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Find video protocols related to scientific articles indexed in Pubmed.
ABCG1 Is Required for Pulmonary B-1 B Cell and Natural Antibody Homeostasis.
J. Immunol.
PUBLISHED: 10-22-2014
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Many metabolic diseases, including atherosclerosis, type 2 diabetes, pulmonary alveolar proteinosis, and obesity, have a chronic inflammatory component involving both innate and adaptive immunity. Mice lacking the ATP-binding cassette transporter G1 (ABCG1) develop chronic inflammation in the lungs, which is associated with the lipid accumulation (cholesterol, cholesterol ester, and phospholipid) and cholesterol crystal deposition that are characteristic of atherosclerotic lesions and pulmonary alveolar proteinosis. In this article, we demonstrate that specific lipids, likely oxidized phospholipids and/or sterols, elicit a lung-specific immune response in Abcg1(-/-) mice. Loss of ABCG1 results in increased levels of specific oxysterols, phosphatidylcholines, and oxidized phospholipids, including 1-palmitoyl-2-(5'-oxovaleroyl)-sn-glycero-3-phosphocholine, in the lungs. Further, we identify a niche-specific increase in natural Ab (NAb)-secreting B-1 B cells in response to this lipid accumulation that is paralleled by increased titers of IgM, IgA, and IgG against oxidation-specific epitopes, such as those on oxidized low-density lipoprotein and malondialdehyde-modified low-density lipoprotein. Finally, we identify a cytokine/chemokine signature that is reflective of increased B cell activation, Ab secretion, and homing. Collectively, these data demonstrate that the accumulation of lipids in Abcg1(-/-) mice induces the specific expansion and localization of B-1 B cells, which secrete NAbs that may help to protect against the development of atherosclerosis. Indeed, despite chronic lipid accumulation and inflammation, hyperlipidemic mice lacking ABCG1 develop smaller atherosclerotic lesions compared with controls. These data also suggest that Abcg1(-/-) mice may represent a new model in which to study the protective functions of B-1 B cells/NAbs and suggest novel targets for pharmacologic intervention and treatment of disease.
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Atheroprotective immunization with malondialdehyde-modified LDL is hapten specific and dependent on advanced MDA adducts: implications for development of an atheroprotective vaccine.
J. Lipid Res.
PUBLISHED: 08-20-2014
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Immunization with homologous malondialdehyde (MDA)-modified LDL (MDA-LDL) leads to atheroprotection in experimental models supporting the concept that a vaccine to oxidation-specific epitopes (OSEs) of oxidized LDL could limit atherogenesis. However, modification of human LDL with OSE to use as an immunogen would be impractical for generalized use. Furthermore, when MDA is used to modify LDL, a wide variety of related MDA adducts are formed, both simple and more complex. To define the relevant epitopes that would reproduce the atheroprotective effects of immunization with MDA-LDL, we sought to determine the responsible immunodominant and atheroprotective adducts. We now demonstrate that fluorescent adducts of MDA involving the condensation of two or more MDA molecules with lysine to form malondialdehyde-acetaldehyde (MAA)-type adducts generate immunodominant epitopes that lead to atheroprotective responses. We further demonstrate that a T helper (Th) 2-biased hapten-specific humoral and cellular response is sufficient, and thus, MAA-modified homologous albumin is an equally effective immunogen. We further show that such Th2-biased humoral responses per se are not atheroprotective if they do not target relevant antigens. These data demonstrate the feasibility of development of a small-molecule immunogen that could stimulate MAA-specific immune responses, which could be used to develop a vaccine approach to retard or prevent atherogenesis.
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Is Radical-by-Radical Reading Strategy Effective for Chinese Pure Alexia? A Case Study.
Neuropsychology
PUBLISHED: 07-29-2014
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Objective: Currently, it is unclear whether a radical-by-radical (RBR) reading strategy exists in Chinese pure alexia, compared to the letter-by-letter (LBL) reading strategy in alphabetic languages. In this study, we focus on exploring the reading features of a Chinese pure alexic patient during the process of language recovery. Method: We investigated this issue using a series of neuropsychological tests at 4 days, 11 days, and 8 months after admission of a Chinese pure alexic patient. Results: The results showed that most words could not be read aloud initially by the patient. One week later, 2 clinical features were noted: RBR reading strategy and integration impairment. Finally, there were statistically significant differences in the reaction time between the single characters and compound characters, which indicated that a radical number effect may exist. Conclusion: Combined with the results of previous studies, the current case study indicates that there is evidence suggesting that the RBR reading strategy in Chinese pure alexia is of limited benefit, a result that appears fundamentally different from the LBL reading strategy. (PsycINFO Database Record (c) 2014 APA, all rights reserved).
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New region-scalable discriminant and fitting energy functional for driving geometric active contours in medical image segmentation.
Comput Math Methods Med
PUBLISHED: 07-07-2014
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We propose a novel region-based geometric active contour model that uses region-scalable discriminant and fitting energy functional for handling the intensity inhomogeneity and weak boundary problems in medical image segmentation. The region-scalable discriminant and fitting energy functional is defined to capture the image intensity characteristics in local and global regions for driving the evolution of active contour. The discriminant term in the model aims at separating background and foreground in scalable regions while the fitting term tends to fit the intensity in these regions. This model is then transformed into a variational level set formulation with a level set regularization term for accurate computation. The new model utilizes intensity information in the local and global regions as much as possible; so it not only handles better intensity inhomogeneity, but also allows more robustness to noise and more flexible initialization in comparison to the original global region and regional-scalable based models. Experimental results for synthetic and real medical image segmentation show the advantages of the proposed method in terms of accuracy and robustness.
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Comparison of two serpins of Clonorchis sinensis by bioinformatics, expression, and localization in metacercaria.
Pathog Glob Health
PUBLISHED: 05-16-2014
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Clonorchiasis, which has been an important public health problem in China, is caused by ingestion of raw or undercooked fish contaminated by live metacercaria. Therefore, preventing fish from infecting is of great significance for controlling the disease. SERPINs (serine protease inhibitors) are well known as negative regulators of hemostasis, thrombolysis, and innate immune responses. In the present study, two full-length sequences encoding SERPIN were identified from metacercaria cDNA library of Clonorchis sinensis (C. sinensis) and were denominated as CsSERPIN and CsSERPIN3, respectively. Bioinformatics analysis showed that the two sequences shares 35.9% identity to each other. Both of the sequences have SERPIN domain and the greatest difference between the two domains is the reactive centre loop. Transmembrane region was found in CsSERPIN3 while not in CsSERPIN. The expression of the two CsSERPINs was significantly higher at the life stage of metacercaria than that of adult. The transcription levels of CsSERPIN and CsSERPIN3 at metacercaria stage were 3.249- and 11.314-fold of that at adult stage, respectively. Furthermore, the expression of CsSERPIN was 4.32-fold of that of CsSERPIN3 at metacercaria stage. Immunobiochemistry revealed that CsERPIN was dispersed at subtegument and oral sucker of metacercaria, while CsSERPIN3 localized intensely in the tegument of metacercaria of C. sinensis inside of the cyst wall. All these indicated that the CsSERPINs play important roles at metacercaria stage of the parasite. CsSERPIN may take part in regulation of endogenous serine proteinase and CsSERPIN3 may be involved in immune evasion and be a potential candidate for vaccine and drug target for clonorchiasis.
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Developmental tuning of reflexive attentional effect to biological motion cues.
Sci Rep
PUBLISHED: 05-07-2014
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The human visual system is extremely sensitive to the direction information retrieved from biological motion. In the current study, we investigate the functional impact of this sensitivity on attentional orienting in young children. We found that children as early as 4 years old, like adults, showed a robust reflexive attentional orienting effect to the walking direction of an upright point-light walker, indicating that biological motion signals can automatically direct spatial attention at an early age. More importantly, the inversion effect associated with attentional orienting emerges by 4 years old and gradually develops into a similar pattern found in adults. These results provide strong evidence that biological motion cues can guide the distribution of spatial attention in young children, and highlight a critical development from a broadly- to finely-tuned process of utilizing biological motion cues in the human social brain.
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Fine neural tuning for orthographic properties of words emerges early in children reading alphabetic script.
J Cogn Neurosci
PUBLISHED: 05-06-2014
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The left-lateralized N170 component of ERPs for words compared with various control stimuli is considered as an electrophysiological manifestation of visual expertise for written words. To understand the information sensitivity of the effect, researchers distinguish between coarse tuning for words (the N170 amplitude difference between words and symbol strings) and fine tuning for words (the N170 amplitude difference between words and consonant strings). Earlier developmental ERP studies demonstrated that the coarse tuning for words occurred early in children (8 years old), whereas the fine tuning for words emerged much later (10 years old). Given that there are large individual differences in reading ability in young children, these tuning effects may emerge earlier than expected in some children. This study measured N170 responses to words and control stimuli in a large group of 7-year-olds that varied widely in reading ability. In both low and high reading ability groups, we observed the coarse neural tuning for words. More interestingly, we found that a stronger N170 for words than consonant strings emerged in children with high but not low reading ability. Our study demonstrates for the first time that fine neural tuning for orthographic properties of words can be observed in young children with high reading ability, suggesting that the emergent age of this effect is much earlier than previously assumed. The modulation of this effect by reading ability suggests that fine tuning is flexible and highly related to experience. Moreover, we found a correlation between this tuning effect at left occipitotemporal electrodes and children's reading ability, suggesting that the fine tuning might be a biomarker of reading skills at the very beginning of learning to read.
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Proteomic identification of potential Clonorchis sinensis excretory/secretory products capable of binding and activating human hepatic stellate cells.
Parasitol. Res.
PUBLISHED: 03-28-2014
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Epidemiological and experimental evidence demonstrated that Clonorchis sinensis is an important risk factor of hepatic fibrosis and cholangiocarcinoma. C. sinensis excretory/secretory products (CsESPs) are protein complex including proteases, antioxidant enzymes, and metabolic enzymes, which may contribute to pathogenesis of liver fluke-associated hepatobiliary diseases. However, potential CsESP candidates involved into hepatic fibrosis and cholangiocarcinoma still remain to be elucidated. In the present study, we performed proteomic identification of CsESP candidates capable of binding and activating human hepatic stellate cell line LX-2. Immunofluorescence analysis confirmed the interaction of CsESPs with LX-2 cell membrane. LX-2 cells could be stimulated by CsESPs from 24 h post incubation (p?
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A Stroop effect emerges in the processing of complex Chinese characters that contain a color-related radical.
Psychol Res
PUBLISHED: 02-19-2014
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Three experiments examined whether a Stroop effect emerges in the processing of complex Chinese characters that contain a color-related radical. In Experiment 1, a Stroop effect occurred when participants responded to the black or white color of the simple characters (black) and (white) by making a left or right keypress. For Experiment 2, in which the stimuli were complex characters whose meanings were unrelated to color but that contained or as a radical, a Stroop effect also occurred, although it was smaller than in Experiment 1. Furthermore, this Stroop effect as a function of radical meaning was shown again in Experiment 3 for low-frequency complex characters but not high-frequency ones. These results suggest that the semantic representations of the complex characters' color-related radicals are accessed in the context of a Stroop color word task, especially for low-frequency characters. Reduction of the Stroop effect in complex characters composed of one radical with color meaning and one without is similar to dilution of the Stroop effect that occurs when a color word is accompanied by a neutral word. Possible implications of the results for accounts of Stroop dilution are discussed.
