Ubiquitous expression of mRFP1 in transgenic mice

Fluorescent proteins provide a powerful means to track gene expression and cellular behaviors in the study of model organisms such as mice. Among the new generation of fluorescent protein markers, the monomeric red fluorescent protein mRFP1 is particularly attractive because of its rapid maturation and minimal interference with GFP and GFP-derived markers. Here we evaluate the utility of mRFP1 as a marker in transgenic mice. We show that high level and ubiquitous expression of mRFP1 does not affect mouse development, general physiology, or reproduction. mRFP1 expression can be readily detected with unaided eyes under daylight in transgenic mice on the albino background. The intensity of mRFP1 signals can be used to distinguish homozygous and heterozygous transgenic mice. Together, these features make mRFP1 an attractive marker for broad applications in transgenic research.     

Modeling the water use efficiency of soybean and maize plants under environmental stresses: application of a synthetic model of photosynthesis-transpiration based on stomatal behavior

Understanding the variability of plant WUE and its control mechanism can promote the comprehension to the coupling relationship of water and carbon cycle in terrestrial ecosystem, which is the foundation for developing water-carbon coupling cycle model. In this paper, we made clear the differences of net assimilation rate, transpiration rate, and WUE between the two species by comparing the experiment data of soybean (Glycine max Merr.) and maize (Zea mays L.) plants under water and soil nutrient stresses. WUE of maize was about two and a half times more than that of soybean in the same weather conditions. Enhancement of water stresses led to the marked decrease of Am and Em of two species, but water stresses of some degree could improve WUE, and this effect was more obvious for soybean. WUE of the two species changed with psiL in a second-order curve relation, and the WUE at high fertilization was higher than that at low fertilization, this effect was especially obvious for maize. Moreover, according to the synthetic model of photosynthesis-transpiration based on stomatal behavior (SMPTSB) presented by Yu et al. (2001), the WUE model and its applicability were discussed with the data measured in this experiment. The WUE estimated by means of the model accorded well with the measured values. However, this model underestimated the WUE for maize slightly, thus further improvement on the original model was made in this study. Finally, by discussing some physiological factors controlling Am and WUE, we made clear the physiological explanation for differences of the relative contributions of stomata- and mesophyll processes to control of Am and WUE, and the applicability of WUE model between the two species. Because the requirement to stomatal conductance by unit change of net assimilation rate is different, the responses of opening-closing activity of stomata to environmental stresses are different between the two species. To obtain the same level of net assimilation rate, soybean has to open its stomata more widely to keep small stomatal resistance, as compared with maize.     

Increase of PTP levels in vascular injury and in cultured aortic smooth muscle cells treated with specific growth factors

Migration and proliferation of vascular smooth muscle cells are key events in injury-induced neointima formation. Several growth factors and ANG II are thought to be involved in neointima formation. A recent report indicated that vascular injury is associated with increased mRNA levels of protein tyrosine phosphatase (PTP)-1B (PTP-1B). In the present study, we tested the following hypotheses: 1) rat carotid artery injury induces the expression of PTP-1B, Src homology-2 domain phosphatase (SHP-2), and PTP-proline, glutamate, serine, and threonine sequence (PEST) protein; and 2) polypeptide growth factors as well as ANG II increase the levels of tyrosine phosphatases in cultured rat aortic smooth muscle cells. We found that vascular injury induced by balloon catheter increases the protein levels of aforementioned phosphatases and that these effects occur in a PTP specific, as well as temporally and regionally specific, manner. Moreover, treatment of cultured primary rat aortic smooth muscle cells with PDGF or bFGF, but not with IGF1, EGF, or ANG II, increases PTP-1B, SHP-2, and PTP-PEST protein levels. These results suggest that increased PDGF and bFGF levels, occurring after vascular injury, may induce expression of several PTPs.     

A single DH gene segment creates its own unique CDR-H3 repertoire and is sufficient for B cell development and immune function

To test the contribution of individual D gene segments to B cell development and function, we used gene targeting to create mice that contain only DFL16.1 in the DH locus. We term this D-limited IgH allele DeltaD-DFL. Although the absolute number of IgM+IgD- B cells in the bone marrow was decreased, homozygous DeltaD-DFL BALB/c mice contained normal numbers of IgM+IgD+ B cells in bone marrow and spleen and normal numbers of B1a, B1b, and B2 cells in the peritoneal cavity. Bone marrow IgM+IgD+ B cells express a CDR-H3 repertoire similar in length and amino acid composition to the DFL16.1 subset of the wild-type BALB/c repertoire but divergent from sequences that do not contain DFL16.1. This similarity in content is the product of both germline bias and somatic selection, especially in the transition to the mature IgM+IgD+ stage of development. Serum Ig concentrations and the humoral immune response to a T-dependent Ag ([4-hydroxy-3-nitrophenyl]acetyl hapten) were nearly identical to wild-type littermate controls. A greater variance in the immune response to the T-independent Ag (alpha(1-->3)-dextran) was observed in DeltaD-DFL homozygotes, with half of the mice exhibiting levels below the range exhibited by controls. Although limited to a repertoire specific to DFL16.1, the presence of a single DH gene segment of normal sequence was sufficient for development of normal numbers of mature B cells and for robust humoral immune function.     

