Neutrophil-specific granule deficiency (SGD) is a rare disorder characterized by recurrent pyogenic infections, defective neutrophil chemotaxis and bactericidal activity, and lack of neutrophil secondary granule proteins. It has been linked to a defect in the transcription factor CCAAT/enhancer binding protein (CEBP) epsilon. Recently, loss-of-function mutations in SMARCD2 were identified from SGD patients. SMARCD2 is chromatin-remodeling factor, that interacts with CEBP epsilon ...
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Mediators pre-stored in neutrophil azurophilic granules are central to the acute inflammatory response and tissue degradation and damage through their proteolytic activity. Different granule populations mobilize and release their content via distinct and hierarchical molecular mechanisms. The molecular mechanisms by which mediators pre-stored in azurophilic granules are mobilized and released to the extracellular space remain largely unknown. We used a number of complementary techniques including; confocal laser scanning microscopy, subcellular fractionation, flow cytometric analyses, Western blot analyses and electron microscopy to examine the ultrastructural and molecular nature of mediator release in neutrophil azurophilic granules. We found that following IL-8 activation, neutrophil azurophilic granules undergo piecemeal degranulation (selective mediator release) leading to altered granule content. Piecemeal degranulation of azurophilic granules is characterized by budding of small secretory ...
Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease characterized by the selective loss of upper and lower motor neurons, a cell type that is intrinsically more vulnerable than other cell types to exogenous stress. The interplay between genetic susceptibility and environmental exposures to toxins has long been thought to be relevant to ALS. One cellular mechanism to overcome stress is the formation of small dense cytoplasmic domains called stress granules (SG) which contain translationally arrested mRNAs. TDP-43 (encoded by TARDBP) is an ALS-causative gene that we have previously implicated in the regulation of the core stress granule proteins G3BP and TIA-1. TIA-1 and G3BP localize to SG under nearly all stress conditions and are considered essential to SG formation. Here, we report that TDP-43 is required for proper SG dynamics, especially SG assembly as marked by the secondary aggregation of TIA-1. We also show that SG assembly, but not initiation, requires G3BP. Furthermore,
Purified cytoplasmic granules from cytotoxic rat large granular lymphocytes (LGL) tumors were cytolytic to erythrocytes, splenocytes, and a number of different lymphoid tumor cells. Granule concentrations of approximately 1 microgram/ml granule protein were adequate to lyse 100% of the erythrocytes, while the nucleated cells required up to 100 micrograms/ml granule protein to achieve complete lysis. Cytoplasmic granules purified from noncytotoxic lymphoid cells did not contain detectable cytolytic activity; purified granules from rat mast cells and rat liver lysosomes likewise failed to display cytolytic activity. However, granules prepared from normal rat peripheral blood LGL were cytolytic. Granule-mediated lysis of erythrocytes and nucleated cells was complete within 3 min at room temperature. The lytic activity required calcium at concentrations of 10(-4)-10(-2) M; magnesium or barium failed to replace calcium, while strontium could replace calcium at 10(-3)-10(-2) M when nucleated cells ...
article{31e5f1bd-a4e6-4a4d-9882-e8e6db59f82f, abstract = {Glucose-evoked insulin secretion exhibits a biphasic time course and is associated with accelerated intracellular granule movement. We combined live confocal imaging of EGFP-labelled insulin granules with capacitance measurements of exocytosis in clonal INS-1 cells to explore the relation between distinct random and directed modes of insulin granule movement, as well as exocytotic capacity. Reducing the temperature from 34 °C to 24 °C caused a dramatic 81% drop in the frequency of directed events, but reduced directed velocities by a mere 25%. The much stronger temperature sensitivity of the frequency of directed events (estimated energy of activation ~ 135 kJ/mol) than that of the granule velocities (~ 22 kJ/mol) suggests that cooling-induced suppression of insulin granule movement is attributable to factors other than reduced motor protein adenosine 5-triphosphatase activity. Indeed, cooling suppresses random granule diffusion by ~ ...