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Cerebellum engages in automation of verb-generation skill.
J. Integr. Neurosci.
PUBLISHED: 02-05-2014
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Numerous studies have shown cerebellar involvement in item-specific association, a form of explicit learning. However, very few have demonstrated cerebellar participation in automation of non-motor cognitive tasks. Applying fMRI to a repeated verb-generation task, we sought to distinguish cerebellar involvement in learning of item-specific noun-verb association and automation of verb generation skill. The same set of nouns was repeated in six verb-generation blocks so that subjects practiced generating verbs for the nouns. The practice was followed by a novel block with a different set of nouns. The cerebellar vermis (IV/V) and the right cerebellar lobule VI showed decreased activation following practice; activation in the right cerebellar Crus I was significantly lower in the novel challenge than in the initial verb-generation task. Furthermore, activation in this region during well-practiced blocks strongly correlated with improvement of behavioral performance in both the well-practiced and the novel blocks, suggesting its role in the learning of general mental skills not specific to the practiced noun-verb pairs. Therefore, the cerebellum processes both explicit verbal associative learning and automation of cognitive tasks. Different cerebellar regions predominate in this processing: lobule VI during the acquisition of item-specific association, and Crus I during automation of verb-generation skills through practice.
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Quantitative proteomics of Sesuvium portulacastrum leaves revealed that ion transportation by V-ATPase and sugar accumulation in chloroplast played crucial roles in halophyte salt tolerance.
J Proteomics
PUBLISHED: 01-08-2014
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Physiological and proteomic responses of Sesuvium portulacastrum leaves under salinity were investigated. Different from glycophytes, this halophyte had optimal growth at 200-300mM NaCl and accumulated more starch grains in chloroplasts under high salinity. Increased contents of soluble sugars, proline, and Na(+) were observed upon salinity. X-ray microanalysis revealed that Na(+) was mainly compartmentalized into cell vacuole. Quantitative proteomics produced 96 salt responsive proteins, and the majority was chloroplast-located proteins. Gene ontology analysis revealed that proteins involved in ion binding, proton transport, photosynthesis and ATP synthesis were overrepresented. The expressions of a Na(+)/H(+) antiporter and several ATP synthase subunits were activated upon high salinity. ATP hydrolysis assay demonstrated that V-ATPase activity at tonoplast was dramatically increased upon NaCl whereas vacuolar H(+)-pyrophosphatase and plasma membrane P-ATPase activities were not increased, which indicated that sodium compartmentalization was mainly performed by enhancing V-ATPase activity rather than P-ATPase and H(+)-pyrophosphatase. Accumulation of soluble sugars as well as sodium compartmentalization maintained the osmotic balance between vacuole and cytoplasm, which finally established ionic homeostasis in saline cells in true halophytes.
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Genetic diversity of Plasmodium vivax population in Anhui province of China.
Malar. J.
PUBLISHED: 01-08-2014
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Although the numbers of malaria cases in China have been declining in recent years, outbreaks of Plasmodium vivax malaria were still being reported in rural areas south of the Yellow River. To better understand the transmission dynamics of P. vivax parasites in China, the extent of genetic diversity of P. vivax populations circulating in Bozhou of Anhui province of China were investigated using three polymorphic genetic markers: merozoite surface proteins 1 and 3? (pvmsp-1 and pvmsp-3?) and circumsporozoite protein (pvcsp).
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Do preschool children learn to read words from environmental prints?
PLoS ONE
PUBLISHED: 01-01-2014
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Parents and teachers worldwide believe that a visual environment rich with print can contribute to young children's literacy. Children seem to recognize words in familiar logos at an early age. However, most of previous studies were carried out with alphabetic scripts. Alphabetic letters regularly correspond to phonological segments in a word and provide strong cues about the identity of the whole word. Thus it was not clear whether children can learn to read words by extracting visual word form information from environmental prints. To exclude the phonological-cue confound, this study tested children's knowledge of Chinese words embedded in familiar logos. The four environmental logos were employed and transformed into four versions with the contextual cues (i.e., something apart from the presentation of the words themselves in logo format like the color, logo and font type cues) gradually minimized. Children aged from 3 to 5 were tested. We observed that children of different ages all performed better when words were presented in highly familiar logos compared to when they were presented in a plain fashion, devoid of context. This advantage for familiar logos was also present when the contextual information was only partial. However, the role of various cues in learning words changed with age. The color and logo cues had a larger effect in 3- and 4- year-olds than in 5-year-olds, while the font type cue played a greater role in 5-year-olds than in the other two groups. Our findings demonstrated that young children did not easily learn words by extracting their visual form information even from familiar environmental prints. However, children aged 5 begin to pay more attention to the visual form information of words in highly familiar logos than those aged 3 and 4.
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AISAIC: a software suite for accurate identification of significant aberrations in cancers.
Bioinformatics
PUBLISHED: 11-29-2013
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?Accurate identification of significant aberrations in cancers (AISAIC) is a systematic effort to discover potential cancer-driving genes such as oncogenes and tumor suppressors. Two major confounding factors against this goal are the normal cell contamination and random background aberrations in tumor samples. We describe a Java AISAIC package that provides comprehensive analytic functions and graphic user interface for integrating two statistically principled in silico approaches to address the aforementioned challenges in DNA copy number analyses. In addition, the package provides a command-line interface for users with scripting and programming needs to incorporate or extend AISAIC to their customized analysis pipelines. This open-source multiplatform software offers several attractive features: (i) it implements a user friendly complete pipeline from processing raw data to reporting analytic results; (ii) it detects deletion types directly from copy number signals using a Bayes hypothesis test; (iii) it estimates the fraction of normal contamination for each sample; (iv) it produces unbiased null distribution of random background alterations by iterative aberration-exclusive permutations; and (v) it identifies significant consensus regions and the percentage of homozygous/hemizygous deletions across multiple samples. AISAIC also provides users with a parallel computing option to leverage ubiquitous multicore machines.Availability and implementation:?AISAIC is available as a Java application, with a users guide and source code, at https://code.google.com/p/aisaic/.
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Schistosoma japonicum tegumental protein 20.8, role in reproduction through its calcium binding ability.
Parasitol. Res.
PUBLISHED: 10-31-2013
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Schistosomiasis threatens thousands of millions of peoples health every year in the world. Schistosoma japonicum, a pathogen of schistosomiasis, is covered by a lipid bilayer membrane which plays an important role in nutrient transport, signal transduction, interaction with hosts immune system, etc. Thus, molecules in the tegumental membrane have gained more and more interest for understanding biological and pathological processes of schistosoma. In this study, we found a protein from S. japonicum cDNA library which has a 20.8 KDa molecular weight (SjTP20.8). Recombinant SjTP20.8 was produced and purified from Escherichia coli. The recombinant protein could be detected by S. japonicum-infected mice and human sera, and it had been found localizing in the tegumental membrane of S. japonicum in the section using immunofluorescence assay. In electrophoretic mobility shift assay, the protein could bind calcium iron in neutral condition. Result of cercariae challenge experiment indicates antibody against this protein can protect mice from chronic hepatic fibrosis. Our results indicate the S. japonicum tegumental protein 20.8 is crucial for the parasites calcium absorbing and reproduction.
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Helix-loop-helix factor inhibitor of differentiation 3 regulates interleukin-5 expression and B-1a B cell proliferation.
Arterioscler. Thromb. Vasc. Biol.
PUBLISHED: 10-10-2013
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Natural immunity is emerging as an important mediator of protection from atherogenesis. Natural IgM antibodies that recognize oxidation-specific epitopes on low-density lipoprotein or phospholipids and the B-1a B cells that produce them attenuate atherosclerosis. We previously demonstrated that Apoe(-/-) mice globally deficient in the helix-loop-helix protein inhibitor of differentiation 3 (Id3) develop early diet-induced atherosclerosis. Furthermore, B cell-mediated attenuation of atherosclerosis in B cell-deficient mice was dependent on Id3. Here, we sought to determine whether Id3 regulates B-1a B cells and the natural antibodies that they produce and identify mechanisms mediating these effects.
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Comparative proteomics of primary and secondary lutoids reveals that chitinase and glucanase play a crucial combined role in rubber particle aggregation in Hevea brasiliensis.
J. Proteome Res.
PUBLISHED: 09-23-2013
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Lutoids are specific vacuole-based organelles within the latex-producing laticifers in rubber tree Hevea brasiliensis. Primary and secondary lutoids are found in the primary and secondary laticifers, respectively. Although both lutoid types perform similar roles in rubber particle aggregation (RPA) and latex coagulation, they vary greatly at the morphological and proteomic levels. To compare the differential proteins and determine the shared proteins of the two lutoid types, a proteomic analysis of lutoid membranes and inclusions was performed, revealing 169 proteins that were functionally classified into 14 families. Biological function analysis revealed that most of the proteins are involved in pathogen defense, chitin catabolism, and proton transport. Comparison of the gene and protein changed patterns and determination of the specific roles of several main lutoid proteins, such as glucanase, hevamine, and hevein, demonstrated that Chitinase and glucanase appeared to play crucial synergistic roles in RPA. Integrative analysis revealed a protein-based metabolic network mediating pH and ion homeostasis, defense response, and RPA in lutoids. From these findings, we developed a modified regulation model for lutoid-mediated RPA that will deepen our understanding of potential mechanisms involved in lutoid-mediated RPA and consequent latex coagulation.
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Mutation in folate metabolism causes epigenetic instability and transgenerational effects on development.
Cell
PUBLISHED: 06-01-2013
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The importance of maternal folate consumption for normal development is well established, yet the molecular mechanism linking folate metabolism to development remains poorly understood. The enzyme methionine synthase reductase (Mtrr) is necessary for utilization of methyl groups from the folate cycle. We found that a hypomorphic mutation of the mouse Mtrr gene results in intrauterine growth restriction, developmental delay, and congenital malformations, including neural tube, heart, and placental defects. Importantly, these defects were dependent upon the Mtrr genotypes of the maternal grandparents. Furthermore, we observed widespread epigenetic instability associated with altered gene expression in the placentas of wild-type grandprogeny of Mtrr-deficient maternal grandparents. Embryo transfer experiments revealed that Mtrr deficiency in mice lead to two distinct, separable phenotypes: adverse effects on their wild-type daughters uterine environment, leading to growth defects in wild-type grandprogeny, and the appearance of congenital malformations independent of maternal environment that persist for five generations, likely through transgenerational epigenetic inheritance.
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Comparative proteomics of Thellungiella halophila leaves from plants subjected to salinity reveals the importance of chloroplastic starch and soluble sugars in halophyte salt tolerance.
