Occurrence of coexisting dendrite morphologies: immiscible fluid displacement in an anisotropic radial hele-shaw cell under a high flow rate regime

Viscous fingering morphologies during the displacement of a high viscosity fluid by a low viscosity immiscible fluid in a radial fourfold anisotropic Hele-Shaw cell are examined. By using the kerosene-glycerin system for which the µ/T ratio (µ being the relative viscosity and T the interfacial tension between the fluids) is about ten times higher than that for the commonly used air-glycerin system, we have been able to access the hitherto unexplored Nca 1 regime (capillary number Nca=Uµ/T, U being the advancing fingertip velocity). Within the anisotropy-dominated regime, and when flow rates are significantly high (capillary number well beyond Nca=1), a new phase is seen to evolve wherein the dendrites grow simultaneously along the channels and along the directions making an angle of 45° with the channels, both being kinetically driven. This new phase resembles the one observed in a miscible fluid system at all flow rates of the displacing fluid

Viscous fingering of miscible fluids in an anisotropic radial hele-shaw cell: coexistence of kinetic and surface-tension dendrite morphology types and an exploration of small-scale influences

The evolution of viscous fingering morphology is examined for the case of a system of miscible fluids in an anisotropic radial Hele-Shaw cell. It is shown that dendritic morphologies similar to the kinetic and surface-tension morphology types coexist for this case. The critical role of the means of introducing anisotropy in the Hele-Shaw cell is established, and an explanation of the pattern behavior is offered on the basis of shape discontinuities of the individual elements of the lattice used to induce anisotropy. The ramifications of such an explanation are experimentally verified by demonstrating a clear difference in the morphology evolution in two halves of a single Hele-Shaw cell, one half of which contains square lattice elements, and the other half of which contains circular lattice elements

Variation in viscous fingering pattern morphology due to surfactant-mediated interfacial recognition events

The study of the formation of finger-like patterns during displacement of a viscous fluid by a less viscous one is of technological importance. The morphology of the viscous-finger patterns generated is a function of many parameters such as the flow rate, difference in viscosities of the two fluids and the interfacial tension. We demonstrate herein that the morphology of patterns formed during viscous fingering in a Hele-Shaw cell during displacement of paraffin oil by aqueous solutions of the surfactant sodium dodecyl sulphate (SDS), is extremely sensitive to interfacial tension variation brought about by complexation of divalent cations with the surfactant SDS. The variation in morphology of the patterns formed has been quantified by measuring the fractal dimensions of structures formed in a radial Hele-Shaw cell as well as the average finger width in a linear Hele-Shaw cell. This technique shows promise for studying other interfacial phenomena in chemistry such as biorecognition as well as dynamic processes occurring at interfaces

Temporal Dynamics of Spontaneous Ca2+ Transients, ERBB4, vGLUT1, GAD1, Connexin, and Pannexin Genes in Early Stages of Human Stem Cell Neurodifferentiation

Spontaneous Ca2+ transients drive stem cell proliferation and neurodifferentiation. Deciphering the relationship between neuronal and glial human genes on one side and spontaneous Ca2+ activity on the other side is essential for our understanding of normal brain development, and for insights into the pathogenesis of neurodegenerative and neurodevelopmental disorders. In the present study, forebrain neurons were derived from human embryonic and induced pluripotent stem cells (hESC-H9 and iPSC-15; 22q11.2 deletion) over a period of 21 days in vitro (DIV). Every 1–2 days, multisite optical imaging technique was applied to detect populations of cells with spontaneous Ca2+ transients. The expression levels of 14 genes of interest were analyzed by quantitative polymerase chain reaction (qPCR) on the same biological samples where physiological recordings were performed. The genes analyzed include: the schizophrenia candidate gene ERBB4, connexin (Cx) genes Cx26, Cx36, Cx43, Cx45, Cx47, pannexin-1 (PNX1), neuronal markers PAX6, vGLUT1, GAD1, TUBB3, glial lineage markers BLBP, GFAP, and housekeeping gene ACTB. We found that Ca2+ signals decrease in amplitude, decrease in duration, and increase in frequency during the first 21 days of human neurodifferentiation. The expression levels of ERBB4, PAX6, GAD1, vGLUT1, BLBP, Cx36, Cx45, and PNX1 were found to be strongly positively correlated with the percentage of cells exhibiting spontaneous Ca2+ transients (“Active Cells”). While expression of BLBP, Cx45, ERBB4, GAD1, PAX6, PNX1, and vGLUT1 were correlated with short-duration and long-amplitude Ca2+ transients, Cx43, TUBB3, and Cx47 were better correlated with long-duration and short-amplitude transients. The expression dynamics of Cx26 was unrelated to any aspect of spontaneous Ca2+ activity. Four genes showed an exponential time course with a distinct onset on a given DIV. The onset of PNX1, ERBB4, and vGLUT1 occurred before, while the onset of Cx36 occurred after the first action potentials were detected in early differentiating human neurons

Identification of novel associations and localization of signals in idiopathic inflammatory myopathies using genome-wide imputation

