The Virulence-Related MYR1 Protein of Toxoplasma gondii as a Novel DNA Vaccine Against Toxoplasmosis in Mice

Toxoplasma gondii causes serious public health problems, but there is no effective treatment strategy against it currently. DNA vaccines have shown promising findings in this regard. MYR1 is a new virulence factor identified in T. gondii that may have potential as a DNA vaccine candidate. We constructed a recombinant eukaryotic plasmid, pVAX1-MYR1, as a DNA vaccine, injected it intramuscularly into BALB/c mice, and evaluated its immunoprotective effects. pVAX1-MYR1 immunization induced a sequential Th1 and Th2 T-cell response, as indicated by high levels of Th1 and mixed Th1/Th2 cytokines at 2 and 6 weeks after immunization, respectively. These findings were corroborated by the antibody assays too. In addition, increased levels of antigen-specific lymphocyte proliferation, CD4+ and CD8+ T lymphocytes, cytotoxic T lymphocyte activity and cytokine (IFN-γ, IL-12, and IL-10) production were also observed in the immunized mice. These findings showed that pVAX1-MYR1 stimulated humoral and cellular immune responses in the immunized mice. The increased production of IFN-γ and IL-12 was correlated with increased expression of the T-bet and p65 genes of the NF-κB pathway. However, no significant increase was observed in the level of IL-4. The survival of mice immunized with pVAX1-MYR1 was also significantly prolonged compared with the control group mice. Based on all the above findings, the current study proposes that pVAX1-MYR1 can induce a T. gondii-specific immune response and should therefore be considered as a promising vaccine candidate against toxoplasmosis. To the best of our knowledge, this is the first report to evaluate the immunoprotective value of an MYR1-based DNA vaccine against T. gondii

Raf kinase activation of adenylyl cyclases: isoform-selective regulation

ABSTRACT Adenylyl cyclases (AC), a family of enzymes that catalyze the synthesis of cyclic AMP, are critical regulators of cellular functions. The activity of adenylyl cyclase is stimulated by a range of hormone receptors, primarily via interactions with G-proteins; however, recently we identified an alternate mechanism by which growth factors sensitize adenylyl cyclase activation. We suggested that this mechanism might involve a Raf kinasemediated serine phosphorylation of adenylyl cyclase. However, the direct involvement of a specific form of Raf kinase is yet to be demonstrated. Furthermore, whether this mechanism is generalized to other isoforms of adenylyl cyclase is unknown. In human embryonic kidney 293 cells, we now demonstrate that in reconstitution studies, c-Raf kinase can mediate phosphorylation of AC VI. Furthermore, AC VI coimmunoprecipitates with c-Raf. Raf kinase-dependent regulation of adenylyl cyclase VI is dependent on the integrity of Ser750 in the fourth intracellular loop of the enzyme and Ser603/Ser608 in the C1b region of the molecule. To examine how generalized this effect is, we studied representative isoforms of the major subfamilies of adenylyl cyclase viz., AC I, AC II, and AC V. Raf kinase-dependent sensitization/ phosphorylation of adenylyl cyclases is common to AC VI, AC V, and AC II isoforms but not AC I. In aggregate, these studies indicate that Raf kinase associates with adenylyl cyclases. Furthermore, Raf kinase regulation of adenylyl cyclase is isoform-selective. These functional interactions (as well as the physical association) between adenylyl cyclases and Raf kinases suggest an important but previously unrecognized interaction between these two key regulatory enzymes

Associations of hair cortisol levels with violence, poor mental health, and harmful alcohol and other substance use among female sex workers in Nairobi, Kenya.

Violence, poor mental health, and harmful substance use are commonly experienced by female sex workers (FSWs) in sub-Saharan Africa, all of which are associated with increased HIV susceptibility. We aimed to investigate the associations between violence, poor mental health and harmful alcohol/substance use with hair cortisol concentration (HCC) levels as a potential biological pathway linking the experiences of these stressors and HIV vulnerability. We used the baseline data of the Maisha Fiti study of FSWs in Nairobi, Kenya. Participants reported recent violence, poor mental health, and harmful alcohol/substance use. Hair samples proximal to the scalp were collected to measure cortisol levels determined by ELISA. We analysed the data of 425 HIV-negative respondents who provided at least 2 cm of hair sample. The prevalence of recent violence was 89.3% (physical 54.6%; sexual 49.4%; emotional 77.0% and financial 66.5%), and 29.1% had been arrested due to sex work. 23.7% of participants reported moderate/severe depression, 11.6% moderate/severe anxiety, 13.5% PTSD and 10.8% recent suicidal thoughts and/or attempts. About half of the participants (48.8%) reported recent harmful alcohol and/or other substance use. In multivariable linear regression analyses, both physical and/or sexual violence (adjusted geometric mean ratio (aGMR) = 1.28; 95% CI 1.01-1.62) and harmful alcohol and/or other substance use (aGMR = 1.31; 95% CI 1.03-1.65) were positively and independently associated with increased HCC levels. Findings suggest a role of violence and substance use in elevated HCC levels, which could increase HIV risk due to cortisol-related T cell activation. However, longitudinal and mechanistic studies are needed to confirm this hypothesis