Mol. Cell Proteomics
PUBLISHED: 05-08-2013
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Thellungiella halophila, a close relative of Arabidopsis, is a model halophyte used to study plant salt tolerance. The proteomic/physiological/transcriptomic analyses of Thellungiella plant leaves subjected to different salinity levels, reported herein, indicate an extraordinary ability of Thellungiella to adapt to large concentrations of exogenous saline by compartmentalizing Na(+) into cell vacuoles and accumulating proline and soluble sugars as organic osmolytes. Salinity stress stimulated the accumulation of starch in chloroplasts, which resulted in a greatly increased content of starch and total sugars in leaves. Comparative proteomics of Thellungiella leaves identified 209 salt-responsive proteins. Among these, the sequences of 108 proteins were strongly homologous to Arabidopsis protein sequences, and 30 had previously been identified as Thellungiella proteins. Functional classification of these proteins into 16 categories indicated that the majority are involved in carbohydrate metabolism, followed by those involved in energy production and conversion, and then those involved in the transport of inorganic ions. Pathway analysis revealed that most of the proteins are involved in starch and sucrose metabolism, carbon fixation, photosynthesis, and glycolysis. Of these processes, the most affected were starch and sucrose metabolism, which might be pivotal for salt tolerance. The gene expression patterns of the 209 salt-responsive proteins revealed through hierarchical clustering of microarray data and the expression patterns of 29 Thellungiella genes evaluated via quantitative RT-PCR were similar to those deduced via proteomic analysis, which underscored the possibility that starch and sucrose metabolism might play pivotal roles in determining the salt tolerance ability of Thellungiella. Our observations enabled us to propose a schematic representation of the systematic salt-tolerance phenotype in Thellungiella and suggested that the increased accumulation of starch, soluble sugars, and proline, as well as subcellular compartmentalization of sodium, might collectively denote important mechanisms for halophyte salt tolerance.
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The orthographic sensitivity to written Chinese in the occipital-temporal cortex.
Exp Brain Res
PUBLISHED: 04-08-2013
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Previous studies have identified an area in the left lateral fusiform cortex that is highly responsive to written words and has been named the visual word form area (VWFA). However, there is disagreement on the specific functional role of this area in word recognition. Chinese characters, which are dramatically different from Roman alphabets in the visual form and in the form to phonological mapping, provide a unique opportunity to investigate the properties of the VWFA. Specifically, to clarify the orthographic sensitivity in the mid-fusiform cortex, we compared fMRI response amplitudes (Exp. 1) as well as the spatial patterns of response across multiple voxels (Exp. 2) between Chinese characters and stimuli derived from Chinese characters with different orthographic properties. The fMRI response amplitude results suggest the existence of orthographic sensitivity in the VWFA. The results from multi-voxel pattern analysis indicate that spatial distribution of the responses across voxels in the occipitotemporal cortex contained discriminative information between the different types of character-related stimuli. These results together suggest that the orthographic rules are likely represented in a distributed neural network with the VWFA containing the most specific information regarding a stimulus orthographic regularity.
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The serological diagnosis of human clonorchiasis by time-resolved fluoroimmunoassay based on GST2-specific IgG4 detection.
Parasitol. Res.
PUBLISHED: 03-01-2013
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Due to its delayed fluorescence of a lanthanide chelate, high accuracy and low background the broad linear range, long fluorescent life-time and large Stokes shift of europium chelates, the time-resolved fluorescence has been developed for higher sensitive immunoassay. In this article, a simple, sensitive and specific method-time-resolved fluoroimmunoassay (TRFIA) was adopted for immunoassay of clonorchiasis, and recombinant glutathione transferases 2 of Clonorchis sinensis (rCsGST2) was used as a diagnostic antigen. To evaluate this novel assay for clinical applications, 409 serum samples were investigated. The diagnostic accuracy of the antigen was evaluated by receiver-operating characteristic (ROC) analysis. The area under the ROC curve (AUC) was 0.965, 95 % confidence interval (CI, 0.946, 0.985). To eliminate the random influence of ambient temperature, test parameters, photometric instruments and so on, the cut-off value was expressed as ratios between the fluorescence of sample and that of a well-defined negative control serum, and the deduced cut-off value was 9.3605. At the optimum cut-off criteria, the technique has a sensitivity of 95.80 %, specificity of 93.60 %. And the cross reactivity revealed that its cross reactivity with Schistosoma japonicum, round worm, hook worm, whip worm, and Toxoplasma gondii was 9.3, 8.3, 7.6, 9.8, and 5.0 %, respectively. Kappa score of agreement between TRFIA and microscopic examination of stools was 0.892, P?
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Expression, immunolocalization, and serological reactivity of a novel sphingomyelin phosphodiesterase-like protein, an excretory/secretory antigen from Clonorchis sinensis.
Parasitol. Res.
PUBLISHED: 03-01-2013
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Clonorchiasis, caused by Clonorchis sinensis infection, is a zoonotic parasitic disease of hepatobiliary system in which the proteins released by adult are major pathogenetic factors. In this study, we first characterized a putative sphingomyelin phosphodiesterase (CsSMPase) A-like secretory protein, which was highly expressed in the adult worm. The full-length gene was cloned. The putative protein is of relatively low homology comparing with SMPase from other species, and of rich T cell and B cell epitopes, suggesting that it is an antigen of strong antigenicity. The complete coding sequence of the gene was expressed in the Escherichia coli. The recombinant CsSMPase (rCsSMPase) can be recognized by C. sinensis-infected serum, and the protein immunoserum can recognize a specific band in excretory/secretory products (ESPs) of C. sinensis adult by western blotting. Immunolocalization revealed that CsSMPase was not only localized on tegument, ventral sucker of metacercaria, and the intestine of adult but also on the nearby epithelium of bile duct of the infected Sprague-Dawley rats, implying that CsSMPase was mainly secreted and excreted through adult intestine and directly interacted with bile duct epithelium. Although immunized rats evoked high level antibody response, the antigen level was low in clonorchiasis patients. And the sensitivity and specificity of rCsSMPase were 50.0 % (12/24) and 88.4 % (61/69), in sera IgG-ELISA, respectively. It is likely due to the fact that CsSMPase binding to the plasma membrane of biliary epithelium decreases the antigen immune stimulation.
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The self and its resting state in consciousness: An investigation of the vegetative state.
Hum Brain Mapp
PUBLISHED: 02-21-2013
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Recent studies have demonstrated resting-state abnormalities in midline regions in vegetative state/unresponsive wakefulness syndrome and minimally conscious state patients. However, the functional implications of these resting-state abnormalities remain unclear. Recent findings in healthy subjects have revealed a close overlap between the neural substrate of self-referential processing and the resting-state activity in cortical midline regions. As such, we investigated task-related neural activity during active self-referential processing and various measures of resting-state activity in 11 patients with disorders of consciousness (DOC) and 12 healthy control subjects. Overall, the results revealed that DOC patients exhibited task-specific signal changes in anterior and posterior midline regions, including the perigenual anterior cingulate cortex (PACC) and posterior cingulate cortex (PCC). However, the degree of signal change was significantly lower in DOC patients compared with that in healthy subjects. Moreover, reduced signal differentiation in the PACC predicted the degree of consciousness in DOC patients. Importantly, the same midline regions (PACC and PCC) in DOC patients also exhibited severe abnormalities in the measures of resting-state activity, that is functional connectivity and the amplitude of low-frequency fluctuations. Taken together, our results provide the first evidence of neural abnormalities in both the self-referential processing and the resting state in midline regions in DOC patients. This novel finding has important implications for clinical utility and general understanding of the relationship between the self, the resting state, and consciousness. Hum Brain Mapp, 2013. © 2013 Wiley Periodicals, Inc.
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Neural competition as a developmental process: early hemispheric specialization for word processing delays specialization for face processing.
Neuropsychologia
PUBLISHED: 02-01-2013
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Little is known about the impact of learning to read on early neural development for word processing and its collateral effects on neural development in non-word domains. Here, we examined the effect of early exposure to reading on neural responses to both word and face processing in preschool children with the use of the Event Related Potential (ERP) methodology. We specifically linked childrens reading experience (indexed by their sight vocabulary) to two major neural markers: the amplitude differences between the left and right N170 on the bilateral posterior scalp sites and the hemispheric spectrum power differences in the ? band on the same scalp sites. The results showed that the left-lateralization of both the word N170 and the spectrum power in the ? band were significantly positively related to vocabulary. In contrast, vocabulary and the word left-lateralization both had a strong negative direct effect on the face right-lateralization. Also, vocabulary negatively correlated with the right-lateralized face spectrum power in the ? band even after the effects of age and the word spectrum power were partialled out. The present study provides direct evidence regarding the role of reading experience in the neural specialization of word and face processing above and beyond the effect of maturation. The present findings taken together suggest that the neural development of visual word processing competes with that of face processing before the process of neural specialization has been consolidated.
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The carcinogenic liver fluke, Clonorchis sinensis: new assembly, reannotation and analysis of the genome and characterization of tissue transcriptomes.
PLoS ONE
PUBLISHED: 01-30-2013
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Clonorchis sinensis (C. sinensis), an important food-borne parasite that inhabits the intrahepatic bile duct and causes clonorchiasis, is of interest to both the public health field and the scientific research community. To learn more about the migration, parasitism and pathogenesis of C. sinensis at the molecular level, the present study developed an upgraded genomic assembly and annotation by sequencing paired-end and mate-paired libraries. We also performed transcriptome sequence analyses on multiple C. sinensis tissues (sucker, muscle, ovary and testis). Genes encoding molecules involved in responses to stimuli and muscle-related development were abundantly expressed in the oral sucker. Compared with other species, genes encoding molecules that facilitate the recognition and transport of cholesterol were observed in high copy numbers in the genome and were highly expressed in the oral sucker. Genes encoding transporters for fatty acids, glucose, amino acids and oxygen were also highly expressed, along with other molecules involved in metabolizing these substrates. All genes involved in energy metabolism pathways, including the ?-oxidation of fatty acids, the citrate cycle, oxidative phosphorylation, and fumarate reduction, were expressed in the adults. Finally, we also provide valuable insights into the mechanism underlying the process of pathogenesis by characterizing the secretome of C. sinensis. The characterization and elaborate analysis of the upgraded genome and the tissue transcriptomes not only form a detailed and fundamental C. sinensis resource but also provide novel insights into the physiology and pathogenesis of C. sinensis. We anticipate that this work will aid the development of innovative strategies for the prevention and control of clonorchiasis.
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Identification and biochemical characterization of adenylate kinase 1 from Clonorchis sinensis.
Parasitol. Res.
