RecA protein promoted homologous pairing in vitro. Pairing between linear duplex DNA bound to HU Protein (nucleosome cores) and nucleoprotein filaments of recA protein-single-stranded DNA

RecA protein promotes two distinct types of synaptic structures between circular single strands and duplex DNA; paranemic joints, where true intertwining of paired strands is prohibited and the classically intertwined plectonemic form of heteroduplex DNA. Paranemic joints are less stable than plectonemic joints and are believed to be the precursors for the formation of plectonemic joints. We present evidence that under strand exchange conditions the binding of HU protein, from Escherichia coli, to duplex DNA differentially affects homologous pairing in vitro. This conclusion is based on the observation that the formation of paranemic joint molecules was not affected, whereas the formation of plectonemic joint molecules was inhibited from the start of the reaction. Furthermore, introduction of HU protein into an ongoing reaction stalls further increase in the rate of the reaction. By contrast, binding of HU protein to circular single strands has neither stimulatory nor inhibitory effect. Since the formation of paranemic joint molecules is believed to generate positive supercoiling in the duplex DNA, we have examined the ability of positive superhelical DNA to serve as a template in the formation of paranemic joint molecules. The inert positively supercoiled DNA could be converted into an active substrate, in situ, by the action of wheat germ topoisomerase I. Taken collectively, these results indicate that the structural features of the bacterial chromosome which include DNA supercoiling and organization of DNA into nucleosome-like structures by HU protein modulate homologous pairing promoted by the nucleoprotein filaments of recA protein single-stranded DNA

Pilot scale microbial production and optimization of Serratia peptidase from Serratia marcescens

Serratia peptidase is active proteolytic enzyme which has the potential of cleaving peptide bond.  Present investigation deals about the Microbial production of serratia peptidase using Serratia marcescens in small scale fermentor. Batch fermentor has been run continuously throughout the night to analyze the production of protein as well as kinetics. Culture broth was maintained at 150rpm for 72 hrs. Protein sample was isolated by centrifuging at 3000rpm for 10mints. The result revealed that Serratia marcescens showed the enormous production of protein in fed batch fermentor compared to the small scale level.  Different substrates were been used for the production of enzyme. Among all cysteine showed the better activity as 2 units/ml of enzyme. Enzymatic assay of Serratia peptidase was done at different time interval of crude broth. Enzyme activity showed that maximum at 40ºC for 72hrs. It was observed that 0.65 units/ml of enzyme. Fed batch pilot scale production of Serratia peptidase was done at 0.5%cystein and 700rpm for 48hrs of run time.Â

Lifetimes of states in 19Ne above the 15 O + alpha breakup threshold

The 15O(alpha,gamma)19Ne reaction plays a role in the ignition of Type I x-ray bursts on accreting neutron stars. The lifetimes of states in 19Ne above the 15O + alpha threshold of 3.53 MeV are important inputs to calculations of the astrophysical reaction rate. These levels in 19Ne were populated in the 3He(20Ne,alpha)19Ne reaction at a 20Ne beam energy of 34 MeV. The lifetimes of six states above the threshold were measured with the Doppler shift attenuation method (DSAM). The present measurements agree with previous determinations of the lifetimes of these states and in some cases are considerably more precise

Measurement of two-halo neutron transfer reaction p(11^{11}Li,9^{9}Li)t at 3AA MeV

The p(\nuc{11}{Li},\nuc{9}{Li})t reaction has been studied for the first time at an incident energy of 3$A$ MeV delivered by the new ISAC-2 facility at TRIUMF. An active target detector MAYA, build at GANIL, was used for the measurement. The differential cross sectionshave been determined for transitions to the \nuc{9}{Li} ground andthe first excited states in a wide range of scattering angles. Multistep transfer calculations using different \nuc{11}{Li} model wave functions, shows that wave functions with strong correlations between the halo neutrons are the most successful in reproducing the observation.Comment: 6 pages, 3 figures, submitted to Physical Review Letter

