Desiccation and zinc binding induce transition of tomato abscisic acid stress ripening 1, a water stress- and salt stress-regulated plant-specific protein, from unfolded to folded state

Abscisic acid stress ripening 1 (ASR1) is a low molecular weight plant-specific protein encoded by an abiotic stress-regulated gene. Overexpression of ASR1 in transgenic plants increases their salt tolerance. The ASR1 protein possesses a zinc-dependent DNA-binding activity. The DNA-binding site was mapped to the central part of the polypeptide using truncated forms of the protein. Two additional zinc-binding sites were shown to be localized at the amino terminus of the polypeptide. ASR1 protein is presumed to be an intrinsically unstructured protein using a number of prediction algorithms. The degree of order of ASR1 was determined experimentally using nontagged recombinant protein expressed in Escherichia coli and purified to homogeneity. Purified ASR1 was shown to be unfolded using dynamic light scattering, gel filtration, microcalorimetry, circular dichroism, and Fourier transform infrared spectrometry. The protein was shown to be monomeric by analytical ultracentrifugation. Addition of zinc ions resulted in a global change in ASR1 structure from monomer to homodimer. Upon binding of zinc ions, the protein becomes ordered as shown by Fourier transform infrared spectrometry and microcalorimetry, concomitant with dimerization. Tomato (Solanum lycopersicum) leaf soluble ASR1 is unstructured in the absence of added zinc and gains structure upon binding of the metal ion. The effect of zinc binding on ASR1 folding and dimerization is discussed.     

The partial contribution of specific airborne pollen to pollen induced allergy

The air that we inhale contains simultaneously a multiple array of allergenic pollen. It is well known that such allergens cause allergic reactions in some 15 of the population of the Western World. However little is known about the quantitative aspect of this phenomenon. What is the lowest concentration of pollen that might trigger allergic responses? As people are exposed to heterogeneous and variable environments, clarification of the partial contribution of each of the major airborne pollen allergens and determination of its role in invoking allergy are of prime importance. Objectives: (1) Assessment of a possible correlation between the concentration of airborne pollen and incidence of allergy. (2) Estimation of the lowest average concentrations for various species of airborne pollen that elicit allergic symptoms when exceeded. (3) Determination of the extent of the variations in manifestation of allergy symptoms that can be explained by fluctuations in the concentration of individual species of airborne pollen. Methods: The study was conducted during 14months with a rural population in Israel. The participants completed a detailed questionnaire and were skin prick tested with the common airborne allergens. The appearance of clinical symptoms, i.e. nasal, bronchial, ocular or dermal, were reported daily by the patients. Concentrations of the airborne pollen and spores were monitored in the center of activity of the residents during one day every week, using three Rotorod pollen traps. The pollen grains were identified by light microscopy. Results: The pollen spectrum was divided into time-blocks presenting the main pollination periods of the investigated species. The correlation between the concentration of airborne pollen of the relevant species and the clinical symptoms of the patients was determined for each time block. The correlation differed for different clinical symptoms and for different pollen allergens. Highest correlation with airborne pollen counts was found for patients with nasal and bronchial symptoms. The onset of the clinical symptoms by sensitive patients started, in each of the relevant groups, once the weekly average concentration of the airborne pollen crossed a threshold level. Under the limitations of the present study, this level was estimated to be 2–4 pollen m^–3 air for olive, 3–5 pollen m^–3 air for grasses, 4–5 pollen m^–3 air for Artemisia, 10–20 pollen m^–3 air for pecan and 50–60 pollen m^–3 air for cypress. Conclusions: Fluctuations in specific airborne pollen grains explained up to 2/3 of the variation in clinical allergy responses. Those were: 69 of the variation for cypress (March–April), 66 for the grasses (March–April), 49 for the pecan (May–June) and 62 for Artemisia (Autumn).     