Objective: The idiopathic inflammatory myopathies (IIMs) are heterogeneous diseases thought to be initiated by immune activation in genetically predisposed individuals. We imputed variants from the ImmunoChip array using a large reference panel to fine-map associations and identify novel associations in IIM. Methods: We analyzed 2,565 Caucasian IIM patient samples collected through the Myositis Genetics Consortium (MYOGEN) and 10,260 ethnically matched control samples. We imputed 1,648,116 variants from the ImmunoChip array using the Haplotype Reference Consortium panel and conducted association analysis on IIM and clinical and serologic subgroups. Results: The HLA locus was consistently the most significantly associated region. Four non-HLA regions reached genome-wide significance, SDK2 and LINC00924 (both novel) and STAT4 in the whole IIM cohort, with evidence of independent variants in STAT4, and NAB1 in the polymyositis (PM) subgroup. We also found suggestive evidence of association with loci previously associated with other autoimmune rheumatic diseases (TEC and LTBR). We identified more significant associations than those previously reported in IIM for STAT4 and DGKQ in the total cohort, for NAB1 and FAM167A-BLK loci in PM, and for CCR5 in inclusion body myositis. We found enrichment of variants among DNase I hypersensitivity sites and histone marks associated with active transcription within blood cells. Conclusion: We found novel and strong associations in IIM and PM and localized signals to single genes and immune cell types

Development and evaluation of a real-time one step Reverse-Transcriptase PCR for quantitation of Chandipura Virus

<p>Abstract</p> <p>Background</p> <p>Chandipura virus (CHPV), a member of family <it>Rhabdoviridae </it>was attributed to an explosive outbreak of acute encephalitis in children in Andhra Pradesh, India in 2003 and a small outbreak among tribal children from Gujarat, Western India in 2004. The case-fatality rate ranged from 55–75%. Considering the rapid progression of the disease and high mortality, a highly sensitive method for quantifying CHPV RNA by real-time one step reverse transcriptase PCR (real-time one step RT-PCR) using TaqMan technology was developed for rapid diagnosis.</p> <p>Methods</p> <p>Primers and probe for P gene were designed and used to standardize real-time one step RT-PCR assay for CHPV RNA quantitation. Standard RNA was prepared by PCR amplification, TA cloning and run off transcription. The optimized real-time one step RT-PCR assay was compared with the diagnostic nested RT-PCR and different virus isolation systems [<it>in vivo </it>(mice) <it>in ovo </it>(eggs), <it>in vitro </it>(Vero E6, PS, RD and Sand fly cell line)] for the detection of CHPV. Sensitivity and specificity of real-time one step RT-PCR assay was evaluated with diagnostic nested RT-PCR, which is considered as a gold standard.</p> <p>Results</p> <p>Real-time one step RT-PCR was optimized using <it>in vitro </it>transcribed (IVT) RNA. Standard curve showed linear relationship for wide range of 10²-10¹⁰(r²= 0.99) with maximum Coefficient of variation (CV = 5.91%) for IVT RNA. The newly developed real-time RT-PCR was at par with nested RT-PCR in sensitivity and superior to cell lines and other living systems (embryonated eggs and infant mice) used for the isolation of the virus. Detection limit of real-time one step RT-PCR and nested RT-PCR was found to be 1.2 × 10⁰PFU/ml. RD cells, sand fly cells, infant mice, and embryonated eggs showed almost equal sensitivity (1.2 × 10²PFU/ml). Vero and PS cell-lines (1.2 × 10³PFU/ml) were least sensitive to CHPV infection. Specificity of the assay was found to be 100% when RNA from other viruses or healthy individual was used.</p> <p>Conclusion</p> <p>On account of the high sensitivity, reproducibility and specificity, the assay can be used for the rapid detection and quantitation of CHPV RNA from clinical samples during epidemics and from endemic areas. The assay may also find application in screening of antiviral compounds, understanding of pathogenesis as well as evaluation of vaccine.</p

Androgen receptor signalling in Vascular Endothelial cells is dispensable for spermatogenesis and male fertility

<p>Abstract</p> <p>Background</p> <p>Androgen signalling is essential both for male development and function of the male reproductive system in adulthood. Within the adult testis, Germ cells (GC) do not express androgen receptor (AR) suggesting androgen-mediated promotion of spermatogenesis must act via AR-expressing somatic cell-types. Several recent studies have exploited the Cre/lox system of conditional gene-targeting to ablate AR function from key somatic cell-types in order to establish the cell-specific role of AR in promotion of male fertility. In this study, we have used a similar approach to specifically ablate AR-signalling from Vascular Endothelial (VE) cells, with a view to defining the significance of androgen signalling within this cell-type on spermatogenesis.</p> <p>Findings</p> <p>AR expression in VE cells of the testicular vasculature was confirmed using an antibody against AR. A Cre-inducible fluorescent reporter line was used to empirically establish the utility of a mouse line expressing Cre Recombinase driven by the Tie2-Promoter, for targeting VE cells. Immunofluorescent detection revealed expression of YFP (and therefore Cre Recombinase function) limited to VE cells and an interstitial population of cells, believed to be macrophages, that did not express AR. Mating of Tie2-Cre males to females carrying a floxed AR gene produced Vascular Endothelial Androgen Receptor Knockout (VEARKO) mice and littermate controls. Ablation of AR from all VE cells was confirmed; however, no significant differences in bodyweight or reproductive tissue weights could be detected in VEARKO animals and spermatogenesis and fertility was unaffected.</p> <p>Conclusions</p> <p>We demonstrate the successful generation and empirical validation of a cell-specific knockout of AR from VE cells, and conclude that AR expression in VE cells is not essential for spermatogenesis or male fertility.</p