Carbon Isotopes Near Drip Lines in the Relativistic Mean-Field Theory

We have investigated the ground-state properties of carbon isotopes in the framework of the relativistic mean-field (RMF) theory. RMF calculations have been performed with the non-linear scalar self-coupling of the $\sigma$ meson using an axially symmetric deformed configuration. We have also introduced the vector self-coupling of the $\omega$ meson for the deformed mean-field calculations. The results show that the RMF predictions on radii and deformations are in good agreement with the available experimental data. It is shown that several carbon isotopes possess a highly deformed shape akin to a superdeformation. The single-particle structure of nuclei away from the stability line has been discussed with a view to understand the properties near the neutron drip line. Predictions of properties of carbon isotopes away from the stability line are made.Comment: Revtex, 29 pages, 11 postscript figures include

Shell Effects in Nuclei with Vector Self-Coupling of Omega Meson in Relativistic Hartree-Bogoliubov Theory

Shell effects in nuclei about the stability line are investigated within the framework of the Relativistic Hartree-Bogoliubov (RHB) theory with self-consistent finite-range pairing. Using 2-neutron separation energies of Ni and Sn isotopes, the role of $\sigma$- and $\omega$-meson couplings on the shell effects in nuclei is examined. It is observed that the existing successful nuclear forces (Lagrangian parameter sets) based upon the nonlinear scalar coupling of $\sigma$-meson exhibit shell effects which are stronger than suggested by the experimental data. We have introduced nonlinear vector self-coupling of $\omega$-meson in the RHB theory. It is shown that the inclusion of the vector self-coupling of $\omega$-meson in addition to the nonlinear scalar coupling of $\sigma$-meson provides a good agreement with the experimental data on shell effects in nuclei about the stability line. A comparison of the shell effects in the RHB theory is made with the Hartree-Fock Bogoliubov approach using the Skyrme force SkP. It is shown that the oft-discussed shell quenching with SkP is not consistent with the available experimental data.Comment: 34 pages latex, 18 ps figures, replaced with minor corrections in some figures, accepted for publication in Phys. Rev.

DDoS defense by offense

This article presents the design, implementation, analysis, and experimental evaluation of speak-up, a defense against application-level distributed denial-of-service (DDoS), in which attackers cripple a server by sending legitimate-looking requests that consume computational resources (e.g., CPU cycles, disk). With speak-up, a victimized server encourages all clients, resources permitting, to automatically send higher volumes of traffic. We suppose that attackers are already using most of their upload bandwidth so cannot react to the encouragement. Good clients, however, have spare upload bandwidth so can react to the encouragement with drastically higher volumes of traffic. The intended outcome of this traffic inflation is that the good clients crowd out the bad ones, thereby capturing a much larger fraction of the server's resources than before. We experiment under various conditions and find that speak-up causes the server to spend resources on a group of clients in rough proportion to their aggregate upload bandwidths, which is the intended result.National Science Foundation (U.S.) (NSF grant CNS-0225660)National Science Foundation (U.S.) (NSF grant CNS-0520241)United States. Dept. of Defense (National Security Science and Engineering Faculty Fellowship

Dysregulated Nephrin in Diabetic Nephropathy of Type 2 Diabetes: A Cross Sectional Study

Podocyte specific proteins are dysregulated in diabetic nephropathy, though the extent of their expression loss is not identical and may be subject to different regulatory factors. Quantifying the degree of loss may help identify the most useful protein to use as an early biomarker of diabetic nephropathy.Protein expression of synaptopodin, podocin and nephrin were quantified in 15 Type 2 diabetic renal biopsies and 12 control patients. We found statistically significant downregulation of synaptopodin (P<0.0001), podocin (P = 0.0002), and nephrin (P<0.0001) in kidney biopsies of diabetic nephropathy as compared with controls. Urinary nephrin levels (nephrinuria) were then measured in 66 patients with Type 2 diabetes and 10 healthy controls by an enzyme-linked immunosorbent assay (Exocell, Philadelphia, PA). When divided into groups according to normo-, micro-, and macroalbuminuria, nephrinuria was found to be present in 100% of diabetic patients with micro- and macroalbuminuria, as well as 54% of patients with normoalbuminuria. Nephrinuria also correlated significantly with albuminuria (rho = 0.89, p<0.001), systolic blood pressure (rho = 0.32, p = 0.007), and correlated negatively with serum albumin (rho = -0.48, p<0.0001) and eGFR (rho = -0.33, p = 0.005).These data suggest that key podocyte-specific protein expressions are significantly and differentially downregulated in diabetic nephropathy. The finding that nephrinuria is observed in a majority of these normoalbuminuric patients demonstrates that it may precede microalbuminuria. If further research confirms nephrinuria to be a biomarker of pre-clinical diabetic nephropathy, it would shed light on podocyte metabolism in disease, and raise the possibility of new and earlier therapeutic targets

