JoVE Visualize What is visualize?
Stop Reading. Start Watching.
Advanced Search
Stop Reading. Start Watching.
Regular Search
Find video protocols related to scientific articles indexed in Pubmed.
Diamagnetic Levitation Promotes Osteoclast Differentiation from RAW264.7 Cells.
IEEE Trans Biomed Eng
PUBLISHED: 11-15-2014
Show Abstract
Hide Abstract
The superconducting magnet with a high magnetic force field can levitate diamagnetic materials. In this study, a specially designed superconducting magnet with large gradient high magnetic field (LGHMF), which provides three apparent gravity levels (? g, 1 g and 2 g), was used to study its influence on receptor activator of nuclear factor-?B ligand (RANKL)-induced osteoclast differentiation from pre-osteoclast cell line RAW264.7. The effects of LGHMF on the viability, nitric oxide (NO) production, morphology in RAW264.7 cells were detected by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) method, Griess method and immunofluorescence staining, respectively. The changes induced by LGHMF in osteoclast formation, mRNA expression and bone resorption were determined by tartrate-resistant acid phosphatase staining, semi-quantity PCR and bone resorption test, respectively. The results showed that: 1) LGHMF had no lethal effect on osteoclast precursors but attenuated NO release in RAW264.7 cells. 2) Diamagnetic levitation (? g) enhanced both the formation and bone resorption capacity of osteoclast. Moreover, diamagnetic levitation up-regulated mRNA expression of RANK, Cathepsin K, MMP-9 and NFATc1 while down-regulated RunX2 in comparison with controls. Furthermore, diamagnetic levitation induced obvious morphological alterations in osteoclast, including active cytoplasmic peripheral pseudopodial expansion, formation of pedosome belt and aggregation of actin ring. 3) Magnetic field produced by LGHMF attenuated osteoclast resorption activity. Collectively, LGHMF with combined effects has multiple effects on osteoclast, which attenuated osteoclast resorption with magnetic field whereas promoted osteoclast differentiation with diamagnetic levitation. Therefore, these findings indicate that diamagnetic levitation could be used as a novel ground-based microgravity simulator which facilitates bone cell research of weightlessness condition.
Related JoVE Video
De Novo Chemoattractants Form Supramolecular Hydrogels for Immunomodulating Neutrophils In Vivo.
Bioconjug. Chem.
PUBLISHED: 11-15-2014
Show Abstract
Hide Abstract
Most of immunomodulatory materials (e.g., vaccine adjuvants such as alum) modulate adap-tive immunity, and yet little effort has focused on developing materials to regulate innate im-munity, which get mentioned only when inflammation affects the biocompatibility of bio-materials. Traditionally considered as short-lived effector cells from innate immunity primarily for the clearance of invading microorganisms without specificity, neutrophils exhibit key role in launching and shaping the immune response. Here we show that the incorporation of un-natural amino acids into a well-known chemoattractant-N-formyl-L-methionyl-L-leucyl-L-phenylalanine (fMLF)-offers a facile approach to create a de novo, multifunctional chemoat-tractant that self-assembles to form supramolecular nanofibrils and hydrogels. This de novo chemoattractant not only exhibits preserved cross-species chemoattractant activity to human and murine neutrophils, but also effectively resists proteolysis. Thus, its hydrogel, in vivo, re-leases the chemoattractant and attracts neutrophils to the desired location in a sustainable manner. As a novel and general approach to generate a new class of biomaterials for modu-lating innate immunity, this work offers a prolonged acute inflammation model for developing various new applications.
Related JoVE Video
8q24 rs4242382 Polymorphism is a Risk Factor for Prostate Cancer among Multi-Ethnic Populations: Evidence from Clinical Detection in China and a Meta-analysis.
Asian Pac. J. Cancer Prev.
PUBLISHED: 10-24-2014
Show Abstract
Hide Abstract
Evidence supporting an association between the 8q24 rs4242382-A polymorphism and prostate cancer (PCa) risk has been reported in North American and Europe populations, though data from Asian populations remain limited. We therefore investigated this association by clinical detection in China, and meta-analysis in Asian, Caucasian and African-American populations.
Related JoVE Video
Is Elapsing Time Really Recoded Into Spatial Linear Representation in Working Memory?
Exp Psychol
PUBLISHED: 10-02-2014
Show Abstract
Hide Abstract
A growing body of evidence suggested that elapsing time is tightly associated with space in a specific way (e.g., Spatial Temporal Association of Response Codes or STARC effect). However, existing findings cannot justify a hypothesis that elapsing time is recoded directly into a spatial linear representation in working memory. The present study addresses this fundamental question by using three modified STARC-related working memory paradigms. In different experiments, participants were asked to give order judgment, order-irrelevant STM recognition judgment, or motor-related free-choice judgment, immediately after successive presentation of a set of disparate stimuli. Results show that responses to early stimuli were faster or more often with the left key and responses to late stimuli were faster or more often with the right key. These findings clearly support the hypothesis that elapsing time is directly and automatically recoded into a spatial linear representation in working memory.
Related JoVE Video
[Identification of original plants of uyghur medicinal materials fructus elaeagni using morphological characteristics and DNA barcode].
Zhongguo Zhong Yao Za Zhi
PUBLISHED: 09-24-2014
Show Abstract
Hide Abstract
Morphology and molecular identification technology were used to identify 3 original plants of Fructus Elaeagni which was commonly used in Uygur medicine. Leaves, flowers and fruits from different areas were selected randomly for morphology research. ITS2 sequence as DNA barcode was used to identify 17 samples of Fructus Elaeagni. The genetic distances were computed by kimura 2-parameter (K2P) model, and the Neighbor-Joining (NJ) and Maximum Likelihood phylogenetic trees were constructed using MEGA5.0. The results showed that Elaeagnus angustifolia, E. oxycarpa and E. angustifolia var. orientalis cannot be distinguished by morphological characteristics of leaves, flowers and fruits. The sequence length of ITS2 ranged from 220 to 223 bp, the average GC content was 61.9%. The haplotype numbers of E. angustifolia, E. oxycarpa and E. angustifolia var. orientals were 4, 3, 3, respectively. The results from the NJ tree and ML tree showed that the 3 original species of Fructus Elaeagni cannot be distinguished obviously. Therefore, 3 species maybe have the same origin, and can be used as the original plant of Uygur medicineal material Fructus Elaeagni. However, further evidence of chemical components and pharmacological effect were needed.
Related JoVE Video
G-protein ?q participates in the steroid hormone 20-hydroxyecdysone nongenomic signal transduction.
J. Steroid Biochem. Mol. Biol.
PUBLISHED: 08-12-2014
Show Abstract
Hide Abstract
The nuclear receptor-mediated genomic pathways of the animal steroid hormones are well known. However, the cell membrane receptor-mediated nongenomic pathways of the animal steroid hormones are little understood. In this study, we report the participation of a G-protein alpha q (G?q)(1) subunit in the 20E nongenomic pathway in the cell membrane and regulating gene expression during molting and metamorphosis in a lepidopteran insect, Helicoverpa armigera. 20E-induced phosphorylation of G?q was detected using two-dimensional electrophoresis techniques. Knockdown of G?q by injecting double-stranded RNA suppressed the development of larvae, delayed metamorphosis, and inhibited 20E-induced gene expression. G?q was distributed throughout the cell, and migrated toward the plasma membrane upon 20E induction. G?q was necessary in the 20E-induced intracellular Ca(2+) release and extracellular Ca(2+) influx. The protein kinase C (PKC) inhibitor could repress 20E-induced phosphorylation of cyclin-dependent kinase 10 (CDK10) and transcription factor ultraspiracle (USP1). PKC inhibitor could repress the G?q phosphorylation and membrane trafficking. These results suggest that G?q participates in 20E signaling in the cell membrane at the pre-genomic stage by modulating the increase of the intracellular Ca(2+) and phosphorylation of CDK10 and USP1 in 20E transcription complex to regulate gene transcription.
Related JoVE Video
Juvenile hormone prevents 20-hydroxyecdysone-induced metamorphosis by regulating the phosphorylation of a newly identified broad protein.
J. Biol. Chem.
PUBLISHED: 08-05-2014
Show Abstract
Hide Abstract
The steroid hormone 20-hydroxyecdysone (20E) initiates insect molting and metamorphosis. By contrast, juvenile hormone (JH) prevents metamorphosis. However, the mechanism by which JH inhibits metamorphosis remains unclear. In this study, we propose that JH induces the phosphorylation of Broad isoform Z7 (BrZ7), a newly identified protein, to inhibit 20E-mediated metamorphosis in the lepidopteran insect Helicoverpa armigera. The knockdown of BrZ7 in larvae inhibited metamorphosis by repressing the expression of the 20E response gene. BrZ7 was weakly expressed and phosphorylated during larval growth but highly expressed and non-phosphorylated during metamorphosis. JH regulated the rapid phosphorylation of BrZ7 via a G-protein-coupled receptor-, phospholipase C-, and protein kinase C-triggered pathway. The phosphorylated BrZ7 bound to the 5'-regulatory region of calponin to regulate its expression in the JH pathway. Exogenous JH induced BrZ7 phosphorylation to prevent metamorphosis by suppressing 20E-related gene transcription. JH promoted non-phosphorylated calponin interacting with ultraspiracle protein to activate the JH pathway and antagonize the 20E pathway. This study reveals one of the possible mechanisms by which JH counteracts 20E-regulated metamorphosis by inducing the phosphorylation of BrZ7.
Related JoVE Video
Infrared image detail enhancement based on the gradient field specification.
Appl Opt
PUBLISHED: 08-05-2014
Show Abstract
Hide Abstract
Human vision is sensitive to the changes of local image details, which are actually image gradients. To enhance faint infrared image details, this article proposes a gradient field specification algorithm. First we define the image gradient field and gradient histogram. Then, by analyzing the characteristics of the gradient histogram, we construct a Gaussian function to obtain the gradient histogram specification and therefore obtain the transform gradient field. In addition, subhistogram equalization is proposed based on the histogram equalization to improve the contrast of infrared images. The experimental results show that the algorithm can effectively improve image contrast and enhance weak infrared image details and edges. As a result, it can give qualified image information for different applications of an infrared image. In addition, it can also be applied to enhance other types of images such as visible, medical, and lunar surface.
