heme oxygenase (HO)-1 mRNA, intramuscular triglyceride content, and the maximal activities of β-hyroxyacyl-CoA dehydrogenase (β-HAD) and citrate synthase (CS). Basal expression of both HSP72 and HO-1 mRNA were lower (P < 0.05) by 33 and 55%, respectively, when comparing diabetic patients with age-matched and young control subjects, with no differences between the latter groups. Both basal HSP72 (r = 0.75, P < 0.001) and HO-1 (r = 0.50, P < 0.05) mRNA expression correlated with the glucose infusion rate during the clamp. Significant correlations were also observed between HSP72 mRNA and both β-HAD (r = 0.61, P < 0.01) and CS (r = 0.65, P < 0.01). HSP72 mRNA was induced (P < 0.05) by the clamp in all groups. Although HO-1 mRNA was unaffected by the clamp in both the young and age-matched control subjects, it was increased (P < 0.05) ∼70-fold in the diabetic patients after the clamp. These data demonstrate that genes involved in providing cellular protection against oxidative stress are defective in patients with type 2 diabetes and correlate with insulin-stimulated glucose disposal and markers of muscle oxidative capacity. The data provide new evidence that the pathogenesis of type 2 diabetes involves perturbations to the antioxidant defense mechanism within skeletal muscle.

Modeling the propagation and defense study of internet malicious information

 Dr. Wen's research includes modelling the propagation dynamics of malicious information, exposing the most influential people and source identification of epidemics in social networks. His research is beneficial to both academia and industry in the field of Internet social networks.

Defense against packet dropping attacks in opportunistic networks

Opportunistic networks (OppNets) are an interesting topic that are seen to have a promising future. Many protocols have been developed to accommodate the features of OppNets such as frequent partitions, long delays, and no end-to-end path between the source and destination nodes. Embedding security into these protocols is challenging and has taken a lot of attention in research. One of the attacks that OppNets are exposed to is the packet dropping attack, where the malicious node attempts to drop some packets and forwards an incomplete number of packets which results in the distortion of the message. To increase the security levels in OppNets, this paper presents an algorithm developed to detect packet dropping attacks, and finds the malicious node that attempted the attack. The algorithm detects the attack by using an indicative field in the header section of each packet; the indicative field has 3 sub fields - the identification field, the flag field, and the offset field. These 3 fields are used to find if a node receives the complete original number of packets from the previous node. The algorithm will have the advantage of detecting packets dropped by each intermediate node, this helps solve the difficulties of finding malicious nodes by the destination node only.

Detection and defense of application-layer DDoS attacks in backbone web traffic

Web servers are usually located in a well-organized data center where these servers connect with the outside Internet directly through backbones. Meanwhile, the application-layer distributed denials of service (AL-DDoS) attacks are critical threats to the Internet, particularly to those business web servers. Currently, there are some methods designed to handle the AL-DDoS attacks, but most of them cannot be used in heavy backbones. In this paper, we propose a new method to detect AL-DDoS attacks. Our work distinguishes itself from previous methods by considering AL-DDoS attack detection in heavy backbone traffic. Besides, the detection of AL-DDoS attacks is easily misled by flash crowd traffic. In order to overcome this problem, our proposed method constructs a Real-time Frequency Vector (RFV) and real-timely characterizes the traffic as a set of models. By examining the entropy of AL-DDoS attacks and flash crowds, these models can be used to recognize the real AL-DDoS attacks. We integrate the above detection principles into a modularized defense architecture, which consists of a head-end sensor, a detection module and a traffic filter. With a swift AL-DDoS detection speed, the filter is capable of letting the legitimate requests through but the attack traffic is stopped. In the experiment, we adopt certain episodes of real traffic from Sina and Taobao to evaluate our AL-DDoS detection method and architecture. Compared with previous methods, the results show that our approach is very effective in defending AL-DDoS attacks at backbones. © 2013 Elsevier B.V. All rights reserved.

Effects of omega-3 PUFA on the vitamin E and glutathione antioxidant defense system in individuals at ultra-high risk of psychosis

BACKGROUND: Oxidative stress and impaired antioxidant defenses are reported in schizophrenia and are associated with disturbed neurodevelopment, brain structural alterations, glutamatergic imbalance, increased negative symptoms, and cognitive impairment. There is evidence that oxidative stress predates the onset of acute psychotic illness. Here, we investigate the effects of omega-3 PUFA on the vitamin E and glutathione antioxidant defense system (AODS). METHOD: In 64 help-seeking UHR-individuals (13-25 years of age), vitamin E levels and glutathione were investigated before and after 12 weeks of treatment with either 1.2g/d omega-3 (PUFA-E) or saturated fatty acids (SFA-E), with each condition also containing 30.4mg/d alpha-tocopherol to ensure absorption without additional oxidative risk. RESULTS: In multivariate tests, the effects on the AODS (alpha-tocopherol, total glutathione) were not significantly different (p=0.13, p=0.11, respectively) between treatment conditions. According to univariate findings, only PUFA-E caused a significant alpha-tocopherol increase, while PUFA-E and SFA-E caused a significant gamma- and delta-tocopherol decrease. Total glutathione (GSHt) was decreased by PUFA-E supplementation. CONCLUSION: Effects of the PUFA-E condition on the vitamin E and glutathione AODS could be mechanisms underlying its clinical effectiveness. In terms of the vitamin E protection system, PUFA-E seems to directly support the antioxidative defense at membrane level. The effect of PUFA-E on GSHt is not yet fully understood, but could reflect antioxidative effects, resulting in decreased demand for glutathione. It is still necessary to further clarify which type of PUFA/antioxidant combination, and in which dose, is effective at each stage of psychotic illness.

Catabolism attack and anabolism defense: a novel attack and traceback mechanism in opportunistic networks

Security is a major challenge in Opportunistic Networks (OppNets) because of its characteristics, such as open medium, dynamic topology, no centralized management and absent clear lines of defense.A packet dropping attack is one of the major security threats in OppNets since neither source nodes nor destination nodes have the knowledge of where or when the packet will be dropped. In this paper, we present a novel attack and traceback mechanism against a special type of packet dropping where the malicious node drops one or more packets and then injects new fake packets instead. We call this novel attack a Catabolism Attack and we call our novel traceback mechanism against this attack Anabolism Defense. Our novel detection and traceback mechanism is very powerful and has very high accuracy. Each node can detect and then traceback the malicious nodes based on a solid and powerful idea that is, hash chain techniques. In our defense techniques we have two stages. The first stage is to detect the attack, and the second stage is to find the malicious nodes. Simulation results show this robust mechanism achieves a very high accuracy and detection rate.

Defense and traceback mechanisms in opportunistic wireless networks

 In this thesis, we have identified a novel attack in OppNets, a special type of packet dropping attack where the malicious node(s) drops one or more packets (not all the packets) and then injects new fake packets instead. We name this novel attack as the Catabolism attack and propose a novel attack detection and traceback approach against this attack referred to as the Anabolism defence. As part of the Anabolism defence approach we have proposed three techniques: time-based, Merkle tree based and Hash chain based techniques for attack detection and malicious node(s) traceback. We provide mathematical models that show our novel detection and traceback mechanisms to be very effective and detailed simulation results show our defence mechanisms to achieve a very high accuracy and detection rate.