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Articles by Xiangrui Li in JoVE

 JoVE Neuroscience

Enabling High Grayscale Resolution Displays and Accurate Response Time Measurements on Conventional Computers


JoVE 3312 2/29/2012

1Center for Cognitive and Behavioral Brain Imaging, The Ohio State University, 2Department of Psychology, University of Southern California, 3Biomedical Engineering, University of Southern California, 4Neuroscience Graduate Program, University of Southern California, 5Department of Psychology, The Ohio State University

Conventional computer hardware can not generate visual stimuli with sufficiently high grayscale resolution and measure response times with sufficient accuracy. We describe how to use the VideoSwitcher to produce high-resolution monochromatic displays, and the RTbox to measure response times with high accuracy on conventional computer hardware.

Other articles by Xiangrui Li on PubMed

Short-term Depression at Thalamocortical Synapses Contributes to Rapid Adaptation of Cortical Sensory Responses in Vivo

In vivo whole-cell recordings revealed that during repeated stimulation, synaptic responses to deflection of facial whiskers rapidly adapt. Extracellular recordings in the somatosensory thalamus revealed that part of the adaptation occurs subcortically, but because cortical adaptation is stronger and recovers more slowly, cortical mechanisms must also contribute. Trains of sensory stimuli that produce profound sensory adaptation did not alter intrinsic membrane properties, including resting membrane potential, input resistance, and current-evoked firing. Synaptic input evoked via intracortical stimulation was also unchanged; however, synaptic input from the somatosensory thalamus was depressed by sensory stimulation, and this depression recovered with a time course matching that of the recovery of sensory responsiveness. These data strongly suggest that synaptic depression of thalamic input to the cortex contributes to the dynamic regulation of neuronal sensitivity during rapid changes in sensory input.

Generating High Gray-level Resolution Monochrome Displays with Conventional Computer Graphics Cards and Color Monitors

Display systems based on conventional computer graphics cards are capable of generating images with about 8-bit luminance resolution. However, most vision experiments require more than 12 bits of luminance resolution. Pelli and Zhang [Spatial Vis. 10 (1997) 443] described a video attenuator for generating high luminance resolution displays on a monochrome monitor, or for driving just the green gun of a color monitor. Here we show how to achieve a white display by adding video amplifiers to duplicate the monochrome signal to drive all three guns of any color monitor. Because of the lack of the availability of high quality monochrome monitors, our method provides an inexpensive way to achieve high-resolution monochromatic displays using conventional, easy-to-get equipment. We describe the design principles, test results, and a few additional functionalities.

Genome-wide Analysis of MRNAs Regulated by the Nonsense-mediated and 5' to 3' MRNA Decay Pathways in Yeast

Transcripts regulated by the yeast nonsense-mediated and 5' to 3' mRNA decay pathways were identified by expression profiling of wild-type, upf1Delta, nmd2Delta, upf3Delta, dcp1Delta, and xrn1Delta cells. This analysis revealed that inactivation of Upf1p, Nmd2p, or Upf3p has identical effects on global RNA accumulation; inactivation of Dcp1p or Xrn1p exhibits both common and unique effects on global RNA accumulation but causes upregulation of only a small fraction of transcripts; and the majority of transcripts upregulated in upf/nmd strains are also upregulated to similar extents in dcp1Delta and xrn1Delta strains. Our results define the core transcripts regulated by NMD, identify several novel structural classes of NMD substrates, demonstrate that nonsense-containing mRNAs are primarily degraded by the 5' to 3' decay pathway even in the absence of functional NMD, and indicate that 3' to 5' decay, not 5' to 3' decay, may be the major mRNA decay activity in yeast cells.

Effects of Chinese Herbal Medicinal Ingredients on Peripheral Lymphocyte Proliferation and Serum Antibody Titer After Vaccination in Chicken

The purpose of these experiments is to study the effects of Chinese herbal medicinal ingredients (CHMIs) on peripheral lymphocyte proliferation and serum antibody titer in chicken vaccinated with Newcastle disease. Nine CHMIs were chosen for the experiments. Astragalus polysaccharide (APS), Isatis root polysaccharide (IRPS), Epimedium flavone (EF), Propolis flavone (PF), Astragalosides (AS) and Ginsenosides (GS) could promote lymphocyte proliferation and antibody titer, while Epimedium polysaccharide (EPS) mainly stimulated cellular immune responses. Chinese angelica polysaccharide (CAPS) and Propolis polysaccharide (PPS) exerted weaker effects on promoting immune responses. APS, IRPS, PPS and PF in promoting lymphocyte proliferation, and IRPS, PPS, EF and PF in promoting humoral immunity in higher dose were significantly stronger than in lower dose. Our results indicated that almost all of the nine CHMIs could promote both humoral and cellular immune responses and would be expected as the component drug of a new-type immunopotentiator.

Degradation of Response Modulation of Visual Cortical Cells in Cats with Chronic Exposure to Morphine

The primary visual cortex (V1) plays an important role in vision and visual perception. Studies in many brain regions demonstrate that opiate abuse can change excitatory and inhibitory neurotransmission. To investigate the effect of chronic morphine exposure on the response modulation of V1 simple and complex neurons, we carried out in vivo extracellular recordings in V1 of morphine- and saline-treated (control) cats. Response modulation was quantified as the ratio of first Fourier components (F1) to the mean response (F0). Compared with saline-treated cats, V1 neurons in morphine-treated cats exhibited weaker response modulation and a longer time course of response. The decrease of response modulation was caused by an increase of F0. Further, morphine re-exposure significantly improved the response properties of V1 neurons in morphine-treated cats. These results suggest that chronic morphine treatment leads to a significant degradation of response modulation of V1 neurons and a morphine dependence of primary visual cortical function.

Chronic Morphine Exposure Affects the Visual Response Properties of V1 Neurons in Cat

Chronic opiate exposure leads to maladaptive changes in brain function. In view of the localization of opiate receptors in mammalian visual system, chronic opiate exposure is likely to affect the visual responses properties of V1 neurons. Using in vivo single-unit recording, we here showed that chronic morphine treatment resulted in the functional abnormality of primary visual cortical cells. When compared with saline-treated (as control) cats, cortical neurons in morphine-treated cats exhibited higher spontaneous activity, lower signal-to-noise ratios and weaker orientation and direction selectivity. However, re-exposure with morphine could significantly improve the function of V1 neurons in morphine-treated cats. These findings demonstrated that chronic morphine treatment could significantly degrade the response properties of V1 neurons and may lead to a function dependence on morphine in visual cortical cells.

Functional Degradation of Visual Cortical Cells in Old Cats

Visual function declines with age. Using extracellular single-unit in vivo recordings, we compared the function of primary visual cortical (area 17) cells in young and old paralyzed, anesthetized cats. The results reveal that cortical neurons in old cats exhibit higher visually evoked responses, higher spontaneous activities, lower signal-to-noise ratios, and weaker orientation and direction selectivity than do cells in young adult cats. These findings are consistent with previously reported age related declines in cortical function in senescent macaque monkeys. Thus, similar declines in cortical function accompany old age in different mammalian species with well developed cortices.