Related JoVE Video
L-Rhamnose-containing supramolecular nanofibrils as potential immunosuppressive materials.
Org. Biomol. Chem.
PUBLISHED: 08-01-2014
Show Abstract
Hide Abstract
An l-rhamnose-based hydrogelator self-assembles to form nanofibrils, which, in contrast to the properties of monomeric l-rhamnose, suppress the antibody response of mice to phycoerythrin (PE), a fluorescent protein antigen. As the first example of the supramolecular assemblies of a saccharide to suppress immunity, this work illustrates a new approach of immunomodulation.
Related JoVE Video
A new group of anti-lipopolysaccharide factors from Marsupenaeus japonicus functions in antibacterial response.
Dev. Comp. Immunol.
PUBLISHED: 07-28-2014
Show Abstract
Hide Abstract
Anti-lipopolysaccharide factors (ALFs) are a group of critical effector molecules with a broad spectrum of antimicrobial activities in crustaceans. Four groups of ALFs (A, B, C, and D) have been identified in peneaid shrimp. In the study, we identified a new group of ALFs (designated as MjALF-E) from Marsupenaeus japonicus. This new group (group E) included MjALF-E1 and E2. MjALF-E1 was highly expressed in hemocytes, heart, and intestine, whereas E2 was highly expressed in gills, stomach, and intestine. Expressions of both MjALF-E1 and E2 were upregulated by bacterial challenge. Synthesized LPS-binding domain peptides of MjALF-E1 and E2 strongly bind to bacterial cell wall components lipopolysaccharide (LPS) and peptidoglycan (PGN). The recombinant rMjALF-E2 showed relatively weak binding activity to LPS and PGN. Both synthesized peptides and rMjALF-E2 exhibited antimicrobial activity against Gram-negative bacteria, whereas rMjALF-E2 could promote the clearance of bacteria in vivo. After knockdown of MjALF-E2 and infection with Vibrio anguillarum, shrimp showed high and rapid mortality compared with GFPi shrimp. These results suggest that MjALF-Es serves a protective function against bacterial infection in shrimp.
Related JoVE Video
Collaboration between a soluble C-type lectin and calreticulin facilitates white spot syndrome virus infection in shrimp.
J. Immunol.
PUBLISHED: 07-28-2014
Show Abstract
Hide Abstract
White spot syndrome virus (WSSV) mainly infects crustaceans through the digestive tract. Whether C-type lectins (CLs), which are important receptors for many viruses, participate in WSSV infection in the shrimp stomach remains unknown. In this study, we orally infected kuruma shrimp Marsupenaeus japonicus to model the natural transmission of WSSV and identified a CL (designated as M. japonicus stomach virus-associated CL [MjsvCL]) that was significantly induced by virus infection in the stomach. Knockdown of MjsvCL expression by RNA interference suppressed the virus replication, whereas exogenous MjsvCL enhanced it. Further analysis by GST pull-down and coimmunoprecipitation showed that MjsvCL could bind to viral protein 28, the most abundant and functionally relevant envelope protein of WSSV. Furthermore, cell-surface calreticulin was identified as a receptor of MjsvCL, and the interaction between these proteins was a determinant for the viral infection-promoting activity of MjsvCL. The MjsvCL-calreticulin pathway facilitated virus entry likely in a cholesterol-dependent manner. This study provides insights into a mechanism by which soluble CLs capture and present virions to the cell-surface receptor to facilitate viral infection.
Related JoVE Video
Role of microRNAs in osteoblasts differentiation and bone disorders.
Curr. Med. Chem.
PUBLISHED: 06-20-2014
Show Abstract
Hide Abstract
Advanced studies of single stranded endogenous ~22 nt microRNAs (miRNAs) have demonstrated their diverse biological functions including control of cell differentiation, cell cycle and pathological conditions. Recent studies suggest the potential application of miRNAs in stem cell engineering. miRNAs play a vital role as post-transcriptional regulators of gene expression which controls osteoblasts-mediated bone formation and osteoclasts related bone remodeling. Transcriptional and post-transcriptional mechanisms regulate the differentiation of osteoblasts and osteogenesis. The differentiation of osteoblasts is a key step in the development of skeletal muscles and it is involved in triggering the signaling pathways. Signaling pathways like TGF?, BMP and Wnt are regulated by miRNAs which in turn are shown to be associated with bone dynamics and bone disorders. This recap highlights the role of miRNAs in osteoblasts differentiation and emphasizes their potential therapeutic role in metabolic bone disorders.
Related JoVE Video
Methoprene-tolerant 1 regulates gene transcription to maintain insect larval status.
J. Mol. Endocrinol.
PUBLISHED: 05-28-2014
Show Abstract
Hide Abstract
Insect molting and metamorphosis are regulated by two hormones: 20-hydroxyecdysone (20E) and juvenile hormone (JH). The hormone 20E regulates gene transcription via the nuclear receptor EcR to promote metamorphosis, whereas JH regulates gene transcription via its intracellular receptor methoprene-tolerant (Met) to prevent larval-pupal transition. However, the function and mechanism of Met in various insect developments are not well understood. We propose that Met1 plays a key role in maintaining larval status not only by promoting JH-responsive gene transcription but also by repressing 20E-responsive gene transcription in the Lepidopteran insect Helicoverpa armigera. Met1 protein is increased during feeding stage and decreased during molting and metamorphic stages. Met1 is upregulated by JH III and a low concentration of 20E independently, but is downregulated by a high concentration of 20E. Knockdown of Met1 in larvae causes precocious pupation, decrease in JH pathway gene expression, and increase in 20E pathway gene expression. Met1 interacts with heat shock protein 90 and binds to JH response element to regulate Krüppel homolog 1 transcription in JH III induction. Met1 interacts with ultraspiracle protein 1 (USP1) to repress 20E transcription complex EcRB1/USP1 formation and binding to ecdysone response element. These data indicate that JH via Met1 regulates JH pathway gene expression and represses 20E pathway gene expression to maintain the larval status.
Related JoVE Video
[Prediction models for the 15 years risk of new-onset hypertension in Chinese people aged from 35 to 64 years old].
Zhonghua Nei Ke Za Zhi
PUBLISHED: 05-27-2014
Show Abstract
Hide Abstract
To set up prediction models for the risk of new-onset hypertension in Chinese people and explore the risk scores to facilitate the clinical application.
Related JoVE Video
[DNA methylation of prostate cancer and clinical application].
Yi Chuan
PUBLISHED: 05-22-2014
Show Abstract
Hide Abstract
It is well-known that the interaction of both genetic and epigenetic mechanisms results in the formation of malignant tumor. Epigenetic mechanism includes DNA methylation, histone modifications, and miRNA regulation. DNA aberrant methylation (hypermethylation and hypomethylation), which leads to genomic instability and inappropriate gene expression, is the best-characterized alteration in prostate cancer. It plays an important role in the initiation and development of prostate cancer. Meanwhile, DNA methylation, as a hotspot in researches of epigenetics, would provide a new methodology and approach for early clinical diagnosis, prognosis and medication treatment of prostate cancer. According to recent studies on DNA hypermethylation and DNA hypomethylation, this review highlights the potential epigenetic mechanism of prostate cancer and discusses the latest research progress in clinical translation.
Related JoVE Video
rs10505474 and rs7837328 at 8q24 cumulatively confer risk of prostate cancer in Northern Han Chinese.
Asian Pac. J. Cancer Prev.
PUBLISHED: 05-13-2014
Show Abstract
Hide Abstract
Genome-wide association studies (GWAS) have identified several risk variants for prostate cancer (pCa) mainly in Europeans, which need to be further verified in other racial groups. We selected six previously identified variants as candidates and to define the association with PCa in Northern Han Chinese.
Related JoVE Video
Impact of cerebrovascular disease mortality on life expectancy in China.
Biomed. Environ. Sci.
PUBLISHED: 04-09-2014
Show Abstract
Hide Abstract
To evaluate the impact of cerebrovascular disease mortality on life expectancy (LE) in China in 2010 compared with 2005, and to identify the high-risk population (age, sex, and region) where cerebrovascular disease mortality has had a major impact on LE.
Related JoVE Video
Impact of cardiovascular disease deaths on life expectancy in Chinese population.
Biomed. Environ. Sci.
PUBLISHED: 04-09-2014
Show Abstract
Hide Abstract
We aimed to analyze the impact of cardiovascular disease (CVD) deaths on life expectancy (LE) in Chinese population and estimate the percentage reduction in CVD mortality needed to increase LE by 1 year from the current level, a national target of health improvement.
Related JoVE Video
BurnCalc assessment study of computer-aided individual three-dimensional burn area calculation.
J Transl Med
PUBLISHED: 04-04-2014
Show Abstract
Hide Abstract
BackgroundAccurate estimation of a burned area is crucial to decisions about fluid resuscitation, surgical options, nutritional support, and prognosis. Widely used clinical methods to estimate a burn area are two-dimensional. They do not consider age, sex, body mass, physical deformities, or other relevant factors. Computer-aided methods have improved the accuracy of estimating burned areas by including data analysis and reducing subjective differences. Three-dimensional (3D) scanning allows us to determine body dimensions rapidly and reproducibly. We describe an individualized, cost-efficient, portable 3D scanning system, BurnCalc, that can create an individual 3D model and then calculate body surface area (BSA) and the burn area accurately and quickly.MethodsThe BurnCalc system was validated by verifying the accuracy and stability of BSA calculation. We measured 10 regular objects in experiment 1, using Student¿s t-test and the intraclass correlation coefficient (ICC) in the analysis. In experiment 2, artificial paper patches of known dimensions were attached to various parts of the body of 40 volunteers. Their sizes were then calculated using BurnCalc. The BurnCalc data were compared to actually measured values to verify accuracy and stability. Total BSAs of these 40 volunteers were also calculated by BurnCalc and compared to those derived from an accepted formula. In experiment 3, four experts using Chinese Rule-of-Nines or Rule-of-Palms methods calculated the percentages of the total BSA in 17 volunteers. Student¿s t-test and ICC, respectively, were used to compare the results obtained with the BurnCalc technique.ResultsStatistically, in experiment 1, p = 0.834 and ICC = 0.999, demonstrating that there was no difference between the BurnCalc and real measurements. Also, the hypothesis of null difference among measures (experiment 2) was true because p¿>¿0.05 and ICC = 0.999, indicating that calculations of the total BSA and the burn area were more accurate using the BurnCalc technology. The reliability of the BurnCalc program was 99.9%. In experiment 3, only the BurnCalc method exhibited values of p¿>¿0.05 (p = 0.774) and ICC = 0.999.ConclusionsBurnCalc technology produced stable, accurate readings, suggesting that BurnCalc could be regarded as a new standard clinical method.