A 252-bp insertion in <Emphasis Type="Italic">BoCER1</Emphasis> is responsible for the glossy phenotype in cabbage (<Emphasis Type="Italic">Brassica oleracea</Emphasis> L. var. <Emphasis Type="Italic">capitata</Emphasis>)

Bright green of leaf head is an important characteristic in cabbage breeding. Wax-less cabbage shows glossy phenotype on leaf surface, which facilitates the brilliant green cabbage breeding. In this study, we identified a spontaneous glossy mutant g21-3 in cabbage. Genetic analyses showed that its glossy phenotype is controlled by a single recessive gene. Further analysis indicated that the glossy phenotype of g21-3 and a known glossy cabbage mutant 10Q-961 was controlled by a same locus. According to the fine-mapping of glossy-controlled gene in 10Q-961, BoCER1 was identified as a candidate gene which was found to be closely related to the glossy phenotype in g21-3. Sub-cellular localization showed that BoCER1 protein is localized to the endoplasmic reticulum. Sequence analysis revealed that a 252-bp insertion was included in the fourth intron of BoCER1 in g21-3, but not in the wild-type 21-3. The insertion significantly inhibited the expression of BoCER1. A marker designed to distinguish between the BoCER1 alleles in g21-3 and wild-type cabbage co-segregated perfectly with glossy/waxy phenotypes in backcross population, confirming that the insertion mutation of BoCER1 is responsible for the glossy phenotype. The allele-specific marker is effective for marker-assisted selection of the glossy cabbage.     

Automatic infection detection based on electronic medical records

Background

Making accurate patient care decision, as early as possible, is a constant challenge, especially for physicians in the emergency department. The increasing volumes of electronic medical records (EMRs) open new horizons for automatic diagnosis. In this paper, we propose to use machine learning approaches for automatic infection detection based on EMRs. Five categories of information are utilized for prediction, including personal information, admission note, vital signs, diagnose test results and medical image diagnose.

Results

Experimental results on a newly constructed EMRs dataset from emergency department show that machine learning models can achieve a decent performance for infection detection with area under the receiver operator characteristic curve (AUC) of 0.88. Out of all the five types of information, admission note in text form makes the most contribution with the AUC of 0.87.

Conclusions

This study provides a state-of-the-art EMRs processing system to automatically make medical decisions. It extracts five types of features associated with infection and achieves a decent performance on automatic infection detection based on machine learning models.

     

Microbial pathways for nitrogen loss in an upland soil

The distribution and importance of anaerobic ammonium oxidation (anammox) and nitrite‐dependent anaerobic methane oxidation (n‐damo) have been identified in aquatic ecosystems; their role in agricultural upland soils however has not yet been well investigated. In this study, we examined spatio‐temporal distributions of anammox and n‐damo bacteria in soil profiles (300 cm depth) from an agricultural upland. Monitoring nitrogen (N) conversion activity using isotope‐tracing techniques over the course of one year showed denitrification (99.0% N‐loss in the winter and 85.0% N‐loss in the summer) predominated over anammox (1.0% N‐loss in the winter and 14.4% N‐loss in the summer) and n‐damo (0.6% N‐loss in the winter) in surface soils (0–20 cm). While below 20 cm depth, N‐loss was dominated by anammox (79.4 ± 14.3% in the winter and 65.4 ± 12.5% in the summer) and n‐damo was not detected. Phylogenetic analysis showed that Candidatus Brocadia anammoxidans dominated the anammox community in the surface soil and Candidatus Brocadia fulgida dominated below 20 cm depth. Dissimilatory nitrate reduction to ammonium (DNRA), another nitrite reduction process, was found to play a limited role (4.9 ± 3.5%) in the surface soil compared with denitrification; below 80 cm DNRA rates were much higher than rates of anammox and denitrification. Ammonium oxidation was the main source of above 80 cm (70.9 ± 23.3%), the key influencing factor on anammox rates, and nitrate reduction (100%) was the main source below 80 cm. Considering the anammox, n‐damo and denitrification rates as a whole in the sampled soil profile, denitrification is still the main N‐loss process in upland soils.