Detection of Ralstonia solanacearum in Soil and Weeds from Commercial Tomato Fields Using Immunocapture and the Polymerase Chain Reaction

A detection assay for Ralstonia solanacearum in soil and weeds was developed by combining immunocapture and the polymerase chain reaction (IC‐PCR). Anti‐R. solanacearum polyclonal antibodies were produced in a white female rabbit and Dynal^® super‐paramagnetic beads were coated with purified immunoglobulinG (IgG). Using IC‐PCR, the 718 bp target DNA was amplified at a detection threshold of approximately 10⁴ colony‐forming units (CFU) bacteria per millilitre of suspension. DNA was not amplified in soil suspensions derived from autoclaved and non‐autoclaved soils, which contained R. solanacearum at 1–10⁵ CFU/g soil. However, a positive PCR result was obtained when bacteria in the soil suspensions were first enriched in nutrient broth. IC‐PCR detected R. solanacearum in tomato stems 24 h after inoculation by stem puncture with a suspension containing approximately 10⁵ CFU/ml. IC‐PCR detected the bacterium in 28 of 55 (51%) weeds and 10 of 32 (31%) soil samples. Of the weeds, Physalis minima, Amaranthus spinosus and Euphorbia hirta had the highest incidence of infection. R. solanacearum was not detected in soil taken from fallow fields, but it was discovered in some weed species. Symptomless tomato and pepper plants collected from the fields in which tomato bacterial wilt had previously occurred were found to contain R. solanacearum. These discoveries suggest that weeds and latent hosts may play a role in the survival of R. solanacearum between cropping cycles.     

Rapid prototyping of 3D DNA-origami shapes with caDNAno

DNA nanotechnology exploits the programmable specificity afforded by base-pairing to produce self-assembling macromolecular objects of custom shape. For building megadalton-scale DNA nanostructures, a long ‘scaffold’ strand can be employed to template the assembly of hundreds of oligonucleotide ‘staple’ strands into a planar antiparallel array of cross-linked helices. We recently adapted this ‘scaffolded DNA origami’ method to producing 3D shapes formed as pleated layers of double helices constrained to a honeycomb lattice. However, completing the required design steps can be cumbersome and time-consuming. Here we present caDNAno, an open-source software package with a graphical user interface that aids in the design of DNA sequences for folding 3D honeycomb-pleated shapes A series of rectangular-block motifs were designed, assembled, and analyzed to identify a well-behaved motif that could serve as a building block for future studies. The use of caDNAno significantly reduces the effort required to design 3D DNA-origami structures. The software is available at http://cadnano.org/, along with example designs and video tutorials demonstrating their construction. The source code is released under the MIT license.     

Zinc and copper intake and sources in healthy adults living in Bangkok and surrounding districts

The levels of intake and food sources of zinc and copper in 231 20–80 yr old healthy adults living in Bangkok and surrounding districts were determined. Dietary data from 3-d food records collected and validated at Research Center, Ramathibodi Hospital were analyzed. Levels of zinc and copper intake were compared between sexes and age groups (20–39, 40–59, and 60–80 yr). Mean daily zinc and copper intakes (±SEM) in men were 6.3 ±0.2 and 1.9 ±0.1 mg, respectively. Mean daily zinc and copper intakes (±SEM) estimated in women (5.5 ±0.2 and 1.6 +-0.1 mg) were significantly lower. Higher zinc and copper intakes were found in the younger (20-39 yr) age group of both sexes. This could be explained by higher density and percentage from animal source of both nutrients. Consumption of various types of meat, fish, egg, and milk accounted for 42 and 22%, and rice, the staple food of Thai people for 9 and 23% of total dietary zinc and copper, respectively.     

Insect-resistant transgenic brinjal plants

A synthetic cry1Ab gene coding for an insecticidal crystal protein (ICP) of Bacillus thuringiensis (Bt) was transferred to brinjal (eggplant) by cocultivating cotyledonary explants with Agrobacterium tumefaciens. Transformant plants resistant to kanamycin were regenerated. Hybridization experiments demonstrated gene integration and mRNA expression. Double-antibody sandwich ELISA analysis revealed Bt toxin protein expression in the transgenic plants. The expression resulted in a significant insecticidal activity of transgenic brinjal fruits against the larvae of fruit borer (Leucinodes orbonalis). The results also demonstrated that a synthetic gene based on monocot codon usage can be expressed in dicotyledonous plants for insect control.     

Periostin as a Tissue and Urinary Biomarker of Renal Injury in Type 2 Diabetes Mellitus

Background

Improving the early detection of diabetic nephropathy remains a great challenge in disease management. Periostin is a marker of renal tubular injury and related to progressive kidney injury in animal models of chronic kidney disease. The clinical implications of urinary periostin activities in patients with type 2 diabetes have not been evaluated.

Methods

Urine samples were obtained from 30 healthy volunteers and 328 type 2 diabetic patients with normoalbuminuria (n=114), microalbuminuria (n=100) and macroalbuminuria (n=114). The excretion levels of urinary periostin were quantified with enzyme-linked immunosorbent assay. Immunohistochemical periostin expression was determined in kidney tissues from overt diabetic nephropathy.

