Frequency-modulated nuclear localization bursts coordinate gene regulation

In yeast, the transcription factor Crz1 is dephosphorylated and translocates into the nucleus in response to extracellular calcium. Here we show, using time-lapse microscopy, that Crz1 exhibits short bursts of nuclear localization (typically lasting 2 min) that occur stochastically in individual cells and propagate to the expression of downstream genes. Strikingly, calcium concentration controls the frequency, but not the duration, of localization bursts. Using an analytic model, we also show that this frequency modulation of bursts ensures proportional expression of multiple target genes across a wide dynamic range of expression levels, independent of promoter characteristics. We experimentally confirm this theory with natural and synthetic Crz1 target promoters. Another stress-response transcription factor, Msn2, exhibits similar, but largely uncorrelated, localization bursts under calcium stress suggesting that frequency-modulation regulation of localization bursts may be a general control strategy used by the cell to coordinate multi-gene responses to external signals

Thermal Degradation of Pigments and Relative Biochemical Changes in Cherries and Apricots

The extent and nature of biochemical changes that take place in canned fruits during storage temperatures above freezing have been reviewed and discussed by Pederson, et al. (1947). These changes include loss in nutritive value, e.g. ascorbic acid, thiamine (Brenner, et al., 1948) and deterioration of color (Tressler, et al., 1955). Bauernfeind (1953) reported that canned peaches, apricots, and sweet cherries, after a few months of storage at 70°F, frequently undergo changes such as destruction of anthocyanin and carotenoid pigments with the subsequent formation of brown colored compounds. Darkening of fruit-color eventually results in their unacceptability at consumer level. Preference for fruit is mainly based upon the attractive appearance of the products. Thus, color is an important factor governing the quality of fruits and fruit products. In earlier studies, conducted elsewhere, emphasis was placed on effects of low storage temperatures on the quality of canned apricots and cherries. Paucity of scientific literature on the stability of processed apricots and cherries gave impetus to a study of the comparative influence of high storage temperatures and their duration, as such tests will have considerable economic bearing upon storing and shipping processed products to tropical countries. This thesis presents the effects of storage temperatures (40, 70, 100, and 120° F) and their duration (16 weeks) on colors (anthocyanins and carotenoids), total titratable acidity, pH, viscosity, carbohydrates (total 2 and free reducing sugars, pectins), volatile reducing substances, hydroxymethyl furfural, and organoleptic quality of canned apricots and cherries

Investigations Into Flavor Chemistry With Special Reference to Synthesis of Volatiles in Developing Tomato Fruit (\u3cem\u3eLycopersicon esculentum\u3c/em\u3e Mill.) Under Field and Glass Greenhouse Growing Conditions

The common tomato of our garden belongs to the natural order Solanaceae and the genus Lycopersicum. The name from lykos, a wolf, and persica a peach, is given to it because of the supposed aphrodisiacal qualities, and the beauty of the fruit. By culture and use it is a vegetable, botanically it is a fruit and among the fruits, it is a berry being indehiscent, pulpy, with one or more seeds that are not stones. Though the tomato was not recognized as a valuable food until about a century ago, its merit is now universally accepted. Often it is referred to as the poor man\u27s orange for it is rich in vitamins and in malic and citric acids, possessing besides, a fine appetizing flavor. The popularity of the tomato in man\u27s diet is due to the fact that it is a most rewarding crop for the home garden. It grows well practically everywhere, affording high nutritional values. The demand for and acceptance of fresh tomato fruit is based largely on its nutritional value, flavor, aroma, taste, and other characteristics, such as color and texture. These quality criteria are dependent primarily on the structure and chemical composition of the fruit. The importance of quality in tomatoes beyond that which can be expressed in calories per gram, or even in vitamin content, is generally accepted in the United States. In order to meet this increasing demand throughout the year tomatoes often have to be grown in the greenhouses. Therefore, in commercial greenhouses, the tomato has replaced lettuce as the principal crop and it is likely to remain as an important underglass crop. Flavor is a composite of taste and odor. Odor is produced by many aromatic substances which are present in fruit. Flavor itself is a very complex sensation. The physiological basis of flavor perception is extremely complex and not clearly understood. Flavor chemistry is a comparatively new field of research. Tomato fruit quality is determined mainly by the sugar acid ratio, pectins, color, and flavor. Among these color and flavor are probably the most useful criteria for estimating maturity of tomato fruit. Higher quality is associated with redness of color and prominence of flavor. The flavor of a fruit becomes pronounced when the sugar content is at its maximum and the color of the skin acquires the richest shade. Isolation of volatile components from natural products is often difficult. Typical flavor and aroma of tomato fruit is primarily due to its volatile components. Neither complete analysis for nor synthesis of tomato flavor has been accomplished due to the marathon of problems associated with the extraction, separation, and identification techniques. The primary aim of this investigation was to separate and identify some of the major flavor and aroma components in the developing tomato fruit and also to assess the influence of certain physiological and biochemical changes on the biosynthesis of these components during fruit growth