Related JoVE Video
Phospholipase C?1 connects the cell membrane pathway to the nuclear receptor pathway in insect steroid hormone signaling.
J. Biol. Chem.
PUBLISHED: 04-01-2014
Show Abstract
Hide Abstract
In addition to the classical nuclear receptor pathway, there is a nongenomic pathway in the cell membrane that regulates gene expression in animal steroid hormone signaling; however, this mechanism is unclear. Here, we report that the insect steroid hormone 20-hydroxyecdysone (20E) regulates calcium influx via phospholipase C?1 (PLCG1) to modulate the protein kinase C phosphorylation of the transcription factor ultraspiracle (USP1) in the lepidopteran insect Helicoverpa armigera. The PLCG1 mRNA levels are increased during the molting and metamorphic stages. The depletion of PLCG1 by RNA interference can block 20E-enhanced pupation, cause larvae death and pupation defects, and repress 20E-induced gene expression. 20E may induce the tyrosine phosphorylation of PLCG1 at the cytosolic tyrosine kinase (Src) homology 2 domains and then determine the migration of PLCG1 toward the plasma membrane. The G-protein-coupled receptor (GPCR) inhibitor suramin, Src family kinase inhibitor PP2, and the depletions of ecdysone-responsible GPCR (ErGPCR) and G?q restrain the 20E-induced tyrosine phosphorylation of PLCG1. PLCG1 participates in the 20E-induced Ca(2+) influx. The inhibition of GPCR, PLC, inositol 1,4,5-trisphosphate receptor, and calcium channels represses the 20E-induced Ca(2+) influx. Through calcium signaling, PLCG1 mediates the transcriptional activation driven by the ecdysone-response element. Through PLCG1 and calcium signaling, 20E regulates PKC phosphorylation of USP1 at Ser-21 to determine its ecdysone-response element binding activity. These results suggest that 20E activates PLCG1 via the ErGPCR and Src family kinases to regulate Ca(2+) influx and PKC phosphorylation of USP1 to subsequently modulate gene transcription for metamorphosis.
Related JoVE Video
Physiological effects of microgravity on bone cells.
Calcif. Tissue Int.
PUBLISHED: 03-12-2014
Show Abstract
Hide Abstract
Life on Earth developed under the influence of normal gravity (1g). With evidence from previous studies, scientists have suggested that normal physiological processes, such as the functional integrity of muscles and bone mass, can be affected by microgravity during spaceflight. During the life span, bone not only develops as a structure designed specifically for mechanical tasks but also adapts for efficiency. The lack of weight-bearing forces makes microgravity an ideal physical stimulus to evaluate bone cell responses. One of the most serious problems induced by long-term weightlessness is bone mineral loss. Results from in vitro studies that entailed the use of bone cells in spaceflights showed modification in cell attachment structures and cytoskeletal reorganization, which may be involved in bone loss. Humans exposed to microgravity conditions experience various physiological changes, including loss of bone mass, muscle deterioration, and immunodeficiency. In vitro models can be used to extract valuable information about changes in mechanical stress to ultimately identify the different pathways of mechanotransduction in bone cells. Despite many in vivo and in vitro studies under both real microgravity and simulated conditions, the mechanism of bone loss is still not well defined. The objective of this review is to summarize the recent research on bone cells under microgravity conditions based on advances in the field.
Related JoVE Video
A shrimp C-type lectin inhibits proliferation of the hemolymph microbiota by maintaining the expression of antimicrobial peptides.
J. Biol. Chem.
PUBLISHED: 03-11-2014
Show Abstract
Hide Abstract
Some aquatic invertebrates such as shrimp contain low albeit stable numbers of bacteria in the circulating hemolymph. The proliferation of this hemolymph microbiota in such a nutrient-rich environment is tightly controlled in healthy animals, but the mechanisms responsible had remained elusive. In the present study, we report a C-type lectin (MjHeCL) from the kuruma shrimp (Marsupenaeus japonicus) that participates in restraining the hemolymph microbiota. Although the expression of MjHeCL did not seem to be modulated by bacterial challenge, the down-regulation of its expression by RNA interference led to proliferation of the hemolymph microbiota, ultimately resulting in shrimp death. This phenotype was rescued by the injection of recombinant MjHeCL, which restored the healthy status of the knockdown shrimp. A mechanistic analysis revealed that MjHeCL inhibited bacterial proliferation by modulating the expression of antimicrobial peptides. The key function of MjHeCL in the shrimp immune homeostasis might be related to its broader recognition spectrum of the hemolymph microbiota components than other lectins. Our study demonstrates the role of MjHeCL in maintaining the healthy status of shrimp and provides new insight into the biological significance of C-type lectins, a diversified and abundant lectin family in invertebrate species.
Related JoVE Video
Protective immunity against recurrent Staphylococcus aureus skin infection requires antibody and interleukin-17A.
Infect. Immun.
PUBLISHED: 03-10-2014
Show Abstract
Hide Abstract
Although many microbial infections elicit an adaptive immune response that can protect against reinfection, it is generally thought that Staphylococcus aureus infections fail to generate protective immunity despite detectable T and B cell responses. No vaccine is yet proven to prevent S. aureus infections in humans, and efforts to develop one have been hampered by a lack of animal models in which protective immunity occurs. Our results describe a novel mouse model of protective immunity against recurrent infection, in which S. aureus skin and soft tissue infection (SSTI) strongly protected against secondary SSTI in BALB/c mice but much less so in C57BL/6 mice. This protection was dependent on antibody, because adoptive transfer of immune BALB/c serum or purified antibody into either BALB/c or C57BL/6 mice resulted in smaller skin lesions. We also identified an antibody-independent mechanism, because B cell-deficient mice were partially protected against secondary S. aureus SSTI and adoptive transfer of T cells from immune BALB/c mice resulted in smaller lesions upon primary infection. Furthermore, neutralization of interleukin-17A (IL-17A) abolished T cell-mediated protection in BALB/c mice, whereas neutralization of gamma interferon (IFN-?) enhanced protection in C57BL/6 mice. Therefore, protective immunity against recurrent S. aureus SSTI was advanced by antibody and the Th17/IL-17A pathway and prevented by the Th1/IFN-? pathway, suggesting that targeting both cell-mediated and humoral immunity might optimally protect against secondary S. aureus SSTI. These findings also highlight the importance of the mouse genetic background in the development of protective immunity against S. aureus SSTI.
Related JoVE Video
In a nongenomic action, steroid hormone 20-hydroxyecdysone induces phosphorylation of cyclin-dependent kinase 10 to promote gene transcription.
Endocrinology
PUBLISHED: 02-11-2014
Show Abstract
Hide Abstract
The insect steroid hormone 20-hydroxyecdysone (20E) regulates gene transcription via a genomic pathway by forming a transcription complex that binds to DNA with the help of the chaperone proteins, heat shock proteins (Hsps) Hsc70 and Hsp90. However, the nongenomic mechanisms by which 20E regulates gene expression remain unclear. In this study, we found that 20E regulated the phosphorylation of serine/threonine protein kinase cyclin-dependent kinase 10 (CDK10) through a nongenomic pathway to mediate gene transcription in the lepidopteran Helicoverpa armigera. The down-regulation of CDK10 by RNA interference in larvae and the epidermal cell line delayed development and suppressed 20E-induced gene transcription. CDK10 was localized to the nucleus via its KKRR motif, and this nuclear localization and the ATPase motif were necessary for the efficient expression of the 20E-inducible gene. The rapid phosphorylation of CDK10 was induced by 20E, whereas it was repressed by the inhibitors of G-protein-coupled receptors, phospholipase C, and Ca²? channels. Phosphorylated CDK10 exhibited increased interactions with Hsps Hsc70 and Hsp90 and then promoted the interactions between Hsps and ecdysone receptor EcRB1 and the binding of the Hsps-EcRB1 complex to the 20E response element for the regulation of gene transcription. CDK10 depletion suppressed the formation of the Hsps-EcRB1 complex at the hormone receptor 3 promoter. These results suggest that 20E induces CDK10 phosphorylation via a nongenomic pathway to regulate gene transcription in the nucleus.
Related JoVE Video
Five-year change in systolic blood pressure is independently associated with carotid atherosclerosis progression: a population-based cohort study.
Hypertens. Res.
PUBLISHED: 02-07-2014
Show Abstract
Hide Abstract
The aim of this study was to investigate whether long-term changes in traditional risk factors affect the progression of carotid atherosclerosis in a Chinese population. This study included 1590 individuals (aged 56.9±8.1 years) with no evidence of carotid plaque at baseline (2002). In 2007, these individuals completed the second risk factors survey and underwent carotid plaque measurement. The incidence of carotid plaque and the total plaque area of maximum plaques (TPA) were used to evaluate the progression of carotid atherosclerosis. In addition to baseline age, systolic blood pressure (SBP), total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), triglyceride (TG), common carotid intima-media thickness (CIMT) and current smoking, a 5-year change in SBP was also associated with the incidence of carotid plaque (odds ratio=1.01, 95% confidence interval: 1.01-1.02, P=0.029). Furthermore, multiple linear regression analysis revealed that a 5-year change in SBP had a linear association with TPA after adjusting for baseline risk factors (standardized regression coefficient=0.071, P=0.014). TPA increased both by increasing baseline SBP and by a 5-year SBP change when adjusted for sex, baseline age, TC, HDL-C, CIMT and current smoking (P for trend <0.001 and 0.004). Our study demonstrates that a 5-year change in SBP is independently associated with the progression of carotid atherosclerosis in the Chinese population. These findings underline the importance of early detection and control of SBP for the prevention of atherosclerosis progression. The progression of atherosclerosis is not only associated with hypertension but can also progress silently with the development of SBP.
