Detection of gfp expression from gfp-labelled bacteria spot inoculated onto sugarcane tissues

Green fluorescent protein (GFP) as a marker gene has facilitated biological research in plant-microbe interactions. However, there is one major limiting factor in the detection of GFP in living organisms whose cells emit background autofluorescence. In this study, Herbaspirillum sp. B501gfp1 bacterial cells were spot inoculated onto 5 month-old sterile micro-propagated sugarcane tissues to detect if the GFP fluorescence expression could be distinguished from the tissue’s background fluorescence. Stem tissues and leaf sections mounted on glass slides were directly inoculated with a single touch using the tip of a syringe previously dipped into the inoculum containing 108 bacterial cells/ml. We observed that GFP fluorescence could be easily distinguished in the stem than in the leaf tissues. However, the brightness level of the fluorescence varied with time as a result of fluctuations in the bacterial celldensity. The presence of chloroplasts in the leaf tissues of sugarcane requires the use of bright GFP variants when monitoring bacteria-plant interactions using GFP labelled bacteria

Effectiveness of highly active antiretroviral therapy in HIV-positive children: evaluation at 12 months in a routine program in Cambodia.

OBJECTIVE: Increasing access to highly active antiretroviral therapy to reach all those in need in developing countries (scale up) is slowly expanding to HIV-positive children, but documented experience remains limited. We aimed to describe the clinical, immunologic, and virologic outcomes of pediatric patients with >12 months of highly active antiretroviral therapy in 2 routine programs in Cambodia. METHODS: Between June 2003 and March 2005, 212 children who were younger than 13 years started highly active antiretroviral therapy. Most patients started a standard first-line regimen of lamivudine, stavudine, and nevirapine, using split adult fixed-dosage combinations. CD4 percentage and body weight were monitored routinely. A cross-sectional virologic analysis was conducted in January 2006; genotype resistance testing was performed for patients with a detectable viral load. RESULTS: Mean age of the subjects was 6 years. Median CD4 percentage at baseline was 6. Survival was 92% at 12 months and 91% at 24 months; 13 patients died, and 4 were lost to follow-up. A total of 81% of all patients had an undetectable viral load. Among the patients with a detectable viral load, most mutations were associated with resistance to lamivudine and non-nucleoside reverse-transcriptase inhibitor drugs. Five patients had developed extensive antiretroviral resistance. Being an orphan was found to be a predictor of virologic failure. CONCLUSIONS: This study provides additional evidence of the effectiveness of integrating HIV/AIDS care with highly active antiretroviral therapy for children in a routine setting, with good virologic suppression and immunologic recovery achieved by using split adult fixed-dosage combinations. Viral load monitoring and HIV genotyping are valuable tools for the clinical follow-up of the patients. Orphans should receive careful follow-up and extra support

Colonization ability of Herbaspirillum spp. B501gfp1 in sugarcane, a non-host plant in the presence of indigenous diazotrophic endophytes

Inoculating sugarcane with a mixture of diazotrophic endophytic bacteria has shown that they can provide substantial amount of biologically fixed nitrogen to the plant. The genera of diazotrophic endophytes previously isolated from sugarcane have been reported associating with other nonleguminousplants showing a broad host range. This study examined the colonization ability of a wild rice isolate, Herbaspirillum spp., in sugarcane plants in the presence of indigenous endophytes using two inoculum concentrations (102 and 108 bacterial cells ml-1). Internal tissue colonization was observed in plants inoculated with both the 102 and 108 B501gfp1 bacterial cells ml-1 inoculum concentrations. However, extensive colonization and higher bacterial numbers were determined only in the basal stem tissues of plants inoculated with the 108 bacterial cells ml-1

Vortex Loop Phase Transitions in Liquid Helium, Cosmic Strings, and High-T_c Superconductors

The distribution of thermally excited vortex loops near a superfluid phase transition is calculated from a renormalized theory. The number density of loops with a given perimeter is found to change from exponential decay with increasing perimeter to algebraic decay as T_c is approached, in agreement with recent simulations of both cosmic strings and high-T_c superconductors. Predictions of the value of the exponent of the algebraic decay at T_c and of critical behavior in the vortex density are confirmed by the simulations, giving strong support to the vortex-folding model proposed by Shenoy.Comment: Version to appear in Phys. Rev. Lett, with a number of corrections and addition

International Collaborative Partnership for the Study of Atrial Fibrillation (INTERAF): Rationale, Design, and Initial Descriptives.