Short-term Synaptic Plasticity in the Rat Geniculo-cortical Pathway During Development in Vivo

The critical period for visual system development in rats normally peaks at postnatal three weeks and ends at postnatal five weeks. However, the change of short-term synaptic plasticity during this period has rarely been investigated. In the present study, we compared the short-term plasticity of visual cortical responses to lateral geniculate nucleus stimulation in rats at different development stages (P20, P30 and adult) in vivo. The results show that paired-pulse depression (PPD) and frequency-dependent depression of evoked field potentials (FP) are present in P20 rats and increase in magnitude with development. The time course of this maturation of synaptic depression parallels that of the visual critical period. The weak synaptic depression observed in juvenile rats may be important in enhancing excitatory neurotransmission at a time when synapses are immature; this could endow immature synapses with wide integrative capabilities. In contrast, suppressive temporal interactions could provide an important substrate for neuronal processing of visual information in the mature cortex.

Characterization of HC58cDNA, a Putative Cysteine Protease from the Parasite Haemonchus Contortus

Because of the complexity of the cathepsin B-like (CBL) family, an information on the biological and biochemical characteristics of individual CBL genes is lacking. In this study, we investigated the degradative effects of the recombinant HC58 protein isolated from Haemonchus contortus parasites on protein substrates over a broad pH range in vitro. This protein, which hydrolyzed the synthetic peptide substrates Z-FR-AMC and Z-RR-AMC, had characteristics of the cysteine protease class of proteins. In the acidic pH range, the isolated protein actively degraded hemoglobin (Hb), the heavy chain of goat immunoglobulin G, and azocasein. By contrast, it degraded fibrinogen in the alkaline pH range. These activities were strongly inhibited in the presence of the cysteine protease inhibitor E-64. While the protein digested Hb, it did not induce the agglutination of erythrocytes from its natural host. These results suggest that the HC58 protein may play a role in the nutrition of this parasite.

Immunologic Synergism with IL-2 and Effects of CCHMIs on MRNA Expression of IL-2 and IFN-gamma in Chicken Peripheral T Lymphocyte

Two compound Chinese herbal medicinal ingredients (cCHMIs) were prepared, respectively, with epimedium polysaccharide plus propolis flavone (cCHMIs 1) and astragalus polysaccharide plus ginsenoside (cCHMIs 2). In animal immune experiment, Newcastle disease vaccine was mixed, respectively, with two cCHMIs and IL-2 to vaccinate 15-day-old chicken in experimental groups. On days 7, 14, 21 and 28 after vaccination, the dynamic changes of serum antibody titers were tested by micro-method. In gene expression experiment, each cCHMIs, at three concentrations, was added into cultured chicken peripheral T lymphocyte. After cultivation of 7h, the expression of IL-2 mRNA in the cell, 24h, IFN-gamma mRNA, were determined by semi-quantitative RT-PCR assay. The results showed that two cCHMIs, whether single or cooperative injection with IL-2, could significantly raise the antibody titers, while immunologic synergisms of two cCHMIs with IL-2 were unconspicuous. cCHMs 1 at three doses and cCHMIs 2 at high dose could remarkably promote the expression of IL-2 mRNA in chicken T lymphocyte, two cCHMIs at three doses, the expression of IFN-gamma mRNA, which could explain why the immunologic synergism of cCHMIs with IL-2 was not obvious.

Functional Degradation of Visual Cortical Cells in Aged Rats

Functional degradation of mammalian visual cortex is associated with aging. It has been hypothesized that much of the decline might be mediated by a degradation of cortical inhibitory system during senescence. In the present work, we compared the properties of adaptation, onset latency and signal-to-noise ratio in primary visual cortex of young and old rats using extracellular single-unit techniques. The short-term synaptic plasticity of young and old rats was also studied using field potential recording techniques. We found significant increased adaptation, prolonged onset latency, lower signal-to-noise ratio and decreased short-term synaptic plasticity in aged rats. The results are in accordance with previously reported functional declines in old monkeys and old cats, indicating a universal mechanism of degradation in cortical function that accompanies old age in different mammalian species.

Perceptual Learning Improves Contrast Sensitivity and Visual Acuity in Adults with Anisometropic Amblyopia

To evaluate the effects of perceptual learning on contrast-sensitivity function and visual acuity in adult observers with amblyopia, 23 anisometropic amblyopes with a mean age of 19.3 years were recruited and divided into three groups. Subjects in Group I were trained in grating detection in the amblyopic eye near pre-training cut-off spatial frequency. Group II received a training regimen of repeated contrast-sensitivity function measurements in the amblyopic eye. Group III received no training. We found that training substantially improved visual acuity and contrast-sensitivity functions in the amblyopic eyes of all the observers in Groups I and II, although no significant performance improvement was observed in Group III. For observers in Group I, performance improvements in the amblyopic eyes were broadly tuned in spatial frequency and generalized to the fellow eyes. The latter result was not found in Group II. In a few cases tested, improvements in visual acuity following training showed about 90% retention for at least 1 year. We concluded that the visual system of adult amblyopes might still retain substantial plasticity. Perceptual learning shows potential as a clinical tool for treating child and adult amblyopia.

Recombinant Galectins of Male and Female Haemonchus Contortus Do Not Hemagglutinate Erythrocytes of Their Natural Host

Recombinant galectins of female and male adult worms of Haemonchus contortus were expressed in Escherichia coli and their hemagglutinating activities to human and different animal erythrocytes were analyzed. The results showed that female and male galectins could be highly expressed in E. coli using a temperature-sensitive plasmid, with the recombinant protein being mainly appeared in inclusion bodies. Hemagglutinating activity assays showed that both of the galectins hemagglutinated human A, B, O type, dog, rabbit, chicken and mouse erythrocytes at the high concentration of 40 microg/well, but did not hemagglutinate erythrocytes of the natural host of H. contortus, the goat. Sugar inhibition assays confirmed that, out of eight sugars tested, only lactose was effective to inhibit agglutination of human type B erythrocytes by the recombinant galectins.

Cloning and Sequence Analysis of Hemonchus Contortus HC58cDNA

The complete coding sequence of Hemonchus contortus HC58cDNA was generated by rapid amplification of cDNA ends and polymerase chain reaction using primers based on the 5' and 3' ends of the parasite mRNA, accession no. AF305964. The HC58cDNA gene was 851 bp long, with open reading frame of 717 bp, precursors to 239 amino acids coding for approximately 27 kDa protein. Analysis of amino acid sequence revealed conserved residues of cysteine, histidine, asparagine, occluding loop pattern, hemoglobinase motif and glutamine of the oxyanion hole characteristic of cathepsin B like proteases (CBL). Comparison of the predicted amino acid sequences showed the protein shared 33.5-58.7% identity to cathepsin B homologues in the papain clan CA family (family C1). Phylogenetic analysis revealed close evolutionary proximity of the protein sequence to counterpart sequences in the CBL, suggesting that HC58cDNA was a member of the papain family.

