Rice LHS1/OsMADS1 Controls Floret Meristem Specification by Coordinated Regulation of Transcription Factors and Hormone Signaling Pathways

SEPALLATA (SEP) MADS box transcription factors mediate floral development in association with other regulators. Mutants in five rice (Oryza sativa) SEP genes suggest both redundant and unique functions in panicle branching and floret development. LEAFY HULL STERILE1/OsMADS1, from a grass-specific subgroup of LOFSEP genes, is required for specifying a single floret on the spikelet meristem and for floret organ development, but its downstream mechanisms are unknown. Here, key pathways and directly modulated targets of OsMADS1 were deduced from expression analysis after its knockdown and induction in developing florets and by studying its chromatin occupancy at downstream genes. The negative regulation of OsMADS34, another LOFSEP gene, and activation of OsMADS55, a SHORT VEGETATIVE PHASE-like floret meristem identity gene, show its role in facilitating the spikelet-to-floret meristem transition. Direct regulation of other transcription factor genes like OsHB4 (a class III homeodomain Leu zipper member), OsBLH1 (a BEL1-like homeodomain member), OsKANADI2, OsKANADI4, and OsETTIN2 show its role in meristem maintenance, determinacy, and lateral organ development. We found that the OsMADS1 targets OsETTIN1 and OsETTIN2 redundantly ensure carpel differentiation. The multiple effects of OsMADS1 in promoting auxin transport, signaling, and auxin-dependent expression and its direct repression of three cytokinin A-type response regulators show its role in balancing meristem growth, lateral organ differentiation, and determinacy. Overall, we show that OsMADS1 integrates transcriptional and signaling pathways to promote rice floret specification and development.

A general methodology for calculating mixed mode stress intensity factors and fracture toughness of solder joints with interfacial cracks

Solder joints in electronic packages undergo thermo-mechanical cycling, resulting in nucleation of micro-cracks, especially at the solder/bond-pad interface, which may lead to fracture of the joints. The fracture toughness of a solder joint depends on material properties, process conditions and service history, as well as strain rate and mode-mixity. This paper reports on a methodology for determining the mixed-mode fracture toughness of solder joints with an interfacial starter-crack, using a modified compact mixed mode (CMM) specimen containing an adhesive joint. Expressions for stress intensity factor (K) and strain energy release rate (G) are developed, using a combination of experiments and finite element (FE) analysis. In this methodology, crack length dependent geometry factors to convert for the modified CMM sample are first obtained via the crack-tip opening displacement (CTOD)-based linear extrapolation method to calculate the under far-field mode I and II conditions (f(1a) and f(2a)), (ii) generation of a master-plot to determine a(c), and (iii) computation of K and G to analyze the fracture behavior of joints. The developed methodology was verified using J-integral calculations, and was also used to calculate experimental fracture toughness values of a few lead-free solder-Cu joints. (C) 2014 Elsevier Ltd. All rights reserved.

Infection of human amniotic and endothelial cells by Japanese encephalitis virus: Increased expression of HLA-F

Productive infection of human amniotic and endothelial cell lines with Japanese encephalitis virus (JEV) was established leading to the induction of NF kappa B and HLA-F, a non-classical MHC molecule. Induction of the HLA-F gene and protein in JEV-infected cells was shown to be NF kappa B dependent since it was blocked by inhibitors of NF kappa B activation. ShRNA targeting lentivirus-mediated stable knockdown of the p65 subunit of NF kappa B inhibited JEV-mediated induction of HLA-F both in the amniotic cell line, AV-3 as well as the human brain microendothelial cell line, HBMEC. The induction of HLA-F by treatment of AV-3 with TNF-alpha was also inhibited by ShRNA mediated knockdown of NF kappa B. TNF-alpha treatment of HEK293T cells that were transfected with reporter plasmids under the control of HLA-F enhancer A elements resulted in significant transactivation of the luciferase reporter gene. NF kappa B-mediated induction of HLA-F following JEV infection and TNF-alpha exposure is being suggested for the first time. (C) 2014 Elsevier Inc. All rights reserved.

A discussion about a class of stress intensity factors and its verification

In this paper, new formulae of a class of stress intensity factors for an infinite plane with two collinear semi-infinite cracks are presented. The formulae differ from those gathered in several handbooks used all over the world. Some experiments and finite element calculations have been developed to verify the new formulae and the results have shown its reliability. Finally, the new formulae and the old are listed to show the differences between them.

The Politics of Internet Regulation in Brazil : dynamics, factors, and actors in broadband policy

In May 2010, Brazil joined the roll of nations with a National Broadband Plan. The Decree nº 7,175/2010 had implemented a program that aimed to offer 30 million permanent broadband accesses until 2014 and established its main goals, such as accelerating economic and social development, promoting digital inclusion, reducing social and regional inequalities, promoting a generation of employment and income, and expanding electronic government services. However, the broadband access in Brazil is limited, expensive, and centralized in the main urban centres. Despite the fast growth in the past years due to mobile internet access, the market is still concentrated in the local incumbent operators that currently provide mobile services, landline services and Paid-TV services, resulting in a high level of market verticalization. The following dissertation investigates the constraint of broadband access development, the dynamics, the actors, and the factors that have delayed the roll-out of broadband services in Brazil. The study also promotes reflections about the challenge posed by the media, by costumers associations and by public opinion as critical observers of the policy making process. This research examines on the political influence towards regulation to determine the way policy will benefit interest groups. Many interviews have been conducted in order to understand the forces which have been acting in the telecommunications in Brazil after privatization, in 1998. This study aims to provide a better understanding of telecommunications regulatory process in Brazil, in order to help the country finding an adequate policy which can lead to the implementation of a broadband roll-out. The universal broadband access is the only way to benefit the whole society in Brazil with a satisfactory level of education and create more jobs and economic development regarding the plenty use of Information and Communications Technology (ICT).