PUBLISHED: 01-29-2013
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Adenylate kinase 1 is responsible for the conversion of AMP into ADP involved in purine metabolism. In the present study, adenylate kinase 1 gene (CsADK1) was isolated from an adult cDNA library of Clonorchis sinensis, and the recombinant protein was expressed in Escherichia coli. Bioinformatics analysis implied that the putative protein contained 197 amino acids, and some residues in conservative binding sites of CsADK1 were substituted. The structure modeling analysis showed that CsADK1 was composed of a core domain, an NMP-binding domain, and a LID domain, which was just a small loop. It demonstrated that CsADK1 was a short isoform of ADKs. Moreover, CsADK1 was identified as an excretory/secretory product by western blot analysis. Real-time quantitative PCR showed that expression level of CsADK1 at the stage of excysted metacercaria was higher than those of adult worm (18.8-folds, P<0.01), metacercariae (1.5-folds, P<0.01), and eggs (5.6-folds, P<0.01). In addition, histochemistry analysis showed that CsADK1 was extensively distributed in metacercariae and in the vitellaria and eggs of adult worms. The Km and Vmax value for substrate ADP were 2.2 mM and 0.9 mM/min, respectively. The optimal temperature and pH value were 37 °C and from 7.5 to 8.0, respectively. The enzyme activity was highly dependent on Mg2+, and the optimal concentration of Mg2+ was 2 mM. However, the enzyme activity was slightly activated by Ca2+, and Mn2+ has no effect on activity. For monovalent ions, activity was highly activated by K+ and NH4+, but slightly by Li+. Taken together, CsADK1 was a metal ion-dependent enzyme involved in purine metabolism, which was important for development and reproduction, and might be a potential candidate for drug target for clonorchiasis.
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Biochemical characterization and functional analysis of fructose-1,6-bisphosphatase from Clonorchis sinensis.
Mol. Biol. Rep.
PUBLISHED: 01-12-2013
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Fructose-1,6-bisphosphatase (FBPase), a key regulatory enzyme of gluconeogenesis, plays an essential role in metabolism and development of most organisms. To the wealth of available knowledge about FBPase from Clonorchis sinensis (CsFBPase), in this study, the characteristics of CsFBPase and its potential role in pathogenesis of clonorchiasis were investigated. The Km value of CsFBPase was calculated to be 41.9 uM. The optimal temperature and pH of CsFBPase were 37 °C and pH 7.5-8.0, respectively. In addition, Mg(2+) or K(+) played a regulatory role in enzyme activity of CsFBPase. Both transcriptional and translational level of CsFBPase were higher in metacercariae (one of larva stages) than those in adult worm (P < 0.05). CsFBPase were observed to extensively express in the intestine, vitellaria and tegument of adult worms and ubiquitously in metacercariae. Moreover, CsFBPase was confirmed as a component of excretory/secretory products. Consequently, the translocation of CsFBPase could be detected on epithelial cells of bile duct in liver of C. sinensis infected rat. Recombinant CsFBPase can specifically bind to the membrane of human hepatic stellate cell line LX-2 by immunofluorescence analysis and stimulated proliferation and activation of LX-2 which demonstrated by Cell Counting Kit-8 and upregulation of key fibrosis-related factors, such as ?-smooth muscle actin, collagen I and collagen III using qRT-PCR. Thus, we predicated that CsFBPase might be a multifunctional enzyme which played as both regulatory enzyme and virulence factor in pathogenesis of C. sinensis infection.
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Left hemiparalexia of Chinese characters: neglect dyslexia or disruption of pathway of visual word form processing?
Brain Struct Funct
PUBLISHED: 01-09-2013
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The objective of the study was to further elucidate the potential mechanisms underlying left hemiparalexia induced by a splenium lesion in corpus callosum. A patient KY, who had infarctions in the splenium and the left ventral medial occipitotemporal area, was examined with neuropsychological tests and fMRI. KY presented left hemiparalexia when he read aloud characters presented in central foveal field tachistoscopically as well as in free-view field. KY also showed left hemialexia for characters in left visual field, while no left hemiparalexia occurred when characters were presented in the right visual field. KY performed poorly in lexical decision tasks. He could judge the directions of Landolts rings gaps in the left or right visual field equally. The result of fMRI indicated that characters in the left visual field could not activate the visual word form area (VWFA), such as left mid-fusiform cortex. All the above neuropsychological and fMRI findings have provided evidences against the assumption of left hemineglect dyslexia. Instead, they support the mechanism of disconnection of visual word form processing pathway. In conclusion, the evidences suggested that the visual information transmission of characters in the left visual field from right occipital area to the VWFA in the left hemisphere was interrupted by the splenium lesion.
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Unimodal and multimodal regions for logographic language processing in left ventral occipitotemporal cortex.
Front Hum Neurosci
PUBLISHED: 01-01-2013
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The human neocortex appears to contain a dedicated visual word form area (VWFA) and an adjacent multimodal (visual/auditory) area. However, these conclusions are based on functional magnetic resonance imaging (fMRI) of alphabetic language processing, languages that have clear grapheme-to-phoneme correspondence (GPC) rules that make it difficult to disassociate visual-specific processing from form-to-sound mapping. In contrast, the Chinese language has no clear GPC rules. Therefore, the current study examined whether native Chinese readers also have the same VWFA and multimodal area. Two cross-modal tasks, phonological retrieval of visual words and orthographic retrieval of auditory words, were adopted. Different task requirements were also applied to explore how different levels of cognitive processing modulate activation of putative VWFA-like and multimodal-like regions. Results showed that the left occipitotemporal sulcus (LOTS) responded exclusively to visual inputs and an adjacent region, the left inferior temporal gyrus (LITG), showed comparable activation for both visual and auditory inputs. Surprisingly, processing levels did not significantly alter activation of these two regions. These findings indicated that there are both unimodal and multimodal word areas for non-alphabetic language reading, and that activity in these two word-specific regions are independent of task demands at the linguistic level.
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IGHV1-69-encoded antibodies expressed in chronic lymphocytic leukemia react with malondialdehyde-acetaldehyde adduct, an immunodominant oxidation-specific epitope.
PLoS ONE
PUBLISHED: 01-01-2013
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The immunoglobulins expressed by chronic lymphocytic leukemia (CLL) B cells are highly restricted, suggesting they are selected for binding either self or foreign antigen. Of the immunoglobulin heavy-chain variable (IGHV) genes expressed in CLL, IGHV1-69 is the most common, and often is expressed with little or no somatic mutation, and restricted IGHD and IGHJ gene usage. We found that antibodies encoded by one particular IGHV1-69 subset, designated CLL69C, with the HCDR3 encoded by the IGHD3-3 gene in reading frame 2 and IGHJ6, specifically bound to oxidation-specific epitopes (OSE), which are products of enhanced lipid peroxidation and a major target of innate natural antibodies. Specifically, CLL69C bound immunodominant OSE adducts termed MAA (malondialdehyde-acetaldehyde-adducts), which are found on apoptotic cells, inflammatory tissues, and atherosclerotic lesions. It also reacted specifically with MAA-specific peptide mimotopes. Light chain shuffling indicated that non-stochastically paired L chain of IGLV3-9 contributes to the antigen binding of CLL69C. A nearly identical CLL69C Ig heavy chain was identified from an MAA-enriched umbilical cord phage displayed Fab library, and a derived Fab with the same HCDR3 rearrangement displayed identical MAA-binding properties. These data support the concept that OSE (MAA-epitopes), which are ubiquitous products of inflammation, may play a role in clonal selection and expansion of CLL B cells.
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Low Divergence of Clonorchis sinensis in China Based on Multilocus Analysis.
PLoS ONE
PUBLISHED: 01-01-2013
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Clonorchis sinensis, an ancient parasite that infects a number of piscivorous mammals, attracts significant public health interest due to zoonotic exposure risks in Asia. The available studies are insufficient to reflect the prevalence, geographic distribution, and intraspecific genetic diversity of C. sinensis in endemic areas. Here, a multilocus analysis based on eight genes (ITS1, act, tub, ef-1a, cox1, cox3, nad4 and nad5 [4.986 kb]) was employed to explore the intra-species genetic construction of C. sinensis in China. Two hundred and fifty-six C. sinensis isolates were obtained from environmental reservoirs from 17 provinces of China. A total of 254 recognized Multilocus Types (MSTs) showed high diversity among these isolates using multilocus analysis. The comparison analysis of nuclear and mitochondrial phylogeny supports separate clusters in a nuclear dendrogram. Genetic differentiation analysis of three clusters (A, B, and C) showed low divergence within populations. Most isolates from clusters B and C are geographically limited to central China, while cluster A is extraordinarily genetically diverse. Further genetic analyses between different geographic distributions, water bodies and hosts support the low population divergence. The latter haplotype analyses were consistent with the phylogenetic and genetic differentiation results. A recombination network based on concatenated sequences showed a concentrated linkage recombination population in cox1, cox3, nad4 and nad5, with spatial structuring in ITS1. Coupled with the history record and archaeological evidence of C. sinensis infection in mummified desiccated feces, these data point to an ancient origin of C. sinensis in China. In conclusion, we present a likely phylogenetic structure of the C. sinensis population in mainland China, highlighting its possible tendency for biogeographic expansion. Meanwhile, ITS1 was found to be an effective marker for tracking C. sinensis infection worldwide. Thus, the present study improves our understanding of the global epidemiology and evolution of C. sinensis.
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Serological diagnosis of clonorchiasis: using a recombinant propeptide of cathepsin L proteinase from Clonorchis sinensis as a candidate antigen.
Parasitol. Res.
PUBLISHED: 10-29-2011
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Clonorchiasis is a common zoonosis in southern and northeastern parts of China, especially in Guangdong, Guangxi and Jilin province. Anti-Clonorchis sinensis antibody detection by enzyme-linked immunosorbent assay (ELISA) has been used for epidemiological surveys of clonorchiasis for its convenience and celerity, but it is still a meaningful work to screen ideal diagnostic antigen or antibody subtype for improvement of diagnostic sensitivity and specificity and for judgement of curative effect. In the present study, recombinant CsCatL-propeptide (rCsCatL-propeptide) was highly expressed in form of inclusion body in Escherichia coli. Soluble rCsCatL-propeptide with high purity were obtained after purification in denatured condition by using His Bind Purification kit, and then renatured. The major antibody subtypes responding to rCsCatL-propeptide in sera from clonorchiasis patients were IgG1 and IgG4, but the level of IgG4 was more predominant (P??0.05). Cross-reactions took place when we detected IgG1 of sera from patients infected with Schistosoma japonicum, Paragonimus westermani, hookworm, Trichuris trichiura and Ascaris lumbricoides with rCsCatL-propeptide, while cross-reactions only took place in sera from patients infected with S. japonicum and P. westermani when we detected specific IgG4. The positive rate of IgG4 detection in sera from clonorchiasis patients with <1,000, 1,000-4,999, 5,000-9,999, and ?10,000 eggs per gram faeces (EPG) were 76.9%, 89.3%, 95.6%, and 100.0%, respectively. The positive rates of serodiagnosis correlated well with the EPG (r?=?0.93). Overall, rCsCatL-propeptide is a valuable candidate for specific IgG4 detection in sera from clonorchiasis patients by the method of ELISA for its few cross-reaction and acceptable sensitivity. In addition, specific IgG4 detection can be used to valuate infected degree and therapeutic effect of clonorchiasis patients.
