Miracle-dz/UniFAR-FTUs huggingface dataset:
Drosophila twinfilin is required for cell migration and synaptic endocytosis

Precise actin regulation is essential for diverse cellular processes such as axonal growth, cell migration and endocytosis. twinfilin (twf) encodes a protein that sequesters actin monomers, but its in vivo functions are unclear. In this study, we characterized twf-null mutants in a metazoan for the first time and found that Drosophila twf negatively regulates F-actin formation in subcellular regions of rapid actin turnover in three different systems, namely postsynaptic neuromuscular junction (NMJ) synapses, migratory border cells and epithelial follicle cells. Loss of twf function results in defects in axonal growth in the brain and border cell migration in the ovary. Additionally, we found that the actin-dependent postsynaptic localization of glutamate receptor GluRIIA, but not GluRIIB, was specifically reduced in twf mutants. More importantly, we showed that twf mutations caused significantly reduced presynaptic endocytosis at NMJ synapses, as detected using the fluorescent dye FM1-43 uptake assay. Furthermore, electrophysiological analysis under high-frequency stimulation showed compromised neurotransmission in twf mutant synapses, confirming an insufficient replenishment of synaptic vesicles. Together, our results reveal that twinfilin promotes actin turnover in multiple cellular processes that are highly dependent on actin dynamics.
Plasma Cytokine Levels in Tanzanian HIV-1-Infected Adults and the Effect of Antiretroviral Treatment

Objective:To evaluate the role immune activation leading to the production and circulation of cytokines has in the pathogenesis of HIV infection in sub-Saharan Africa and the effect of antiretroviral treatment (ART) on these parameters. Methods:Plasma concentrations of tumor necrosis factor (TNF)-α, interleukin (IL)-6, IL-8, monocyte chemotactic protein (MCP)-1, IL-10, and IL-1 receptor antagonist; plasma HIV RNA; hemoglobin concentration; and white blood cells were measured in 229 HIV-infected, 54 HIV-uninfected, and after 2 and 4 months, respectively, of ART in 35 eligible individuals in northeastern Tanzania. Results:Plasma levels of tumor necrosis factor-α, IL-6, IL-8, monocyte chemotactic protein-1, and IL-1 receptor antagonist were significantly higher in HIV-infected individuals compared with HIV-uninfected individuals and also significantly higher in HIV-infected individuals with CD4 cell counts below 200 cells per microliter than individuals with CD4 cell counts above 200 cells per microliter. HIV RNA was the strongest predictor of all cytokine expression in multivariate analysis. ART leads to a decrease in all cytokines to levels close to those of HIV-uninfected individuals. Conclusions:Our findings support the role of HIV viral replication as the most important promoter of immune activation and prove the importance of ART in reducing immune activation and viral replication even in sub-Saharan Africa where patients are exposed to an abundance of other infectious agents.
Distinct mobilization of leukocytes and hematopoietic stem cells by CXCR4 peptide antagonist LY2510924 and monoclonal antibody LY2624587

Stromal cell-derived factor-1 (SDF-1) and its receptor CXCR4 play a critical role in mobilization and redistribution of immune cells and hematopoietic stem cells (HSCs). We evaluated effects of two CXCR4-targeting agents, peptide antagonist LY2510924 and monoclonal antibody LY2624587, on mobilizing HSCs and white blood cells (WBCs) in humans, monkeys, and mice. Biochemical analysis showed LY2510924 peptide blocked SDF-1/CXCR4 binding in all three species; LY2624587 antibody blocked binding in human and monkey, with minimal activity in mouse. Cellular analysis showed LY2624587 antibody, but not LY2510924 peptide, down-regulated cell surface CXCR4 and induced hematological tumor cell death; both agents have been shown to inhibit SDF-1/CXCR4 interaction and downstream signaling. In animal models, LY2510924 peptide induced robust, prolonged, dose- and time-dependent WBC and HSC increases in mice and monkeys, whereas LY2624587 antibody induced only moderate, transient increases in monkeys. In clinical trials, similar pharmacodynamic effects were observed in patients with advanced cancer: LY2510924 peptide induced sustained WBC and HSC increases, while LY2624587 antibody induced only minimal, transient WBC changes. These distinct pharmacodynamic effects in two different classes of CXCR4 inhibitors are clinically important and should be carefully considered when designing combination studies with immune checkpoint inhibitors or other agents for cancer therapy.
Ethylene- and Shade-Induced Hypocotyl Elongation Share Transcriptome Patterns and Functional Regulators1[OPEN]