The partial contribution of specific airborne pollen to pollen induced allergy

The air that we inhale contains simultaneously a multiple array of allergenic pollen. It is well known that such allergens cause allergic reactions in some 15 of the population of the Western World. However little is known about the quantitative aspect of this phenomenon. What is the lowest concentration of pollen that might trigger allergic responses? As people are exposed to heterogeneous and variable environments, clarification of the partial contribution of each of the major airborne pollen allergens and determination of its role in invoking allergy are of prime importance. Objectives: (1) Assessment of a possible correlation between the concentration of airborne pollen and incidence of allergy. (2) Estimation of the lowest average concentrations for various species of airborne pollen that elicit allergic symptoms when exceeded. (3) Determination of the extent of the variations in manifestation of allergy symptoms that can be explained by fluctuations in the concentration of individual species of airborne pollen. Methods: The study was conducted during 14?months with a rural population in Israel. The participants completed a detailed questionnaire and were skin prick tested with the common airborne allergens. The appearance of clinical symptoms, i.e. nasal, bronchial, ocular or dermal, were reported daily by the patients. Concentrations of the airborne pollen and spores were monitored in the center of activity of the residents during one day every week, using three ‘Rotorod’ pollen traps. The pollen grains were identified by light microscopy. Results: The pollen spectrum was divided into time-blocks presenting the main pollination periods of the investigated species. The correlation between the concentration of airborne pollen of the relevant species and the clinical symptoms of the patients was determined for each time block. The correlation differed for different clinical symptoms and for different pollen allergens. Highest correlation with airborne pollen counts was found for patients with nasal and bronchial symptoms. The onset of the clinical symptoms by sensitive patients started, in each of the relevant groups, once the weekly average concentration of the airborne pollen crossed a threshold level. Under the limitations of the present study, this level was estimated to be 2–4 pollen m^?3 air for olive, 3–5 pollen m^?3 air for grasses, 4–5 pollen m^?3 air for Artemisia, 10–20 pollen m^?3 air for pecan and 50–60 pollen m^?3 air for cypress. Conclusions: Fluctuations in specific airborne pollen grains explained up to 2/3 of the variation in clinical allergy responses. Those were: 69 of the variation for cypress (March–April), 66 for the grasses (March–April), 49 for the pecan (May–June) and 62 for Artemisia (Autumn).     

Disruption of a Quorum Sensing mechanism triggers tumorigenesis: a simple discrete model corroborated by experiments in mammary cancer stem cells

Background  

The balance between self-renewal and differentiation of stem cells is expected to be tightly controlled in order to maintain tissue homeostasis throughout life, also in the face of environmental hazards. Theory, predicting that homeostasis is maintained by a negative feedback on stem cell proliferation, implies a Quorum Sensing mechanism in higher vertebrates.     

Protein glycosylation in Haloferax volcanii: partial characterization of a 98-kDa glycoprotein

Eubacterium ramulus, a flavonoid-degrading anaerobic bacterium from the human gastrointestinal tract, was tested for its ability to transform the isoflavonoids genistein-7-O-glucoside (genistin), genistein and daidzein. Genistein was completely degraded by E. ramulus via 6'-hydroxy-O-desmethylangolensin to 2-(4-hydroxyphenyl)-propionic acid. Dihydrogenistein was neither observed as an intermediate in this transformation nor converted itself by growing cells or cell-free extracts of E. ramulus. Genistein-7-O-glucoside was partially transformed by way of genistein to the product 2-(4-hydroxyphenyl)-propionic acid. Daidzein was in part degraded to O-desmethylangolensin, the corresponding metabolite to 6'-hydroxy-O-desmethylangolensin. The hydroxyl group in position 6' of O-desmethylangolensin is crucial for further degradation.     

Maturation of the humoral immune response as an optimization problem.