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Sodium instead of potassium and chloride is an important macronutrient to improve leaf succulence and shoot development for halophyte Sesuvium portulacastrum.
Plant Physiol. Biochem.
PUBLISHED: 08-21-2011
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Soil salinity is contributed largely by NaCl but some halophytes such as Sesuvium portulacastrum have evolved to adapt salinity environment and demonstrate optimal development under moderate salinity. To elucidate the detail mechanisms of the great salt tolerance and determine the respective contributions of Na(+), K(+) and Cl(-) on the development of S. portulacastrum, morphological and physiological analysis were performed using plants supplied with 200 mM of different ions including cations (Na(+), K(+), Li(+)) and anions (Cl(-), NO(3)(-), Ac(-)) respectively. The results revealed that the salt-treated plants accumulated large amounts of sodium in both leaf and stem. There was a greater shoot growth in presence of external Na(+) compared to K(+) and Cl(-). Na(+) was found more effective than K(+) and Cl(-) in cell expansion, leaf succulence, and shoot development. Flame emission and X-Ray microanalysis revealed the relative Na(+) content was much higher than K(+) and Cl(-) in both leaf and stem of well developed S. portulacastrum, leading to a higher Na(+)/K(+) ratio. The effects of different ions on the development of S. portulacastrum were listed as the following: Na(+) > NO(3)(-) > CK > Cl(-) > K(+) > Ac(-) > Li(+). These results demonstrated NaCl toxicity is attributable largely to the effect of Cl(-) but rarely to Na(+), and thus sodium is concluded as a more important macronutrient than potassium and chloride for improving leaf succulence and shoot development of halophyte S. portulacastrum.
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Robust and task-independent spatial profile of the visual word form activation in fusiform cortex.
PLoS ONE
PUBLISHED: 08-17-2011
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Written language represents a special category of visual information. There is strong evidence for the existence of a cortical region in ventral occipitotemporal cortex for processing the visual form of written words. However, due to inconsistent findings obtained with different tasks, the level of specialization and selectivity of this so called visual word form area (VWFA) remains debated. In this study, we examined category selectivity for Chinese characters, a non-alphabetic script, in native Chinese readers. In contrast to traditional approaches of examining response levels in a restricted predefined region of interest (ROI), a detailed distribution of the BOLD signal across the mid-fusiform cortical surface and the spatial patterns of responses to Chinese characters were obtained. Results show that a region tuned for Chinese characters could be consistently found in the lateral part of the left fusiform gyrus in Chinese readers, and this spatial pattern of selectivity for written words was not influenced by top-down tasks such as phonological or semantic modulations. These results provide strong support for the robust spatial coding of category selective response in the mid-fusiform cortex, and demonstrate the utility of the spatial distribution analysis as a more meaningful approach to examine functional magnetic resonance imaging (fMRI) data.
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Molecular expression and characterization of a novel protein phosphatase 2A gene from Clonorchis sinensis.
Parasitol. Res.
PUBLISHED: 08-01-2011
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Reversible phosphorylation of proteins is a critical mechanism involved in physiological function of organisms, including Clonorchis sinensis. In the present study, One cDNA clone encoding protein phosphatase 2A (CsPP2A) was isolated from a C. sinensis adult cDNA plasmid library. The open reading frame of the novel gene contains 924 bp and encoded a putative protein of 307 amino acids. A similarity analysis showed high homology with Schistosoma japonicum (76.3%) and Homo sapiens (84.4%), respectively. Recombinant CsPP2A (rCsPP2A) was expressed and purified from Escherichia coli BL21 using pET28a (+) as an expression vector. CsPP2A showed higher transcript level in adult worm but excysted metacercaria (P > 0.05), metacercaria (P < 0.05), and egg (P < 0.05) using real-time RT-PCR. Western blotting analysis showed that rCsPP2A could be identified by anti-rCsPP2A rat serum, C. sinensis-infected rat serum, and the serum from the rats immunized with excretory-secretory products of C. sinensis. Immunohistochemical assay showed that CsPP2A was deposited at the egg, the vitellarium of adult worm, and the excretory bladder of metacercaria. Collectively, the results of this study suggested that CsPP2A may be involved in the development of adult and metacercaria of C. sinensis.
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Multiple compartmentalization of sodium conferred salt tolerance in Salicornia europaea.
Plant Physiol. Biochem.
PUBLISHED: 07-20-2011
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Euhalophyte Salicornia europaea L., one of the most salt-tolerant plant species in the world, can tolerate more than 1000 mM NaCl. To study the salt tolerance mechanism of this plant, the effects of different NaCl concentrations on plant growth, as well as Na(+) accumulation and distribution at organ, tissue, and subcellular levels, were investigated. Optimal growth and an improved photosynthetic rate were observed with the plant treated with 200-400 mM NaCl. The Na(+) content in the shoots was considerably higher than that in the roots of S. europaea. The Na(+) in S. europaea cells may act as an effective osmotic adjuster to maintain cell turgor, promoting photosynthetic competence and plant growth. The results from the SEM-X-ray and TEM-X-ray microanalyses demonstrate that Na(+) was compartmentalized predominantly into the cell vacuoles of shoot endodermis tissues. Accordingly, the transcript amounts of SeNHX1, SeVHA-A, and SeVP1 increased significantly with increased NaCl concentration, suggesting their important roles in Na(+) sequestration into the vacuoles. Therefore, a multiple sodium compartmentalization mechanism is proposed to enhance further the salt tolerance of S. europaea.
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Gene cloning, expression, and localization of antigen 5 in the life cycle of Echinococcus granulosus.
Parasitol. Res.
PUBLISHED: 06-30-2011
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Antigen 5 (Ag5) has been identified as a dominant component of cyst fluid of Echinococcus granulosus and is considered as a member of serine proteases family, which in other helminth, plays an important role in the egg hatch and larva invasion. However, whether Ag5 is expressed and secreted in all life stages is unknown. In this study, according to the sequence in GenBank, we cloned and sequenced the open reading frame (ORF) of Ag5 gene from the protoscolices of E. granulosus isolated from the sheep in Qinhai Province of China, and found several substitutions and a base insert and deletion in a short region near the stop code, leading to a frameshift mutation which is conserved with the homologue of other cestode. The ORF is 1,455 bp in length, encoding 484 amino acids with a secretory signal peptide. Bioinformatics analysis predicted several phosphorylation and myristoylation sites and a N-glycosylation site and a species-specific linear B epitope in the protein. The ORF was cloned into the plasmid pET28a(+) vector and expressed in Escherichia coli . The recombinant protein was purified by affinity chromatography. Anti-rEgAg5 antiserum was prepared in rats and used to analyze the localization of Ag5 in protoscolex and adult worm by immunofluorescence technique. Results demonstrated that the Ag5 is strongly expressed in the tegument of protoscolex and the embryonic membrane of egg and surface of oncosphere; meanwhile, it is also weakly expressed in tegument of the adult. This study showed that Ag5 is expressed in all stages of life cycle, secreted from the surface of the worm and may be anchored in membrane by its myristoylation sites; these characteristics make it a candidate antigen for diagnosis and vaccine for both intermediate and definitive hosts.
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PET Imaging and Biodistribution of Silicon Quantum Dots in Mice.
ACS Med Chem Lett
PUBLISHED: 05-07-2011
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Investigation of nanomaterial disposition and fate in the body is critical before such material can be translated into clinical application. Herein a new macrocyclic ligand-(64)Cu(2+) complex was synthesized and used to label dextran-coated silicon quantum dots (QD), with an average hydrodynamic diameter of 15.1 ± 7.6 nm. The chelate showed exceptional stability, demonstrated by no loss radiolabel under a ligand competition reaction with EDTA. The QDs biodistribution in mice was quantitatively evaluated by in vivo positron emission tomography (PET) imaging and ex vivo gamma counting. Results showed that they were excreted via renal filtration shortly postinjection and also accumulated in the liver.
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Minimal effects on human memory following long-term living at moderate altitude.
High Alt. Med. Biol.
PUBLISHED: 04-02-2011
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A number of studies describe memory deficits at extremely high altitudes. However, little is known about the effect of long-term living at moderate altitude (MA). The subjects for this study were 52 college students originally from sea level (SL), but studying at a MA of 2260?m over a 7-month period, with a return to SL for 30 days in the middle of the period. Fifty-two matched college students who stayed at SL all the time were the control group. The neuropsychological battery of assessments included the Chinese revised version of Wechsler Memory Scale tests (WMS-CR), verbal and spatial two-back working memory tests, long-term explicit memory (word recall and recognition of words, faces, and pictures) tests, Rey Auditory Verbal Learning test, Rey-Osterrieth Complex Figure (ROCF) test, degraded picture naming test, and the Serial Reaction Time Test. We found that the MA subjects showed significantly poorer performances than SL controls only in short-term visual construction assessed in the visual reproduction test from WMS-CR and in the ROCF immediate test. There were no significant differences in all other tasks between the MA group and SL group. These findings suggest that long-term hypoxic exposure at moderate altitude has minimal effects on human memory.
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The protective effect of the recombinant 53-kDa protein of Trichinella spiralis on experimental colitis in mice.
Dig. Dis. Sci.
PUBLISHED: 03-21-2011
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Helminth infection has been proven to reduce the severity of experimental inflammatory bowel disease (IBD). The excretory-secretory proteins of helminths play an important role in the process of immunomodulation.
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Cerebrovascular reactivity among native-raised high altitude residents: an fMRI study.
BMC Neurosci
PUBLISHED: 03-11-2011
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The impact of long term residence on high altitude (HA) on human brain has raised concern among researchers in recent years. This study investigated the cerebrovascular reactivity among native-born high altitude (HA) residents as compared to native sea level (SL) residents. The two groups were matched on the ancestral line, ages, gender ratios, and education levels. A visual cue guided maximum inspiration task with brief breath holding was performed by all the subjects while Blood-Oxygenation-Level-Dependent (BOLD) functional Magnetic Resonance Imaging (fMRI) data were acquired from them.
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Altered negative unconscious processing in major depressive disorder: an exploratory neuropsychological study.
PLoS ONE
PUBLISHED: 03-10-2011
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Major depressive disorder (MDD) has been characterized by abnormalities in emotional processing. However, what remains unclear is whether MDD also shows deficits in the unconscious processing of either positive or negative emotions. We conducted a psychological study in healthy and MDD subjects to investigate unconscious emotion processing and its valence-specific alterations in MDD patients.
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Dissociation between anterior and posterior cortical regions during self-specificity and familiarity: a combined fMRI-meta-analytic study.