Ethylene and shade share a conserved set of molecular regulators to control the plasticity of hypocotyl elongation in Arabidopsis. Plants have evolved shoot elongation mechanisms to escape from diverse environmental stresses such as flooding and vegetative shade. The apparent similarity in growth responses suggests a possible convergence of the signaling pathways. Shoot elongation is mediated by passive ethylene accumulating to high concentrations in flooded plant organs and by changes in light quality and quantity under vegetation shade. Here, we study hypocotyl elongation as a proxy for shoot elongation and delineate Arabidopsis (Arabidopsis thaliana) hypocotyl length kinetics in response to ethylene and shade. Based on these kinetics, we further investigated ethylene- and shade-induced genome-wide gene expression changes in hypocotyls and cotyledons separately. Both treatments induced a more extensive transcriptome reconfiguration in the hypocotyls compared with the cotyledons. Bioinformatics analyses suggested contrasting regulation of growth promotion- and photosynthesis-related genes. These analyses also suggested an induction of auxin, brassinosteroid, and gibberellin signatures and the involvement of several candidate regulators in the elongating hypocotyls. Pharmacological and mutant analyses confirmed the functional involvement of several of these candidate genes and physiological control points in regulating stress-escape responses to different environmental stimuli. We discuss how these signaling networks might be integrated and conclude that plants, when facing different stresses, utilize a conserved set of transcriptionally regulated genes to modulate and fine-tune growth.
[Roles of miR-590-5p and miR-590-3p in the development of hepatocellular carcinoma].

OBJECTIVE To study the effect of the two arms of miR-590, miR-590-5p and miR-590-3p, on hepatoma cell proliferation and their roles in tumor development. METHODS We analyzed and verified the expression pattern of miR-590 in liver cancer specimens and cell lines by miRNA microarrays and QPCR. MiR-590 mimic or inhibitor was transfected into normal liver cells or liver cancer cells via liposome, and the changes in cell proliferation and survival were determined by MTT assay and soft agar colony formation assay. The target genes of miR-590-5p and miR-590-3p were predicated with Targetscan and validated by luciferase reporter system and Western blotting. RESULTS The expressions of miR-590-5p and miR-590-3ps were up-regulated in 3 hepatocellular carcinoma (HCC) tissues and their synchronization was significantly up-regulated in 8 out of 10 HCC tissues as compared with the adjacent tissues. QPCR further showed that miR-590-5p/3p was up-regulated in 3 HCC cell lines (HepG2, Hep3B, and Huh7) in comparison with the normal liver cell line L-O2. L-O2 cells over-expressing miR-590-5p and miR-590-3p exhibited significantly increased proliferation (P<0.05), while down-regulation of miR-590-5p and miR-590-3p caused significantly suppressed proliferation in HepG2, Hep3B, and Huh7 cells. Targetscan predicted PDCD4 and PTEN as the potential target genes of miR-590-5p and miR-590-3p, which was verified by luciferase reporter system and Western blotting. miR-590-3p was found to activate PI3K-AKT signaling pathway by down-regulating PTEN to promote AKT1-S473 phosphorylation. CONCLUSION MiR-590 is an important tumorigenic factor for HCC, and its two arms can both promote tumorigenesis by regulating the expression of their target tumor suppressor gene, PDCD4 and PTEN, to promote HCC cell proliferation and survival and activate the core tumor signal pathway PI3K-AKT.
Binding properties of indocyanine green in human blood.

PURPOSE Binding properties of indocyanine green (ICG) to human plasma proteins were identified using electrophoresis and a fundus video system. METHODS Blood samples were obtained from three healthy volunteers after intravenous administration of ICG. The resulting plasma samples were fractionated by agarose gel immunoelectrophoresis and polyacrylamide gel DISC electrophoresis. In the former, antisera, anti-apolipoprotein (Apo)-A, and anti-Apo-B antibodies were used to identify all classes of plasma proteins, high-density lipoprotein (HDL), and low-density lipoprotein (LDL), respectively. In the latter method, plasma samples could be separated into chylomicron, very low-density lipoprotein, LDL, and HDL. The electrophoretic pattern obtained by each method was observed with an ICG fundus video system. Furthermore, we studied the affinity of ICG for lipids that were common molecular components of HDL and LDL. Four kinds of ICG solutions mixed with phospholipid, free cholesterol, esterified cholesterol, and triacylglycerol were observed with the ICG fundus video system. RESULTS Both electrophoretic studies showed that ICG bound intensely to HDL and moderately to LDL, and only the solution with phospholipid fluoresced brightly when observed with the ICG fundus video system. CONCLUSIONS These findings indicated low vascular or tissue permeability of ICG, which is caused by the larger molecular size of HDL and LDL. Also noted was that the ICG fluorescence observed in the angiogram may be equivalent to the hemodynamics of HDL alone or in combination with LDL in the bloodstream. This biochemical consideration may be a basis for the further understanding of ICG angiography.
Using tools and technology to promote education and adherence to oral agents for cancer.