Identification and Molecular Cloning of Putative Odorant-binding Proteins and Chemosensory Protein from the Bethylid Wasp, Scleroderma Guani Xiao Et Wu

Two putative odorant-binding proteins (OBPs) and one putative chemosensory protein (CSP) from females of the ant-like bethylid wasp, Scleroderma guani Xiao et Wu (Hymenoptera: Bethylidae), were identified and cloned. The putative OBPs and CSP were identified by nondenaturing polyacrylamide gel electrophoresis (native-PAGE). 3' rapid amplification of cDNA ends (3'RACE) was performed to obtain the sequences of the mature proteins by using degenerate primers designed from N-terminal sequences. Gene-specific primers for 5' rapid amplification of cDNA ends (5'RACE) were designed according to 3'RACE results and used in polymerase chain reaction (PCR) to obtain full-length sequences. The proteins (Sgua-OBP1, Sgua-OBP2, and Sgua-CSP1) encode 133, 142, and 129 amino acid-deduced sequences, respectively. Prediction of signal peptide sequences matches the N-terminal amino acid sequence of the isolated proteins. Database searches suggest that the Sgua-OBP1 and Sgua-OBP2 are homologs of OBPs from other insects, and Sgua-CSP1 shares a high level of identity with previously described CSPs.

The Second Glutamic Acid in the C-terminal CRD Affects the Carbohydrate-binding Properties of Recombinant Galectins of Haemonchus Contortus

The effects of the second glutamic acid (E) in the C-terminal CRDs on the hemagglutination and lactose-binding characteristics of the recombinant galectins of nematode Haemonchus contortus were observed using two isoforms of recombinant galectins as models, and the sugar-binding abilities of the N-terminal and C-terminal CRDs of the galectins were also compared. The second E in the CRD, WGNEER, of Hco-GAL-m was mutated to glycine acid (G) and resulted in a recombinant galectin (MG mutate) with a CRD of WGNEGR, identical to that of Hco-GAL-f. The G in Hco-GAL-f CRD, WGNEGR, was mutated to E and produced a recombinant galectin (FE mutate) equal to that of Hco-GAL-m. At the same time, the CRDs of the N-terminal (FNh,MNh) and C-terminal (FCh,MCh) of Hco-GAL-f, Hco-GAL-m were amplified by PCR. The abilities of carbohydrate binding and hemagglutination of the four galectins and the four CRDs were analysed, respectively, by alpha-lactose-agarose affinity chromatography and hemagglutination assay. The results showed that Hco-GAL-m and FE mutate bound effectively to alpha-lactose-agarose compared to Hco-GAL-f and MG mutate, which almost could not bind to the conjugate column. The hemagglutinating abilities of the Hco-GAL-m and FE mutate to human B type red blood cells were similar and were nearly two times higher than that of the Hco-GAL-f and MG mutate. The hemagglutinating ability of the MCh was five times to that of the MNh and FNh and almost two times to that of the FCh. The binding ability of the MCh and FCh were significantly reduced compared to that of the Hco-GAL-m and FE mutate, but still remained. As for the MNh and FNh, no elution peak was observed in the lactose-agarose affinity chromatography. These results suggested that the second amino acid E in the C-terminal CRD motif of H. contortus galectin was involved in carbohydrate binding and hemagglutination, and C-terminal CRDs had stronger carbohydrate ability than N-terminal CRDs.

Decreased Proportion of GABA Neurons Accompanies Age-related Degradation of Neuronal Function in Cat Striate Cortex

Electrophysiological studies indicate that a decline of GABAergic inhibition in the visual cortex may underlie age-related degradation of visual function [A.G. Leventhal, Y. Wang, M. Pu, Y. Zhou, Y. Ma, GABA and its agonists improved visual cortical function in senescent monkeys, Science 300 (2003) 812-815; M.T. Schmolesky, Y. Wang, M. Pu, A.G. Leventhal, Degradation of stimulus selectivity of visual cortical cells in senescent rhesus monkeys, Nat. Neurosci. 3 (2000) 384-390]. To date, there is little direct evidence to support this hypothesis. Using Nissl staining and immunohistochemical techniques, we quantitatively compared the density of total neurons (Nissl-stained neurons) and GABA-immunoreactive neurons as well as the proportion of GABA-immunoreactive neurons to total neurons in the primary visual cortex between 4 young adult (1-3 year old) cats and 4 old (12 year old) cats, which had been previously examined in a single-unit recording study [T. Hua, X. Li, L. He, Y. Zhou, Y. Wang, A.G. Leventhal, Functional degradation of visual cortical cells in old cats, Neurobiol. Aging 27 (2006) 155-162]. In that study, we found the function of V(1) (area 17) neurons in the old cats was significantly degraded relative to young adult cats. Our present results indicate that the density of total neurons in each cortical layer of V(1) exhibit no significant difference in the two age groups of cats. However, the density of GABA-immunoreactive neurons in old cats is significantly lower than in young adults. Further, the ratio of GABA-immunoreactive neurons to total neurons in each layer of V(1) in old cats is also significantly decreased when compared to young adult cats. These results provide direct morphological evidence of decreased GABAergic inhibition in the striate visual cortex of old animals, which accompany the functional degradation of visual cortical neurons.

Blood Oxygenation Level-dependent Contrast Response Functions Identify Mechanisms of Covert Attention in Early Visual Areas

Covert attention can lead to improved performance in perceptual tasks. The neural and functional mechanisms of covert attention are still under investigation. Using both rapid event-related and mixed designs, we measured the blood oxygenation level-dependent functional MRI contrast response functions over the full range of contrast (0-100%) in the retinotopically defined early visual areas (V1, V2, V3, V3A, and V4) in humans. Covert attention increased both the baseline activities and contrast gains in the five cortical areas. The effect on baseline can be decomposed into a transient trial-by-trial component and a component across an entire attention block. On average, increase in contrast gain accounted for approximately 88.0%, 28.5%, 12.7%, 35.9%, and 25.2% of the trial-by-trial effects of attention in the five areas, respectively, and 22.2%, 12.8%, 7.4%, 19.7%, and 17.3% of the total effects of attention in those areas, consistent with single-unit findings in V4 and MT. The results provide strong evidence for a stimulus enhancement mechanism of attention as demonstrated in various behavioral studies.

