CDPK-mediated abiotic stress signaling

Calcium-dependent protein kinases (CDPKs) constitute a large multigene family in various plant species. CDPKs have been shown to have important roles in various physiological processes, including plant growth and development and abiotic and biotic stress responses in plants. Functional analysis using gain-of-function and loss-of-function mutants has revealed the biological function of CDPKs in planta. Several CDPKs have been shown to be essential factors in abiotic stress tolerance, positively or negatively regulating stress tolerance by modulating ABA signaling and reducing the accumulation of reactive oxygen species (ROS). This review summarizes recent results describing the biological function of CDPKs that are involved in abiotic stress tolerance.     

c-Abl Inhibition Delays Motor Neuron Degeneration in the G93A Mouse,an Animal Model of Amyotrophic Lateral Sclerosis

Background

Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease characterized by progressive death of motor neurons. Although the pathogenesis of ALS remains unclear, several cellular processes are known to be involved, including apoptosis. A previous study revealed the apoptosis-related gene c-Abl to be upregulated in sporadic ALS motor neurons.

Methodology/Findings

We investigated the possibility that c-Abl activation is involved in the progression of ALS and that c-Abl inhibition is potentially a therapeutic strategy for ALS. Using a mouse motor neuron cell line, we found that mutation of Cu/Zn-superoxide dismutase-1 (SOD1), which is one of the causative genes of familial ALS, induced the upregulation of c-Abl and decreased cell viability, and that the c-Abl inhibitor dasatinib inhibited cytotoxicity. Activation of c-Abl with a concomitant increase in activated caspase-3 was observed in the lumbar spine of G93A-SOD1 transgenic mice (G93A mice), a widely used model of ALS. The survival of G93A mice was improved by oral administration of dasatinib, which also decreased c-Abl phosphorylation, inactivated caspase-3, and improved the innervation status of neuromuscular junctions. In addition, c-Abl expression in postmortem spinal cord tissues from sporadic ALS patients was increased by 3-fold compared with non-ALS patients.

Conclusions/Significance

The present results suggest that c-Abl is a potential therapeutic target for ALS and that the c-Abl inhibitor dasatinib has neuroprotective properties in vitro and in vivo.     

RG-II from Panax ginseng C.A. Meyer suppresses asthmatic reaction

In asthma, T helper 2 (T(H)2)-type cytokines such as interleukin (IL)-4, IL-5, and IL-13 are produced by activated CD4(+) T cells. Dendritic cells played an important role in determining the fate of naive T cells into either T(H)1 or T(H)2 cells. We determined whether RG-II regulates the T(H)1/T(H)2 immune response by using an ovalbumin-induced murine model of asthma. RG-II reduced IL-4 production but increased interferon- gamma production, and inhibited GATA-3 gene expression. RG-II also inhibited asthmatic reactions including an increase in the number of eosinophils in bronchoalveolar lavage fluid, an increase in inflammatory cell infiltration in lung tissues, airway luminal narrowing, and airway hyperresponsiveness. This study provides evidence that RG-II plays a critical role in ameliorating the pathogenic process of asthmatic inflammation in mice. These findings provide new insights into the immunotherapeutic role of RG-II in terms of its effects in a murine model of asthma.     

Receptor Diversity and Host Interaction of Bacteriophages Infecting Salmonella enterica Serovar Typhimurium

Background

Salmonella enterica subspecies enterica serovar Typhimurium is a Gram-negative pathogen causing salmonellosis. Salmonella Typhimurium-targeting bacteriophages have been proposed as an alternative biocontrol agent to antibiotics. To further understand infection and interaction mechanisms between the host strains and the bacteriophages, the receptor diversity of these phages needs to be elucidated.

Methodology/Principal Findings

Twenty-five Salmonella phages were isolated and their receptors were identified by screening a Tn5 random mutant library of S. Typhimurium SL1344. Among them, three types of receptors were identified flagella (11 phages), vitamin B₁₂ uptake outer membrane protein, BtuB (7 phages) and lipopolysaccharide-related O-antigen (7 phages). TEM observation revealed that the phages using flagella (group F) or BtuB (group B) as a receptor belong to Siphoviridae family, and the phages using O-antigen of LPS as a receptor (group L) belong to Podoviridae family. Interestingly, while some of group F phages (F-I) target FliC host receptor, others (F-II) target both FliC and FljB receptors, suggesting that two subgroups are present in group F phages. Cross-resistance assay of group B and L revealed that group L phages could not infect group B phage-resistant strains and reversely group B phages could not infect group L SPN9TCW-resistant strain.

Conclusions/Significance

In this report, three receptor groups of 25 newly isolated S. Typhimurium-targeting phages were determined. Among them, two subgroups of group F phages interact with their host receptors in different manner. In addition, the host receptors of group B or group L SPN9TCW phages hinder other group phage infection, probably due to interaction between receptors of their groups. This study provides novel insights into phage-host receptor interaction for Salmonella phages and will inform development of optimal phage therapy for protection against Salmonella.     