Related JoVE Video
G-protein-coupled receptor participates in 20-hydroxyecdysone signaling on the plasma membrane.
Cell Commun. Signal
PUBLISHED: 02-03-2014
Show Abstract
Hide Abstract
Animal steroid hormones are conventionally known to initiate signaling via a genomic pathway by binding to the nuclear receptors. The mechanism by which 20E initiates signaling via a nongenomic pathway is unclear.
Related JoVE Video
THADA gene polymorphism and prostate cancer risk: a meta-analysis.
Oncol Res Treat
PUBLISHED: 01-29-2014
Show Abstract
Hide Abstract
The single nucleotide polymorphism (SNP) rs1465618 in THADA at 2p21 has been identified as being associated with prostate cancer (PCa) risk in Europeans; however, it is not clear whether the SNP is related to PCa risk in multiple populations. We investigated the association of rs1465618 in THADA with PCa in a Chinese population and carried out a meta-analysis in multiple populations, testing the relevance of this SNP for PCa risk.
Related JoVE Video
Calnexin functions in antibacterial immunity of Marsupenaeus japonicus.
Dev. Comp. Immunol.
PUBLISHED: 01-22-2014
Show Abstract
Hide Abstract
Calnexin (Cnx) is an endoplasmic reticulum membrane-bound lectin chaperone that comprises a dedicated maturation system with another lectin chaperone calreticulin (Crt). This maturation system is known as the Cnx/Crt cycle. The main functions of Cnx are Ca(2+) storage, glycoprotein folding, and quality control of synthesis. Recent studies have shown that Cnx is important in phagocytosis and in optimizing dendritic cell immunity. However, the functions of Cnx in invertebrate innate immunity remain unclear. In this research, we characterized Cnx in the kuruma shrimp Marsupenaeus japonicus (designated as MjCnx) and detected its function in shrimp immunity. The expression of MjCnx was upregulated in several tissues challenged with Vibrio anguillarum. Recombinant MjCnx could bind to bacteria by binding polysaccharides. MjCnx protein existed in the cytoplasm and on the membrane of hemocytes and was upregulated by bacterial challenge. The recombinant MjCnx enhanced the clearance of V. anguillarum in vivo, and the clearance effects were impaired after silencing MjCnx with RNA interference assay. Recombinant MjCnx promoted phagocytosis efficiency of hemocytes. These results suggest that MjCnx functions as one of the pattern recognition receptors and has crucial functions in shrimp antibacterial immunity.
Related JoVE Video
A fibrinogen-related protein (FREP) is involved in the antibacterial immunity of Marsupenaeus japonicus.
Fish Shellfish Immunol.
PUBLISHED: 01-18-2014
Show Abstract
Hide Abstract
Fibrinogen-related proteins (FREPs) in invertebrates have important functions in innate immunity. In this study, the cDNA of FREP was identified from the kuruma shrimp Marsupenaeus japonicus (MjFREP2). The full-length cDNA of MjFREP2 is 1138 bp with an open reading frame of 954 bp that encodes a 317-amino acid protein comprising a signal peptide and a fibrinogen-like domain. MjFREP2 could be detected in hemocytes, heart, hepatopancreas, gills, stomach, and intestines. MjFREP2 could also be upregulated in hemocytes after Vibrio anguillarum and Staphylococcus aureus challenge. Agglutination and binding assay results revealed that the recombinant MjFREP2 bound to bacteria and polysaccharides. Immunocytochemical analysis results showed that MjFREP2 proteins were mainly distributed in the cytoplasm of hemocytes from unchallenged shrimp and transported to the membrane or secreted out of the cell after V. anguillarum or S. aureus challenge. The secreted MjFREP2 bound to the bacteria presented in shrimp hemolymph. The overexpression of MjFREP2 could enhance bacterial clearance by inducing the phagocytosis of hemocytes. This ability was impaired by knockdown of MjFREP2 with RNA interference. The cumulative mortality of MjFREP2-silenced shrimp was significantly higher than that of the control shrimp. These results suggested that MjFREP2 has an important function in the antibacterial immunity of M. japonicus.
Related JoVE Video
L-Type lectin from the kuruma shrimp Marsupenaeus japonicus promotes hemocyte phagocytosis.
Dev. Comp. Immunol.
PUBLISHED: 01-17-2014
Show Abstract
Hide Abstract
L-Type lectins (LTLs) contain a luminal carbohydrate recognition domain, which exhibits homology to leguminous lectins. These type I membrane proteins are involved in the early secretory pathway of animals, and have functions in glycoprotein sorting, trafficking and targeting. Recent studies suggest that LTLs may be involved in immune responses in vertebrates, but no functional studies have been reported. This study reports an LTL, designated as MjLTL1, from the kuruma shrimp Marsupenaeus japonicus. MjLTL consists of a signal peptide, leguminous lectin domain, and transmembrane region. It was upregulated following challenge of shrimp with Vibrio anguillarum. MjLTL1 could agglutinate several bacteria with the presence of calcium, and bind to several Gram-positive and Gram-negative bacteria through lipopolysaccharide and peptidoglycan binding. MjLTL1 could enhance the clearance of V. anguillarum in vivo. MjLTL1 silencing by RNA interference could impair bacterial clearance ability. Further study suggested that MjLTL1 promoted hemocyte phagocytosis. To analyze the possible mechanism, a disintegrin and metalloprotease-like protein (MjADAM) mediating the proteolytic release of extracellular domains from the membrane-bound precursors was also studied in the shrimp. MjADAM exhibited similar tissue location and expression profiles to MjLTL1. After knockdown of MjADAM, the hemocyte phagocytosis rate also declined significantly. ADAM was reported to have an ectodomain shedding function to LTL and release the ectodomain of the lectin from cell membrane. Therefore, our results suggest that the extracellular domain of MjLTL1 might be released from the cell surface as a soluble protein by MjADAM, and function as an opsonin involved in the antibacterial immune responses in shrimp.
Related JoVE Video
Effect of Nrf2 on rat ovarian tissues against atrazine-induced anti-oxidative response.
Int J Clin Exp Pathol
PUBLISHED: 01-01-2014
Show Abstract
Hide Abstract
The environmental persistence and bioaccumulation of herbicide atrazine may pose a significant threat to human health. In this experiment, Wistar rats were treated by 5, 25 and 125 mg·kg(-1) atrazine respectively for 28 days, and the oxidative stress responses as well as the activations of Nrf2 signaling pathway in ovarian tissues induced by atrazine were observed. The results showed that after be treated by atrazine, the proportion of atretic follicles in the rat ovary were increased, the contents of NO and MDA in the tissue homogenates were increased, the over-expressed Nrf2 transferred into the nuclei and played an antioxidant role by up-regulated the expression of II phase detoxifying enzymes such as HO1 and NQO1 and the expression of antioxidant enzymes such as CAT, SOD and GSH-PX.
Related JoVE Video
Identification of susceptibility variants in ADIPOR1 gene associated with type 2 diabetes, coronary artery disease and the comorbidity of type 2 diabetes and coronary artery disease.
PLoS ONE
PUBLISHED: 01-01-2014
Show Abstract
Hide Abstract
Adiponectin receptor 1 (encoded by ADIPOR1) is one of the major adiponectin receptors, and plays an important role in glucose and lipid metabolism. However, few studies have reported simultaneous associations between ADIPOR1 variants and type 2 diabetes (T2D), coronary artery disease (CAD) and T2D with CAD. Based on the "common soil" hypothesis, we investigated whether ADIPOR1 polymorphisms contributed to the etiology of T2D, CAD, or T2D with CAD in a Northern Han Chinese population.
Related JoVE Video
Capacity limit of simultaneous temporal processing: how many concurrent 'clocks' in vision?
PLoS ONE
PUBLISHED: 01-01-2014
Show Abstract
Hide Abstract
A fundamental ability for humans is to monitor and process multiple temporal events that occur at different spatial locations simultaneously. A great number of studies have demonstrated simultaneous temporal processing (STP) in human and animal participants, i.e., multiple 'clocks' rather than a single 'clock'. However, to date, we still have no knowledge about the exact limitation of the STP in vision. Here we provide the first experimental measurement to this critical parameter in human vision by using two novel and complementary paradigms. The first paradigm combines merits of a temporal oddball-detection task and a capacity measurement widely used in the studies of visual working memory to quantify the capacity of STP (CSTP). The second paradigm uses a two-interval temporal comparison task with various encoded spatial locations involved in the standard temporal intervals to rule out an alternative, 'object individuation'-based, account of CSTP, which is measured by the first paradigm. Our results of both paradigms indicate consistently that the capacity limit of simultaneous temporal processing in vision is around 3 to 4 spatial locations. Moreover, the binding of the 'local clock' and its specific location is undermined by bottom-up competition of spatial attention, indicating that the time-space binding is resource-consuming. Our finding that the capacity of STP is not constrained by the capacity of visual working memory (VWM) supports the idea that the representations of STP are likely stored and operated in units different from those of VWM. A second paradigm confirms further that the limited number of location-bound 'local clocks' are activated and maintained during a time window of several hundreds milliseconds.
Related JoVE Video
A galectin from the kuruma shrimp (Marsupenaeus japonicus) functions as an opsonin and promotes bacterial clearance from hemolymph.
PLoS ONE
PUBLISHED: 01-01-2014
Show Abstract
Hide Abstract
Galectins are a lectin family characterized by a conserved sequence motif in the carbohydrate recognition domain, which preferential binds to galactosyl moieties. However, few studies about the biological roles of galectins in invertebrates have been reported except for the galectin (CvGal1) from the eastern oyster Crassostrea virginica. Furthermore, galectins have been described in only a few crustacean species, and no functional studies have been reported so far. In this study, we identified and functionally characterized a galectin from the kuruma shrimp Marsupenaeus japonicus, which we designated MjGal. Upon Vibrio anguillarum challenge, expression of MjGal was up-regulated mostly in hemocytes and hepatopancreas, and the protein bound to both Gram-positive and Gram-negative bacteria through the recognition of lipoteichoic acid (LTA) or lipopolysaccharide (LPS), respectively. By also binding to the shrimp hemocyte surface, MjGal functions as an opsonin for microbial pathogens, promoting their phagocytosis. Further, as shown by RNA interference, MjGal participates in clearance of bacteria from circulation, and thereby contributes to the shrimp's immune defense against infectious challenge. Elucidation of functional and mechanistic aspects of shrimp immunity will enable the development of novel strategies for intervention in infectious diseases currently affecting the shrimp farming industry worldwide.