Length weight relationship, condition factors and fecundity of Chrysichthys nigrodigitatus and Chrysichthys walkeri in Asejire Lake

Sex ratio and fecundity variations of Chrysichthys nigrodigitatus and Chrysichthys walkeri from Asejire Lake (Nigeria) were examined. The Logarithm transformation of weight (W) against standard length (SL) gave a straight-line graph represented by the following equations: 1) C. nigrodigitatus LogW =-0.66 + 2.13 Log SL; = 0.854; (P < 0.001) n = 209; 2) C. walkeri LogW = -1.23 + 2.63 Log SL; = 0.759; (P < 0.001) n = 237. Males were generally more than females in both species. The ratio of males:females was higher in C. nigrodigitatus (1:0.18) than in C. walkeri (1:0.8). C. walkeri attained sexual maturity at a smaller size of 20.0 g (12.0 cm Standard Length) compared with C. nigrodigitatus maturity size of 45.0 g (14.0 cm Standard Length). Relative fecundity was not dependent on body weight and standard length for C. walkeri but it was significant at P < 0.05 and P < 0.01 respectively for C. nigrodigitatus

A cocktail of thermally stable, chemically synthesized capture agents for the efficient detection of anti-gp41 antibodies from human sera and techniques

This thesis reports on a method to improve in vitro diagnostic assays that detect immune response, with specific application to HIV-1. The inherent polyclonal diversity of the humoral immune response was addressed by using sequential in situ click chemistry to develop a cocktail of peptide-based capture agents, the components of which were raised against different, representative anti-HIV antibodies that bind to a conserved epitope of the HIV-1 envelope protein gp41. The cocktail was used to detect anti-HIV-1 antibodies from a panel of sera collected from HIV-positive patients, with improved signal-to-noise ratio relative to the gold standard commercial recombinant protein antigen. The capture agents were stable when stored as a powder for two months at temperatures close to 60°C.

In vivo analysis of interactions between trans-acting factors and their target genes

The investigations presented in this thesis use various in vivo techniques to understand how trans-acting factors control gene expression. The first part addresses the transcriptional regulation of muscle creatine kinase (MCK). MCK expression is activated during the course of development and is found only in differentiated muscle. Several in vivo footprints are observed at the enhancer of this gene, but all of these interactions are limited to cell types that express MCK. This is interesting because two of the footprints appear to represent muscle specific use of general transcription factors, while the other two correspond to sites that can bind the myogenic regulator, MyoD1, in vitro. MyoD1 and these general factors are present in myoblasts, but can bind to the enhancer only in myocytes. This suggests that either the factors themselves are post-translationally modified (phosphorylation or protein:protein interactions), or the accessibility of the enhancer to the factors is limited (changes in chromatin structure). The in vivo footprinting study of MCK was performed with a new ligation mediated, single-sided PCR (polymerase chain reaction) technique that I have developed.

The second half of the thesis concerns the regulation of mouse metallothionein (MT). Metallothioneins are a family of highly conserved housekeeping genes whose expression can be induced by heavy metals, steroids, and other stresses. By adapting a primer extension method of genomic sequencing to in vivo footprinting, I've observed both metal inducible and noninducible interactions at the promoter of MT-I. From these results I've been able to limit the possible mechanisms by which metal responsive trans-acting factors induce transcription. These interpretations correlate with a second line of experiments involving the stable titration of positive acting factors necessary for induction of MT. I've amplified the promoter of MT to 10^2-10^3 copies per cell by fusing the 5' and 3' ends of the MT gene to the coding region of DHFR and selecting cells for methotrexate resistance. In these cells, there is a metal-specific titration effect, and although it acts at the level of transcription, it appears to be independent of direct DNA binding factors.

Physico-chemical factors and bacteria in fish ponds

Analyses of pond water and mud samples show that nitrifying bacteria (including ammonifying bacteria, nitrite bacteria, nitrobacteria and denitrifying bacteria) are in general closely correlated with various physico-chemical factors, ammonifying bacteria are mainly correlated with dissolved oxygen; denitrifying bacteria are inversely correlated with phosphorus; nitrite bacteria are closely correlated with nitrites, nitrobacteria are inversely correlated with ammoniac nitrogen. The nitrifying bacteria are more closely correlated with heterotrophic bacteria. Nitrobacteria are inversely correlated with anaerobic heterotrophic bacteria. The correlation is quite weak between all the nitrite bacteria which indicates that the nitrite bacteria have a controlling and regulating function in water quality and there is no interdependence as each plays a role of its own. The paper also discusses how the superficial soil (pond mud down to 3.5 cm deep) and different layers of the mud affect the biomass of bacteria. The study shows that the top superficial layer (down to 1.5 cm deep) is the major area for decomposing and converting organic matter.