Influence of operating parameters on concentration and purification of L-lactic acid using electrodialysis

An experimental investigation was presented to determine the optimum configuration of influential parameters (concentrate volume, flow rate, temperature, initial lactate concentration, voltage, and impurities) for the best performance. AMX-CMX ion-exchange membranes were used in all experiments. Temperature was found to possess an important role in the increase of lactate recovery, and it was not increased to high values due to membrane destruction. When the higher voltage was applied to electrodialyzer, the better performance of electrodialysis was observed due to enhancement of the driving force. It was found that, to achieve an optimum operating condition, feed volume, concentrate volume, flow rate, temperature, initial lactate concentration in concentrate, and voltage should be 2 L, 1 L, 0.8 L/min, 32°C, 1 g/L, and 15.0 V, respectively. Under these optimized conditions, 97% of lactate was successfully recovered from the fermentation broth, where the lactate flux and energy cost were 7.2 ± 0.6 moles/m²/h and 0.25 kWh/kg, respectively. The results of experiments indicate that the lactic acid in the fermentation broth can be economically purified by electrodialysis.     

The innervation and adaptive significance of extensively distributed neuromasts in <Emphasis Type="Italic">Glossogobius olivaceus</Emphasis> (Perciformes: Gobiidae)

Components of the lateral line system and their innervation were examined in Glossogobius olivaceus (Gobiidae), with almost all of the trunk scales bearing a row of superficial neuromasts, the latter comprising some 2,900 of the total (ca. 4,800) neuromasts on the body. The relationship between orientation and innervation of the superficial neuromasts on the head showed the buccal and mandibular rami to be clearly separated. On the trunk, the lateral ramus detached a number of branches, typically comprising dorsal, lateral and ventral ramules, to innervate neuromasts. Extensively distributed neuromasts were considered as an adaptation to a nocturnal habit, compensating for reduced vision.     

Polymorphisms of the 5′ regulatory region of the porcine <Emphasis Type="Italic">PPARGC1A</Emphasis> gene and the effects on muscle fiber characteristics and meat quality

The purpose of this study was to determine the structure of the porcine PPARGC1A 5′ upstream region, and to find suitable molecular markers for improved meat quality and good lean meat production. Ten DNA polymorphisms, including 7 SNPs, 2 microsatellites, and 1 insertion or deletion were newly found in the 5′ upstream region of PPARGC1A. Three SNPs that had restriction enzyme site were evaluated for associations with muscle fiber characteristics and production traits. Two hundred fifty-two pigs (Yorkshire and Landrace) were used in this analysis. The c.-2894G>A genotypes was significantly associated with muscle fiber characteristics, including the number of fiber type I and IIb composition (P < 0.05), mean cross-sectional area of fibers (P < 0.01), and fiber number per unit area (P < 0.05). The animals with the GG genotype had a higher percentage of type I fibers and a lower percentage of type IIb fibers with better meat quality [higher pH value (P < 0.05) and lower drip loss (P < 0.05)] and lean meat production [larger loin eye area (P < 0.05)]. Moreover, the mRNA expression levels of PPARGC1A among genotypes were significantly different with the highest level of GG genotype. The c.-2885G>T and c.-1402A>T sites showed similar results that had significant effects on the mean cross-sectional area (CSA; P < 0.05), fiber number per unit area (P < 0.05) and loin eye area (P < 0.01). Therefore, we suggest that the c.-2894G>A polymorphism in the 5′ upstream region of the porcine PPARGC1A gene can be used as a meaningful molecular marker for simultaneous improvement of lean meat production and quality traits.     

Novel reporter systems for facile evaluation of I-SceI-mediated genome editing

Two major limitations to achieve efficient homing endonuclease-stimulated gene correction using retroviral vectors are low frequency of gene targeting and random integration of the targeting vectors. To overcome these issues, we developed a reporter system for quick and facile testing of novel strategies to promote the selection of cells that undergo targeted gene repair and to minimize the persistence of random integrations and non-homologous end-joining events. In this system, the gene target has an I-SceI site upstream of an EGFP reporter; and the repair template includes a non-functional EGFP gene, the positive selection transgene MGMTP140K tagged with mCherry, and the inducible Caspase-9 suicide gene. Using this dual fluorescent reporter system it is possible to detect properly targeted integration. Furthermore, this reporter system provides an efficient approach to enrich for gene correction events and to deplete events produced by random integration. We have also developed a second reporter system containing MGMTP140K in the integrated target locus, which allows for selection of primary cells with the integrated gene target after transplantation. This system is particularly useful for testing repair strategies in primary hematopoietic stem cells. Thus, our reporter systems should allow for more efficient gene correction with less unwanted off target effects.