BACKGROUND The use of oral agents for cancer (OACs) is increasing, and oncology nurses are in an ideal position to educate patients about them and suggest methods to improve adherence. Once an OAC is ordered, the administration is the responsibility of the patient. Oncology nurses can use tools and technology to assist with education, which may promote adherence, and suggest reminder tools that can be used. Many electronic tools have been developed, such as smartphone applications, text messaging, electronic alarms, and glowing pill bottles. OBJECTIVES The researchers reviewed electronic devices, as well as traditional methods such as calendars and pillboxes, that can assist patients in remembering to take the medication they are administering at home. METHODS A literature search was compiled and websites were searched for patient education tools, reminder tools (electronic and manual), and smartphone applications. The project was part of the Oncology Nursing Society Putting Evidence Into Practice effort on oral adherence. FINDINGS Education alone is insufficient to promote adherence to oral medication regimens. Multicomponent interventions have demonstrated improved adherence, and tools and technology directed at improving adherence to oral agents can be used. The researchers found multiple reminder aids to assist patients in adhering to an oral regimen. They are highlighted in this article.
Protein kinase C can inhibit TRPC3 channels indirectly via stimulating protein kinase G

There are two known phosphorylation‐mediated inactivation mechanisms for TRPC3 channels. Protein kinase G (PKG) inactivates TRPC3 by direct phosphorylation on Thr‐11 and Ser‐263 of the TRPC3 proteins, and protein kinase C (PKC) inactivates TRPC3 by phosphorylation on Ser‐712. In the present study, we explored the relationship between these two inactivation mechanisms of TRPC3. HEK cells were first stably transfected with a PKG‐expressing construct and then transiently transfected with a TRPC3‐expressing construct. Addition of 1‐oleoyl‐2‐acetyl‐sn‐glycerol (OAG), a membrane‐permeant analog of diacylglycerol (DAG), elicited a TRPC3‐mediated [Ca2+]i rise in these cells. This OAG‐induced rise in [Ca2+]i could be inhibited by phorbol 12‐myristate 13‐acetate (PMA), an agonist for PKC, in a dose‐dependent manner. Importantly, point mutations at two PKG phosphorylation sites (T11A‐S263Q) of TRPC3 markedly reduced the PMA inhibition. Furthermore, inhibition of PKG activity by KT5823 (1 µM) or H8 (10 µM) greatly reduced the PMA inhibition of TRPC3. These data strongly suggest that the inhibitory action of PKC on TRPC3 is partly mediated through PKG in these PKG‐overexpressing cells. The importance of this scheme was also tested in vascular endothelial cells, in which PKG plays a pivotal functional role. In these cells, OAG‐induced [Ca2+]i rise was inhibited by PMA, which activates PKC, and by 8‐BrcGMP and S‐nitroso‐N‐acetylpenicillamine (SNAP), both of which activate PKG. Importantly, the PMA inhibition on OAG‐induced [Ca2+]i rise was significantly reduced by PKG inhibitor KT5823 (1 µM) or DT‐3 (500 nM), suggesting an important role of PKG in the PMA‐induced inhibition of TRPC channels in native endothelial cells. J. Cell. Physiol. 207: 315–321, 2006. © 2005 Wiley‐Liss, Inc.
Hepatitis B virus infection initiates with a large surface protein-dependent binding to heparan sulfate proteoglycans.