Initial Report of a Phase I Study of LY2510924, Idarubicin, and Cytarabine in Relapsed/Refractory Acute Myeloid Leukemia

Background: The CXCR4/SDF-1α axis plays a vital role in the retention of stem cells within the bone marrow and downstream activation of cell survival signaling pathways. LY2510924, a second generation CXCR4, showed significant anti-leukemia activity in a murine AML model.Methods: We conducted a phase I study to determine the safety and toxicity of LY2510924, idarubicin and cytarabine (IA) combination therapy in relapsed/refractory (R/R) AML. Eligible patients were 18–70 years of age receiving up to salvage 3 therapy. A peripheral blood absolute blast count of &lt; 20,000/μL was required for inclusion. LY2510924 was administered daily for 7 days followed by IA from day 8. Two dose escalation levels (10 and 20 mg) were evaluated, with a plan to enroll up to 12 patients in the phase I portion.Results: The median age of the enrolled patients (n = 11) was 55 years (range, 19–70). Median number of prior therapies was 1 (1–3). Six and five patients were treated at dose-levels “0” (10 mg) and “1” (20 mg), respectively. Only one patient experiencing a dose limiting toxicity (grade 3 rash and myelosuppression). Three and one complete responses were observed at dose-levels “0” and “1,” respectively; the overall response rate (ORR) was 36% (4 of 11 patients). A ≥ 50% decrease in CXCR4 mean fluorescence intensity was observed in 4 of 9 patients by flow cytometry, indicating incomplete suppression of CXCR4-receptor occupancy.Conclusions: The combination of LY2510924 with IA is safe in R/R AML. Dose-escalation to a 30 mg LY2510924 dose is planned to achieve complete blockade of CXCR4 receptor occupancy, followed by expansion phase at the recommended phase 2 dose-level

Early and extended erythropoietin monotherapy after hypoxic ischaemic encephalopathy:a multicentre double-blind pilot randomised controlled trial

Objective: To examine the feasibility of early and extended erythropoietin monotherapy after hypoxic ischaemic encephalopathy (HIE). Design: Double-blind pilot randomised controlled trial.Setting: Eight neonatal units in South Asia. Patients: Neonates (≥36 weeks) with moderate or severe HIE admitted between 31 December 2022 and 3 May 2023. Interventions: Erythropoietin (500 U/kg daily) or to the placebo (sham injections using a screen) within 6 hours of birth and continued for 9 days. MRI at 2 weeks of age. Main outcomes and measures: Feasibility of randomisation, drug administration and assessment of brain injury using MRI. Results: Of the 154 neonates screened, 56 were eligible; 6 declined consent and 50 were recruited; 43 (86%) were inborn. Mean (SD) age at first dose was 4.4 (1.2) hours in erythropoietin and 4.1 (1.0) hours in placebo. Overall mortality at hospital discharge occurred in 5 (19%) vs 11 (46%) (p=0.06), and 3 (13%) vs 9 (40.9%) (p=0.04) among those with moderate encephalopathy in the erythropoietin and placebo groups. Moderate or severe injury to basal ganglia, white matter and cortex occurred in 5 (25%) vs 5 (38.5%); 14 (70%) vs 11 (85%); and 6 (30%) vs 2 (15.4%) in the erythropoietin and placebo group, respectively. Sinus venous thrombosis was seen in two (10%) neonates in the erythropoietin group and none in the control group. Conclusions: Brain injury and mortality after moderate or severe HIE are high in South Asia. Evaluation of erythropoietin monotherapy using MRI to examine treatment effects is feasible in these settings. Trial registration number: NCT05395195