Targeting of the granule membrane or its cargo with a photoactivatable/photoconvertible protein allows in-depth visualisation and tracking of single insulin granules in dependence upon glucose. This technique may also open the way to elucidating the regulation of granule movement velocity within the …
Author summary Zika virus (ZIKV) is transmitted to humans primarily through mosquito bites, but there have also been cases of sexual, perinatal, and suspected blood transfusion transmission. It has been associated with fetal malformations and neurological disorders in adults. The rising concern about this pathogen led the World Health Organization to declare it as a public health emergency of international concern regarding neurological disorders. There is an urgent global scientific effort underway to better understand ZIKV biology and define interactions that occur between the virus and the host cell. We evaluated how ZIKV infection counteracts the assembly of dynamic aggregates of RNA and proteins called stress granules (SGs). We observed that ZIKV blocks SG assembly induced by sodium arsenite (Ars), but not by sodium selenite or Pateamine A. We demonstrate that this difference is related to the ability of ZIKV to modulate the dephosphorylation of eIF2α via its phosphatase. Our work demonstrates
Background: The proportion of granule types in wheat starch is an important characteristic that can affect its functionality. It is widely accepted that granule types are either large, disc-shaped A-type granules or small, spherical B-type granules. Additionally, there are some reports of the tiny C-type granules. The differences between these granule types are due to its carbohydrate composition and crystallinity which is highly, but not perfectly, correlated with the granule size. A majority of the studies that have considered granule types analyse them based on a size threshold rather than chemical composition ...
Lately, we have identified two polypeptides of 92-94 kDa (GRL1) and 45-60 kDa (GRL2), expressed in cytoplasmic granules of chicken granulocytes and thrombocytes. Here, we report that GRL1 and GRL2 are widely distributed in all exocrine and several endocrine cell types, but not in neurons of the central nervous system, during late stages of embryonic development, as well as in newly hatched and two-month-old chickens. Immunogold studies in ultrathin frozen sections of pancreatic acinar cells show that GRL1 and GRL2 are co-localized at the periphery of zymogen granules, in granules fused with apical acinar membranes and on apical membranes of acini, while the pregranular compartments of the secretory pathway are weakly or not labeled. Semiquantitative morphometric studies indicate that GRL1 and GRL2 are equally distributed in secretory granules. A variety of physical and metabolic studies reveal that GRL2, a highly N-glycosylated polypeptide, is an intrinsic membrane protein, while GRL1 is a ...
An azurophilic granule is a cellular object readily stainable with a Romanowsky stain. In white blood cells and hyperchromatin, staining imparts a burgundy or merlot coloration. Neutrophils in particular are known for containing azurophils loaded with a wide variety of anti-microbial defensins that fuse with phagocytic vacuoles. Azurophils may contain myeloperoxidase, phospholipase A2, acid hydrolases, elastase, defensins, neutral serine proteases, bactericidal/permeability-increasing protein, lysozyme, cathepsin G, proteinase 3, and proteoglycans. Azurophil granules are also known as "primary granules". Furthermore, the term "azurophils" may refer to a unique type of cells, identified only in reptiles. These cells are similar in size to so-called heterophils with abundant cytoplasm that is finely to coarsely granular and may sometimes contain vacuoles. Granules may impart a purplish hue to the cytoplasm, particularly to the outer region. Occasionally, azurophils are observed with vacuolated ...
Indicated (A) P-body or (B) stress granule markers were grown for 4 days in Tein E (apoE) gene to families with a higher risk of glucose-containing media and
The prototypic long PTX3 has long been known to be produced by diverse cell types on demand, i.e., in a gene expression-dependent fashion in response to extracellular signals (e.g., LPS, IL-1β, TNFα, and TLR agonists) (3). The finding that PTX3 is stored in neutrophil granules is therefore unexpected. PTX3 is not stored in MPO+ granules (primary or azurophilic). By confocal analysis among the MPO− granules, PTX3 was found to localize in lactoferrin+ and in lactoferrin+/gelatinase+ (specific) but not in gelatinase+ (tertiary) granules. Storage of PTX3 in neutrophil granules is selective, inasmuch as short PTXs are absent and other granulated circulating elements (eosinophils, basophils, and NK cells) do not contain preformed PTX3. In addition to the diversity generated during granulopoiesis, granules are secreted in a targeted manner, with a timing hierarchy in granule exocytosis (50, 51). PTX3 is localized in granules that are rapidly mobilized and secreted upon stimulation, in agreement ...