Vaccination of Chickens with a Chimeric DNA Vaccine Encoding Eimeria Tenella TA4 and Chicken IL-2 Induces Protective Immunity Against Coccidiosis

A fusion DNA vaccine co-expressed Eimeria tenella TA4 and chicken IL-2 (chIL-2) was constructed and its efficacy against E. tenella challenge was observed. TA4 gene of E. tenella and chIL-2 gene were cloned into expression vector pcDNA3.1 and pcDNA4.0c in different forms, producing vaccines pcDNA3.1-TA4-IL-2, pcDNA3.1-TA4 and pcDNA4.0c-IL-2. The expression of aim genes in vivo was detected by RT-PCR and western blot. Animal experiment was carried out to evaluate the immune efficacy of the vaccines. Results indicated these DNA vaccines were successfully constructed and the antigen genes could be expressed effectively in vivo. The animal experimental results showed that DNA vaccines could obviously alleviate cecal lesions, body weight loss and increase oocyst decrease ratio. The ACI of pcDNA3.0-TA4-IL-2 group was 192, higher than that of pcDNA3.1-TA4 group. The results suggested that TA4 was an effective candidate antigen for vaccine and co-expression of cytokine with antigen was an alternative method to enhance DNA vaccine immunity.

Functional Dissociations of Risk and Reward Processing in the Medial Prefrontal Cortex

Making a risky decision is a complex process that involves evaluation of both the value of the options and the associated risk level. Yet the neural processes underlying these processes have not so far been clearly identified. Using functional magnetic resonance imaging and a task that simulates risky decisions, we found that the dorsal region of the medial prefrontal cortex (MPFC) was activated whenever a risky decision was made, but the degree of this activity across subjects was negatively correlated with their risk preference. In contrast, the ventral MPFC was parametrically modulated by the received gain/loss, and the activation in this region was positively correlated with an individual's risk preference. These results extend existing neurological evidence by showing that the dorsal and ventral MPFC convey different decision signals (i.e., aversion to uncertainty vs. approach to rewarding outcomes), where the relative strengths of these signals determine behavioral decisions involving risk and uncertainty.

The Optimal Immunization Procedure of DNA Vaccine PcDNA-TA4-IL-2 of Eimeria Tenella and Its Cross-immunity to Eimeria Necatrix and Eimeria Acervulina

The immunization procedure of DNA vaccine pcDNA-TA4-IL-2 of Eimeria tenella, including route, dose, time of immunization and age of primary immunization of chicken, was optimized. The stability and the cross-species protection of the vaccine were also analyzed. Efficacy of immunization was evaluated on the basis of oocyst decrease ratio, lesion score, body-weight gain and the anti-coccidial index (ACI). Chinese Yellow chickens were randomly distributed into corresponding groups (30/group). The challenged, unchallenged and vector control groups were designed. The results illustrated that 25 microg was the optimal dose and intramuscular injection was the most effective route to induce protective immunity. There were no significant differences of ACIs between boosting and non-boosting groups. Storage time and temperature had little effect on the immunizing efficacy of the vaccine. The vaccine could provide partial cross-protection against the challenge with E. necatrix and E. acervulina, but not with E. maxima.

Neural Correlates of Risk Prediction Error During Reinforcement Learning in Humans

Behavioral studies have shown for decades that humans are sensitive to risk when making decisions. More recently, brain activities have been shown to be correlated with risky choices. But an important gap needs to be filled: How does the human brain learn which decisions are risky? In cognitive neuroscience, reinforcement learning has never been used to estimate reward variance, a common measure of risk in economics and psychology. It is thus unknown which brain regions are involved in risk learning. To address this question, participants completed a decision-making task during fMRI. They chose repetitively from four decks of cards and each selection was followed by a stochastic payoff. Expected reward and risk differed among the decks. Participants' aim was to maximize payoffs. Risk and reward prediction errors were calculated after each payoff based on a novel reinforcement learning model. For reward prediction error, the strongest correlation was found with the BOLD response in the striatum. For risk prediction error, the strongest correlation was found with the BOLD responses in the insula and inferior frontal gyrus. We conclude that risk and reward prediction errors are processed by distinct neural circuits during reinforcement learning. Additional analyses revealed that the BOLD response in the inferior frontal gyrus was more pronounced for risk aversive participants, suggesting that this region also serves to inhibit risky choices.

Immunoproteomic Analysis of the Second-generation Merozoite Proteins of Eimeria Tenella

Whole proteins of the second-generation merozoite of Eimeria tenella were analyzed by two-dimensional gel electrophoresis (2-DE) and western blot using the chicken sera infected artificially with E. tenella. Approximately 640 spots were detected on proteome map of the second-generation merozoite stained by Coomassie brilliant blue G-250 and 85 spots were recognized on western blot map as antigens. Forty four spots of the antigens were identified by matrix-assisted laser desorption ionization time-of-flight MS (MALDI-TOF-MS) and MALDI-TOF-TOF-MS. Twenty six proteins of E. tenella and three homologous proteins to other apicomplexan parasites or protozoan were identified using 'Mascot' server. These proteins included lactate dehydrogenase, enolase, 14-3-3 protein, microneme proteins, tubulin beta chain, SERPIN1 protein precursor, large subunit ribosomal protein L23 and surface antigens of E. tenella, heat shock protein (HSP70) of Eimeria acervulina, protein phosphatase type 1 of Toxoplasma gondii and hypothetical protein GSPATT00020155001 of Paramecium tetraurelia. The rest proteins were searched against the E. tenella protein sequence database using 'Mascot in-house' (version 2.1) search engine in automated mode and 11 unknown proteins were identified. After the amino acid sequence of the unknown proteins were searched using BLAST against non-redundant sequence databases (NCBI), surface antigen 12 of E. tenella and six homologous proteins to other apicomplexa parasites were found. They were membrane skeleton protein IMC2A, mitochondrial F1-ATP synthase beta subunit precursor, 3-oxoacyl-acyl-carrier-protein synthase and catalase of T. gondii, Vps26 of Plasmodium falciparum 3D7, and hypothetical protein TRIADDRAFT_60424 of Trichoplax adhaerens. No homologues of 8 spots of the 44 analyzed proteins were found. These proteins enriched the data of immunogenic proteins of the second-generation merozoite of E. tenella.

The Iowa Gambling Task in FMRI Images

The Iowa Gambling Task (IGT) is a sensitive test for the detection of decision-making impairments in several neurological and psychiatric populations. Very few studies have employed the IGT in functional magnetic resonance imaging (fMRI) investigations, in part, because the task is cognitively complex. Here we report a method for exploring brain activity using fMRI during performance of the IGT. Decision-making during the IGT was associated with activity in several brain regions in a group of healthy individuals. The activated regions were consistent with the neural circuitry hypothesized to underlie somatic marker activation and decision-making. Specifically, a neural circuitry involving the dorsolateral prefrontal cortex (for working memory), the insula and posterior cingulate cortex (for representations of emotional states), the mesial orbitofrontal and ventromedial prefrontal cortex (for coupling the two previous processes), the ventral striatum and anterior cingulate/SMA (supplementary motor area) for implementing behavioral decisions was engaged. These results have implications for using the IGT to study abnormal mechanisms of decision making in a variety of clinical populations.