Results

Increased periostin expression in glomeruli and tubular epithelium in diabetic renal pathology was observed. Urinary periostin levels were significantly elevated in the patients of the normoalbuminuria [3.06 (IQR: 1.12, 6.77) ng/mgCr], microalbuminuria [8.71 (IQR: 5.09, 19.29) ng/mgCr] and macroalbuminuria [13.58 (IQR: 3.99, 16.19) ng/mgCr] compared with healthy controls [1.15 (IQR: 0.60, 1.63) ng/mgCr] (P<0.01).Increased urine periostin level significantly correlated with aging, high albuminuria and decline of GFR. Urine periostin ELISA also demonstrated high performance for the diagnosis of established normoalbuminuric, microalbuminuric and macroalbuminuric type 2 diabetes (AUC 0.78 (95%CI, 0.71 to 0.86), 0.99 (95%CI, 0.98 to 1.00) and 0.95 (95%CI, 0.91 to 0.98), respectively).

Conclusion

The study indicates that increased urine periostin levels can be detected in patients with type 2 diabetes before the onset of significant albuminuria. Urinary periostin is an associated renal derangement in patients with established diabetic nephropathy and it may be used as an early marker of diabetic renal injury.     

Local,cell-nonautonomous feedback regulation of myosin dynamics patterns transitions in cell behavior: a role for tension and geometry?

How robust patterns of tissue dynamics emerge from heterogeneities, stochasticities, and asynchronies in cell behavior is an outstanding question in morphogenesis. A clear understanding of this requires examining the influence of the behavior of single cells on tissue patterning. Here we develop single-cell manipulation strategies to uncover the origin of patterned cell behavior in the amnioserosa during Drosophila dorsal closure. We show that the formation and dissolution of contractile, medial actomyosin networks previously shown to underlie pulsed apical constrictions in the amnioserosa are apparently asynchronous in adjacent cells. We demonstrate for the first time that mechanical stresses and Rho1 GTPase control myosin dynamics qualitatively and quantitatively, in amplitude and direction, both cell autonomously and nonautonomously. We then demonstrate that interfering with myosin-dependent contractility in single cells also influences pulsed constrictions cell nonautonomously. Our results suggest that signals and stresses can feedback regulate the amplitude and spatial propagation of pulsed constrictions through their influence on tension and geometry. We establish the relevance of these findings to native closure by showing that cell delamination represents a locally patterned and collective transition from pulsed to unpulsed constriction that also relies on the nonautonomous feedback control of myosin dynamics.     

Inhibition of the Water Oxidizing Complex of Photosystem II and the Reoxidation of the Quinone Acceptor QA ? by Pb2+

The action of the environmental toxic Pb²⁺ on photosynthetic electron transport was studied in thylakoid membranes isolated from spinach leaves. Fluorescence and thermoluminescence techniques were performed in order to determine the mode of Pb²⁺ action in photosystem II (PSII). The invariance of fluorescence characteristics of chlorophyll a (Chl a) and magnesium tetraphenylporphyrin (MgTPP), a molecule structurally analogous to Chl a, in the presence of Pb²⁺ confirms that Pb cation does not interact directly with chlorophyll molecules in PSII. The results show that Pb interacts with the water oxidation complex thus perturbing charge recombination between the quinone acceptors of PSII and the S₂ state of the Mn₄Ca cluster. Electron transfer between the quinone acceptors Q_A and Q_B is also greatly retarded in the presence of Pb²⁺. This is proposed to be owing to a transmembrane modification of the acceptor side of the photosystem.     

Lucidafuranocoumarins B and C from the twigs of Feroniella lucida: Absolute configurations of lucidafuranocoumarin C

Two new furanocoumarins, lucidafuranocoumarins B (1) and C (2), were isolated from the twigs of Feroniella lucida, together with five known compounds (3–7). The structures of these compounds were identified on the basis of extensive spectroscopic methods. The absolute configurations of lucidafuranocoumarin C at C-2″ and C-5″ were established as R- and S-configurations, respectively, by applying Mosher's method. Some isolates were also evaluated for their cytotoxicity against KB, MCF-7 and NCI-H187 cell lines.     

Potential of Sonchus Arvensis for the Phytoremediation of Lead-Contaminated Soil

Sonchus arvensis is one of the pioneer plant species that were found in the abandoned Bo Ngam Pb mine in Thailand. S. arvensis was collected from three sites. The highest Pb shoot concentration was 9317 mg kg^?1 and the highest translocation factor (TF) and bioaccumulation factor (BF) values were 2.5 and 6.0, respectively. To investigate Pb uptake capacity of S. arvensis, a hydroponic experiment was performed for 15 d. S. arvensis exposed to 5 mg L^?1 Pb solution had the highest Pb shoot accumulation (849 mg kg^?1). In a pot study, S. arvensis was grown in Pb mine soils amended with organic and inorganic fertilizers for 2 mo. The addition of organic fertilizer to the soil increased plant dry biomass sharply. All treatments with ethylene-diamine-tetra-acetic acid (EDTA) had Pb accumulation in shoots greater than 1000 mg kg^?1 and the highest Pb shoot accumulation was found in S. arvensis grown in soil amended with organic fertilizer and EDTA (1397 mg kg^?1). In a field trial study, S. arvensis was grown at three sites in the mine area for 6 mo. S. arvensis could tolerate a total Pb of 100,000 mg kg^?1 in the soil and accumulated Pb in the shoots up to 3664 mg kg^?1 with high TF (2.19) and BF (2.38) values. These results suggest that S. arvensis is a good candidate for Pb phytoremediation.