Regulated, stimulus-dependent secretion via the exocytosis of dense core granule content provides a mechanism for controlling delivery of hormones, enzymes, neuropeptides, and neurotransmitters to a target cell or organ in a timely and quantified manner (Burgoyne and Morgan, 2003; Loh et al., 2004; Lou et al., 2005). In this study, we have investigated the contributions of cholesterol, 7-DHC and lathosterol to granule biogenesis and regulated secretion in exocrine and endocrine tissues of mouse models of SLOS and lathosterolosis. We report here that cholesterol is necessary for the correct formation of granules in vivo. In the formation of regulated secretory vesicles in animals with inborn errors of cholesterol synthesis, cholesterol cannot be substituted by other lipids with structural similarity. We also show deregulated secretion of cargo in acinar cells of the exocrine pancreas, owing to the absence of cholesterol, which suggests impairment of cellular function.. It has been reported that ...
Concentrated deposits of certain substances which are presented/located in the cytoplasm of certain bacteria are known as cytoplasmic granules or inclusion bodies.They serves as storage areas for nutrients, e.g. volutin granules are reserve of high energy stored in the form of polymerized metaphosphate. ...
A key cellular stress granule protein, G3BP1, is critical for efficient norovirus infection, representing the first pan-norovirus, pro-viral factor identified to date.
The dense granules of human platelets contain adenosine diphosphate (ADP), adenosine triphosphate (ATP), ionized calcium (which is necessary for several steps of the coagulation cascade), and serotonin.[6] Dense granules are similar to lysosomes with an acidic pH and even some lysosomal proteins like CD63.[7] There is a granular adenine nucleotide pool within the dense granule. It is thought that it is made up of system of insoluble calcium. This pool is likely to be different than that of the cytoplasmic nucleotides. In some animals it has been shown that the platelets contain histamine.[3] During exocytosis, the pool of ATP within the dense granule is released. Serotonin is picked up by the dense granules where it interacts with ATP and calcium.[3] The serotonin that is then released by the dense granule, recruits other platelets and helps play a major role in stoping the loss of blood at the injury. The calcium from a dense granule accounts for the majority of the calcium within the platelets ...
View more ,Immune-modulating therapies have revolutionized the treatment of chronic diseases, particularly cancer. However, their success is restricted and there is a need to identify new therapeutic targets. Here, we show that natural killer cell granule protein 7 (NKG7) is a regulator of lymphocyte granule exocytosis and downstream inflammation in a broad range of diseases. NKG7 expressed by CD4+ and CD8+ T cells played key roles in promoting inflammation during visceral leishmaniasis and malaria-two important parasitic diseases. Additionally, NKG7 expressed by natural killer cells was critical for controlling cancer initiation, growth and metastasis. NKG7 function in natural killer and CD8+ T cells was linked with their ability to regulate the translocation of CD107a to the cell surface and kill cellular targets, while NKG7 also had a major impact on CD4+ T cell activation following infection. Thus, we report a novel therapeutic target expressed on a range of immune cells with functions in ...
An autosomal recessive disorder characterized by recurrent pyogenic infections, defective neutrophil chemotaxis and bactericidal activity, and lack of neutrophil secondary granule proteins. Neutrophils of affected individuals lack lactoferrin and show abnormal nuclear segmentation, bilobed nuclei, low alkaline phosphatase, and increased number of neutrophil mitochondria and ribosomes ...
cytoplasmic stress granule, extracellular exosome, extracellular matrix, extracellular space, nucleus, plasma membrane, secretory granule, heparin binding, peptidase activity, serine-type endopeptidase activity
Reactome is pathway database which provides intuitive bioinformatics tools for the visualisation, interpretation and analysis of pathway knowledge.
Reactome is pathway database which provides intuitive bioinformatics tools for the visualisation, interpretation and analysis of pathway knowledge.