Cross Immunity of DNA Vaccine PVAX1-cSZ2-IL-2 to Eimeria Tenella, E. Necatrix and E. Maxima

The study describes cross protection experiments with chimeric DNA vaccine pVAX1-cSZ2-IL-2 to determine its efficacy against four important Eimeria species. Seven-day-old chickens were randomly divided into nine groups; group 1 negative control, groups 2, 3, 4, 5 positive controls; and groups 6, 7, 8 and 9 experimental groups. On days 7 and 14, groups 1-5 were injected with TE buffer, and groups 6-9 with the vaccine. At 21 days of age, all chickens were inoculated with 5 x 10(4) sporulated oocysts except for the negative control. Groups 2 and 6 were inoculated with Eimeria tenella, groups 3 and 7 with Eimerianecatrix, groups 4 and 8 with Eimeria acervulina and groups 5 and 9 with Eimeria maxima. Seven days later, all chickens were weighed and slaughtered to obtain intestinal samples. Efficacy of immunization was evaluated on the basis of oocyst decrease ratio, lesion score, body-weight gain and anti-coccidial index. The results indicated that the recombinant plasmid can induce host immune responses by alleviating intestinal lesions, body weight loss and oocyst ratio and imparting good protection against E. tenella and E.acervulina, medium protection against E. necatrix but little effect against E. maxima. It is concluded that the conserved antigen can provide cross protection and should be explored further.

A Recombinant DNA Vaccine Encoding Eimeria Acervulina CSZ-2 Induces Immunity Against Experimental E. Tenella Infection

The study describes vaccination experiments with highly immunogenic sporozoite Eimeria acervulina cSZ-2 to determine its efficacy against E. tenella challenge. Chickens were randomly divided into six groups. The chickens in the vaccinated groups were vaccinated with pVAX1-cSZ2, pVAX1-IL-2 and pVAX1-cSZ2-IL-2 twice 'at days 14 and 21' with dose rate of 100microg intramuscularly. At 28 days of age, all chickens except the non-vaccinated non-challenged control group were inoculated orally with 5x10(4) sporulated oocysts of E. tenella. Seven days later, all chickens were weighed and slaughtered for caecum examination. Our findings make it obvious that cSZ-2 DNA immunisation can induce host immune responses by decreasing intestinal lesions, body weight loss and oocyst ratio, imparting partial protection. It is concluded that co-administration of interleukin 2 can further enhance its immunity, leading towards further research.

RTbox: a Device for Highly Accurate Response Time Measurements

Although computer keyboards and mice are frequently used in measuring response times (RTs), the accuracy of these measurements is quite low. Specialized RT collection devices must be used to obtain more accurate measurements. However, all the existing devices have some shortcomings. We have developed and implemented a new, commercially available device, the RTbox, for highly accurate RT measurements. The RTbox has its own microprocessor and high-resolution clock. It can record the identities and timing of button events with high accuracy, unaffected by potential timing uncertainty or biases during data transmission and processing in the host computer. It stores button events until the host computer chooses to retrieve them. The asynchronous storage greatly simplifies the design of user programs. The RTbox can also receive and record external signals as triggers and can measure RTs with respect to external events. The internal clock of the RTbox can be synchronized with the computer clock, so the device can be used without external triggers. A simple USB connection is sufficient to integrate the RTbox with any standard computer and operating system.

Haemonchus Contortus: Cloning and Characterization of Serpin

The serpin gene of Haemonchus contortus (hc-serpin) was cloned and characterized in this study. Specific primers for rapid amplification cDNA ends (RACE) were designed based on the expression sequence tag (EST, BM173953) to amplify the 3'- and 5'-ends of hc-serpin. The full length of the cDNA of this gene was obtained by overlapping the sequences of 3'- and 5'-extremities and amplification by reverse transcription-PCR. The biochemical activities of the recombinant protein (rHc-Serpin), which was expressed in prokaryotic cells and purified by affinity chromatography and size-exclusion chromatography, were analyzed by assays of trypsin inhibition, anti-coagulation activity, and stability to temperature and pH. The results showed that the cloned full-length cDNA comprised 1317bp and encoded a peptide with 367 amino acid residues which showed sequence similarity to several known serpins. The rHc-Serpin inhibited trypsin activity effectively and prolonged the coagulation time of rabbit blood in vitro. The rHc-Serpin was stable from pH 2.0-10.0 and kept activity at high temperature until 75 degrees C. Optimal pH of rHc-Serpin protein to inhibit trypsin activity was at pH 7.6. The natural serpin of H. contortus detected by immunoblot assay was about 63kDa, and the rHc-Serpin was recognized strongly by serum from naturally infected goats. By immunohistochemistry, the serpin was localised exclusively in the epithelial cells of gastrointestinal tract in adult H. contortus. The results indicated that the cloned gene was serpin and that the protein may play important roles in the biological functions of H. contortus.

Efficacy of DNA Vaccines Carrying Eimeria Acervulina Lactate Dehydrogenase Antigen Gene Against Coccidiosis

The efficacies of DNA vaccines encoding either Eimeria acervulina lactate dehydrogenase (LDH) antigen or a combination of LDH antigen and chicken IL-2 or IFN-gamma were evaluated against chicken coccidiosis. Three vaccine plasmids pVAX-LDH, pVAX-LDH-IFN-gamma and pVAX-LDH-IL-2 were constructed using the eukaryotic expression vector pVAX1. Expressions of proteins encoded by plasmids DNA in vivo were detected by reverse transcription-polymerase chain reaction (RT-PCR) and western blot assay. Average body weight gain, oocyst output, survival rate and lesion scores were measured to evaluate the protective effects of vaccination on challenge infection. The results showed that DNA vaccines could obviously alleviate body weight loss, duodenal lesions, oocyst output and enhance oocyst decrease ratio. Anti-coccidial indexes (ACIs) of pVAX-LDH-IFN-gamma and pVAX-LDH-IL-2 groups were higher than that of other groups. Flow cytometric analysis of T lymphocytes in spleen and cecal tonsil demonstrated that DNA vaccines had significantly increased percentages of CD3(+) T cells compared with pVAX1 alone or TE buffer. The results provided the first proof that DNA vaccine carrying E. acervulina LDH antigen gene induced protective immunity against homologous infection and its effect could be enhanced by co-expression of chicken IL-2 or IFN-gamma.

