Peptide Antigen Concentration Modulates Digital NFAT1 Activation in Primary Mouse Naive CD8(+) T Cells as Measured by Flow Cytometry of Isolated Cell Nuclei

Circulating naive T cells exist in a quiescent state. After TCR contact with the cognate peptide presented by APCs in secondary lymphoid structures, T cells undergo a period of rapid transcriptional changes that set the stage for fate-determining effector or memory programming. We describe a novel method to analyze TCR signaling pathway activation in nuclei isolated from primary mouse naive T cells after stimulation with natural peptide Ags. We prelabeled cells with cell tracking dye to easily distinguish CD8(+) T cell nuclei from APC nuclei by conventional flow cytometry. Using this approach, we observed clear digital activation of NFAT1 transcription factor in OT-I T cells stimulated with OVA peptide presented by bulk splenocytes. OVA concentration had discrete control over the fraction of the cells that translocated NFAT1, indicating that a distinct threshold amount of TCR signaling is required to switch on NFAT1 in naive T cells. This behavior was cell contact dependent and qualitatively more exact than the NFAT1 response in ionomycin-stimulated naive T cells. These data contribute to our understanding of the digital behavior of TCR signaling components documented in other studies and indicate how T cells might discriminate log-fold changes in Ag availability during an actual infection. Overall, these results highlight the potential of this coculture nuclei isolation protocol to address stimulation-dependent translocation of proteins in primary lymphocytes

Fine Scale Influence of Weather on Northern Bobwhite Abundance, Breeding Success, and Harvest in South Texas

Weather plays a substantial role in annual changes in populations of northern bobwhite (Colinus virginianus) within and among ecological regions. Few studies have tested this relationship within the confines of specific sites. We examined the fine scale influence of annual (12-month), seasonal (6-month), and monthly Modified Palmer Drought Severity Indices (PMDI) and raw precipitation on abundance, breeding success, and harvest of northern bobwhites on 2 sites in south Texas. We used 18 years (1984–01) of roadside census, juvenile : adult ratios, and harvest records from the Chaparral Wildlife Management Area (CWMA) in La Salle County and 15 years (1984–99) of juvenile : adult ratios and harvest records from a private property in Brooks County (BCP) to examine relationships and trends with weather variables. Bobwhite abundance was correlated (r \u3e= 0.50, P \u3c= 0.035) with 12- and 6-month sums of precipitation and PMDI. Breeding success was correlated (r \u3e= 0.53, P \u3c= 0.023) with 12-month precipitation for both sites and was correlated (r = 0.53, P = 0.040) with 6-month precipitation for BCP only. Harvest variables for CWMA were correlated (r \u3e= 0.54, P \u3c= 0.022) with 12- and 6-month PMDI, while BCP harvest/ha was correlated (r = 0.54, P = 0.027) with the 12-month precipitation sum. Monthly correlates with precipitation increased from spring to summer until July when they became negatively related to rainfall on both sites. Monthly PMDI correlates became increasingly important from spring through summer including July. Our findings account for at least part of the annual variation in northern bobwhite abundance in south Texas and provide information useful in understanding of the influence of weather at fine spatial scales

Ratcheting synthesis

Synthetic chemistry has traditionally relied on reactions between reactants of high chemical potential and transformations that proceed energetically downhill to either a global or local minimum (thermodynamic or kinetic control). Catalysts can be used to manipulate kinetic control, lowering activation energies to influence reaction outcomes. However, such chemistry is still constrained by the shape of one-dimensional reaction coordinates. Coupling synthesis to an orthogonal energy input can allow ratcheting of chemical reaction outcomes, reminiscent of the ways that molecular machines ratchet random thermal motion to bias conformational dynamics. This fundamentally distinct approach to synthesis allows multi-dimensional potential energy surfaces to be navigated, enabling reaction outcomes that cannot be achieved under conventional kinetic or thermodynamic control. In this Review, we discuss how ratcheted synthesis is ubiquitous throughout biology and consider how chemists might harness ratchet mechanisms to accelerate catalysis, drive chemical reactions uphill and programme complex reaction sequences.<br/

Observations and simulations of nova Vul 1984 no. 2: A nova with ejecta rich in oxygen, neon, and magnesium

Nova Vul 1984 no. 2 was observed with IUE from Dec. 1984 through Nov. 1987. The spectra are characterized by strong lines from Mg, Ne, C, Si, O, N, and other elements. Data obtained in the ultraviolet, infrared, and optical show that this nova is ejecting material rich in oxygen, neon, and magnesium

Cobalt Complexes of Bulky PNP Ligand: H2 Activation and Catalytic Two-Electron Reactivity in Hydrogenation of Alkenes and Alkynes

The reactivity of cobalt pincer complexes supported by the bulky tetramethylated PNP ligands Me4PNPR(R = iPr, tBu) has been investigated. In these ligands, the undesired H atom loss reactivity observed earlier in some classical CH2-arm PNP cobalt complexes is blocked, allowing them to be utilized for promoting two-electron catalytic transformations at the cobalt center. Accordingly, reaction of the formally CoIMe complex 3 with H2 under ambient pressure and temperature afforded the CoIII trihydride 4-H, in a reaction cascade reasoned to proceed by two-electron oxidative addition and reductive eliminations. This mechanistic proposal, alongside the observance of alkene insertion and ethane production upon sequential exposure of 3 to ethylene and H2, prompted an exploration into 3 as a catalyst for hydrogenation. Complex 4-H, formed in situ from 3 under H2, was found to be active in the catalytic hydrogenation of alkenes and alkynes. The proposed two-electron mechanism is reminiscent of the platinum group metals and demonstrates the utility of the bulky redox-innocent Me4PNPR ligand in the avoidance of one-electron reactivity, a concept that may show broad applicability in expanding the scope of earth-abundant first-row transition-metal catalysis.journal articl

