Targeted single-cell gene induction by optimizing the dually regulated CRE/loxP system by a newly defined heat-shock promoter and the steroid hormone in Arabidopsis thaliana

Multicellular organisms rely on intercellular communication systems to organize their cellular functions. In studies focusing on intercellular communication, the key experimental techniques include the generation of chimeric tissue using transgenic DNA recombination systems represented by the CRE/loxP system. If an experimental system enables the induction of chimeras at highly targeted cell(s), it will facilitate the reproducibility and precision of experiments. However, multiple technical limitations have made this challenging. The stochastic nature of DNA recombination events, especially, hampers reproducible generation of intended chimeric patterns. Infrared laser-evoked gene operator (IR-LEGO), a microscopic system that irradiates targeted cells using an IR laser, can induce heat shock-mediated expression of transgenes, for example, CRE recombinase gene, in the cells. In this study, we developed a method that induces CRE/loxP recombination in the target cell(s) of plant roots and leaves in a highly specific manner. We combined IR-LEGO, an improved heat-shock-specific promoter, and dexamethasone-dependent regulation of CRE. The optimal IR-laser power and irradiation duration were estimated via exhaustive irradiation trials and subsequent statistical modeling. Under optimized conditions, CRE/loxP recombination was efficiently induced without cellular damage. We also found that the induction efficiency varied among tissue types and cellular sizes. The developed method offers an experimental system to generate a precisely designed chimeric tissue, and thus, will be useful for analyzing intercellular communication at high resolution in roots and leaves

Semi-supervised discovery of differential genes

BACKGROUND: Various statistical scores have been proposed for evaluating the significance of genes that may exhibit differential expression between two or more controlled conditions. However, in many clinical studies to detect clinical marker genes for example, the conditions have not necessarily been controlled well, thus condition labels are sometimes hard to obtain due to physical, financial, and time costs. In such a situation, we can consider an unsupervised case where labels are not available or a semi-supervised case where labels are available for a part of the whole sample set, rather than a well-studied supervised case where all samples have their labels. RESULTS: We assume a latent variable model for the expression of active genes and apply the optimal discovery procedure (ODP) proposed by Storey (2005) to the model. Our latent variable model allows gene significance scores to be applied to unsupervised and semi-supervised cases. The ODP framework improves detectability by sharing the estimated parameters of null and alternative models of multiple tests over multiple genes. A theoretical consideration leads to two different interpretations of the latent variable, i.e., it only implicitly affects the alternative model through the model parameters, or it is explicitly included in the alternative model, so that the interpretations correspond to two different implementations of ODP. By comparing the two implementations through experiments with simulation data, we have found that sharing the latent variable estimation is effective for increasing the detectability of truly active genes. We also show that the unsupervised and semi-supervised rating of genes, which takes into account the samples without condition labels, can improve detection of active genes in real gene discovery problems. CONCLUSION: The experimental results indicate that the ODP framework is effective for hypotheses including latent variables and is further improved by sharing the estimations of hidden variables over multiple tests

A New Soldier-Producing Aphid Species, Pseudoregma baenzigeri, sp. nov., from Northern Thailand

Pseudoregma baenzigeri, sp. nov., is described from northern Thailand. This species forms dense, huge colonies on shoots of the bamboo Dendrocalamus sp., and produces many first-instar, pseudoscorpion-like soldiers. Alate sexuparae were found from the end of September to mid October. Two syrphids, Eupeodes sp. A (allied to E. confrater) and Dideoides chrysotoxoides, and the pyralid Dipha aphidivora were recorded as predators of P. baenzigeri. The aphids were also likely to be eaten by some rodents. The apterous adult, nymphs, soldier and alate sexupara of P. baenzigeri can be distinguished from those of the other congeners by the longer, conical ultimate rostral segment. A tentative key to the species of Pseudoregma living on bamboo is provided

General intelligence is associated with working memory-related brain activity: new evidence from a large sample study

Psychometric intelligence is closely related to working memory capacity. Here we aim to determine the associations of neural activation patterns during the N-back working memory paradigm with psychometric intelligence and working memory performance. We solved the statistical problems of previous studies using (1) a large cohort of 1235 young adults and (2) robust voxel-by-voxel permutation-based statistics at the whole-brain level. Many of the significant correlations were we ak, and our findings were not consistent with those of previous studies. We observed that many of the significant correlations involved brain areas in the periphery or boundaries between the task-positive network (TPN) and task-negative network (TNN), suggesting that the expansion of the TPN or TNN is associated with greater cognitive ability. Lower activity in TPN and less task-induced deactivation (TID) in TNN were associated with greater cognitive ability. These findings indicate that subjects with greater cognitive ability have a lower brain response to task demand, consistent with the notion that TID in TNN reflects cognitive demand but partly inconsistent with the prevailing neural efficiency theory. One exception was the pre-supplementary motor area, which plays a key role in cognitive control and sequential processing. In this area, intelligent subjects demonstrated greater activity related to working memory, suggesting that the pre-supplementary motor area plays a unique role in the execution of working memory tasks in intelligent subjects

Patient Knowledge on Malaria Symptoms Is a Key to Promoting Universal Access of Patients to Effective Malaria Treatment in Palawan, the Philippines

<div><p>Introduction</p><p>Palawan, where health care facilities are still limited, is one of the most malaria endemic provinces in the Philippines. Since 1999, microscopists (community health workers) have been trained in malaria diagnosis and feasibility of early diagnosis and treatments have been enhanced throughout the province. To accelerate the universal access of malaria patients to diagnostic testing in Palawan, positive health seeking behavior should be encouraged when malaria infection is suspected.</p><p>Methods</p><p>In this cross-sectional study, structured interviews were carried out with residents (N = 218) of 20 remote malaria-endemic villages throughout Palawan with a history of suspected malaria from January to February in 2012. Structural equation modeling (SEM) was conducted to determine factors associated with appropriate treatment, which included: (1) socio-demographic characteristics; (2) proximity to a health facility; (3) health seeking behavior; (4) knowledge on malaria; (5) participation in community awareness-raising activities.</p><p>Results</p><p>Three factors independently associated with appropriate treatment were identified by SEM (CMIN = 10.5, df = 11, CFI = 1.000, RMSEA = .000): “living near microscopist” (<i>p</i> < 0.001), “not living near private pharmacy” (<i>p</i> < 0.01), and “having severe symptoms” (<i>p</i> < 0.01). “Severe symptoms” were positively correlated with more “knowledge on malaria symptoms” (<i>p</i> < 0.001). This knowledge was significantly increased by attending “community awareness-raising activities by microscopists” (<i>p</i> < 0.001).</p><p>Conclusions</p><p>In the resource-limited settings, microscopists played a significant role in providing appropriate treatment to all participants with severe malaria symptoms. However, it was considered that knowledge on malaria symptoms made participants more aware of their symptoms, and further progressed self-triage. Strengthening this recognition sensitivity and making residents aware of nearby microscopists may be the keys to accelerating universal access to effective malaria treatment in Palawan.</p></div

Map of Annual Parasite Incidence (API) (confirmed infections/1,000 population at risk) by municipalities, 2010.

<p>This map was constructed by the authors based on the data of provincial health report 2010 [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0127858#pone.0127858.ref005" target="_blank">5</a>].</p