The DNA-induced Protective Immunity with Chicken Interferon Gamma Against Poultry Coccidiosis

The immunogenicity of Eimeria acervulina cSZ-2 and chicken interferon gamma was observed against Eimeria tenella challenge. The chickens were randomly divided into six groups of 24 chicks each. Three groups of chickens were injected with DNA vaccines pVAX1-cSZ2, pVAX1-chIFN-gamma and pVAX1-cSZ2-chIFN-gamma two times (at days 14 and 21) at a dose of 100 microg intramuscularly. Three other groups were kept as control and injected with TE buffer (10 mM Tris-HCl pH 7.6 and 1 mM EDTA). One week following the booster dose, all chickens except the non-infected, non-vaccinated control group were inoculated orally with 5 x 10(4) sporulated oocysts of E. tenella. Seven days post challenge, all chickens were weighted and slaughtered for cecal lesion scoring and oocyst counts. The results demonstrated that cSZ-2 in combination with interferon gamma can protect chickens from coccidiosis by significantly decreasing body weight loss and oocyst excretion reflecting partial protection against E. tenella infection, and further studies are necessary to test for protection against other Eimeria species.

[Cloning, Expression and Characterization of a Gene Encoding Alpha2 Subunit of the Proteasome in Schistosoma Japonicum]

The 26S proteasome is a proteolytic complex responsible for the degradation of the vast majority of eukaryotic proteins. Regulated proteolysis by the proteasome is thought to influence cell cycle progression, transcriptional control, and other critical cellular processes. A novel Schistosoma japonicum gene (GenBank Accession No. AY813725) proteasome alpha2 subunit (SjPSMA2) was cloned. Sequence analysis revealed that the ORF of SjPSMA2 gene contains 708 nucleotides encoding 235 amino acids, and the molecular weight was estimated to be 25.84 kDa. Real-time PCR analysis showed that this gene expressed in 7 d, 13 d, 18 d, 23 d, 32 d and 42 d schistosoma. The mRNA level of SjPSMA2 was lower in 7 d and 23 d schistosomulum than that in other stages. The SjPSMA2 cDNA fragment was subcloned into an expression vector pET28a(+) and transformed into E. coli BL21 (DE3) cells. After induction with IPTCQ the 30 kDa fusion protein was produced as included bodies. Western-blotting revealed that the fusion protein could be recognized by the rabbit serum anti-Schistosoma japonicum adult worm antigen preparation, and the protein in native could be detected. After immunization of BALB/c mice with the fusion protein, the reduction rates of worm counts and liver egg counts were 12.33% and 35.23%. ELISA results revealed that the vaccinated group showed a significant increase in the level of IgG antibody. This study provided an important basis for investigating the regulation mechanism of the proteasome during the development of Schistosoma japonicum.

Schistosoma Japonicum: Cloning, Expression and Characterization of a Gene Encoding the α5-subunit of the Proteasome

The development of an effective vaccine against the schistosome is thought to be the most desirable means to control schistosomiasis, even though there is an effective means of chemotherapy with praziquantel. A full-length cDNA encoding the Schistosoma japonicum proteasome subunit alpha type 5 protein (SjPSMA5) was first isolated from 18-day-schistosomulum cDNAs. The cDNA had an open reading frame (ORF) of 747bp and encoded 248 amino acids. Real-time quantitative RT-PCR analysis revealed that SjPSMA5 is up-regulated in 18-day and 32-day schistosomes, and the level of expression in male is around fourfold higher than that in female worms at 42 days. The SjPSMA5 was subcloned into pET28a(+) and expressed as inclusion bodies in Escherichia coli BL21 (DE3) cells. Western blotting showed that the recombinant SjPSMA5 (rSjPSMA5) was immunogenic. After immunization of BALB/c mice with rSjPSMA5, reductions of 23.29% and 35.24% were obtained in the numbers of worms and eggs in the liver, respectively. The levels of specific IgG antibodies and CD(4)(+) cells were significantly higher (P<0.01) in the group vaccinated with rSjPSMA5 combined with Seppic 206 adjuvant than in the other groups, as detected by enzyme linked immunosorbent assay (ELISA) and flow cytometry. The study suggested that rSjPSMA5 induced partial immunoprotection against S. japonicum in BALB/c mice, and it could be a potential vaccine candidate against schistosomiasis.

Changes of Cytokines and IgG Antibody in Chickens Vaccinated with DNA Vaccines Encoding Eimeria Acervulina Lactate Dehydrogenase

The aim of this study was to investigate the changes of cytokines and specific serum IgG in chickens following vaccination with DNA vaccines encoding either Eimeria acervulina (E. acervulina) lactate dehydrogenase (LDH) antigen or LDH and chicken IL-2 or IFN-γ. Two-week-old chickens were randomly divided into five groups. Experimental group of chickens were immunized with DNA vaccines while control group of chickens were injected with pVAX1 plasmid alone or sterile water. All immunizations were boosted 2 weeks later. The LDH-specific IgG antibody response was measured at weeks 1-6 post-second immunization. The result showed that the antibody titers in chickens vaccinated with DNA vaccines were significantly different from those of the control groups 1 week after the second immunization (P<0.05) and reached the maximum values 3 weeks post-second immunization. The systemic and local cytokine mRNA expression was determined by quantitative RT-PCR 7 days post-second immunization. The specific IgG antibody levels against LDH of all chickens vaccinated with vaccines were increased compared to those of sterile water (H(2)O) and plasmid (pVAX1) control chickens 1-6 weeks post-second immunization (P<0.05). The mRNA levels of IFN-γ, IL-2, TNFSF15, IL-17D as well as TGF-β4 in both spleen and cecal tonsil were also increased in experimental chickens. In contrast, the only significant change of IL-4 mRNA level was observed in spleen of chickens immunized with pVAX-LDH-IL-2 compared with pVAX-LDH and control groups (P<0.05). These results suggested that DNA vaccines could increase the IgG antibody level and induce the expressions of cytokines.

Immunoproteomic Analysis of Whole Proteins from Male and Female Adult Haemonchus Contortus

Whole proteins of male and female adult Haemonchus contortus were analysed by immunoproteomic techniques. Approximately 662 and 680 spots were detected on proteome maps of male and female nematodes, respectively, stained with Coomassie brilliant blue G-250. There were 609 shared spots. Approximately 193 and 196 spots were recognised on Western blot maps of male and female nematodes, respectively, using antiserum from naturally infected goats as the source of primary antibodies. There were 129 gender-specific spots in male nematodes and 132 in females. Twenty-three shared immunogenic spots were identified by MALDI-TOF or MALDI-TOF-TOF mass spectrometry. These proteins included glutamate dehydrogenase (GDH), homologues of Dim-1, actin, globin-like excretory/secretory protein F6, glutathione S-transferase (GST), ATPase and glyceraldehyde-3-phosphate dehydrogenase. GDH and GST have been identified as immunogenic proteins of H. contortus previously, whereas the other proteins are newly recognised immunogenic proteins in this nematode.

