Characteristics of natural and planted sandalwood-supporting soils in Seoni district, Madhya Pradesh

Two sandalwood-supporting pedons, one planted (P1) and other natural growing, were characterized in Seoni district of Madhya Pradesh. The solum depth of both the pedon was 30 cm underlain by weathered basalt beyond 50 cm. Pedon was dark reddish brown while P2 was characterized by dark reddish brown to dusky red. P2 had sandy clay texture (A1 horizon) and other horizons of both the pedons was associated with clay texture. These soils had more than 35 cmol(p+)kg-1 CEC and dominance of smectite (>20%) followed by vermiculite. There was more than 20 per cent feldspar in Bw horizon of P1 while underlying horizon of P2 had 25 per cent quartz. The nearly neutral soils had more than 1.0 per cent organic carbon barring A2 horizon of P2. These calcareous soils were dominated by exchangeable Ca++ followed by Mg++, Na+, K+ on exchange complex and grouped as Typic Haplustepts and Typic Ustorthents at subgroup level

Evaluation of Different Extractants for Boron Estimation in Soils of Odisha and Andhra Pradesh

The present study was conducted to compare the four different extractants widely used in boron (B) determination in soils and to screen the most suitable extractant for acidic (Alfisols) and alkaline (Vertisols) soils of Odisha and Andhra Pradesh. A total of 200 surface soil samples were collected across two sites (100 from each site) representing different pH ranges. Hot-water-soluble boron (HWS-B) extraction procedure being the most widely used B determination procedure was kept as a benchmark in order to compare the B extracting efficiency by the extractants viz., 0.01 M Calcium Chloride (CaCl2), 1 N Ammonium acetate (NH4OAc) pH-7.0 and 0.01 M Barium Chloride. The mean values of hot water extractable B for acidic soils were 0.49 (mean between 0.18 and 1.50 mg kg-1), CaCl2 – 0.42 (mean between 0.14 and 1.52 mg kg-1), BaCl2 – 0.45 (mean between 0.10 and 1.68 mg kg-1) and NH4OAc – 0.60 (mean between 0.17 and 2.43 mg kg-1). The mean values of hot water extractable B for alkaline soils were 1.87 (mean between 0.71 and 4.79 mg kg-1), CaCl2 – 1.57 (mean between 0.45 and 5.43 mg kg-1), BaCl2 – 1.37 (mean between 0.52 and 4.15 mg kg-1) and NH4OAc – 1.92 (mean between 0.85 and 8.33 mg kg-1) in acidic and alkaline soils respectively. The coefficient of variation for extractable B varied from 53.0 to 66.6 and 42.7 to 55.7 in acidic and alkaline soils respectively. The variation in B extracting efficiency in both the types of soils was found in the order: Hot water > Hot CaCl2 > BaCl2 > NH4OAc. Authors conclude that amongst all the compared extractants, 0.01 M CaCl2 extraction may be an adequate procedure for B determination in both the acidic as well as alkaline soils

Shigella IpaB and IpaD displayed on L. lactis bacterium-like particles induce protective immunity in adult and infant mice

Shigella spp. are among the enteric pathogens with the highest attributable incidence of moderate-to-severe diarrhea in children under 5 years of age living in endemic areas. There are no vaccines available to prevent this disease. In this work, we investigated a new Shigella vaccine concept consisting of non-living, self-adjuvanted, Lactococcus lactis bacterium-like particles (BLP) displaying Shigella invasion plasmid antigen (Ipa) B and IpaD and examined its immunogenicity and protective efficacy in adult and newborn/infant mice immunized via the nasal route. Unique advantages of this approach include the potential for broad protection due to the highly conserved structure of the Ipas and the safety and practicality of a probiotic-based mucosal/adjuvant delivery platform. Immunization of adult mice with BLP-IpaB and BLP-IpaD (BLP-IpaB/D) induced high levels of Ipa-specific serum IgG and stool IgA in a dose-dependent manner. Immune responses and protection were enhanced by BLP delivery. Vaccine-induced serum antibodies exhibited opsonophagocytic and cytotoxic neutralizing activity, and IpaB/D IgG titers correlated with increased survival post-challenge. Ipa-specific antibody secreting cells were detected in nasal tissue and lungs, as well as IgG in bronchoalveolar lavage. Bone marrow cells produced IpaB/D-specific antibodies and contributed to protection after adoptive transfer. The BLP-IpaB/D vaccine conferred 90% and 80% protection against S. flexneri and S. sonnei, respectively. Mice immunized with BLP-IpaB/D as newborns also developed IpaB and IpaD serum antibodies; 90% were protected against S. flexneri and 44% against S. sonnei. The BLP-IpaB/D vaccine is a promising candidate for safe, practical and potentially effective immunization of children against shigellosis

Biophysical Characterization of the Type III Secretion Tip Proteins and the Tip Proteins Attached to Bacterium-Like Particles