Hi there, I have run an experiment which is about stress granules formation in yeast using leica sp8 confocal microscope and microfluidics device custom made for the confocal images - its a time lapse with z slices, …
The localization of mRNA forms a key facet of the post-transcriptional control of gene expression and recent evidence suggests that it may be considerably more widespread than previously anticipated. For example, defined mRNA-containing granules can be associated with translational repression or activation. Furthermore, mRNA P-bodies (processing bodies) harbour much of the mRNA decay machinery and stress granules are thought to play a role in mRNA storage. In the present review, we explore the process of mRNA localization in the yeast Saccharomyces cerevisiae, examining connections between organellar mRNA localization and the response to stress. We also review recent data suggesting that even where there is a global relocalization of mRNA, the specificity and kinetics of this process can be regulated. ...
We use high-field NMR spectroscopy, scattering methods, AUC, fluorescence methods, light microscopy, biochemical assays and cell biology. (But not all of these in a single project!). We are collaborating with several other groups in the field. Please contact me for more information at. [email protected] A. Molliex, J. Temirov, J. Lee, M. Coughlin, A.P. Kanagaraj, H.J. Kim, T. Mittag*, J.P. Taylor*. (2015) Phase Separation by Low Complexity Domains Promotes Stress Granule Assembly and Drives Pathological Fibrillization. Cell. 163 (1): 123-33. PMID: 26406374. M. Marzahn, S. Marada, J. Lee, A. Nourse, S. Kenrick, H. Zhao, G. Ben-Nissan, R.M. Kolaitis, J.L. Peters, S. Pounds, W.J. Errington, G.G. Prive, J.P. Taylor, M. Sharon, P. Schuck, S.K. Ogden*, T. Mittag*. (2016) Higher-order oligomerization promotes localization of SPOP to liquid nuclear speckles. EMBO J. 35 (12): 1254-75.. ...
The first decision step separates most granule cells from the other cell types. Since a number of morphologically identified granule cells did not, or hardly, displayed spontaneous activity, we used as one criterion a firing frequency ,0.5 Hz. Other granule cells with spontaneous activity were very irregular, with both very short intervals during high-frequency bursts and very long intervals between bursts. Because of this type of firing, such granule cells usually had CVlog values ,0.38, which served as an alternative criterion to the average frequency. In this way, 82% (n = 9) of the 11 granule cells were properly classified. Cells with a CVlog ,0.34 and with an average firing frequency ,0.6 Hz (n = 75) were moved to the next decision step. The remaining two cells (a basket and a stellate cell) were designated as border cells.. The second decision step takes advantage of the regular firing pattern of UB cells. We found that the regularity of firing is best characterized by the CV2 statistic ...
... contain specific proteins and other macromolecules that are destined for secretion into the extracellular space. This slide shows secretory granules in pancreatic acinar cells. Their size is approximately 1 µm in diameter, and they accumulate on the apical side of the cell above the nucleus. ...
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Deregulation of gene expression contributes to aberrant phenotypes and behaviors of cancer cells. Acquiring a new profile of expressed proteins and their subsequent activation enable cancer cells to re-enter the cell cycle, or give them survival and migratory advantages over those of the normal cells. Alterations in cis-acting sequences, RNA binding proteins, or in upstream signaling pathways affect the stability and/or translational efficiency of mRNAs encoding proto-oncogenes, cytokines, cell cycle regulators and other regulatory proteins to promote tumorigenesis and cancer progression, especially during cellular stress induced by hypoxia or reactive oxygen species (ROS). In addition, these environments can also induce stress granule formation, a consequence of translational suppression and eIF21 phosphorylation, which can inhibit apoptosis. In this application we will focus on developing technology to characterize the deregulation of gene expression by: 1) evaluating the transcriptional ...
The use of granule ready-to-use formulas is based on a style of prescribing developed in Taiwan. It is used frequently and with much success. The most common formulas are used according to their actions for a particular disease pattern and are modified according to the patients individual diagnosis. The widespread opinion in Taiwan is that herbs that have been boiled together and then granulated will have a better effect than formulas built from combining single granules.. ...