UNLABELLED Contrary to many other viruses, the initial steps of the hepatitis B virus (HBV) infection, including attachment to hepatocytes, specific receptor interactions, and membrane fusion, are unsolved. Using HepaRG cells as an in vitro cell culture system, we here report that HBV entry into hepatocytes depends on the interaction with the glycosaminoglycan (GAG) side chains of cell-surface-associated heparan sulfate proteoglycans. Binding to GAGs requires the integrity of the pre-S domain as a part of the large (L-) viral envelope protein. HBV infection was abrogated by incubation of virions with heparin, but not the structurally related GAGs chondroitin sulfate A, B, and C. Infection was also abolished by suramin, a known inhibitor of duck hepatitis B virus infection or highly sulfated dextran sulfate. Polycationic substances such as poly-L-lysine, polybrene, and protamine also prevented infection, however, by addressing cellular components. Enzymatic removal of defined acidic carbohydrate structures from the cell surface using heparinase I/III or the obstruction of GAG synthesis by sodium chlorate inhibited HBV infection of HepaRG cells and, moreover, led to a reduction of HBV cell surface binding sites. The biochemical analysis showed selective binding of L-protein-enriched viral particles (virions or filaments) to heparin. GAG-dependent binding of HBV was improved by polyethylene glycol, a substance that specifically enhances HBV infection. CONCLUSION HBV infection requires the initial attachment to the carbohydrate side chains of hepatocyte-associated heparan sulfate proteoglycans as attachment receptors. This interaction initializes the multistep entry process of HBV and cannot be bypassed by alternative routes.
Ultrastructural Immunogold Localization of Osteopontin in Human Gastric Mucosa

We performed ultrastructural immunogold localization of osteopontin in the mucosa of human stomach. This adhesive glycoprotein was present in mucous and chief cells of the epithelial layer and in macrophages in the lamina propria. Parietal and endocrine cells of the epithelial layer and mast cells and plasma cells in the lamina propria did not contain osteopontin, serving as internal negative controls. Subcellular localizations of osteopontin included secretory granules and synthetic organelles in mucous and chief cells and phagolysosomes in macrophages. Extracellular concentrations of osteopontin were present in the glycocalyx and in an electron-lucent band between epithelial surface cells and the gastric lumen. Paracellular edema between the epithelium of the same cells was devoid of osteopontin. Immunogold localization of pepsinogen II was done to identify cells with mixed granule populations and contents of multicompartmental secretory granules. These studies revealed mucous cell granules and chief cell granules, each containing compartmentalized storage products, which included osteopontin and mucigen in mucous cells and osteopontin and pepsinogen II in chief cells. Cytochemical controls for the immunogold localizations were negative. The subcellular distribution of osteopontin in human gastric mucosa suggests possible roles for this glycoprotein in barrier function, host defense, and/or secretion.
Distribution and Appearance of Tumor Necrosis Factor-&agr; in the Dorsal Root Ganglion Exposed to Experimental Disc Herniation in Rats

Study Design. Distribution and appearance of tumor necrosis factor-alpha (TNF-&agr;) in the dorsal root ganglion (DRG) exposed to experimental disc herniation were investigated using an immunohistochemical method in rats. Objectives. To study the distribution and appearance of TNF-&agr; in the DRG following experimental disc herniation in rats. Summary of Background Data. Nucleus pulposus in the epidural space induces spinal nerve root injury not only by mechanical but also chemical mechanisms. Cytokines may play a key role in the chemical damage. There is, however, no report on the distribution and appearance of TNF-&agr; in the DRG exposed to nucleus pulposus. Methods. Nucleus pulposus from the discs was smeared on the glass slides and processed for immunohistochemistry by the avidin-biotinylated peroxidase complex technique using rabbit antisera to TNF-&agr; in rats. A herniation of the nucleus pulposus was made by incision of the L4–L5 disc in rats. The L4 and L5 DRGs were resected 1, 3, 7, 14, and 21 days after surgery. The specimens were processed for immunohistochemistry using rabbit antisera to TNF-&agr;. The TNF-&agr;-positive cells were observed and counted using light microscopy. Distribution of the TNF-&agr; products was compared on each day after surgery. Results. A positive staining was seen in the cell bodies and in the matrix between the cells in the smeared nucleus pulposus. In the L4 DRG sections, the number of positive cells was significantly higher in the disc incision group than in the sham group at 1, 3, 7, and 14 days after surgery (P < 0.05). The positive cells showed a decrease in number day by day after surgery. On the contrary, in the L5 DRG, only a few positive cells were observed in the disc incision group after surgery. There was no statistically significant difference between disc incision and the sham groups at each day after surgery for the L5 DRGs. Conclusions. The immunoreactivity of TNF-&agr; in the DRG directly exposed to nucleus pulposus increases during 2 weeks. A collapse of the positive cells was seen in the DRG directly exposed to the nucleus pulposus.
Daily mood and sleep: reciprocal relations and links with adjustment problems