Bacterium-like particles (BLPs), derived from Lactococcus lactis, offer a self-adjuvanting delivery vehicle for subunit protein vaccines. Proteins can be specifically loaded onto the BLPs via a peptidoglycan anchoring domain (PA). In this study, the tip proteins IpaD, SipD and LcrV belonging to type three secretion systems of Shigella flexneri, Salmonella enterica and Yersinia enterocolitica, respectively, were fused to the PA and loaded onto the BLPs. Herein, we biophysically characterized these nine samples and condensed the spectroscopic results into three-index empirical phase diagrams (EPDs). The EPDs show distinctions between the IpaD/SipD and LcrV subfamilies of tip proteins, based on their physical stability, even upon addition of the PA. Upon attachment to the BLPs, the BLPs become defining moiety in the spectroscopic measurements, leaving the tip proteins to have a subtle yet modulating effect on the structural integrity of the tip proteins-BLPs binding. In summary, this work provides a comprehensive view of physical stability of the tip proteins and tip protein-BLPs and serves as a baseline for screening of excipients to increase the stability of the tip protein-BLPs for future vaccine formulation

Characterization of a Novel Fusion Protein from IpaB and IpaD of Shigella spp. and Its Potential as a Pan-Shigella Vaccine

Shigellosis is an important disease in the developing world, where about 90 million people become infected with Shigella spp. each year. We previously demonstrated that the type three secretion apparatus (T3SA) proteins IpaB and IpaD are protective antigens in the mouse lethal pulmonary model. In order to simplify vaccine formulation and process development, we have evaluated a vaccine design that incorporates both of these previously tested Shigella antigens into a single polypeptide chain. To determine if this fusion protein (DB fusion) retains the antigenic and protective capacities of IpaB and IpaD, we immunized mice with the DB fusion and compared the immune response to that elicited by the IpaB/IpaD combination vaccine. Purification of the DB fusion required coexpression with IpgC, the IpaB chaperone, and after purification it maintained the highly α-helical characteristics of IpaB and IpaD. The DB fusion also induced comparable immune responses and retained the ability to protect mice against Shigella flexneri and S. sonnei in the lethal pulmonary challenge. It also offered limited protection against S. dysenteriae challenge. Our results show the feasibility of generating a protective Shigella vaccine comprised of the DB fusion

Impact of Detergent on the Biophysical Properties and Immune Response of the IpaDB Fusion Protein, a Candidate Subunit Vaccine against Shigella spp.

Shigella spp. are causative agents of bacillary dysentery, a human illness with high global morbidity levels, particularly among elderly and infant populations. Shigella infects via the fecal-oral route, and its virulence is dependent upon a type III secretion system (T3SS). Two components of the exposed needle tip complex of the Shigella T3SS, invasion plasmid antigen D (IpaD) and IpaB, have been identified as broadly protective antigens in the mouse lethal pneumonia model. A recombinant fusion protein (DB fusion) was created by joining the coding sequences of IpaD and IpaB. The DB fusion is coexpressed with IpaB\u27s cognate chaperone, IpgC, for proper recombinant expression. The chaperone can then be removed by using the mild detergents octyl oligooxyethelene (OPOE) or N,N-dimethyldodecylamine N-oxide (LDAO). The DB fusion in OPOE or LDAO was used for biophysical characterization and subsequent construction of an empirical phase diagram (EPD). The EPD showed that the DB fusion in OPOE is most stable at neutral pH below 55°C. In contrast, the DB fusion in LDAO exhibited remarkable thermal plasticity, since this detergent prevents the loss of secondary and tertiary structures after thermal unfolding at 90°C, as well as preventing thermally induced aggregation. Moreover, the DB fusion in LDAO induced higher interleukin-17 secretion and provided a higher protective efficacy in a mouse challenge model than did the DB fusion in OPOE. These data indicate that LDAO might introduce plasticity to the protein, promoting thermal resilience and enhanced protective efficacy, which may be important in its use as a subunit vaccine