The first objective was to use voltage-clamp recordings from single granule cells (holding potential, Vhold = −77 mV) to characterize the classes of synaptic inputs received by granule cells during patterned ON stimulation. In all experiments, granule cells displayed barrages of inward current events that persisted for the duration of the stimulus (Fig. 1). The exact kinetic characteristics of the events varied significantly between cells, in what appeared to be an age-dependent manner. In young animals [postnatal day 8 (P8) or younger], the events tended to have "moderate" decay kinetics (Fig. 1a-c, GC1) with a 20-80% rise time of 1.43 ± 0.02 ms and decay time constants of 5.7 ± 0.3 ms (n = 17). Approximately 44% of the granule cells (8 of 18) in young animals consisted entirely of such events (termed "type 1" events). In contrast, granule cells in bulb slices taken from older animals (older than P14) generally showed more rapid events (Fig. 1a-c, GC2) (20-80% rise time, 0.34 ± 0.01 ms; ...
Define myelocyte: a bone-marrow cell; especially : a motile cell with cytoplasmic granules that gives rise to the granulocytes of the blood and…
Organization is key to an efficient workplace, and cells are no exception to this rule. New evidence from Johns Hopkins researchers suggests that, in addition to membranes, cells have another way to keep their contents and activities separate: with ribbons of spinning proteins. A summary of their findings appears today in the journal eLife.. Each cell is a busy warehouse of activity. To keep things orderly, protein workers are "assigned" to specific areas of the cell where other workers are collaborating on the same project. Most of the project areas, or organelles, in the cell are cordoned off by flexible membranes that let things in and out on an as-needed basis, but some organelles, like RNA granules, do not seem to have clear boundaries.. RNA granules float throughout the watery space inside the cell and are responsible for transporting, storing and controlling RNA - DNAs chemical cousin - which holds blueprints for proteins. Until now, researchers thought that the granules didnt have ...
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GP2 overexpression lysate, 0.1 mg. Transient overexpression lysate of glycoprotein 2 (zymogen granule membrane) (GP2), transcript variant 2
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Hey, do osmcate plus root tabs dissolve in your substrate or do they stay in granule form even after they have been exhausted??? Thanks.
TY - JOUR. T1 - Core formation and the acquisition of fusion competence are linked during secretory granule maturation in Tetrahymena. AU - Bowman, Grant R.. AU - Elde, Nels C.. AU - Morgan, Garry. AU - Winey, Mark. AU - Turkewitz, Aaron P.. PY - 2005/4/1. Y1 - 2005/4/1. N2 - The formation of dense core secretory granules is a multi-stage process beginning in the trans Golgi network and continuing during a period of granule maturation. Direct interactions between proteins in the membrane and those in the forming dense core may be important for sorting during this process, as well as for organizing membrane proteins in mature granules. We have isolated two mutants in dense core granule formation in the ciliate Tetrahymena thermophila, an organism in which this pathway is genetically accessible. The mutants lie in two distinct genes but have similar phenotypes, marked by accumulation of a set of granule cargo markers in intracellular vesicles resembling immature secretory granules. Sorting to ...
RNA granules play diverse role in controlling the turnover and translation of specific mRNAs. FMRP is one of the major components of RNA granules including neuronal granules (Barbee et al., 2006), stress granules (Mazroui et al., 2002), and in vitro assembled granules (Han et al., 2012; Kato et al., 2012). Overexpression of FMRP induces formation of FMRP granules that are reminiscent of RNA granules. Here we used FMRP granules formed in Drosophila cells as a model to investigate how FMRP could promote formation of RNA granules and to define the determinants of FMRP involved in its trafficking in and out RNA granules.. During the past few years, several mammalian SG nucleating factors have been shown to induce the formation of SG-like RNA granules based on the effect of their overexpression. Formation of these SG-like granules was reported to occur through mechanisms which involve the phosphorylation of eIF2α, a key event essential for the triggering of SG formation upon stress (Kedersha et al., ...