Pulsatile Dynamics in the Yeast Proteome

The activation of transcription factors in response to environmental conditions is fundamental to cellular regulation. Recent work has revealed that some transcription factors are activated in stochastic pulses of nuclear localization, rather than at a constant level, even in a constant environment. In such cases, signals control the mean activity of the transcription factor by modulating the frequency, duration, or amplitude of these pulses. Although specific pulsatile transcription factors have been identified in diverse cell types, it has remained unclear how prevalent pulsing is within the cell, how variable pulsing behaviors are between genes, and whether pulsing is specific to transcriptional regulators or is employed more broadly. To address these issues, we performed a proteome-wide movie-based screen to systematically identify localization-based pulsing behaviors in Saccharomyces cerevisiae. The screen examined all genes in a previously developed fluorescent protein fusion library of 4,159 strains in multiple media conditions. This approach revealed stochastic pulsing in ten proteins, all transcription factors. In each case, pulse dynamics were heterogeneous and unsynchronized among cells in clonal populations. Pulsing is the only dynamic localization behavior that we observed, and it tends to occur in pairs of paralogous and redundant proteins. Taken together, these results suggest that pulsatile dynamics play a pervasive role in yeast and may be similarly prevalent in other eukaryotic species

Combinatorial gene regulation by modulation of relative pulse timing

Studies of individual living cells have revealed that many transcription factors activate in dynamic, and often stochastic, pulses within the same cell. However, it has remained unclear whether cells might exploit the dynamic interaction of these pulses to control gene expression. Here, using quantitative single-cell time-lapse imaging of Saccharomyces cerevisiae, we show that the pulsatile transcription factors Msn2 and Mig1 combinatorially regulate their target genes through modulation of their relative pulse timing. The activator Msn2 and repressor Mig1 showed pulsed activation in either a temporally overlapping or non-overlapping manner during their transient response to different inputs, with only the non-overlapping dynamics efficiently activating target gene expression. Similarly, under constant environmental conditions, where Msn2 and Mig1 exhibit sporadic pulsing, glucose concentration modulated the temporal overlap between pulses of the two factors. Together, these results reveal a time-based mode of combinatorial gene regulation. Regulation through relative signal timing is common in engineering and neurobiology, and these results suggest that it could also function broadly within the signalling and regulatory systems of the cell

The impact of scaling up cervical cancer screening and treatment services among women living with HIV in Kenya: a modelling study

Introduction: We aimed to quantify health outcomes and programmatic implications of scaling up cervical cancer (CC) screening and treatment options for women living with HIV in care aged 18–65 in Kenya. Methods: Mathematical model comparing from 2020 to 2040: (1) visual inspection with acetic acid (VIA) and cryotherapy (Cryo); (2) VIA and Cryo or loop excision electrical procedure (LEEP), as indicated; (3) human papillomavirus (HPV)-DNA testing and Cryo or LEEP; and (4) enhanced screening technologies (either same-day HPV-DNA testing or digitally enhanced VIA) and Cryo or LEEP. Outcomes measured were annual number of CC cases, deaths, screening and treatment interventions, and engaged in care (numbers screened, treated and cured) and five yearly age-standardised incidence. Results: All options will reduce CC cases and deaths compared with no scale-up. Options 1–3 will perform similarly, averting approximately 28 000 (33%) CC cases and 7700 (27%) deaths. That is, VIA screening would yield minimal losses to follow-up (LTFU). Conversely, LTFU associated with HPV-DNA testing will yield a lower care engagement, despite better diagnostic performance. In contrast, option 4 would maximise health outcomes, averting 43 200 (50%) CC cases and 11 800 (40%) deaths, given greater care engagement. Yearly rescreening with either option will impose a substantial burden on the health system, which could be reduced by spacing out frequency to three yearly without undermining health gains. Conclusions: Beyond the specific choice of technologies to scale up, efficiently using available options will drive programmatic success. Addressing practical constraints around diagnostics’ performance and LTFU will be key to effectively avert CC cases and deaths

Role of dipolar and exchange interactions in the positions and widths of EPR transitions for the single-molecule magnets Fe8 and Mn12