Hum Brain Mapp
PUBLISHED: 03-09-2011
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The familiarity to the subject of any potential stimuli presents one of the major difficulties for the investigation of the self; the separation of effects resulting from familiarity from self-effects being extremely problematic. The aim of this study was thus to investigate the neural distinction between self and familiarity by combining two sets of fMRI data with a meta-analysis. In the first fMRI experiment, regions responding to self/familiarity were investigated using the subjects own name and names of familiar others. These effects were confirmed and extended in a second fMRI experiment using the subjects own name and a strangers name, as spoken by familiar and unfamiliar voices. Finally, a meta-analysis of self- and familiarity-related studies was conducted. Neural activity in the anterior brain regions, such as the anterior cingulate (ACC) and anterior insula (AI), was found to be specific for self-specific stimuli. In contrast, posterior brain regions, such as the posterior cingulate, were activated by familiar stimuli. Finally, the distinction between anterior and posterior regions for self and familiarity was confirmed by meta-analytic data. This study demonstrates a clear anterior-posterior cortical partition between self-specificity and familiarity.
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Chinese and Korean characters engage the same visual word form area in proficient early Chinese-Korean bilinguals.
PLoS ONE
PUBLISHED: 03-02-2011
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A number of recent studies consistently show an area, known as the visual word form area (VWFA), in the left fusiform gyrus that is selectively responsive for visual words in alphabetic scripts as well as in logographic scripts, such as Chinese characters. However, given the large difference between Chinese characters and alphabetic scripts in terms of their orthographic rules, it is not clear at a fine spatial scale, whether Chinese characters engage the same VWFA in the occipito-temporal cortex as alphabetic scripts. We specifically compared Chinese with Korean script, with Korean script serving as a good example of alphabetic writing system, but matched to Chinese in the overall square shape. Sixteen proficient early Chinese-Korean bilinguals took part in the fMRI experiment. Four types of stimuli (Chinese characters, Korean characters, line drawings and unfamiliar Chinese faces) were presented in a block-design paradigm. By contrasting characters (Chinese or Korean) to faces, presumed VWFAs could be identified for both Chinese and Korean characters in the left occipito-temporal sulcus in each subject. The location of peak response point in these two VWFAs were essentially the same. Further analysis revealed a substantial overlap between the VWFA identified for Chinese and that for Korean. At the group level, there was no significant difference in amplitude of response to Chinese and Korean characters. Spatial patterns of response to Chinese and Korean are similar. In addition to confirming that there is an area in the left occipito-temporal cortex that selectively responds to scripts in both Korean and Chinese in early Chinese-Korean bilinguals, our results show that these two scripts engage essentially the same VWFA, even at the level of fine spatial patterns of activation across voxels. These results suggest that similar populations of neurons are engaged in processing the different scripts within the same VWFA in early bilinguals.
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Proteomic analysis of excretory secretory products from Clonorchis sinensis adult worms: molecular characterization and serological reactivity of a excretory-secretory antigen-fructose-1,6-bisphosphatase.
Parasitol. Res.
PUBLISHED: 02-23-2011
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Clonorchis sinensis is a food-borne zoonotic parasite that resides in bile ducts and causes clonorchiasis, which may result in cholelithiasis, cholecystitis, hepatic fibrosis, and liver tumors. Although total excretory secretory products (ESP) of C. sinensis adults induce hepatic fibrosis in vivo in rats, the causative mechanism is not well understood. To study components of the ESP, C. sinensis culture medium was collected and analyzed using shotgun LC-MS/MS. We identified a total of 110 proteins, including glycometabolic enzymes (such as fructose-1,6-bisphosphatase (FBPase) and enolase), detoxification enzymes (such as glutamate dehydrogenase, dihydrolipoamide dehydrogenase and cathepsin B endopeptidase), and a number of RAB family proteins. To identify a potential causative agent for hepatic fibrosis, we expressed and purified a recombinant FBPase, a 1,041-bp gene product that encodes a 41.7-kDa protein with prototypical FBPase domains and that can form a tetramer with a molecular mass of 166.8 kDa. In addition, we found that FBPase is an antigen present in the ESP and in circulation. Immunofluorescence showed that FBPase localizes to the intestinal cecum and vitellarium in C. sinensis adults. Our results describe the components of the excretory secretory products from C. sinensis adult worms and suggest that FBPase may be an important antigen present in the ESP of C. sinensis and may lay the foundation for additional studies on the development of clonorchiasis-associated hepatic fibrosis.
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Clonorchis sinensis enolase: identification and biochemical characterization of a glycolytic enzyme from excretory/secretory products.
Mol. Biochem. Parasitol.
PUBLISHED: 02-21-2011
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Enolase plays a key role in energy metabolism and development of most organisms. We isolated a gene encoding enolase from Clonorchis sinensis (C. sinensis) adult cDNA library and expressed the recombinant protein in Escherichia coli. C. sinensis enolase (Csenolase) was identified as both an excretory/secretory product and a tegumental component of C. sinensis by western blot analysis. The transcriptional level of Csenolase was examined at adult worm, metacercaria, cercaria and egg of C. sinensis, and results showed that Csenolase is transcribed at the four life stages of C. sinensis while showing a significant higher expression level at the stage of adult worm. Immunohistochemical localization indicated that Csenolase was specifically deposited on the tegument of adult worm and cyst wall of metacercaria. Ligand blot assay revealed a specific characteristic of dose-dependent plasminogen-binding activity of Csenolase and kinetic parameters were explored using 2-phospho-D-glycerate (2-PGA) as the primary substrate by monitoring the conversion of nicotinamide-adenine dinucleotide (NADH) into nicotinamide adenine dinucleotide (NAD). In addition, Csenolase exhibited active enzyme activity in catalytic reactions while the anti-Csenolase serum inhibited the enzyme activity. In vitro incubation experiments revealed that Csenolase might play key roles in the growth of the parasites. In conclusion, Csenolase is an important glycolytic enzyme required for the development of C. sinensis, and may be a potential vaccine candidate and drug target against C. sinensis infection.
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The full-length transcripts and promoter analysis of intergenic microRNAs in Drosophila melanogaster.
Genomics
PUBLISHED: 02-04-2011
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MicroRNA (miRNA) transcription is still not well understood until now. To increase the miRNA abundance, we stimulated miRNA transcription with CuSO(4) and knocked down Drosha enzyme using dsRNA in Drosophila S2 cells. The full length transcripts of bantam, miR-276a and miR-277, the 5-end of miR-8, the 3-end of miR-2b and miR-10 were obtained. We also conducted a series of miRNA promoter analysis to prove the reliability of RACE results. Luciferase-reporter assays proved that both bantam and miR-276a promoters successfully drove the expressions of downstream luciferase genes. The promoter activities were impaired by introducing one or multiple mutations at predicted transcription factor binding sites. Chromatin immunoprecipitation analysis confirmed that hypophosphorylated RNA polymerase II and transcription factor c-Myc physically bind at miRNA promoter. RNA interference of transcription factors Mad and Prd led to down-expression of bantam, miR-277 and miR-2b but not miR-276a, whereas RNAi of Dorsal had the opposite effect.
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Adaptive influence of long term high altitude residence on spatial working memory: an fMRI study.
Brain Cogn
PUBLISHED: 02-02-2011
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With an increasing population living at a high altitude (HA), the impact of HA residence on human cognitive function has raised concerns. We recruited two groups of college students with one group born and grew up at HA until early adulthood and the control group born and grew up at near sea level (SL); the two groups were matched at age, gender ratio, educational level, the ancestral lines, and peripheral physiology (especially the hemoglobin concentration). A 2-back spatial working memory task was performed by each subject in the scanner while fMRI data were acquired. Compared to the SL control group, the HA group showed equal response accuracy, with more variance in reaction time and a larger average value. fMRI data indicated that both groups showed common activation patterns in the neural pathway typically associated with working memory. The HA group had greater activation at the left pyramis, the left superior temporal gyrus and less activation at the left middle occipital gyrus. Significant correlations were found within each group between the reaction times and BOLD signal change amplitudes at the frontal cortex and the precentral cortex.
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Molecular cloning, characterization, and immunolocalization of two lactate dehydrogenase homologous genes from Taenia solium.
Parasitol. Res.
PUBLISHED: 02-02-2011
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Two novel genes encoding lactate dehydrogenase A (LDHA) and B (LDHB) homologues, respectively, were identified from the cDNA libraries of adult Taenia solium (T. solium). The two deduced amino acid sequences both show more than 50% identity to the homologues for Danio rerio, Xenopus laevis, Schistosoma japonicum, Sus scrofa, Homo sapiens, et al. The identity of the amino acid sequence between TsLDHA and TsLDHB is 57.4%, and that of the nucleotide sequence is 61.5%. Recombinant TsLDHA homologue (rTsLDHA) and TsLDHB homologue (rTsLDHB) were expressed in Escherichia coli BL21/DE3 and purified. Though there were some differences in the sequence, the two LDH isozyme homologues show similarity in the conserved LDH domain, topological structure, primary immunological traits, localization on the tegument of T. solium adult, and partial physicochemical properties. The linear B-cell epitope analysis of TsLDHA and TsLDHB discovered a TsLDHA specific epitope. The purified rTsLDHA and rTsLDHB could be recognized by rat immuno-sera, serum from swine, or a patient infected with T. solium, respectively, but Western blot analysis showed cross-reactions, not only between these two LDH members but also with other common human tapeworms or helminths. The results suggested that the two LDH homologues are similar in the characteristics of LDH family, and they are not specific antigens for immunodiagnosis.
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The draft genome of the carcinogenic human liver fluke Clonorchis sinensis.
Genome Biol.
PUBLISHED: 01-31-2011
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Clonorchis sinensis is a carcinogenic human liver fluke that is widespread in Asian countries. Increasing infection rates of this neglected tropical disease are leading to negative economic and public health consequences in affected regions. Experimental and epidemiological studies have shown a strong association between the incidence of cholangiocarcinoma and the infection rate of C. sinensis. To aid research into this organism, we have sequenced its genome.
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Oxidation-specific epitopes are danger-associated molecular patterns recognized by pattern recognition receptors of innate immunity.
Circ. Res.
PUBLISHED: 01-22-2011
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Oxidation reactions are vital parts of metabolism and signal transduction. However, they also produce reactive oxygen species, which damage lipids, proteins and DNA, generating "oxidation-specific" epitopes. In this review, we discuss the hypothesis that such common oxidation-specific epitopes are a major target of innate immunity, recognized by a variety of "pattern recognition receptors" (PRRs). By analogy with microbial "pathogen-associated molecular patterns" (PAMPs), we postulate that host-derived, oxidation-specific epitopes can be considered to represent "danger (or damage)-associated molecular patterns" (DAMPs). We also argue that oxidation-specific epitopes present on apoptotic cells and their cellular debris provided the primary evolutionary pressure for the selection of such PRRs. Furthermore, because many PAMPs on microbes share molecular identity and/or mimicry with oxidation-specific epitopes, such PAMPs provide a strong secondary selecting pressure for the same set of oxidation-specific PRRs as well. Because lipid peroxidation is ubiquitous and a major component of the inflammatory state associated with atherosclerosis, the understanding that oxidation-specific epitopes are DAMPs, and thus the target of multiple arcs of innate immunity, provides novel insights into the pathogenesis of atherosclerosis. As examples, we show that both cellular and soluble PRRs, such as CD36, toll-like receptor-4, natural antibodies, and C-reactive protein recognize common oxidation-specific DAMPs, such as oxidized phospholipids and oxidized cholesteryl esters, and mediate a variety of immune responses, from expression of proinflammatory genes to excessive intracellular lipoprotein accumulation to atheroprotective humoral immunity. These insights may lead to improved understanding of inflammation and atherogenesis and suggest new approaches to diagnosis and therapy.