Vaccination of Chickens with DNA Vaccine Expressing Eimeria Tenella MZ5-7 Against Coccidiosis

A chimeric DNA vaccine co-expressing Eimeria tenella MZ5-7 and chicken IL-17 gene was constructed and its efficacy against E. tenella challenge was evaluated. The ORF of MZ5-7 from E. tenella's second generation merozoite and the mature interleukin 17 gene of chicken were cloned into the expression vector of pcDNA4.0 to construct DNA vaccine pcDNA4.0-MZ and pcDNA4.0-MZ-IL17. The expression of aim gene products in vivo was detected by western blotting. The expression of IL-2 and IFN-γ in chicken splenocytes was detected by RT-PCR 7 days post-immunization. The expression levels of the two cytokines in the pcDNA4.0-MZ-IL17 DNA vaccine immunized group were significantly higher than that in the pcDNA4.0-MZ immunized group (p<0.05). Either pcDNA4.0-MZ or pcDNA4.0-MZ-IL17 DNA vaccine could obviously alleviate cecal lesions, body weight loss and increase oocyst decrease ratio. The ACI of pcDNA4.0-MZ-IL17 group was 190, which is higher than that of pcDNA4.0-MZ group and all the three control groups. In short, MZ5-7 was an effective candidate antigen for vaccine and co-expression of cytokine with antigen was an alternative method to enhance the immunity DNA vaccine.

Construction of DNA Vaccines Encoding Eimeria Acervulina CSZ-2 with Chicken IL-2 and IFN-γ and Their Efficacy Against Poultry Coccidiosis

The study describes vaccination experiments with highly immunogenic sporozoite E. acervulina cSZ-2 co-administered with chicken IL-2 (chIL-2) and interferon-γ (chIFN-γ) to determine their efficacies against homologue challenge. The entire coding sequence of cSZ2, chIL-2 and chIFN-γ were cloned into eukaryotic expression vector pVAX1, constructing DNA vaccines pVAX1-cSZ2, pVAX1-chIL-2, pVAX1-chIFN-γ, pVAX1-cSZ2-chIL-2 and pVAX1-cSZ2-chIFN-γ. The expression of target genes in vivo was detected by RT-PCR and Western blot. Chicken experiments were carried out by vaccinating chickens two times at dose rate of 100 μg intramuscularly. At 28 days of age, all chickens were inoculated orally with 1×10(5) sporulated oocysts of E. acervulina except the unchallenged control group. Seven days after challenge, all chickens were weighted and slaughtered for duodenum collection. The results indicated that these DNA vaccines were successfully constructed and the antigen genes could be expressed effectively in vivo. The findings also demonstrated best synergistic effect of IL-2 with this protein which suggested that co-administration of cytokines with this antigen was a powerful method to enhance immunity by alleviating intestinal lesions, body weight loss and oocyst count imparting partial protection against homologous challenge.

Attention Extracts Signal in External Noise: a BOLD FMRI Study

On the basis of results from behavioral studies that spatial attention improves the exclusion of external noise in the target region, we predicted that attending to a spatial region would reduce the impact of external noise on the BOLD response in corresponding cortical areas, seen as reduced BOLD responses in conditions with large amounts of external noise but relatively low signal, and increased dynamic range of the BOLD response to variations in signal contrast. We found that, in the presence of external noise, covert attention reduced the trial-by-trial BOLD response by 15.5-18.9% in low signal contrast conditions in V1. It also increased the BOLD dynamic range in V1, V2, V3, V3A/B, and V4 by a factor of at least three. Overall, covert attention reduced the impact of external noise by about 73-85% in these early visual areas. It also increased the contrast gain by a factor of 2.6-3.8.

Proteomic Analysis of Schistosoma Japonicum Schistosomulum Proteins That Are Differentially Expressed Among Hosts Differing in Their Susceptibility to the Infection

Schistosomiasis is a tropical, parasitic disease affecting humans and several animal species. The aim of this study was to identify proteins involved in the growth and survival of the parasitic forms inside a host. Schistosomula of Schistosoma japonicum were isolated from three different hosts: the susceptible BALB/c mice; the Wistar rats, which have a considerably lower susceptibility; and the resistant reed vole, Microtus fortis. Soluble proteins of the schistosomula collected from the above three hosts 10 days postinfection were subjected to two-dimensional difference gel electrophoresis. Comparative proteomic analyses revealed that 39, 21, and 25 protein spots were significantly differentially expressed between schistosomula from mice and rats, mice and reed voles, or rats and reed voles, respectively (ANCOVA, p < 0.05). Further, the protein spots were identified by liquid chromatography-tandem MS. Bioinformatics analysis showed that the differentially expressed proteins were essentially those involved in the metabolism of proteins, ribonucleotides, or carbohydrates, or in stress response or cellular movement. This study represents the first attempt at profiling S. japonicum living in different states and provides a basis for a better understanding of the molecular mechanisms in the development and survival of S. japonicum in different host environments.

Vaccination of Goats with a Glutathione Peroxidase DNA Vaccine Induced Partial Protection Against Haemonchus Contortus Infection

Due to their critical functions in worm physiology, glutathione peroxidases in Haemonchus contortus are potential candidates for vaccine to control haemonchosis. However, information on the protection potential of these molecules is lacking. In this study, recombinant HC29 antigen was tested for protection against experimental H. contortus infections in goats. Fifteen animals were allocated into three trial groups. The animals of HC29 group was vaccinated with a recombinant HC29 DNA vaccine twice at day 0 and day 14, and then challenged with 5000 infective H. contortus L3 (third larval stage) on the 28th day. An unvaccinated positive control group was challenged with L3 at the same time. An unvaccinated negative control group was not challenged with L3. The results indicated HC29 DNA vaccine was transcribed at local injection sites and expressed in vivo post immunizations, respectively. Following L3 challenge, the mean eggs per gram feces and worm burdens of HC29 group were reduced by 36.1% and 35.6% respectively compared to the positive control group. After immunization with the DNA vaccine, significantly high levels of serum IgG, serum IgA, mucosal IgA and CD4(+) T lymphocytes were detected. These results suggest that recombinant H. contortus HC29 glutathione peroxidase DNA vaccine induced a partial immune response and has protective potentials against caprine haemonchosis.