Efficient immune response often depends on the production of high affinity antibodies. We show analytically that the optimal strategy for a fast production of high affinity antibodies is to utilize a step-function mutation rate, i.e. a minimal mutation rate in early stages of the immune response, followed by a discontinuous switch to the maximal possible rate when the proliferating population of B-cells exceeds a threshold value. Our results are in accordance with the biological observations concerning the time of onset of the hypermutation process, and with the mutation rate during the later stages of the primary immune response. Indeed the hypermutation process plays a crucial role in responding to a prevailing pathogen at each round of immune response, and not only for coping with future infections. Moreover, as the effect of hypermutations is shown to be crucially dependent on the number of proliferating B-cells, its onset is not expected to depend on an external signal, but rather to be related to the clone's age. This suggests that the onset is host species specific, rather than pathogen specific. Another implication of the present results is that activation of hypermutations before the B-cell population has reached the critical size may impede the efficiency of the response.     

Dickkopf1 regulates fate decision and drives breast cancer stem cells to differentiation: an experimentally supported mathematical model

Background

Modulation of cellular signaling pathways can change the replication/differentiation balance in cancer stem cells (CSCs), thus affecting tumor growth and recurrence. Analysis of a simple, experimentally verified, mathematical model suggests that this balance is maintained by quorum sensing (QS).

Methodology/Principal Findings

To explore the mechanism by which putative QS cellular signals in mammary stem cells (SCs) may regulate SC fate decisions, we developed a multi-scale mathematical model, integrating extra-cellular and intra-cellular signal transduction within the mammary tissue dynamics. Preliminary model analysis of the single cell dynamics indicated that Dickkopf1 (Dkk1), a protein known to negatively regulate the Wnt pathway, can serve as anti-proliferation and pro-maturation signal to the cell. Simulations of the multi-scale tissue model suggested that Dkk1 may be a QS factor, regulating SC density on the level of the whole tissue: relatively low levels of exogenously applied Dkk1 have little effect on SC numbers, whereas high levels drive SCs into differentiation. To verify these model predictions, we treated the MCF-7 cell line and primary breast cancer (BC) cells from 3 patient samples with different concentrations and dosing regimens of Dkk1, and evaluated subsequent formation of mammospheres (MS) and the mammary SC marker CD44⁺CD24^lo. As predicted by the model, low concentrations of Dkk1 had no effect on primary BC cells, or even increased MS formation among MCF-7 cells, whereas high Dkk1 concentrations decreased MS formation among both primary BC cells and MCF-7 cells.

Conclusions/Significance

Our study suggests that Dkk1 treatment may be more robust than other methods for eliminating CSCs, as it challenges a general cellular homeostasis mechanism, namely, fate decision by QS. The study also suggests that low dose Dkk1 administration may be counterproductive; we showed experimentally that in some cases it can stimulate CSC proliferation, although this needs validating in vivo.     

Fast and Slow Readers of the Hebrew Language Show Divergence in Brain Response ～200 ms Post Stimulus: An ERP Study

Higher N170 amplitudes to words and to faces were recently reported for faster readers of German. Since the shallow German orthography allows phonological recoding of single letters, the reported speed advantages might have their origin in especially well-developed visual processing skills of faster readers. In contrast to German, adult readers of Hebrew are forced to process letter chunks up to whole words. This dependence on more complex visual processing might have created ceiling effects for this skill. Therefore, the current study examined whether also in the deep Hebrew orthography visual processing skills as reflected by N170 amplitudes explain reading speed differences. Forty university students, native speakers of Hebrew without reading impairments, accomplished a lexical decision task (i.e., deciding whether a visually presented stimulus represents a real or a pseudo word) and a face decision task (i.e., deciding whether a face was presented complete or with missing facial features) while their electroencephalogram was recorded from 64 scalp positions. In both tasks stronger event related potentials (ERPs) were observed for faster readers in time windows at about 200 ms. Unlike in previous studies, ERP waveforms in relevant time windows did not correspond to N170 scalp topographies. The results support the notion of visual processing ability as an orthography independent marker of reading proficiency, which advances our understanding about regular and impaired reading development.