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Molecular cloning, expression, and characterization of cyclophilin A from Clonorchis sinensis.
Parasitol. Res.
PUBLISHED: 01-18-2011
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This study described the recognization, cloning, and recombinant expression of cyclophilin A-like gene from Clonorchis sinensis adult complementary DNA library (CsCyPA) and its expression and secretion in adult. Western blotting demonstrated the recombinant CsCyPA could be recognized by sera of clonorchiasis patients and a sole protein of the same size in the excretory-secretory antigens of in vitro cultured adult could be recognized by antiserum raised against the recombinant CsCyPA. Immunohistochemistry demonstrated that the CsCyPA was secreted in scattered vesicles from subtegumental parenchyma cells to the surface of tegument and mainly released from the tegument. ELISA showed the serum levels of IgG against CsCyPA in clonorchiasis patients negatively correlated with worm loads. This study suggested that C. sinensis adult in biliary ducts could release CsCyPA without signal peptide through nonclassical secretory pathway into the liver and might play a role in inflammation and biliary epithelium proliferation and adenomatoid hyperplasia.
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Impaired directed forgetting in abstinent heroin addicts.
Memory
PUBLISHED: 01-18-2011
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Drug-related memories persist long into abstinence and are potent elicitors of drug craving and relapse. We report two experiments examining whether heroin-dependent individuals are impaired in intentionally suppressing drug-related memories. Experiment 1 adopted the Item paradigm where addicts and healthy controls were presented with a list of words each followed by a remember or forget cue. Experiment 2 adopted the List paradigm where they studied one list of items and were then split into a remember group and a forget group. Both groups studied a second list, except that the forget group was told to forget the first list. Compared with controls, addicts showed a reduced directed forgetting effect in the Item method and a total absence of one measure of directed forgetting in the List method (List 2 benefits). Results indicate that heroin addicts are impaired in directed forgetting and that the deficits are likely associated with memory encoding as opposed to retrieval. Possible problems include reduced ability in actively suppressing/stopping encoding of irrelevant information into memory or inability in changing/resetting encoding strategies. In neither experiment did the addicts show any differential directed forgetting effects between drug-related words and neutral words, indicating the generic nature of their intentional forgetting deficits.
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Functional foveal splitting: evidence from neuropsychological and multimodal MRI investigations in a Chinese patient with a splenium lesion.
PLoS ONE
PUBLISHED: 01-15-2011
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It remains controversial and hotly debated whether foveal information is double-projected to both hemispheres or split at the midline between the two hemispheres. We investigated this issue in a unique patient with lesions in the splenium of the corpus callosum and the left medial occipitotemporal region, through a series of neuropsychological tests and multimodal MRI scans. Behavioral experiments showed that (1) the patient had difficulties in reading simple and compound Chinese characters when they were presented in the foveal but left to the fixation, (2) he failed to recognize the left component of compound characters when the compound characters were presented in the central foveal field, (3) his judgments of the gender of centrally presented chimeric faces were exclusively based on the left half-face and he was unaware that the faces were chimeric. Functional MRI data showed that Chinese characters, only when presented in the right foveal field but not in the left foveal field, activated a region in the left occipitotemporal sulcus in the mid-fusiform, which is recognized as visual word form area. Together with existing evidence in the literature, results of the current study suggest that the representation of foveal stimuli is functionally split at object processing levels.
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Left-lateralized early neurophysiological response for Chinese characters in young primary school children.
Neurosci. Lett.
PUBLISHED: 01-14-2011
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Adult readers consistently show an enhanced early event-related potential (ERP) response, N170, for visual words compared with other stimuli at left posterior electrodes. Developmental studies with words in alphabetic languages showed that this neurophysiological specialization for print develops rapidly from 6 to 10-years of age and becomes established around 10-11 years of age. Here we report for the first time the development of the word-related N170 in Chinese children learning to read Chinese, a logographic writing system radically different from alphabetic scripts in visual and linguistic features. We recorded ERP responses elicited by Chinese characters and line drawings of common objects in three groups of primary school children at 7, 9, and 11 years of age as well as college students. Results showed that the amplitude of N170 evoked by Chinese characters in the 7-year-old group was significantly larger than that in the 11-year-old group and the adult readers. Remarkably, all four age groups - even the youngest group - showed an increased and left-lateralized N170 response for Chinese characters, as compared with line drawings, suggesting that a relatively specialized mechanism for processing Chinese characters is already emergent by as early as 7 years of age. Our results, combined with studies of non-Chinese child readers suggest that the developmental pattern of word-related N170 is highly similar across different scripts, possibly reflecting increased visual processing expertise that children acquire through everyday reading.
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Reward and motivation systems: a brain mapping study of early-stage intense romantic love in Chinese participants.
Hum Brain Mapp
PUBLISHED: 01-14-2011
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Early-stage romantic love has been studied previously in the United States and United Kingdom (Aron et al. [2005]: J Neurophysiol 94:327–337; Bartels and Zeki [2000]: Neuroreport 11:3829–3834; Ortigue et al. [2007]: J Cogn Neurosci 19:1218–1230), revealing activation in the reward and motivation systems of the brain. In this study, we asked what systems are activated for early-stage romantic love in Easterners, specifically Chinese participants? Are these activations affected by individual differences within a cultural context of Traditionality and Modernity? Also, are these brain activations correlated with later satisfaction in the relationship? In Beijing, we used the same procedure used by Aron et al. (Aron et al. [2005]: J Neurophysiol 94:327–337). The stimuli for 18 Chinese participants were a picture of the face of their beloved, the face of a familiar acquaintance, and a countback task. We found significant activations specific to the beloved in the reward and motivation systems, particularly, the ventral tegmental area and the caudate. The mid-orbitofrontal cortex and cerebellum were also activated, whereas amygdala, medial orbitofrontal, and medial accumbens activity were decreased relative to the familiar acquaintance. Self-reported Traditionality and Modernity scores were each positively correlated with activity in the nucleus accumbens, although in different regions and sides of the brain. Activity in the subgenual area and the superior frontal gyrus was associated with higher relationship happiness at 18-month follow-up. Our results show that midbrain dopamine-rich reward/motivation systems were activated by early-stage romantic love in Chinese participants, as found by other studies. Neural activity was associated with Traditionality and Modernity attitudes as well as with later relationship happiness for Chinese participants.
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Thermal denaturation produced degenerative proteins and interfered with MS for proteins dissolved in lysis buffer in proteomic analysis.
Electrophoresis
PUBLISHED: 01-11-2011
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In 1-DE, proteins were traditionally mixed with the standard Laemmli buffer and boiled for several minutes. Recently, proteins dissolved in lysis buffer were used to produce better-resolved 2-DE gels, but thermal denaturation procedure still remained in some proteomic analysis. To determine the detailed effects of thermal denaturation on SDS-PAGE and MS, both 1-DE and 2-DE were performed using proteins heated at 100°C for different periods of time, and 17 protein bands/spots were positively identified by MALDI TOF/TOF MS/MS. Protein profiles on both 1-DE and 2-DE gels changed obviously and more polydisperse bands/spots were observed with increased heating time for over-heated samples. Based on these observations, an alternative protein marker-producing method was designed by directly dissolving protein standards without BSA into lysis buffer. This new kind of protein marker could be stored at room temperature for a long time, thus was more convenient for using and shipping. The identification of 17 proteins via MS and comparison of their identities revealed MASCOT-searched scores, number of both matched peptides, total searched peptides and sequence coverage became progressively lower with increasing denaturation intensity, probably due to the interference of thermal denaturation on trypsin cleavage efficiency and produced redundant modified peptides. Therefore, it was concluded that thermal denaturation not only changed the protein profiles and produced more polydisperse protein bands/spots, but also heavily interfered with the subsequent MS analysis, hence not recommended in future proteomic analysis for proteins dissolved in lysis buffer.
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Appetite at high altitude: an fMRI study on the impact of prolonged high-altitude residence on gustatory neural processing.
Exp Brain Res
PUBLISHED: 11-11-2010
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Regulation of food intake is very important for health. It has been reported that people have decreased appetite at high altitude (HA). The current study recruited long-term HA residents to participate in an fMRI experiment which involved food craving. Result shows that the HA group showed decreased activation in the neural circuit for food craving, accompanied by decreased activation in regions for cognitive control and increased activation in regions for emotional processing. Such results also reflect the decreased gray matter volume and the hypometabolism mechanism under prolonged hypoxia stress at HA.
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Prolonged high-altitude residence impacts verbal working memory: an fMRI study.
Exp Brain Res
PUBLISHED: 10-19-2010
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Oxygen is critical to normal brain functioning and development. In high altitude where the oxygen concentration and pressure are very low, human cognitive capability such as working memory has been found to be jeopardized. Such effect might persist with long-term high-altitude residence. The current study investigated the verbal working memory of 28 high-altitude residents with blood level oxygen dependent (BOLD) functional magnetic resonance imaging (fMRI), in contrast with that of the 30 sea level residents. All of the subjects were healthy college students, matched on their age, gender ratio and social-economic status; they also did not show any difference on their hemoglobin level. The high-altitude subjects showed longer reaction time and decreased response accuracy in behavioral performance. Both groups showed activation in the typical regions associated with the 2-back verbal working memory task, and the behavioral performance of both groups showed significant correlations with the BOLD signal change amplitude and Granger causality values (as a measure of the interregional effective connectivity) between these regions. With group comparison statistics, the high-altitude subjects showed decreased activation at the inferior and middle frontal gyrus, the middle occipital and the lingual gyrus, the pyramis of vermis, as well as the thalamus. In conclusion, the current study revealed impairment in verbal working memory among high-altitude residents, which might be associated with the impact of prolonged chronic hypoxia exposure on the brain functionality.
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Structures of the human GTPase MMAA and vitamin B12-dependent methylmalonyl-CoA mutase and insight into their complex formation.