Morphological diversity of single neurons in molecularly defined cell types.

Dendritic and axonal morphology reflects the input and output of neurons and is a defining feature of neuronal types1,2, yet our knowledge of its diversity remains limited. Here, to systematically examine complete single-neuron morphologies on a brain-wide scale, we established a pipeline encompassing sparse labelling, whole-brain imaging, reconstruction, registration and analysis. We fully reconstructed 1,741 neurons from cortex, claustrum, thalamus, striatum and other brain regions in mice. We identified 11 major projection neuron types with distinct morphological features and corresponding transcriptomic identities. Extensive projectional diversity was found within each of these major types, on the basis of which some types were clustered into more refined subtypes. This diversity follows a set of generalizable principles that govern long-range axonal projections at different levels, including molecular correspondence, divergent or convergent projection, axon termination pattern, regional specificity, topography, and individual cell variability. Although clear concordance with transcriptomic profiles is evident at the level of major projection type, fine-grained morphological diversity often does not readily correlate with transcriptomic subtypes derived from unsupervised clustering, highlighting the need for single-cell cross-modality studies. Overall, our study demonstrates the crucial need for quantitative description of complete single-cell anatomy in cell-type classification, as single-cell morphological diversity reveals a plethora of ways in which different cell types and their individual members may contribute to the configuration and function of their respective circuits

Cellular anatomy of the mouse primary motor cortex.

An essential step toward understanding brain function is to establish a structural framework with cellular resolution on which multi-scale datasets spanning molecules, cells, circuits and systems can be integrated and interpreted1. Here, as part of the collaborative Brain Initiative Cell Census Network (BICCN), we derive a comprehensive cell type-based anatomical description of one exemplar brain structure, the mouse primary motor cortex, upper limb area (MOp-ul). Using genetic and viral labelling, barcoded anatomy resolved by sequencing, single-neuron reconstruction, whole-brain imaging and cloud-based neuroinformatics tools, we delineated the MOp-ul in 3D and refined its sublaminar organization. We defined around two dozen projection neuron types in the MOp-ul and derived an input-output wiring diagram, which will facilitate future analyses of motor control circuitry across molecular, cellular and system levels. This work provides a roadmap towards a comprehensive cellular-resolution description of mammalian brain architecture

A multimodal cell census and atlas of the mammalian primary motor cortex

ABSTRACT We report the generation of a multimodal cell census and atlas of the mammalian primary motor cortex (MOp or M1) as the initial product of the BRAIN Initiative Cell Census Network (BICCN). This was achieved by coordinated large-scale analyses of single-cell transcriptomes, chromatin accessibility, DNA methylomes, spatially resolved single-cell transcriptomes, morphological and electrophysiological properties, and cellular resolution input-output mapping, integrated through cross-modal computational analysis. Together, our results advance the collective knowledge and understanding of brain cell type organization: First, our study reveals a unified molecular genetic landscape of cortical cell types that congruently integrates their transcriptome, open chromatin and DNA methylation maps. Second, cross-species analysis achieves a unified taxonomy of transcriptomic types and their hierarchical organization that are conserved from mouse to marmoset and human. Third, cross-modal analysis provides compelling evidence for the epigenomic, transcriptomic, and gene regulatory basis of neuronal phenotypes such as their physiological and anatomical properties, demonstrating the biological validity and genomic underpinning of neuron types and subtypes. Fourth, in situ single-cell transcriptomics provides a spatially-resolved cell type atlas of the motor cortex. Fifth, integrated transcriptomic, epigenomic and anatomical analyses reveal the correspondence between neural circuits and transcriptomic cell types. We further present an extensive genetic toolset for targeting and fate mapping glutamatergic projection neuron types toward linking their developmental trajectory to their circuit function. Together, our results establish a unified and mechanistic framework of neuronal cell type organization that integrates multi-layered molecular genetic and spatial information with multi-faceted phenotypic properties