Secretory granules are specialized intracellular organelles that serve as a storage pool for selected secretory products. The exocytosis of secretory granules is markedly amplified under physiologically stimulated conditions. While granules have been recognized as post-Golgi carriers for almost 40 years, the molecular mechanisms involved in their formation from the trans-Golgi network are only beginning to be defined. This review summarizes and evaluates current information about how secretory proteins are thought to be sorted for the regulated secretory pathway and how these activities are positioned with respect to other post-Golgi sorting events that must occur in parallel. In the first half of the review, the emerging role of immature secretory granules in protein sorting is highlighted. The second half of the review summarizes what is known about the composition of granule membranes. The numerous similarities and relatively limited differences identified between granule membranes and other ...
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A partially purified secretory granule fraction, isolated from rat islets of Langerhans by differential centrifugation, was used for investigating the stability of the beta granules during incubation in various conditions. Effects of pH, temperature, and time were studied; the granules possessed optimal stability at 4° and pH 6.0, and could be solubilized at pH 4.0 or 8.5, or in the presence of sodium deoxycholate, but not by phospholipase c, ouabain, or alloxan. Incubation with glucose or some of its metabolites, or with tolbutamide, ATP, or cyclic 3,5-AMP did not alter the stability of the beta granules Exogenous insulin-131I was not bound by the isolated granules under the conditions used; no specific insulin-degrading activity could be detected in subcellular fractions of the islets. These findings indicate that intracellular solubilization of the granules with subsequent diffusion of the insulin into the extracellular space is not a likely mode of insulin secretion in vivo, and suggest ...
article{f429a20c-cc6f-41ca-9f07-404d5d006fd6, abstract = {Cysteine-rich secretory protein 3 (CRISP-3; also known as SGP28) was originally discovered in human neutrophilic granulocytes. We have recently developed a sensitive sandwich enzyme-linked immunosorbent assay for CRISP-3 and demonstrated the presence of CRISP-3 in exocrine secretions. To investigate the subcellular localization and mobilization of CRISP-3 in human neutrophils, we performed subcellular fractionation of resting and activated neutrophils on three-layer Percoll density gradients, release-studies of granule proteins in response to different secretagogues, and double-labeling immunogold electron microscopy. CRISP-3 was found to be localized in a subset of granules with overlapping characteristics of specific and gelatinase granules and mobilized accordingly, thus confirming the hypothesis that peroxidase-negative granules exist as a continuum from specific to gelatinase granules regarding protein content and mobilization. ...
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Currently, my lab addresses the following questions:. How does the pseudophophatase MK-STYX inhibit stress granule formation?. Environmental cues such as heat shock, UV irradiation, hypoxia, and oxidative stress initiate many eukaryotic cellular responses, including protective responses to ensure their survival. One of the most rapid of these responses is to prevent mRNA translation to into protein to allow cells to adapt to stress. This stalled mRNA is sequestered to specific cytosolic compartments known as stress granule. Stress granules represent a complex assemblage of translational initiation factors, proteins involved in translational control, the microtubule array, and chaperone proteins such as G3BP-1 (Ras-GTPase-activating protein Src homology 3 domain-binding protein-1). We have shown that MK-STYX binds G3BP-1 and inhibit stress granule formation. However, how MK-STYX accomplishes this inhibition is unclear. Thus, we seek to elucidate the mechanism required by MK-STYX to inhibit stress ...
Background: Grass pollen is one of the most important aeroallergens in Europe. It highly contributes to respiratory allergic diseases, mainly allergic rhinitis. In contact to water or airborne pollutants, pollen grains can release pollen cytoplasmic granules (PCGs) containing allergens. Because of their size (| 5 mu m), PCGs may penetrate deeper into the lungs to induce higher allergic responses, such as asthma. They have been associated with thunderstorm-related asthma. The aim of this study was to evaluate, with Brown Norway rats, the allergenic potential of isolated PCGs and to compare it with the allergenicity of whole timothy grass pollen. Methods: Rats were sensitized (day 0) and challenged (day 21), in controlled comparative conditions, with pollen grains (0.5 mg) or PCGs (4.5 x 10(7) and 0.5 mg). At day 25, blood samples, bronchoalveolar lavage fluid (BALF) and bronchial lymph node were collected. IgE and IgG1 levels in sera were assessed by ELISA. Alveolar cells, protein and cytokine