Related JoVE Video
Association between APOC1 polymorphism and Alzheimer's disease: a case-control study and meta-analysis.
PLoS ONE
PUBLISHED: 01-01-2014
Show Abstract
Hide Abstract
Previous association studies examining the relationship between the APOC1 polymorphism and susceptibility to Alzheimer's disease (AD) have shown conflicting results, and it is not clear if an APOC1 variant acts as a genetic risk factor in AD etiology across multiple populations.
Related JoVE Video
Antibacterial activity of serine protease inhibitor 1 from kuruma shrimp Marsupenaeus japonicus.
Dev. Comp. Immunol.
PUBLISHED: 01-01-2014
Show Abstract
Hide Abstract
Serine protease inhibitors (Serpins) are a large family of protease inhibitors involved in many critical biological processes such as blood coagulation, fibrinolysis, programmed cell death, development, and innate immunity. We identified MjSerp1, a serpin in the kuruma shrimp Marsupenaeus japonicus. The MjSerp1 cDNA has a 1239 bp open reading frame (ORF) that encodes a 412-amino acid protein with a 23 aa signal peptide and a classic serpin domain. MjSerp1 has a calculated molecular mass of 46.3 kDa and a predicted isoelectric point of 5.51. MjSerp1 is mainly expressed in the hepatopancreas and the intestines, and is moderately expressed in hemocytes. Expression pattern analysis indicated that MjSerp1 is upregulated in the hepatopancreas after Vibrio anguillarum challenge. rMjSerp1 inhibits three Gram-positive bacteria and two Gram-negative bacteria, but does not inhibit phenoloxidase activity. The microorganism binding assay showed that rMjSerp1 closely binds to both Gram-positive and Gram-negative bacteria. MjSerp1 also exhibits inhibitory activity against microbial serine proteases, such as subtilisin A and proteinase K, indicating that MjSerp1 acts as a microbial serine protease inhibitor. rMjSerp1 injection into shrimp enhances V. anguillarum clearance, but MjSerp1 knockdown through RNA interference impairs Vibrio clearance in vivo. These results indicate that MjSerp1 functions as a direct effector in the bacterial clearance of M. japonicus. All together, our findings provide novel evidences for the serine protease inhibitor in shrimp immunity.
Related JoVE Video
C-type lectin binds to ?-integrin to promote hemocytic phagocytosis in an invertebrate.
J. Biol. Chem.
PUBLISHED: 12-09-2013
Show Abstract
Hide Abstract
Phagocytosis is a conserved cellular response among metazoans. Opsonins are some molecules that label targets to increase their susceptibility to phagocytosis. Opsonins are usually captured by receptors on the surface of phagocytes. Our previous study found the C-type lectin FcLec4 from Chinese white shrimp Fenneropenaeus chinensis might function as an opsonin to facilitate bacterial clearance. In the present study, we purified the native FcLec4 protein and confirmed its opsonic activity in the near relation, kuruma shrimp Marsupenaeus japonicus. The possible receptor of FcLec4 was identified as ?-integrin by panning a T7 phage display library of shrimp hemocytes and then confirmed by co-immunoprecipitation assay. We further proved that the interaction between FcLec4 and ?-integrin did not rely on the carbohydrates recognition domain but on the N-terminus of FcLec4. In addition, inhibition of FcLec4 expression using RNAi delayed bacterial clearance, and ?-integrin knockdown suppressed the opsonic activity of FcLec4. This study is the first to show the direct interaction between an opsonin and its receptor in crustaceans. Our study provides new insights into invertebrate phagocytosis and the functions of C-type lectins.
Related JoVE Video
Prevalence and determinants of hyperuricemia in type 2 diabetes mellitus patients with central obesity in Guangdong Province in China.
Asia Pac J Clin Nutr
PUBLISHED: 11-16-2013
Show Abstract
Hide Abstract
This study investigated the prevalence and determinants of hyperuricemia in Chinese type 2 diabetes mellitus (T2DM) patients with central obesity. A multicentric hospital-based cross-sectional study was carried out in Guangdong Province between August 2011 and March 2012. At each hospital, Chinese T2DM patients with central obesity who were aged over 20 years, whose serum uric acid levels were measured, and who had lived in Guangdong Province for >=1 year, were recruited. Hyperuricemia was defined as serum uric acid >420 ?mol/L in men and >360 ?mol/L in women. Binary logistic regression was used to assess associated risk factors for hyperu-ricemia. A total of 2,917 T2DM patients with central obesity took part. The overall prevalence of hyperuricemia was 32.6% (36.1% for women, 28.4% for men). Binary logistic regression analyses demonstrated that women (OR: 1.576; 95% confidence interval (CI): 1.231, 2.018), high BMI (OR: 1.228; 95% CI: 1.094, 1.379), waist cir-cumference (OR: 1.135; 95% CI: 1.009, 1.276), hypertension (OR: 1.603; 95% CI: 1.263, 2.035), high total cho-lesterol (OR: 1.133; 95% CI: 1.002, 1.281), triglycerides (OR: 1.134; 95% CI: 1.069, 1.203), low HDL-cholesterol (OR: 0.820; 95% CI: 0.677, 0.995) and low estimated glomerular filtration rate (OR: 0.840; 95% CI: 0.815, 0.866) were risk factors associated with hyperuricemia. Hyperuricemia is prevalent in Chinese T2DM patients with central obesity and is significantly positively associated with women, cardiovascular risk factors such as obesity, hypertension and dyslipidemia, and low eGFR.
Related JoVE Video
Upregulation of the expression of prodeath serine/threonine protein kinase for programmed cell death by steroid hormone 20-hydroxyecdysone.
Apoptosis
PUBLISHED: 10-30-2013
Show Abstract
Hide Abstract
Serine/threonine protein kinases phosphorylate protein substrates to initiate further cellular events. Different serine/threonine protein kinases have varied functions despite their highly conserved homology. We propose prodeath-S/TK, a prodeath serine/threonine protein kinase from the lepidopteran insect Helicoverpa armigera, promotes programmed cell death (PCD) during metamorphosis. Prodeath-S/TK is expressed in various tissues with a high expression level during molting and metamorphosis by 20-hydroxyecdysone (20E) induction. Prodeath-S/TK is localized in the larval midgut during metamorphosis. Prodeath-S/TK knockdown by injecting dsRNA into larval hemocoel suppresses the 20E-induced metamorphosis and PCD, as well as downregulates a set of genes involved in the PCD and 20E signaling pathway. 20E upregulates prodeath-S/TK expression through its nuclear receptor EcR-B1 and USP1. Prodeath-S/TK overexpression in the epidermal cell line leads to PCD with DNA fragmentation and the activation of caspases 3 and 7. Prodeath-S/TK plays role in the cytoplasm. The N-terminal and C-terminal sequences of prodeath-S/TK determine its subcellular location. These data indicate that prodeath-S/TK participates in PCD by regulating gene expression in the 20E signaling pathway.
Related JoVE Video
[Analysis of associations between molecular subtypes and responses to neoadjuvant chemotherapy in primary breast cancer patients].
Zhonghua Yi Xue Za Zhi
PUBLISHED: 10-16-2013
Show Abstract
Hide Abstract
To explore the correlations between molecular subtypes and responses to neoadjuvant chemotherapy in primary breast cancer patients.
Related JoVE Video
Sub-acute exposure to the herbicide atrazine suppresses cell immune functions in adolescent mice.
Biosci Trends
PUBLISHED: 09-24-2013
Show Abstract
Hide Abstract
Atrazine (ATR), one of the most widely used herbicides worldwide, has caused a series of toxicological and environmental problems. This study sought to investigate the effects of ATR on the immune system of mice. Four-week-old female C57B l/6 mice were treated with 5, 25, and 125 mg/kg ATR for 28 days. On day 29, blood was collected and the spleen was harvested to detect lymphocyte transformation, natural killer (NK) cell activity, cellular phenotypes, and cytokines. Results indicated that the thymus and spleen weights decreased after ATR treatment, and the spleen was found to be more sensitive to ATR than the thymus. Decreases in lymphocyte transformation and NK cell activity were also observed in mice treated with 25 mg/kg ATR and 125 mg/kg ATR compared to the control group. In addition, there were also alterations of lymphocyte phenotypes in the spleen, and the percentages of CD3+ and CD4+ cells decreased in mice treated with 25 mg/kg ATR and 125 mg/kg ATR compared to the control group. Moreover, serum interleukin-4 level decreased significantly after treatment with 25 mg/kg and 125 mg/kg ATR, but ATR did not affect the expression of interleukin-2, interferon-?, and tumor necrosis factor-?. These results suggest that ATR may have induced damage in spleen cells. As ATR is an environmental contaminant, its immunosuppressive action raises concerns that it may potentiate clinical conditions such as tumors, inflammation, and infections. Thus, it needs to be carefully monitored and studied.
Related JoVE Video
A new type antimicrobial peptide astacidin functions in antibacterial immune response in red swamp crayfish Procambarus clarkii.
Dev. Comp. Immunol.