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Extreme Type-II Superconductors in a Magnetic Field: A Theory of Critical Fluctuations

A theory of critical fluctuations in extreme type-II superconductors subjected to a finite but weak external magnetic field is presented. It is shown that the standard Ginzburg-Landau representation of this problem can be recast, with help of a novel mapping, as a theory of a new "superconductor", in an effective magnetic field whose overall value is zero, consisting of the original uniform field and a set of neutralizing unit fluxes attached to $N_{\Phi}$ fluctuating vortex lines. The long distance behavior is related to the anisotropic gauge theory in which the original magnetic field plays the role of "charge". The consequences of this "gauge theory" scenario for the critical behavior in high temperature superconductors are explored in detail, with particular emphasis on questions of 3D XY vs. Landau level scaling, physical nature of the vortex "line liquid" and the true normal state, and fluctuation thermodynamics and transport. A "minimal" set of requirements for the theory of vortex-lattice melting in the critical region is also proposed and discussed.Comment: 28 RevTeX pages, 4 .ps figures; appendix A added, additional references, streamlined Secs. IV and V in response to referees' comment

miR-143 Overexpression Impairs Growth of Human Colon Carcinoma Xenografts in Mice with Induction of Apoptosis and Inhibition of Proliferation

BACKGROUND: MicroRNAs (miRNAs) are aberrantly expressed in human cancer and involved in the (dys)regulation of cell survival, proliferation, differentiation and death. Specifically, miRNA-143 (miR-143) is down-regulated in human colon cancer. In the present study, we evaluated the role of miR-143 overexpression on the growth of human colon carcinoma cells xenografted in nude mice (immunodeficient mouse strain: N: NIH(s) II-nu/nu). METHODOLOGY/PRINCIPAL FINDINGS: HCT116 cells with stable miR-143 overexpression (Over-143) and control (Empty) cells were subcutaneously injected into the flanks of nude mice, and tumor growth was evaluated over time. Tumors arose ∼ 14 days after tumor cell implantation, and the experiment was ended at 40 days after implantation. miR-143 was confirmed to be significantly overexpressed in Over-143 versus Empty xenografts, by TaqMan® Real-time PCR (p<0.05). Importantly, Over-143 xenografts displayed slower tumor growth compared to Empty xenografts from 23 until 40 days in vivo (p<0.05), with final volumes of 928±338 and 2512±387 mm(3), respectively. Evaluation of apoptotic proteins showed that Over-143 versus Empty xenografts displayed reduced Bcl-2 levels, and increased caspase-3 activation and PARP cleavage (p<0.05). In addition, the incidence of apoptotic tumor cells, assessed by TUNEL, was increased in Over-143 versus Empty xenografts (p<0.01). Finally, Over-143 versus Empty xenografts displayed significantly reduced NF-κB activation and ERK5 levels and activation (p<0.05), as well as reduced proliferative index, evaluated by Ki-67 immunohistochemistry (p<0.01). CONCLUSIONS: Our results suggest that reduced tumor volume in Over-143 versus Empty xenografts may result from increased apoptosis and decreased proliferation induced by miR-143. This reinforces the relevance of miR-143 in colon cancer, indicating an important role in the control of in vivo tumor progression, and suggesting that miR-143 may constitute a putative novel therapeutic tool for colon cancer treatment that warrants further investigation