Closed-loop control of boundary layer streaks induced by free-stream turbulence

The central aim of the paper is to carry out a theoretical and numerical study of active wall transpiration control of streaks generated within an incompressible boundary layer by free-stream turbulence. The disturbance flow model is based on the linearized unsteady boundary-region (LUBR) equations, studied by Leib, Wundrow, and Goldstein [J. Fluid Mech. 380, 169 (1999)], which are the rigorous asymptotic limit of the Navier-Stokes equations for low-frequency and long-streamwise wavelength. The mathematical formulation of the problem directly incorporates the random forcing into the equations in a consistent way. Due to linearity, this forcing is factored out and appears as a multiplicative factor. It is shown that the cost function (integral of kinetic energy in the domain) is properly defined as the expectation of a random quadratic function only after integration in wave number space. This operation naturally introduces the free-stream turbulence spectral tensor into the cost function. The controller gains for each wave number are independent of the spectral tensor and, in that sense, universal. Asymptotic matching of the LUBR equations with the free-stream conditions results in an additional forcing term in the state-space system whose presence necessitates the reformulation of the control problem and the rederivation of its solution. It is proved that the solution can be obtained analytically using an extension of the sweep method used in control theory to obtain the standard Riccati equation. The control signal consists of two components, a feedback part and a feed-forward part (that depends explicitly on the forcing term). Explicit recursive equations that provide these two components are derived. It is shown that the feed-forward part makes a negligible contribution to the control signal. We also derive an explicit expression that a priori (i.e., before solving the control problem) leads to the minimum of the objective cost function (i.e., the fundamental performance limit), based only on the system matrices and the initial and free-stream boundary conditions. The adjoint equations admit a self-similar solution for large spanwise wave numbers with a scaling which is different from that of the LUBR equations. The controlled flow field also has a self-similar solution if the weighting matrices of the objective function are chosen appropriately. The code developed to implement this algorithm is efficient and has modest memory requirements. Computations show the significant reduction of energy for each wave number. The control of the full spectrum streaks, for conditions corresponding to a realistic experimental case, shows that the root-mean-square of the streamwise velocity is strongly suppressed in the whole domain and for all the frequency ranges examined

The Effect of Acoustic Forcing on Instabilities and Breakdown in Swept-Wing Flow over a Backward-Facing Step

Instability interaction and breakdown were experimentally investigated in the flow over a swept backward-facing step. Acoustic forcing was used to excite the Tollmien-Schlichting (TS) instability and to acquire phase-locked results. The phase-averaged results illustrate the complex nature of the interaction between the TS and stationary cross flow instabilities. The weak stationary cross flow disturbance causes a distortion of the TS wavefront. The breakdown process is characterized by large positive and negative spikes in velocity. The positive spikes occur near the same time and location as the positive part of the TS wave. Higher-order spectral analysis was used to further investigate the nonlinear interactions between the TS instability and the traveling cross flow disturbances. The results reveal that a likely cause for the generation of the spikes corresponds to nonlinear interactions between the TS, traveling cross flow, and stationary cross flow disturbances. The spikes begin at low amplitudes of the unsteady and steady disturbances (2-4% U (sub e) (i.e. boundary layer edge velocity)) but can achieve very large amplitudes (20-30 percent U (sub e) (i.e. boundary layer edge velocity)) that initiate an early, though highly intermittent, breakdown to turbulence

Cross-site comparison of ribosomal depletion kits for Illumina RNAseq library construction

Background Ribosomal RNA (rRNA) comprises at least 90% of total RNA extracted from mammalian tissue or cell line samples. Informative transcriptional profiling using massively parallel sequencing technologies requires either enrichment of mature poly-adenylated transcripts or targeted depletion of the rRNA fraction. The latter method is of particular interest because it is compatible with degraded samples such as those extracted from FFPE and also captures transcripts that are not poly-adenylated such as some non-coding RNAs. Here we provide a cross-site study that evaluates the performance of ribosomal RNA removal kits from Illumina, Takara/Clontech, Kapa Biosystems, Lexogen, New England Biolabs and Qiagen on intact and degraded RNA samples. Results We find that all of the kits are capable of performing significant ribosomal depletion, though there are differences in their ease of use. All kits were able to remove ribosomal RNA to below 20% with intact RNA and identify ~ 14,000 protein coding genes from the Universal Human Reference RNA sample at >1FPKM. Analysis of differentially detected genes between kits suggests that transcript length may be a key factor in library production efficiency. Conclusions These results provide a roadmap for labs on the strengths of each of these methods and how best to utilize them. Keywords: RNAseqr; RNA depletion; Illumina; NGS; ABRF; TranscriptomicsNational Cancer Institute (U.S.) (Grant P30-CA14051)National Institute of Environmental Health Sciences (Grant P30-ES002109

Numerical Evaluation of P-Multigrid Method for the Solution of Discontinuous Galerkin Discretizations of Diffusive Equations

This paper describes numerical experiments with P-multigrid to corroborate analysis, validate the present implementation, and to examine issues that arise in the implementations of the various combinations of relaxation schemes, discretizations and P-multigrid methods. The two approaches to implement P-multigrid presented here are equivalent for most high-order discretization methods such as spectral element, SUPG, and discontinuous Galerkin applied to advection; however it is discovered that the approach that mimics the common geometric multigrid implementation is less robust, and frequently unstable when applied to discontinuous Galerkin discretizations of di usion. Gauss-Seidel relaxation converges 40% faster than block Jacobi, as predicted by analysis; however, the implementation of Gauss-Seidel is considerably more expensive that one would expect because gradients in most neighboring elements must be updated. A compromise quasi Gauss-Seidel relaxation method that evaluates the gradient in each element twice per iteration converges at rates similar to those predicted for true Gauss-Seidel