PCMs heat transfer performance enhancement with expanded graphite and its thermal stability

AbstractThis work focuses on developing a high performance phase change materials (PCMs) adding with a few expanded graphite (EG). A series of composite phase change materials (paraffin) with various EG concentration are formulated in vacuum condition and ambient pressure respectively. The heat fusion of composite PCMs is measured by DSC. A rig for PCMs heat transfer performance test is built in lab. The heat transfer performance test of composites PCMs are carried out. The heat storage and release processed of pure PCMs and composite significantly improved after EG adding. The composited PCMs formulated in vacuum condition also presents better heat transfer performance. It shows composite PCMs can be well mixed in vacuum. Due to the repeated heating and cooling in PCMs application, thermal stability test is carried out in this work. PCMs are putting in water bath with temperature controlling by cyclic phase change to quantify the rate of degradation heat fusion and thermal charge/discharge process. After cycling 60 times, up to 2% heat fusion decrement is observed. Heat transfer performance studies also show that there is little change. In brief, PCMs with EG has high thermal performance for thermal energy storage process

Optimized Parameters and Forecast Analysis of High-position Hole for Goaf Gas Drainage

AbstractIn order to obtain the ideal parameters and drainage quantity of high-position hole for gas drainage, the flow equation of high-position hole for goaf gas drainage is established, and No.1111 Fully Mechanized Coal Mining working face of Zhangji mine was taken as the field for simulating methane flow by Matlab software to gain high-position ideal drainage parameters. It showed that borehole interval should be 0.5 m, the drainage pressure should be in the range from 0.015MPa to 0.025MPa, the borehole diameter should be 108mm. At the same time, in order to control working face air quantity and gas drainage quantity of goaf, the relation of methane quantity drained by wind and methane drainage quantity of high-position hole was analysed, and forecast model of high-position hole drainage quantity was established. The drainage parameters and forecast model of high-position hole can be introduced to the other similar mines of coal seam storage, which is beneficial to safety production for high methane mines

J. Nanopart. Res.

This work concerns rheological and frictional behaviour of lubricating oils containing platelet molybdenum disulfide (MoS2) nanoparticles (average diameter similar to 50 nm; single layer thickness similar to 3 nm). Stable nano-MoS2 lubricants were formulated and measured for their rheological behaviour and tribological performance. Rheological experiments showed that the nano-MoS2 oils were non-Newtonian following the Bingham plastic fluid model. The viscosity data fitted the classic Hinch-Leal (H-L) model if an agglomeration factor of 1.72 was introduced. Tribological experiments indicated that the use of MoS2 nanoparticles could enhance significantly the tribological performance of the base lubricating oil (reduced frictional coefficient, reduced surface wear and increased stability). Scanning electron microscopy, laser confocal microscope and x-ray energy dispersive spectroscopy analyses suggested that the reduced frictional coefficient and surface wear be associated with surface patching effects. Such patching effects were shown to depend on the concentration of MoS2 nanoparticles, and an effective patching required a concentration over approximately 1 wt%. The increased stability could be attributed to the enhanced heat transfer and lubricating oil film strength due to the presence of nanoparticles.This work concerns rheological and frictional behaviour of lubricating oils containing platelet molybdenum disulfide (MoS2) nanoparticles (average diameter similar to 50 nm; single layer thickness similar to 3 nm). Stable nano-MoS2 lubricants were formulated and measured for their rheological behaviour and tribological performance. Rheological experiments showed that the nano-MoS2 oils were non-Newtonian following the Bingham plastic fluid model. The viscosity data fitted the classic Hinch-Leal (H-L) model if an agglomeration factor of 1.72 was introduced. Tribological experiments indicated that the use of MoS2 nanoparticles could enhance significantly the tribological performance of the base lubricating oil (reduced frictional coefficient, reduced surface wear and increased stability). Scanning electron microscopy, laser confocal microscope and x-ray energy dispersive spectroscopy analyses suggested that the reduced frictional coefficient and surface wear be associated with surface patching effects. Such patching effects were shown to depend on the concentration of MoS2 nanoparticles, and an effective patching required a concentration over approximately 1 wt%. The increased stability could be attributed to the enhanced heat transfer and lubricating oil film strength due to the presence of nanoparticles