Characterization of a Hyperthermophilic Redox Protein, Rubredoxin, as a Potential Targeted Cancer Therapeutic

Cancer is an elusive neoplastic disease that claims the lives of many people around the world every year. Though treatments have become more specific to the different types of cancer, the need remains for antineoplastic drugs that target cancer cells and leave normal cells unharmed, with little to no systemic toxicity. The search for a targeted cancer therapeutic is necessary and urgent, and Pyrococcus furiosus rubredoxin might be such a tool. Rubredoxin is a small (53 amino acids), water soluble, non-heme iron electron transfer protein that contains an iron atom cofactor bound by the sulfurs of four cysteine residues, which contribute to the redox activity of this protein. Rubredoxin from the hyperthermophile Pyrococcus furiosus is thermostable and appears to have low immunogenicity. The focus of this project was to incorporate tumor specificbinding sequences at the central loop, express, and purify these recombinantrubredoxin proteins. Next, the wild-type and recombinant rubredoxins werecharacterized based on absorption spectra, thermostability, metal content, andantibody affinity. Lastly, the effect of wild-type and recombinant rubredoxinswas assessed on cancer cells in-vitro. The Pyrococcus furiosus rubredoxin gene was manipulated via site-directed mutagenesis to incorporate two test epitopes (E-tag and RGD-tag) at the central loop (between D20 and N21 position). The mutant proteins (D20-Etag and D20-RGD) were purified and analyzed using absorption spectroscopy, thermostability, SDS-PAGE, electrospray ionization mass spectroscopy (ESI-MS), and inductively coupled mass spectrometry (ICP-MS). Binding studies for the D20-Etag mutant were done using a dot blot. Lastly, integrin-stimulated Jurkat cancer cells were incubated with wild-type rubredoxin, D20-Etag, D20-RGD, and the cells were assayed for apoptosis via gel electrophoresis at 24 and 48 hour time points.The E-tag epitope was successfully incorporated between the D20 and N21 amino acid residues using site directed mutagenesis. The D20-Etag and D20-RGD mutant rubredoxin proteins were successfully expressed, purified, and analyzed. There was an apoptotic effect of D20-RGD rubredoxin on the Jurkat cell line. These results provided a further understanding and appreciation of rubredoxin as a potential targeted therapeutic to cancer cells

Impact of early versus delayed umbilical cord clamping on post partum blood loss: a randomized controlled trial

Background: Evidence published to date has not clearly established the impact of the timing of cord clamping on postpartum blood loss and necessitates further research. The objective was to find out the impact of early and delayed cord clamping on the post partum blood loss and on infant haemoglobin and hematocrit at 90 ±7 days postpartum.Methods: A randomized, controlled trial was performed on 183 consenting women fulfilling eligibility criteria in labour ward of a teaching hospital in India. They were randomly assigned into early cord clamping or delayed cord clamping group; other components of active management of third stage of labour being same for all. The duration of third stage of labour was noted. Quantitative assessment of post partum blood loss was done using BRASSS-V drapes and weighing blood soaked pads. Cord blood haemoglobin and hematocrit at birth and infant haemoglobin and hematocrit at 90 ±7 days were estimated. Statistical analysis done using students unpaired ‘t’ test.Results: No difference was noted in the mean blood loss (203.52 ± 122.74 ml versus 200.74± 104.07 ml in early and delayed cord clamping respectively) and the duration of third stage of labour (Mean: 296.59± 98.97 seconds versus 281.79±104.59 seconds for early and delayed clamping respectively). Infant haemoglobin at 90 ±7 days was 11.07 ± 1.27 gm/dl versus 12.70± 1.41 gm/dl (p=0.0000) and infant hematocrit at 90 ±7 days was 34.13±3.93 % versus 39.33 ±4.88 % in early and delayed clamping respectively (p=0.0000).Conclusion: The timing of cord clamping has no impact on the mean blood loss and duration of third stage of labour. Delayed clamping significantly increases mean venous haemoglobin and hematocrit at 90 ±7 days without increasing NICU admissions

Agent Based Intrusion Detection and Response System for Wireless LANs

Wireless LAN technology, despite the numerous advantages it has over competing technologies, has not seen widespread deployment. A primary reason for markets not adopting this technology is its failure to provide adequate security. Data that is sent over wireless links can be compromised with utmost ease. In this project, we propose a distributed agent based intrusion detection and response system for wireless LANs that can detect unauthorized wireless elements like access points, wireless clients that are in promiscuous mode etc. The system reacts to intrusions by either notifying the concerned personnel, in case of rogue access points and promiscuous nodes, or by blocking unauthorized users from accessing the network resources

Macrophage Activation Syndrome as a Complication of Systemic Lupus Erythematosus in an Adult Male

Macrophage activation syndrome (MAS) is a rare life-threatening complication that sometimes occurs in patients with systemic lupus erythematosus (SLE) and other connective tissue diseases. This syndrome is universally fatal without treatment, and therefore, prompt diagnosis and initiation of treatment is of vital importance

Ultra-thin titanium nitride films for refractory spectral selectivity

We demonstrate a selectively emitting optical Fabry-P\'erot resonator based on a few-nm-thin continuous metallic titanium nitride film, separated by a dielectric spacer from an optically thick titanium nitride back-reflector, which exhibits excellent stability at 1070 K against chemical degradation, thin-film instabilities and melting point depression. The structure paves the way to the design and fabrication of refractory thermal emitters using the well-established processes known from the field of multilayer and rugate optical filters. We demonstrate that a few-nanometer thick films of titanium nitride can be stable under operation at temperatures exceeding 1070 K. This type of selective emitter provides a means towards near-infrared thermal emission that could potentially be tailored to the accuracy level known from rugate optical filters.Comment: 16 pages, 6 figure