PUBLISHED: 09-22-2013
Show Abstract
Hide Abstract
A new antibacterial peptide called astacidin was characterized from hemocytes of red swamp crayfish Procambarus clarkii, and designated as PcAst. The full-length cDNA of PcAst contained 828 nucleotides with a polyadenylation sequence and a poly-A tail. PcAst encoded a peptide of 43 amino acids, with a signal peptide of 23 amino acids. The mature peptide contained 20 amino acids, among which four were proline/arginine amino acids. Similarity analysis showed that PcAst shared high identity with astacidin 2 from freshwater crayfish Pacifastacus leniusculus. Quantitative real-time PCR analysis showed that PcAst transcripts were mainly distributed in hemocytes and gills. The time-course expression analysis showed that after Vibrio anguillarum and Staphylococcus aureus injection, the transcripts of PcAst were upregulated in the gills. The synthetic small peptide for mature PcAst displayed inhibitory activity against the growth of some Gram-positive and Gram-negative bacteria. This peptide also had a binding ability to bacterial cell wall components, including peptidoglycan, lipopolysaccharide and lipoteichoic acid. PcAst functioned in the bacterial clearance immune reaction after V. anguillarum and S. aureus infection. These results indicate that PcAst has an important function in antibacterial innate immune response in red swamp crayfish P. clarkii.
Related JoVE Video
Prohibitin Interacts with envelope proteins of white spot syndrome virus and prevents infection in the red swamp crayfish, Procambarus clarkii.
J. Virol.
PUBLISHED: 09-18-2013
Show Abstract
Hide Abstract
Prohibitins (PHBs) are ubiquitously expressed conserved proteins in eukaryotes that are associated with apoptosis, cancer formation, aging, stress responses, cell proliferation, and immune regulation. However, the function of PHBs in crustacean immunity remains largely unknown. In the present study, we identified a PHB in Procambarus clarkii red swamp crayfish, which was designated PcPHB1. PcPHB1 was widely distributed in several tissues, and its expression was significantly upregulated by white spot syndrome virus (WSSV) challenge at the mRNA level and the protein level. These observations prompted us to investigate the role of PcPHB1 in the crayfish antiviral response. Recombinant PcPHB1 (rPcPHB1) significantly reduced the amount of WSSV in crayfish and the mortality of WSSV-infected crayfish. The quantity of WSSV in PcPHB1 knockdown crayfish was increased compared with that in the controls. The effects of RNA silencing were rescued by rPcPHB1 reinjection. We further confirmed the interaction of PcPHB1 with the WSSV envelope proteins VP28, VP26, and VP24 using pulldown and far-Western overlay assays. Finally, we observed that the colloidal gold-labeled PcPHB1 was located on the outer surface of the WSSV, which suggests that PcPHB1 specifically binds to the envelope proteins of WSSV. VP28, VP26, and VP24 are structural envelope proteins and are essential for attachment and entry into crayfish cells. Therefore, PcPHB1 exerts its anti-WSSV effect by binding to VP28, VP26, and VP24, preventing viral infection. This study is the first report on the antiviral function of PHB in the innate immune system of crustaceans.
Related JoVE Video
[Association of TET2, LMTK2 and FAM84B gene expression with prostate cancer risk in Chinese patients].
Zhonghua Zhong Liu Za Zhi
PUBLISHED: 08-30-2013
Show Abstract
Hide Abstract
To explore the association between the common variations of TET2 (rs7679673, A), MTK2 (rs6465657, T) and FAM84B (rs12543663, C) genes and prostate cancer (Pca) risk in Chinese population in Beijing, and to understand the relationship between genotypes and phenotypes including clinical characteristics and life style, etc. in patients with prostate cancer.
Related JoVE Video
[Study and translation of systemic control technology for the burn and trauma related lung injuries].
Zhonghua Shao Shang Za Zhi
PUBLISHED: 08-30-2013
Show Abstract
Hide Abstract
Burns and traumas are common injuries during both peacetime and wartime. Lung is the earliest organ subjected to dysfunction and the incidence is highest. The systemic protective technology for the burn and trauma related lung injuries is based on evidence-based medicine and translational medicine. It includes a series of effective measures, such as rescue and treatment scheme for massive burn casualties, prophylactic tracheostomy, protective ventilation strategy, sequential cell protection, and prevention and treatment of sequelae, which prevents aggravation of lung injuries caused by ischemia reperfusion, oxidative stress, and iatrogenic factors, as well as reduces the incidence of complications to ensure the recovery after burns and traumas.
Related JoVE Video
Two novel C-type lectins with a low-density lipoprotein receptor class A domain have antiviral function in the shrimp Marsupenaeus japonicus.
Dev. Comp. Immunol.
PUBLISHED: 07-29-2013
Show Abstract
Hide Abstract
C-type lectins (CTLs) are pattern-recognition receptors (PRRs) that play important roles in immune response. In this study, two new CTLs containing a low-density lipoprotein receptor class A domain (LDLR) and a carbohydrate recognition domain (CRD) were identified in Marsupenaeus japonicus and designated as LdlrLec1 and LdlrLec2. The two CTLs expressed in all the tested tissues of shrimp, however, LdlrLec1 was mainly expressed in hemocytes, heart, gill and intestines, whereas LdlrLec2 was expressed in hepatopancreas and heart. The expression patterns of both LdlrLec1 and LdlrLec2 mRNA were obviously upregulated upon white spot syndrome virus (WSSV) challenge. Injection of recombinant LdlrLec1 or LdlrLec2 into shrimp inhibited WSSV replication, whereas knocking down the expression of LdlrLec1 and LdlrLec2 by RNA interference increased WSSV replication in vivo. The infection rates of WSSV incubated with LdlrLecs were reduced significantly compared with the control group. The LdlrLec proteins could interact with VP28, a major envelope protein of WSSV, which is necessary for the attachment and penetration of WSSV into shrimp cells. These results indicate that LdlrLec1 and LdlrLec2 may function in antiviral response by binding to WSSV and inhibiting their pervasion and replication in shrimp.
Related JoVE Video
Deficiency of Smad3 results in enhanced inducible nitric oxide synthase-mediated hypotension in lipopolysaccharide-induced endotoxemia.
J. Surg. Res.
PUBLISHED: 07-23-2013
Show Abstract
Hide Abstract
Smad3 is a principal intracellular mediator of signaling for transforming growth factor ?, a cytokine involved in pleiotropic pathophysiological processes including inflammation and immunity. The function of Smad3 in regulating inducible nitric oxide synthase (iNOS) expression and septic shock has not been characterized.
Related JoVE Video
Bacteria activate sensory neurons that modulate pain and inflammation.
Nature
PUBLISHED: 07-17-2013
Show Abstract
Hide Abstract
Nociceptor sensory neurons are specialized to detect potentially damaging stimuli, protecting the organism by initiating the sensation of pain and eliciting defensive behaviours. Bacterial infections produce pain by unknown molecular mechanisms, although they are presumed to be secondary to immune activation. Here we demonstrate that bacteria directly activate nociceptors, and that the immune response mediated through TLR2, MyD88, T cells, B cells, and neutrophils and monocytes is not necessary for Staphylococcus aureus-induced pain in mice. Mechanical and thermal hyperalgesia in mice is correlated with live bacterial load rather than tissue swelling or immune activation. Bacteria induce calcium flux and action potentials in nociceptor neurons, in part via bacterial N-formylated peptides and the pore-forming toxin ?-haemolysin, through distinct mechanisms. Specific ablation of Nav1.8-lineage neurons, which include nociceptors, abrogated pain during bacterial infection, but concurrently increased local immune infiltration and lymphadenopathy of the draining lymph node. Thus, bacterial pathogens produce pain by directly activating sensory neurons that modulate inflammation, an unsuspected role for the nervous system in host-pathogen interactions.
Related JoVE Video
Characterization of an immune deficiency homolog (IMD) in shrimp (Fenneropenaeus chinensis) and crayfish (Procambarus clarkii).
Dev. Comp. Immunol.
PUBLISHED: 05-03-2013
Show Abstract
Hide Abstract
The immune deficiency (IMD) signal pathway mediates immunity against Gram-negative bacteria in Drosophila. Recent studies show that the IMD pathway also involves in antiviral innate immune responses. The functions of the pathway in crustacean immunity are largely unknown. In this paper, two IMDs (FcIMD and PcIMD), one of the key elements of the IMD pathway, were identified from Chinese white shrimp Fenneropenaeus chinensis and red swamp crayfish Procambarus clarkii. Both proteins have a death domain located at the C-terminal. FcIMD was mainly expressed in the gills and stomach and PcIMD was mainly detected in the heart, hepatopancreas, and stomach. FcIMD peaked in hemocytes at 12 h after white spot syndrome virus (WSSV) challenge and it peaked in the gills at 6 h after WSSV challenge, but it was decreased at 2 h and kept the low level to 24 h in hemocytes and no obviously change in gill after Vibrio anguillarum challenge. PcIMD first decreased in hemocytes at 2 h and peaked at 12 h in hemocytes after V. anguillarum challenge. It was also upregulated in gill after bacterial challenge, peaked at 2 h, and decreased at 6 h, and then gradually increased at 12-24 h. PcIMD has no significant change in hemocytes and gill after WSSV challenge. Western blot analysis detected FcIMD protein in all tissues, and immunocytochemical analysis localized FcIMD in the cytoplasm of hemocytes. RNA interference analysis showed that the IMD pathway was involved in regulating the expression of three kinds AMP genes, including crustins, anti-lipopolysaccharide factors and lysozymes, in shrimp and crayfish. They are Cru 1, Cru 2, ALF 1, ALF 2 and Lys 1 in crayfish, and Cru1, Cru 3, ALF 6, ALF 8, and Lys2 in shrimp. These results suggest that although IMD distribution and expression patterns have some differences, the IMD pathway may have conserved function for AMP regulation in shrimp and crayfish immunity against Gram-negative bacteria.
Related JoVE Video
Influencing factors of the quality of life in Chinese burn patients: Investigation with adapted Chinese version of the BSHS-B.
Burns
PUBLISHED: 05-02-2013
Show Abstract
Hide Abstract
The study aims to evaluate the quality of life (QOL) in burn patients in China and find out principal influencing factors, so as to provide evidence for interventions.
Related JoVE Video
A single whey acidic protein domain containing protein (SWD) inhibits bacteria invasion and dissemination in shrimp Marsupenaeus japonicus.
Fish Shellfish Immunol.