The Tec kinase ITK differentially optimizes NFAT, NF-κB, and MAPK signaling during early T cell activation to regulate graded gene induction [preprint]

The strength of peptide:MHC interactions with the T cell receptor (TCR) is correlated with the time to first cell division, the relative scale of the effector cell response, and the graded expression of activation-associated proteins like IRF4. To regulate T cell activation programming, the TCR and the TCR proximal kinase ITK simultaneously trigger many biochemically separate TCR signaling cascades. T cells lacking ITK exhibit selective impairments in effector T cell responses after activation, but under the strongest signaling conditions ITK activity is dispensable. To gain insight into whether TCR signal strength and ITK activity tune observed graded gene expression through unequal activation of disparate signaling pathways, we examined Erk1/2 activation and NFAT, NF-κB translocation in naive OT-I CD8+ cell nuclei. We observed consistent digital activation of NFAT1 and Erk-MAPK, but NF-κB displayed dynamic, graded activation in response to variation in TCR signal strength and was tunable by treatment with an ITK inhibitor. Inhibitor-treated cells showed dampened induction of AP-1 factors Fos and Fosb, NF-κB response gene transcripts, and survival factor Il2 transcripts. ATAC-seq analysis also revealed genomic regions most sensitive to ITK inhibition were enriched for NF-κB and AP-1 motifs. Specific inhibition of NF-κB during peptide stimulation tuned expression of early gene products like c-Fos. Together, these data indicate a key role for ITK in orchestrating optimal activation of separate TCR downstream pathways, specifically aiding NF-κB activation. More broadly, we revealed a mechanism by which variation in TCR signal strength can produce patterns of graded gene expression in activated T cells

The H alpha Galaxy Survey V. The star formation history of late-type galaxies

This study of 117 low-redshift Im and Sm galaxies investigates the star formation rates of late-type galaxies, to determine whether they are quasi-continuous or dominated by bursts with quiescent interludes. We analyse the distribution of star formation timescales (stellar masses/star formation rates) for the entire sample, and of gas depletion timescales for those galaxies with gas mass measurements. We find that, on average, the late-type galaxies studied could have produced their total stellar masses by an extrapolation of their current star formation activity over a period of just under a Hubble time. This is not the case for a comparison sample of earlier-type galaxies, even those with disk-dominated morphologies and similar total stellar masses to the late-type galaxies. The earlier-type galaxies are on average forming their stars more slowly at present than the average rate over their past histories. No totally quiescent Im or Sm galaxies are found, and although some evidence of intrinsic variation in the star formation rate with time is found, this is typically less than a factor of 2 increase or decrease relative to the mean level. The Im and Sm galaxies have extensive gas reservoirs and can maintain star formation at the current rate for more than another Hubble time. The average spatial distribution of star formation in the Im galaxies, and to a lesser extent the Sm galaxies, is very similar to that of the older stellar population traced by the red light.Comment: 8 pages, 7 figures, accepted for publication in Astronomy & Astrophysic

Quantitative histopathologic assessment of perfusion MRI as a marker of glioblastoma cell infiltration in and beyond the peritumoral edema region

Background: Conventional MRI fails to detect regions of glioblastoma cell infiltration beyond the contrast‐enhanced T1 solid tumor region, with infiltrating tumor cells often migrating along host blood vessels. Purpose: To quantitatively and qualitatively analyze the correlation between perfusion MRI signal and tumor cell density in order to assess whether local perfusion perturbation could provide a useful biomarker of glioblastoma cell infiltration. Study Type: Animal model. Subjects: Mice bearing orthotopic glioblastoma xenografts generated from a patient‐derived glioblastoma cell line. Field Strength/Sequences: 7T perfusion images acquired using a high signal‐to‐noise ratio (SNR) multiple boli arterial spin labeling sequence were compared with conventional MRI (T1/T2 weighted, contrast‐enhanced T1, diffusion‐weighted, and apparent diffusion coefficient). Assessment: Immunohistochemistry sections were stained for human leukocyte antigen (probing human‐derived tumor cells). To achieve quantitative MRI‐tissue comparison, multiple histological slices cut in the MRI plane were stacked to produce tumor cell density maps acting as a “ground truth.” Statistical Tests: Sensitivity, specificity, accuracy, and Dice similarity indices were calculated and a two‐tailed, paired t‐test used for statistical analysis. Results: High comparison test results (Dice 0.62–0.72, Accuracy 0.86–0.88, Sensitivity 0.51–0.7, and Specificity 0.92–0.97) indicate a good segmentation for all imaging modalities and highlight the quality of the MRI tissue assessment protocol. Perfusion imaging exhibits higher sensitivity (0.7) than conventional MRI (0.51–0.61). MRI/histology voxel‐to‐voxel comparison revealed a negative correlation between tumor cell infiltration and perfusion at the tumor margins (P = 0.0004). Data Conclusion: These results demonstrate the ability of perfusion imaging to probe regions of low tumor cell infiltration while confirming the sensitivity limitations of conventional imaging modalities. The quantitative relationship between tumor cell density and perfusion identified in and beyond the edematous T2 hyperintensity region surrounding macroscopic tumor could be used to detect marginal tumor cell infiltration with greater accuracy