Children's sleep problems are common and associated with increased risk for adjustment problems. We examined daily links between children's sleep and mood, using a daily diary method and actigraphy. We also tested children's daily mood as a mediator of relations among sleep and children's broader internalizing and externalizing symptoms. A community sample of 142 children (mean age = 10.69 years; 57% girls; 69% European American, 31% African American) and their parents participated. For 1 week, children wore actigraphs and parents completed a daily telephone interview about their child's mood. Following the week of actigraphy, mothers and fathers reported on their child's adjustment. Multi‐level models indicated within‐person relations between children's mood and subsequent sleep fragmentation (indicated by increased activity) and sleep latency, and between‐person relations between sleep latency and subsequent mood on the next day. Significant indirect effects were found such that a more negative daily mood (aggregated across diary days) mediated relations between poor sleep efficiency and longer sleep latency and parent‐reported internalizing and externalizing symptoms. Findings extend previous research by highlighting disruptions to children's daily mood as a potential mechanism linking sleep problems to children's mental health.
Effect of exercise on 24-month weight loss maintenance in overweight women.

BACKGROUND Debate remains regarding the amount of physical activity that will facilitate weight loss maintenance. METHODS Between December 1, 1999, and January 31, 2003, 201 overweight and obese women (body mass index [calculated as weight in kilograms divided by height in meters squared], 27 to 40; age range, 21-45 years) with no contraindications to weight loss or physical activity were recruited from a hospital-based weight loss research center. Participants were assigned to 1 of 4 behavioral weight loss intervention groups. They were randomly assigned to groups based on physical activity energy expenditure (1000 vs 2000 kcal/wk) and intensity (moderate vs vigorous). Participants also were told to reduce intake to 1200 to 1500 kcal/d. A combination of in-person conversations and telephone calls were conducted during the 24-month study period. RESULTS Weight loss did not differ among the randomized groups at 6 months' (8%-10% of initial body weight) or 24 months' (5% of initial body weight) follow-up. Post-hoc analysis showed that individuals sustaining a loss of 10% or more of initial body weight at 24 months reported performing more physical activity (1835 kcal/wk or 275 min/wk) compared with those sustaining a weight loss of less than 10% of initial body weight (P < .001). CONCLUSIONS The addition of 275 mins/wk of physical activity, in combination with a reduction in energy intake, is important in allowing overweight women to sustain a weight loss of more than 10%. Interventions to facilitate this level of physical activity are needed.
O-Linked β-N-acetylglucosamine (O-GlcNAc) Regulates Stress-induced Heat Shock Protein Expression in a GSK-3β-dependent Manner*

To investigate the mechanisms by which O-linked β-N-acetylglucosamine modification of nucleocytoplasmic proteins (O-GlcNAc) confers stress tolerance to multiple forms of cellular injury, we explored the role(s) of O-GlcNAc in the regulation of heat shock protein (HSP) expression. Using a cell line in which deletion of the O-GlcNAc transferase (OGT; the enzyme that adds O-GlcNAc) can be induced by 4-hydroxytamoxifen, we screened the expression of 84 HSPs using quantitative reverse transcriptase PCR. In OGT null cells the stress-induced expression of 18 molecular chaperones, including HSP72, were reduced. GSK-3β promotes apoptosis through numerous pathways, including phosphorylation of heat shock factor 1 (HSF1) at Ser303 (Ser(P)303 HSF1), which inactivates HSF1 and inhibits HSP expression. In OGT null cells we observed increased Ser(P)303 HSF1; conversely, in cells in which O-GlcNAc levels had been elevated, reduced Ser(P)303 HSF1 was detected. These data, combined with those showing that inhibition of GSK-3β in OGT null cells recovers HSP72 expression, suggests that O-GlcNAc regulates the activity of GSK-3β. In OGT null cells, stress-induced inactivation of GSK-3β by phosphorylation at Ser9 was ablated providing a molecular basis for these findings. Together, these data suggest that stress-induced GlcNAcylation increases HSP expression through inhibition of GSK-3β.