PUBLISHED: 04-04-2013
Show Abstract
Hide Abstract
The single whey acidic protein (WAP) domain containing proteins (SWDs) in crustacean belong to type III crustins and have antiprotease activities and/or antimicrobial activities. Their functions in vivo in crustacean immunity need to be clarify. In this study, a new single WAP domain containing protein (SWD) was obtained from Marsupenaeus japonicus, designated as MjSWD. The full-length cDNA of MjSWD was 522 bp.The open reading frame of MjSWD encoded a protein of 79 amino acids, with a 24 amino acid signal peptide and a WAP domain. Tissue distribution analysis revealed that MjSWD transcripts were generally expressed in all the tested tissues, including hemocytes, heart, hepatopancreas, gill, stomach and intestine. The time course expression of MjSWD was analyzed by quantitative real time PCR, and the results exhibited that MjSWD was upregulated after bacteria (Vibrio anguillarum, Staphylococcus aureus) and white spot syndrome virus (WSSV) challenge in gills and stomach of the shrimp. The purified recombinant protein of MjSWD could bind to several Gram-negative and Gram-positive bacteria though binding to microbial polysaccharides (peptidoglycan). MjSWD could inhibit the activity of Subtilisin A and Proteinase K and bacteria-secreted proteases. The results of natural infection with MjSWD incubated bacteria showed that the inhibition of MjSWD against bacterial secreted proteases was contributed to inhibiting bacteria invasion and dissemination in the shrimp. The MjSWD is, thus, involved in the shrimp antibacterial innate immunity.
Related JoVE Video
The hormone-dependent function of Hsp90 in the crosstalk between 20-hydroxyecdysone and juvenile hormone signaling pathways in insects is determined by differential phosphorylation and protein interactions.
Biochim. Biophys. Acta
PUBLISHED: 04-01-2013
Show Abstract
Hide Abstract
Heat shock protein 90 (Hsp90) interacts with steroid hormone receptors, signaling kinases, and various transcription factors. However, the mechanism by which Hsp90 interacts with different proteins in various pathways remains unclear.
Related JoVE Video
Macrophage migration inhibitory factor counter-regulates dexamethasone-induced annexin 1 expression and influences the release of eicosanoids in murine macrophages.
Immunology
PUBLISHED: 03-29-2013
Show Abstract
Hide Abstract
Macrophage migration inhibitory factor (MIF), a pro-inflammatory cytokine and glucocorticoid (GC) counter-regulator, has emerged as an important modulator of inflammatory responses. However, the molecular mechanisms of MIF counter-regulation of GC still remain incomplete. In the present study, we investigated whether MIF mediated the counter-regulation of the anti-inflammatory effect of GC by affecting annexin 1 in RAW 264.7 macrophages. We found that stimulation of RAW 264.7 macrophages with lipopolysaccharide (LPS) resulted in down-regulation of annexin 1, while GC dexamethasone (Dex) or Dex plus LPS led to significant up-regulation of annexin 1 expression. RNA interference-mediated knockdown of intracellular MIF increased annexin 1 expression with or without incubation of Dex, whereas Dex-induced annexin 1 expression was counter-regulated by the exogenous application of recombinant MIF. Moreover, recombinant MIF counter-regulated, in a dose-dependent manner, inhibition of cytosolic phospholipase A2? (cPLA2?) activation and prostaglandin E2 (PGE2 ) and leukotriene B4 (LTB4 ) release by Dex in RAW 264.7 macrophages stimulated with LPS. Endogenous depletion of MIF enhanced the effects of Dex, reflected by further decease of cPLA2? expression and lower PGE2 and LTB4 release in RAW 264.7 macrophages. Based on these data, we suggest that MIF counter-regulates Dex-induced annexin 1 expression, further influencing the activation of cPLA2? and the release of eicosanoids. These findings will add new insights into the mechanisms of MIF counter-regulation of GC.
Related JoVE Video
BAX inhibitor-1 silencing suppresses white spot syndrome virus replication in red swamp crayfish, Procambarus clarkii.
Fish Shellfish Immunol.
PUBLISHED: 03-27-2013
Show Abstract
Hide Abstract
BAX inhibitor-1 (BI-1) was originally described as an anti-apoptotic protein in both animal and plant cells. BI-1 overexpression suppresses ER stress-induced apoptosis in animal cells. Inhibition of BI-1 activity could induce the cell death in mammals and plants. However, the function of BI-1 in crustacean immunity was unclear. In this paper, the full-length cDNA of a BI-1 protein in red swamp crayfish, Procambarus clarkii (PcBI-1) was cloned and its expression profiles in normal and infected crayfish were analyzed. The results showed that PcBI-1 was expressed in hemocytes, heart, hepatopancreas, gills, stomach, and intestines of the crayfish and was upregulated after challenged with Vibrio anguillarum and with white spot syndrome virus (WSSV). To determine the function of PcBI-1 in the innate immunity of the crayfish, the RNA interference against PcBI-1 was performed and the results indicated the hemocyte programmed cell death rate was increased significantly and WSSV replication was declined after PcBI-1 knocked down. Altogether, PcBI-1 plays an anti-apoptotic role, wherein high PcBI-1 expression suppresses programmed cell death, which is beneficial for WSSW replication in crayfish.
Related JoVE Video
Distribution of Borrelia burgdorferi sensu lato in Ixodes ricinus populations across central Britain.
Vector Borne Zoonotic Dis.
PUBLISHED: 02-19-2013
Show Abstract
Hide Abstract
Lyme borreliosis is rapidly emerging in the United Kingdom, with over 1000 cases per annum now reported. Lyme borreliosis is caused by the Borrelia burgdorferi sensu lato (s.l.) group of spirochetes, which are transmitted by ixodid ticks. In the United Kingdom, Ixodes ricinus is recognized as the principal vector of the spirochetes, and this tick species is widely distributed across the country. However, as yet, it is unclear whether the distribution of B. burgdorferi essentially mirrors that of its vector, or if there are marked differences between the 2. The aim of this survey was to investigate the prevalence of B. burgdorferi in I. ricinus populations across northern England, north Wales, and the Scottish Border region. We surveyed for questing I. ricinus nymphs and adults at 17 sites, encountering ticks at 12. At 8 sites, large numbers (>160) ticks were collected, and at 3 of these sites B. burgdorferi infection prevalence was significantly higher (>7.5%) than the other 5 (<1.0%). Habitat type was associated with B. burgdorferi prevalence, with ticks from deciduous and mixed woodland being significantly more likely to be infected than those from other habitat types. Identification of infecting Borrelia species indicated that Borrelia valaisiana was the most common and widespread species encountered. B. garinii was common at sites where infection prevalence was high, and B. afzelii was also occasionally encountered at these sites. The survey revealed a surprisingly uneven distribution of B. burgdorferi s.l. across the region, thereby indicating that the presence of ticks does not necessarily mean the presence of the pathogen. Indeed, the spirochete appears to be absent or very rare at some sites where ticks are abundant.
Related JoVE Video
A Lysin motif (LysM)-containing protein functions in antibacterial responses of red swamp crayfish, Procambarus clarkii.
Dev. Comp. Immunol.
PUBLISHED: 02-07-2013
Show Abstract
Hide Abstract
Lysin domain (LysM) is a widely spread domain in nature and could bind different peptidoglycans and chitin-like compounds in bacteria and eukaryotes. In plants, Lysin motif containing proteins are one of the major classes of pattern recognition proteins which can recognize GlcNAc-containing glycans and have important functions in plant immunity. However, their functions in animal immunity are still unclear. In this study, a cDNA encoding a LysM containing protein was identified from red swamp crayfish, Procambarus clarkii. The cDNA of PcLysM contained 1200 base pair nucleotides with an open reading frame of 702bp encoding a protein of 233 amino acid residues. The deduced protein had a calculated molecular mass of 25.950kDa and a pI of 6.84. Tissue distribution analysis in mRNA level showed that it was highly expressed in gills, hemocytes, and intestine, and lowly expressed in hearts, hepatopancreas, and stomach. Time course expression pattern analysis showed that PcLysM was upregulated in hemocytes and gills after challenge with Vibrio anguillarum, and it was upregulated at 12h after challenge with Staphylococcus aureus in gills. The recombinant PcLysM could bind to different bacteria, and yeast. Further study revealed that PcLysM could bind to peptidoglycans from different bacteria, and chitin. After PcLysM was knocked down, the upregulation of antimicrobial peptide (AMP) genes (crustins and antilipopolysaccharide factors) was suppressed in response to bacterial infection in gills. These results suggest that PcLysM recognizes different microorganisms through binding to polysaccharides, such as peptidoglycans and chitin and regulates the expression of some antimicrobial peptide genes though unknown pathways and regulates the expression of some antimicrobial peptide genes though unknown pathways. This study might provide a clue to elucidate the roles of PcLysM in the innate immune reaction of crayfish P. clarkii.
Related JoVE Video
Steroid hormone 20-hydroxyecdysone regulation of the very-high-density lipoprotein (VHDL) receptor phosphorylation for VHDL uptake.
Insect Biochem. Mol. Biol.
PUBLISHED: 02-03-2013
Show Abstract
Hide Abstract
During the metamorphic stage of holometabolous insects, the biosynthetic precursors needed for the synthesis of a large number of adult proteins are acquired from the selective absorption of storage proteins. The very-high-density lipoprotein (VHDL), a non-hexameric storage protein, is consumed by the fat body from the hemolymph through VHDL receptor (VHDL-R)-mediated endocytosis. However, the mechanism of the uptake of VHDL by a VHDL-R remains unclear. In this study, a VHDL-R from Helicoverpa armigera was found to be involved in 20E-regulated VHDL uptake through the regulation of steroid hormone 20-hydroxyecdysone (20E). The transcripts of VHDL-R were detected mainly in the fat body and integument during the wandering stage. The transcription of VHDL-R was upregulated by 20E through the ecdysteroid receptor (EcRB1) and Ultraspiracle (USP1). In addition, 20E stimulates the phosphorylation of VHDL-R through protein kinase C for ligand binding. VHDL-R knockdown in larvae results the inhibition of development to adulthood. These data imply that 20E regulates VHDL-R on both transcriptional and posttranslational levels for VHDL absorption.
Related JoVE Video
Allelic variation of the MMP3 promoter affects transcription activity through the transcription factor C-MYB in human brain arteriovenous malformations.