J. Biol. Chem.
PUBLISHED: 09-28-2010
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Vitamin B(12) (cobalamin, Cbl) is essential to the function of two human enzymes, methionine synthase (MS) and methylmalonyl-CoA mutase (MUT). The conversion of dietary Cbl to its cofactor forms, methyl-Cbl (MeCbl) for MS and adenosyl-Cbl (AdoCbl) for MUT, located in the cytosol and mitochondria, respectively, requires a complex pathway of intracellular processing and trafficking. One of the processing proteins, MMAA (methylmalonic aciduria type A), is implicated in the mitochondrial assembly of AdoCbl into MUT and is defective in children from the cblA complementation group of cobalamin disorders. To characterize the functional interplay between MMAA and MUT, we have crystallized human MMAA in the GDP-bound form and human MUT in the apo, holo, and substrate-bound ternary forms. Structures of both proteins reveal highly conserved domain architecture and catalytic machinery for ligand binding, yet they show substantially different dimeric assembly and interaction, compared with their bacterial counterparts. We show that MMAA exhibits GTPase activity that is modulated by MUT and that the two proteins interact in vitro and in vivo. Formation of a stable MMAA-MUT complex is nucleotide-selective for MMAA (GMPPNP over GDP) and apoenzyme-dependent for MUT. The physiological importance of this interaction is highlighted by a recently identified homoallelic patient mutation of MMAA, G188R, which, we show, retains basal GTPase activity but has abrogated interaction. Together, our data point to a gatekeeping role for MMAA by favoring complex formation with MUT apoenzyme for AdoCbl assembly and releasing the AdoCbl-loaded holoenzyme from the complex, in a GTP-dependent manner.
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Implicit reading in Chinese pure alexia.
Brain Lang
PUBLISHED: 07-30-2010
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A number of recent studies have shown that some patients with pure alexia display evidence of implicit access to lexical and semantic information about words that they cannot read explicitly. This phenomenon has not been investigated systematically in Chinese patients. We report here a case study of a Chinese patient who met the criteria for pure alexia and had lesions in the left occipitotemporal region and the splenium of the corpus callosum. His explicit and implicit reading was evaluated with various stimuli in a number of tasks. We found that despite his severe impairment in overt reading and the definition of any characters, his performance was well above chance in various implicit tasks. His accuracy with respect to lexical decisions was so high that his performance was almost normal. These findings provide unequivocal evidence for the existence of implicit reading in Chinese patients with pure alexia and further support the involvement of the right hemisphere.
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Cloning and expression of 21.1-kDa tegumental protein of Clonorchis sinensis and human antibody response to it as a trematode-nematode pan-specific serodiagnosis antigen.
Parasitol. Res.
PUBLISHED: 06-30-2010
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A complete cDNA encoding a 21.1-kDa tegumental protein (CsTP21.1) was recognized from Clonorchis sinensis adult full-length cDNA plasmid library by bioinformatics analysis. Recombinant CsTP21.1 was highly expressed in Escherichia coli, purified by affinity chromatography, and identified by Western blotting. Immunohistochemistry demonstrated that CsTP21.1 is localized in the tegument of the adult worm. The rCsTP21.1-specific IgG1, IgG2, and IgG4 subclasses could be detected in the sera of clonorchiasis patients by ELISA, but their sensitivity was much lower than that of total IgG. The sensitivity and specificity of IgG in 66 serum samples of clonorchiasis patients were 100% and 95.5%, and the sensitivity was independent of worm loads; the cross-reaction rates in 86, 24, and 31 serum samples from patients infected with Fasciola hepatica, Schistosoma japonicum, and nematode were 98.8%, 83.3%, 93.3%, respectively, whereas no cross-reactions with Toxoplasma gondii and sparganum. This study demonstrated that CsTP21.1 is a trematode-nematode pan-specific antigen that is valuable in the development of a universal immunodiagnostic kit for human trematode and nematode infections.
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Molecular characterization and expression of the MYND-ZF gene from Clonorchis sinensis.
Parasitol. Res.
PUBLISHED: 06-26-2010
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The MYND-type zinc finger protein (MYND-ZF) is a large group of proteins containing the MYND domain which play an important role in protein-protein interactions. A cDNA clone encoding a novel MYND-ZF was isolated and identified from a Clonorchis sinensis (C. sinensis) adult cDNA library. The open reading frame of this novel cDNA sequence contains 1,440 base pairs with a putative protein of 479 amino acids showing a high homology with the MYND-ZF identified from other species. Recombinant CsMYND-ZF was expressed and purified from Escherichia coli BL21 (DE3). CsMYND-ZF transcripts were detected in the cDNA of adult worms and metacercariae but not in eggs of C. sinensis. Immunohistochemistry results revealed that CsMYND-ZF was deposited at the tegument of adult worms and metacercariae C. sinensis using anti-recombinant CsMYND-ZF serum. These findings may contribute to the development of a reliable diagnostic method.
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Structural impact of human and Escherichia coli biotin carboxyl carrier proteins on biotin attachment.
Biochemistry
PUBLISHED: 05-07-2010
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Holocarboxylase synthetase (HCS, human) and BirA (Escherichia coli) are biotin protein ligases that catalyze the ATP-dependent attachment of biotin to apocarboxylases. Biotin attachment occurs on a highly conserved lysine residue within a consensus sequence (Ala/Val-Met-Lys-Met) that is found in carboxylases in most organisms. Numerous studies have indicated that HCS and BirA, as well as biotin protein ligases from other organisms, can attach biotin to apocarboxylases from different organisms, indicating that the mechanism of biotin attachment is well conserved. In this study, we examined the cross-reactivity of biotin attachment between human and bacterial biotin ligases by comparing biotinylation of p-67 and BCCP87, the biotin-attachment domain fragments from human propionyl-CoA carboxylase and E. coli acetyl-CoA carboxylase, respectively. While BirA has similar biotinylation activity toward the two substrates, HCS has reduced activity toward bacterial BCCP87 relative to its native substrate, p-67. The crystal structure of a digested form of p-67, spanning a sequence that contains a seven-residue protruding thumb loop in BCCP87, revealed the absence of a similar structure in the human peptide. Significantly, an engineered "thumbless" bacterial BCCP87 could be biotinylated by HCS, with substrate affinity restored to near normal. This study suggests that the thumb loop found in bacterial carboxylases interferes with optimal interaction with the mammalian biotin protein ligase. While the function of the thumb loop remains unknown, these results indicate a constraint on specificity of the bacterial substrate for biotin attachment that is not itself a feature of BirA.
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Implicit reading in Chinese pure alexia.
Brain Lang
PUBLISHED: 05-06-2010
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A number of recent studies have shown that some patients with pure alexia display evidence of implicit access to lexical and semantic information about words that they cannot read explicitly. This phenomenon has not been investigated systematically in Chinese patients. We report here a case study of a Chinese patient who met the criteria for pure alexia and had lesions in the left occipitotemporal region and the splenium of the corpus callosum. His explicit and implicit reading was evaluated with various stimuli in a number of tasks. We found that despite his severe impairment in overt reading and the definition of any characters, his performance was well above chance in various implicit tasks. His accuracy with respect to lexical decisions was so high that his performance was almost normal. These findings provide unequivocal evidence for the existence of implicit reading in Chinese patients with pure alexia and further support the involvement of the right hemisphere.
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Electrophysiological correlates of proactive interference in the Recent Probes verbal working memory task.
Neuropsychologia
PUBLISHED: 04-02-2010
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Using event-related potentials (ERPs), the present study examined the temporal dynamics of proactive interference in working memory using a recent probes task. Participants memorized and retained a target set of four letters over a short retention interval. They then responded to a recognition probe by judging whether it was from the memory set. ERP waveforms elicited by positive probes compared to those from negative probes showed positive shifts in a fronto-central early N2 component and a parietal late positive component (LPC). The LPC was identified as the electrophysiological signature of proactive interference, as it differentiated between two types of negative probes defined based on whether they were recently encountered. These results indicate that the proactive interference we observed arises from a mismatch between familiarity and contextual information during recognition memory. When considered together with related studies in the literature, the results also suggest that there are different forms of proactive interference associated with different neural correlates.
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Anterior cingulate activity and the self in disorders of consciousness.
Hum Brain Mapp
PUBLISHED: 03-26-2010
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The aim of this study was to investigate the relationship between medial cortical activation and the presence of self and consciousness in healthy subjects and patients with vegetative state and minimally conscious state using functional magnetic resonance imaging (fMRI).
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The trait of sensory processing sensitivity and neural responses to changes in visual scenes.
Soc Cogn Affect Neurosci
PUBLISHED: 03-04-2010
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This exploratory study examined the extent to which individual differences in sensory processing sensitivity (SPS), a temperament/personality trait characterized by social, emotional and physical sensitivity, are associated with neural response in visual areas in response to subtle changes in visual scenes. Sixteen participants completed the Highly Sensitive Person questionnaire, a standard measure of SPS. Subsequently, they were tested on a change detection task while undergoing functional magnetic resonance imaging (fMRI). SPS was associated with significantly greater activation in brain areas involved in high-order visual processing (i.e. right claustrum, left occipitotemporal, bilateral temporal and medial and posterior parietal regions) as well as in the right cerebellum, when detecting minor (vs major) changes in stimuli. These findings remained strong and significant after controlling for neuroticism and introversion, traits that are often correlated with SPS. These results provide the first evidence of neural differences associated with SPS, the first direct support for the sensory aspect of this trait that has been studied primarily for its social and affective implications, and preliminary evidence for heightened sensory processing in individuals high in SPS.
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Structural modifications of the brain in acclimatization to high-altitude.
PLoS ONE
PUBLISHED: 02-24-2010
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Adaptive changes in respiratory and cardiovascular responses at high altitude (HA) have been well clarified. However, the central mechanisms underlying HA acclimatization remain unclear. Using voxel-based morphometry (VBM) and diffusion tensor imaging (DTI) with fractional anisotropy (FA) calculation, we investigated 28 Han immigrant residents (17-22 yr) born and raised at HA of 2616-4200 m in Qinghai-Tibetan Plateau for at least 17 years and who currently attended college at sea-level (SL). Their family migrated from SL to HA 2-3 generations ago and has resided at HA ever since. Control subjects were matched SL residents. HA residents (vs. SL) showed decreased grey matter volume in the bilateral anterior insula, right anterior cingulate cortex, bilateral prefrontal cortex, left precentral cortex, and right lingual cortex. HA residents (vs. SL) had significantly higher FA mainly in the bilateral anterior limb of internal capsule, bilateral superior and inferior longitudinal fasciculus, corpus callosum, bilateral superior corona radiata, bilateral anterior external capsule, right posterior cingulum, and right corticospinal tract. Higher FA values in those regions were associated with decreased or unchanged radial diffusivity coinciding with no change of longitudinal diffusivity in HA vs. SL group. Conversely, HA residents had lower FA in the left optic radiation and left superior longitudinal fasciculus. Our data demonstrates that HA acclimatization is associated with brain structural modifications, including the loss of regional cortical grey matter accompanied by changes in the white matter, which may underlie the physiological adaptation of residents at HA.
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What is Visualize?

JoVE Visualize is a tool created to match the last 5 years of PubMed publications to methods in JoVE's video library.

How does it work?

We use abstracts found on PubMed and match them to JoVE videos to create a list of 10 to 30 related methods videos.

Video X seems to be unrelated to Abstract Y...

In developing our video relationships, we compare around 5 million PubMed articles to our library of over 4,500 methods videos. In some cases the language used in the PubMed abstracts makes matching that content to a JoVE video difficult. In other cases, there happens not to be any content in our video library that is relevant to the topic of a given abstract. In these cases, our algorithms are trying their best to display videos with relevant content, which can sometimes result in matched videos with only a slight relation.