Ultrastructural Characterization of Olfactory Sensilla and Immunolocalization of Odorant Binding and Chemosensory Proteins from an Ectoparasitoid Scleroderma Guani (Hymenoptera: Bethylidae)

The three-dimensional structures of two odorant binding proteins (OBPs) and one chemosensory protein (CSP) from a polyphagous ectoparasitoid Scleroderma guani (Hymenoptera: Bethylidae) were resolved bioinformatically. The results show that both SguaOBP1 and OBP2 are classic OBPs, whereas SguaCSP1 belongs to non-classic CSPs which are considered as the "Plus-C" CSP in this report. The structural differences between the two OBPs and between OBP and CSP are thoroughly described, and the structural and functional significance of the divergent C-terminal regions (e.g., the prolonged C-terminal region in SguaOBP2 and the additional pair of cysteines in SguaCSP1) are discussed. The immunoblot analyses with antisera raised against recombinant SguaOBP1, OBP2, and CSP1, respectively, indicate that two SguaOBPs are specific to antennae, whereas SguaCSP1, which are more abundant than OBPs and detected in both male and female wasps, expresses ubiquitously across different tissues.We also describe the ultrastructure of the antennal sensilla types in S. guani and compare them to 19 species of parasitic Hymenoptera. There are 11 types of sensilla in the flagellum and pedicel segments of antennae in both male and female wasps. Seven of them, including sensilla placodea (SP), long sensilla basiconica (LSB), sensilla coeloconica (SC), two types of double-walled wall pore sensilla (DWPS-I and DWPS-II), and two types of sensilla trichodea (ST-I and ST-II), are multiporous chemosensilla. The ultralsturctures of these sensilla are morphologically characterized. In comparison to monophagous specialists, the highly polyphagous generalist ectoparasitoids such as S. guani possess more diverse sensilla types which are likely related to their broad host ranges and complex life styles. Our immunocytochemistry study demonstrated that each of the seven sensilla immunoreacts with at least one antiserum against SguaOBP1, OBP2, and CSP1, respectively. Anti-OBP2 is specifically labeled in DWPS-II, whereas the anti-OBP1 shows a broad spectrum of immunoactivity toward four different sensilla (LSB, SP, ST-I and ST-II). On the other hand, anti-CSP1 is immunoactive toward SP, DWPS-I and SC. Interestingly, a cross co-localization pattern between SguaOBP1 and CSP1 is documented for the first time. Given that the numbers of OBPs and CSPs in many insect species greatly outnumber their antennal sensilla types, it is germane to suggest such phenomenon could be the rule rather than the exception.

Risky Decision-making in Older Adults Without Cognitive Deficits: An FMRI Study of VMPFC Using the Iowa Gambling Task

Some older adults without neurological disease exhibit impaired decision-making in risky, nontransparent situations, like the Iowa Gambling Task (IGT). The prefrontal cortices are particularly vulnerable to age-related decline, and numerous studies implicate the ventromedial prefrontal cortex (VMPFC) in successful IGT performance. However, the relationship between IGT performance and VMPFC function in older adults has not yet been tested by fMRI. In the present study, older adults with seemingly no cognitive impairments performed the IGT and a non-gambling control task during fMRI. Group analyses indicate that in these older adults, regardless of IGT performance level, a right VMPFC subregion is activated during the IGT, while successful IGT performance is correlated with left VMPFC activation, suggesting that bilateral VMPFC during risky, nontransparent situations may contribute to successful decision-making in older adults. Individual subject analyses reveal substantial variation regarding the extent and location of VMPFC activation during the IGT, a finding not captured in the group analysis: There is no correlation between IGT performance and extent of activation in the right VMPFC, although there is such a correlation between left VMPFC activation and IGT performance.

Protective Efficacy Evaluation Induced by Recombinant Protein LHD-Sj23-GST of S. Japonicum Emulsified with Three Different Adjuvants

As a complement to chemotherapy and other control approaches, the development of an effective vaccine is necessary to combat Schistosomiasis japonica that remains a serious public health problem in China. In the present study, mice were vaccinated with purified recombinant protein LHD-Sj23-GST(large hydrophilic domain of 23 kDa antigen of S. japonicum fused with Sj26GST) emulsified with Freund's adjuvant (FA), Montanide ISA 206 and Montanide ISA 70M and challenged with cercariae, the protective efficacy induced by the recombinant protein was evaluated, and the LHD-Sj23-GST specific IgG and it's subtypes were determined. The result revealed that a significant worm burden reduction (58.8%, 26.3%, and 54.3%; P < 0.05) was obtained in mice vaccinated with LHD-Sj23-GST emulsified with three different adjuvants compared to those mice treated with respective adjuvant only. ELISA test suggested that the high level production of LHD-Sj23-GST specific IgG1, IgG2a and IgG3 antibodies may participate in protecting against schistosome infection. © 2012 Blackwell Publishing Ltd.

Vaccination of Goats with DNA Vaccines Encoding H11 and IL-2 Induces Partial Protection Against Haemonchus Contortus Infection

DNA vaccines expressing Haemonchus contortus H11 antigen with or without interleukin (IL)-2 were tested for protection against H. contortus infection in goats. Sixteen goats (8-10 months of age) were allocated into four trial groups. On days 0 and 14, group 1 was immunised with a DNA vaccine expressing H11 and IL-2 and group 2 was immunised with a DNA vaccine expressing H11 only. Group 3 was an unvaccinated positive control group challenged with H. contortus third stage larvae (L3). Group 4 was an unvaccinated negative control group that was not challenged with L3. Animals in groups 1-3 were challenged with 5000 infective H. contortus L3 14 days after the second immunisation. Transcription of H11 and IL-2 was demonstrated in muscle by reverse transcriptase-PCR 10 days after primary immunisation and translation of H11 was detected by Western blot analysis 7 days after the second immunisation. Following immunisation with a DNA vaccine expressing H11 and IL-2, high levels of specific serum immunoglobulin (Ig) G, non-specific serum IgA, mucosal IgA, CD4(+) T lymphocytes, CD8(+) T lymphocytes and B lymphocytes were produced. Following challenge with L3, cumulative mean faecal worm egg counts and worm burdens in group 1 were reduced by 56.6% and 46.7%, respectively, while corresponding reductions in group 2 were 44.8% and 38.0%. There was a small but significant difference in abomasal worm burdens in goats in groups 1 (395.3±37.6) and 2 (459.5±101.6) compared to group 3 (741.5±241.5; P<0.05). Use of a DNA vaccine expressing H11 and IL-2 conferred partial protection against Haemonchus contortus infection in goats.

Molecular Cloning, Expression and Characterization of Enolase from Adult Haemonchus Contortus

Enolase represents a multifunctional protein involved in basic energy metabolism. In the present research, the enolase gene of Haemonchus contortus (HcENO) was cloned and characterized. Specific primers for the rapid amplification of cDNA ends (RACE) were designed based on the expression sequence tag (EST, GenBank Accession No. BF422728) to amplify the 3'- and 5'-ends of HcENO. The full length of cDNA from this gene was obtained by overlapping the sequences of 3'- and 5'-extremities and amplification by reverse transcription PCR. The biochemical activities of the recombinant protein HcENO, which was expressed in prokaryotic cells and purified by affinity chromatography, were analyzed by assays of enzymatic activity, stability to pH. The results showed that the cloned full length cDNA comprised 1583bp and encoded a peptide with 434 amino acid residues which showed sequence similarity to several known enolases. The biochemical assay showed that the protein encoded by the HcENO exhibited enzymatic activity, whilst the HcENO was stable between pH 6 and 8. The natural enolase of H. contortus detected by immunoblot assay was approximately 49kDa in size, and the recombinant HcENO was recognized strongly by serum from experimentally infected goats.

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