PLoS ONE
PUBLISHED: 01-29-2013
Show Abstract
Hide Abstract
MMPs comprise a family of proteolytic enzymes that degrade pericellular substances, which may result in the destabilization of vessels and related to the development of brain arteriovenous malformations (BAVM). MMP3 is a key member of this family, overexpressed in BAVM tissues, and a single nucleotide polymorphism within MMP3, -709A>G (rs522616), is significantly associated with the risk of BAVM. In this study, we aimed to investigate the mechanism through which the polymorphism rs522616 regulates the expression of MMP3. Our results showed that -709A led to a over 2-fold higher transcriptional activity compared with the G allele (P<0.05) and this transcriptional activity can be depressed by co-transfecting cells with competitive DNA fragments containing -709A but not -709G. Bioinformatics analyses suggested that the transcription factor C-MYB might bind to the area around rs522616. Overexpressed C-MYB significantly increased the transcriptional activity of -709A compared with -709G or controls that did not overexpress c-myb (P<0.01) in HEK293 and HUVEC cells. ChIP assays indicated that C-MYB bound to the SNP region in the two cell lines and three BAVM tissue samples. Together, these data indicated that C-MYB can bind to the -709A allele of the MMP3 promoter, activate its transcription and lead to a higher expression of this gene. This novel hypothesis, supported by molecular evidence, explains how this SNP affects MMP3 promoter function and results in a risk of BAVM development.
Related JoVE Video
Overexpression of a C-type lectin enhances bacterial resistance in red swamp crayfish Procambarus clarkii.
Fish Shellfish Immunol.
PUBLISHED: 01-22-2013
Show Abstract
Hide Abstract
C-type lectins play important roles in the innate immune system of crustaceans. In this study, a novel C-type lectin gene, designated as PcLec4, was obtained from the red swamp crayfish (Procambarus clarkii). Quantitative real-time polymerase chain reaction revealed that PcLec4 is mainly expressed in the crayfish hepatopancreas and intestine, and the PcLec4 mRNA expression is upregulated after challenged with the bacteria Vibrio anguillarum. PcLec4 was recombinantly expressed in Escherichia coli and anti-PcLec4 polyclonal antiserum was prepared. Binding experiments revealed that the recombinant PcLec4 binds to various bacteria and polysaccharides on the bacterial surface, which suggests that PcLec4 recognizes bacterial pathogens. Overexpression of PcLec4 in crayfish using the pIeLec4 vector was performed. The results show that the crayfish overexpressing PcLec4 eliminate injected V. anguillarum more quickly than the control, which suggests that PcLec4 elicits further immune response for removing invading bacteria. The results of the survival experiment confirmed the function of PcLec4 in resisting V. anguillarum because PcLec4 overexpression in crayfish significantly increased the crayfish survival rate. These results reveal that PcLec4 has an important role in the antibacterial immunity of crayfish, and in vivo PcLec4 overexpression might be used as a disease control strategy in aquiculture.
Related JoVE Video
Local inflammation exacerbates the severity of Staphylococcus aureus skin infection.
PLoS ONE
PUBLISHED: 01-01-2013
Show Abstract
Hide Abstract
Staphylococcus aureus is the leading cause of skin infections. In a mouse model of S. aureus skin infection, we found that lesion size did not correlate with bacterial burden. Athymic nude mice had smaller skin lesions that contained lower levels of myeloperoxidase, IL-17A, and CXCL1, compared with wild type mice, although there was no difference in bacterial burden. T cell deficiency did not explain the difference in lesion size, because TCR ?? (-/-) mice did not have smaller lesions, and adoptive transfer of congenic T cells into athymic nude mice prior to infection did not alter lesion size. The differences observed were specific to the skin, because mortality in a pneumonia model was not different between wild type and athymic nude mice. Thus, the clinical severity of S. aureus skin infection is driven by the inflammatory response to the bacteria, rather than bacterial burden, in a T cell independent manner.
Related JoVE Video
Lipopolysaccharide-induced M2 to M1 macrophage transformation for IL-12p70 production is blocked by Candida albicans mediated up-regulation of EBI3 expression.
PLoS ONE
PUBLISHED: 01-01-2013
Show Abstract
Hide Abstract
Macrophages are heterogeneous cell populations that are present in all tissues. Macrophages can be divided into classically activated inflammatory macrophages (M1) and alternatively activated anti-inflammatory macrophages (M2). It has been generally accepted that M1 macrophages are polarised in an inflammatory environment to produce pro-inflammatory cytokines, whilst M2 macrophages are involved in anti-inflammation and aid tissue repair in wound healing. Bacterial endotoxin (lipopolysaccharide; LPS) is a potent factor in infection, which induces M1 macrophages resulting in higher levels of iNOS, TNF? and IL-12p70 which dictate inflammatory T cell responses. M2 macrophages can be transformed into M1 macrophages following LPS stimulation to promote inflammation. Candida albicans is a commensal fungal microorganism, which has been suggested to induce immune tolerance; however, the mechanism of C. albicans-induced immune tolerance has not been investigated in detail. IL-35 is a recently identified anti-inflammatory cytokine which is a heterodimeric protein consisting of the Epstein-Barr virus-induced gene 3 (EBI3) and IL-12p35. IL-35 shares the protein subunit p35, with IL-12p70. IL-12p70 is the most potent cytokine to induce Th1 responses during inflammation. In this study, we demonstrate that heat-killed C. albicans (HKC) strongly suppressed LPS-induced IL-12p70 production in M2 macrophages. Candida albicans induced a high level of EBI3 expression in M2 macrophages, which served as a mechanism for IL-12p70 suppression by competitive binding of the common protein subunit (p35) of IL-35 and IL-12p70. To demonstrate that EBI3 expression had the ability to block IL-12p70 production intracellularly, a Chinese Hamster Ovary (CHO) cell line with biscistronic expression of IL-12p40 and p35 was constructed, followed by ectopic over-expression of EBI3. The over-expression of EBI3 in the IL-12p70 producing cell line effectively suppressed IL-12p70 production. IL-35 secretion was also detected in the cell line, with suppressed IL-12p70 production by immune-precipitation Western blotting. However, this secretion was not evident in M2 macrophages following stimulation by HKC. This can be explained by the constitutive expression of IL-35 receptors (gp130 and IL-12R?2) in M2 macrophages for cytokine consumption. Our results have indicated that C. albicans can suppress host inflammatory responses in mucosal skin by suppressing LPS-induced IL-12p70 production. Lower IL-12p70 production may avoid an unnecessary Th1 response in order to retain immune tolerance, which may be one of the mechanisms by which C. albicans achieves a successful commensal lifestyle without having a detrimental effect on the hosts health.
Related JoVE Video
[Impact of response of positive axillary nodes to neoadjuvant chemotherapy on breast cancer survival].
Zhonghua Yi Xue Za Zhi
PUBLISHED: 11-19-2011
Show Abstract
Hide Abstract
To compare the distant disease-free survival between breast cancer patients with nodal pathological complete response (pCR) and those with nodal residual disease (RD) after neoadjuvant chemotherapy.
Related JoVE Video
[Clinicopathologic characteristics and prognosis in young Chinese women with breast cancer].
Zhonghua Yi Xue Za Zhi
PUBLISHED: 11-19-2011
Show Abstract
Hide Abstract
To analyze the clinicopathologic characteristics and evaluate the prognosis in young Chinese women with breast cancer.
Related JoVE Video
Unplanned decannulation of tracheotomy tube in massive burn patients: a retrospective case series study.
Chin. Med. J.
PUBLISHED: 11-18-2011
Show Abstract
Hide Abstract
Unplanned extubation is associated with adverse outcomes in intensive care unit. The massive burn patient differs from other critically ill patients in many ways. However, little is known about the unplanned decannulation (UD) in Burn Intensive Care Unit. This paper describes the special features of the circumstances and outcome of UD of tracheotomy tube in massive burn patients.
Related JoVE Video
Minocycline-induced attenuation of iron overload and brain injury after experimental intracerebral hemorrhage.
Stroke
PUBLISHED: 10-13-2011
Show Abstract
Hide Abstract
Brain iron overload plays a detrimental role in brain injury after intracerebral hemorrhage (ICH). A recent study found that minocycline acts as an iron chelator and reduces iron-induced neuronal death in vitro. The present study investigated if minocycline reduces iron overload after ICH and iron-induced brain injury in vivo.
Related JoVE Video
[The occupational and procreation health of immigrant female workers in electron factory].
Zhonghua Lao Dong Wei Sheng Zhi Ye Bing Za Zhi
PUBLISHED: 09-09-2011
Show Abstract
Hide Abstract
To explore the occupational and reproductive health problems of migrant female workers in electron factory.
Related JoVE Video
Bartonella quintana infections in captive monkeys, China.
Emerging Infect. Dis.
PUBLISHED: 09-06-2011
Show Abstract
Hide Abstract
Bartonella quintana has been considered to be specifically adapted to humans. Our isolation of the organism from 2 of 36 captive rhesus macaques in China and finding antibodies against B. quintana in 12 of 33 indicates that the reservoir hosts of B. quintana may include primates other than humans.
Related JoVE Video
A novel protein with a fibrinogen-like domain involved in the innate immune response of Marsupenaeus japonicus.
Fish Shellfish Immunol.
PUBLISHED: 08-31-2011
Show Abstract
Hide Abstract
Fibrinogen-related proteins play important roles in innate immunity. We isolated a fibrinogen-related protein gene (MjFREP1) in kuruma shrimp Marsupenaeus japonicus. MjFREP1 encoded a protein of 270 amino acids, including a 223 amino acid fibrinogen-like domain. Quantitative real-time polymerase chain reaction analysis shows that MjFREP1 is mainly expressed in the gills and the expression is significantly upregulated by Vibrio anguillarum, Staphylococcus aureus, or white spot syndrome virus (WSSV) challenge. Recombinant MjFREP1 fibrinogen-like domain agglutinates Gram-positive bacteria Bacillus subtilis, Bacillus thuringiensis, Bacillus megaterium, and S. aureus in the presence of calcium ions. The fibrinogen-like domain of MjFREP1 binds peptidoglycans, LPS, bacteria, and the VP28 of WSSV. These results suggest that the MjFREP1 may play an important role in the shrimp immune response against different pathogens.
Related JoVE Video

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.