We examine quantitatively the temperature dependence of the linewidths and line shifts in electron paramagnetic resonance experiments on single crystals of the single-molecule magnets Fe$_8$ and Mn$_{12}$, at fixed frequency, with an applied magnetic field along the easy axis. We include inter-molecular spin-spin interactions (dipolar and exchange) and distributions in both the uniaxial anisotropy parameter $D$ and the Land\'{e} $g$-factor. The temperature dependence of the linewidths and the line shifts are mainly caused by the spin-spin interactions. For Fe$_8$ and Mn$_{12}$, the temperature dependence of the calculated line shifts and linewidths agrees well with the trends of the experimental data. The linewidths for Fe$_8$ reveal a stronger temperature dependence than those for Mn$_{12}$, because for Mn$_{12}$ a much wider distribution in $D$ overshadows the temperature dependence of the spin-spin interactions. For Fe$_8$, the line-shift analysis suggests two competing interactions: a weak ferromagnetic exchange coupling between neighboring molecules and a longer-ranged dipolar interaction. This result could have implications for ordering in Fe$_8$ at low temperatures.Comment: published versio

The MACHO Project HST Follow-Up: The Large Magellanic Cloud Microlensing Source Stars

We present Hubble Space Telescope (HST) WFPC2 photometry of 13 microlensed source stars from the 5.7 year Large Magellanic Cloud (LMC) survey conducted by the MACHO Project. The microlensing source stars are identified by deriving accurate centroids in the ground-based MACHO images using difference image analysis (DIA) and then transforming the DIA coordinates to the HST frame. None of these sources is coincident with a background galaxy, which rules out the possibility that the MACHO LMC microlensing sample is contaminated with misidentified supernovae or AGN in galaxies behind the LMC. This supports the conclusion that the MACHO LMC microlensing sample has only a small amount of contamination due to non-microlensing forms of variability. We compare the WFPC2 source star magnitudes with the lensed flux predictions derived from microlensing fits to the light curve data. In most cases the source star brightness is accurately predicted. Finally, we develop a statistic which constrains the location of the Large Magellanic Cloud (LMC) microlensing source stars with respect to the distributions of stars and dust in the LMC and compare this to the predictions of various models of LMC microlensing. This test excludes at > 90% confidence level models where more than 80% of the source stars lie behind the LMC. Exotic models that attempt to explain the excess LMC microlensing optical depth seen by MACHO with a population of background sources are disfavored or excluded by this test. Models in which most of the lenses reside in a halo or spheroid distribution associated with either the Milky Way or the LMC are consistent which these data, but LMC halo or spheroid models are favored by the combined MACHO and EROS microlensing results.Comment: 28 pages with 10 included PDF figures, submitted to Ap

Fold Lens Flux Anomalies: A Geometric Approach

We develop a new approach for studying flux anomalies in quadruply-imaged fold lens systems. We show that in the absence of substructure, microlensing, or differential absorption, the expected flux ratios of a fold pair can be tightly constrained using only geometric arguments. We apply this technique to 11 known quadruple lens systems in the radio and infrared, and compare our estimates to the Monte Carlo based results of Keeton, Gaudi, and Petters. We show that a robust estimate for a flux ratio from a smoothly varying potential can be found, and at long wavelengths those lenses deviating from from this ratio almost certainly contain significant substructure.Comment: 16 pages, including 8 figure

The MACHO Project Hubble Space Telescope Follow-Up: Preliminary Results on the Location of the Large Magellanic Cloud Microlensing Source Stars

We attempt to determine whether the MACHO microlensing source stars are drawn from the average population of the LMC or from a population behind the LMC by examining the HST color-magnitude diagram (CMD) of microlensing source stars. We present WFPC2 HST photometry of eight MACHO microlensing source stars and the surrounding fields in the LMC. The microlensing source stars are identified by deriving accurate centroids in the ground-based MACHO images using difference image analysis (DIA) and then transforming the DIA coordinates to the HST frame. We consider in detail a model for the background population of source stars based on that presented by Zhao, Graff & Guhathakurta. In this model, the source stars have an additional reddening = 0.13 mag and a slightly larger distance modulus ~ 0.3 mag than the average LMC population. We also investigate a series of source star models, varying the relative fraction of source stars drawn from the average and background populations and the displacement of the background population from the LMC. Due to the small number of analyzed events the distribution of probabilities of different models is rather flat. A shallow maximum occurs at a fraction s_LMC ~ 0.8 of the source stars in the LMC. This is consistent with the interpretation that a significant fraction of observed microlensing events are due to lenses in the Milky Way halo, but does not definitively exclude other models.Comment: revised version, results slightly changed, accepted by Ap