Syndecan-1 regulates αvβ3 and αvβ5 integrin activation during angiogenesis and is blocked by synstatin,a novel peptide inhibitor

Syndecan-1 (Sdc1) is a matrix receptor shown to associate via its extracellular domain with the α_vβ₃ and α_vβ₅ integrins, potentially regulating cell adhesion, spreading, and invasion of cells expressing these integrins. Using Sdc1 deletion mutants expressed in human mammary carcinoma cells, we identified the active site within the Sdc1 core protein and derived a peptide inhibitor called synstatin (SSTN) that disrupts Sdc1''s interaction with these integrins. Because the α_vβ₃ and α_vβ₅ integrins are critical in angiogenesis, a process in which a role for Sdc1 has been uncertain, we used human vascular endothelial cells in vitro to show that the Sdc1 regulatory mechanism is also required for integrin activation on these cells. We found Sdc1 expressed in the vascular endothelium during microvessel outgrowth from aortic explants in vitro and in mouse mammary tumors in vivo. Moreover, we show that SSTN blocks angiogenesis in vitro or when delivered systemically in a mouse model of angiogenesis in vivo, and impairs mammary tumor growth in an orthotopic mouse tumor model. Thus, Sdc1 is a critical regulator of these two important integrins during angiogenesis and tumorigenesis, and is inhibited by the novel SSTN peptide.Angiogenesis, or the sprouting of new blood vessels from existing ones, occurs during development and in diseases such as diabetic retinopathy, endometriosis, psoriasis, rheumatoid arthritis, and tumor-induced angiogenesis (1). Vascular endothelial cells rely on signaling from multiple integrins during the angiogenic process (for review see reference 2), including the α_vβ₃ and α_vβ₅ integrins; signaling by the α_vβ₃ and α_vβ₅ integrin leads to endothelial cell proliferation, migration, matrix metalloprotease activation, and resistance to apoptosis (3).The α_vβ₃ and α_vβ₅ integrins are subject to regulation during angiogenesis. Fibroblast growth factor (FGF) and vascular endothelial growth factor (VEGF), two potent angiogenic factors released by tumors, induce the expression of these two integrins that collaborate with the FGF and VEGF receptors in angiogenic signaling pathways (4); disrupting angiogenic signaling by inactivation of either integrin or growth factor receptor leads to endothelial cell apoptosis (5). The integrins are often up-regulated on metastatic tumors as well, leading to enhanced invasion, proliferation, and tumor survival (6–9) by largely the same mechanisms operative in endothelial cells. For these reasons, the integrins and their regulatory mechanisms are attractive targets for the development of therapeutic drugs. Drugs that are currently being tested range from inhibitory integrin antibodies (e.g., Vitaxin [10], based on the inhibitory antibody LM609 [11]), to cyclic RGD peptides that interfere with ligand binding (e.g., cRGDfV, cilengitide, and ST1646 [12–15]), to peptidomimetics based on the RGD sequence (e.g., S247 [16]). These inhibitors have all been shown to disrupt the growth of solid tumors as well as angiogenesis.We have recently identified a regulatory mechanism by which syndecan-1 (Sdc1), a cell-surface matrix receptor, regulates the activation of the α_vβ₃ and α_vβ₅ integrins on mammary carcinoma cells and fibroblasts (17–20). The syndecans are multifunctional extracellular matrix receptors on the surface of all adherent cells (21–23). They anchor to the matrix via heparan sulfate (HS) glycosaminoglycan chains attached near the distal tips of their core proteins; these chains recognize “heparin-binding” domains present in most matrix ligands, including fibronectin (FN), laminins, vitronectin (VN), thrombospondin, and the fibrillar collagens (21). In addition, mounting evidence suggests that they assemble with and control the signaling of other cell surface receptors, including integrins. McFall et al. first described a “cell-binding domain” in the extracellular domain of Sdc4 (24, 25); this site has recently been shown to regulate β1-containing integrins on mesenchymal cells, although the exact integrin target and regulatory mechanism remain unknown (26, 27). Recombinant Sdc2 extracellular domain alters adhesion mechanisms in colon carcinoma cells, suggesting that a regulatory site also exists in its extracellular domain (28, 29). More recently, we have shown that Sdc1 is necessary for activation of the α_vβ₃ integrin on mammary carcinoma cells (17, 20). Silencing Sdc1 expression, selective deletion of amino acids in its extracellular domain, or targeted competition with domain-specific antibodies or recombinant extracellular domain protein disrupts integrin activation and matrix recognition necessary for cell spreading and invasion. Similar activation of the α_vβ₅ integrin by Sdc1 occurs on B82L fibroblasts, which rely exclusively on this integrin for attachment to VN and FN (19). These extracellular syndecan-specific regulatory sites are readily accessible to therapeutic drugs and may hold promise as targets for combating tumorigenesis and other diseases in which their regulated mechanisms play a role.Given the importance of the α_vβ₃ and α_vβ₅ integrins in angiogenesis, we examined the possibility that Sdc1 regulates these integrins on vascular endothelial cells during tumor-induced angiogenesis. We found that Sdc1 is expressed by mouse and human endothelial cells in vitro, and is expressed during angiogenesis induced by FGF or VEGF in vitro and in vivo. We found that the α_vβ₃ and α_vβ₅ integrins associate with Sdc1 and that this association can be disrupted by a peptide called synstatin (SSTN) that is derived from the active site in the Sdc1 core protein. Furthermore, SSTN is an effective inhibitor of angiogenesis in vitro and in vivo, and of mammary carcinoma formation in nude mice. These results define the Sdc1 regulatory mechanism as a critical component of the angiogenic and tumorigenic process.     

Expression of B7-1 and B7-2 mediated by adenovirus enhanced T cell anti-hepatoma immunity

The ultimate aim of cancer immunotherapy is theinduction of tumor-specific T-lymphocyte responseseffective in eradicating disseminated tumors. There aregenerally tumor infiltrating T lymphocytes in situ oftumor, but most of which are inactive, one of the majorreasons of such Inactivation is that T lymphocytes areexposured to an antigen-specific TCR signal in theabsence of antigen-nonspecific or co-stimulatory signal.The co-stimulatory factors, B7-l and B7-2 on the surfaceof antigen presenting cells provide a key signal for thegeneration of T cell immunity. Sincc hepatoma usually     

Essential roles of sphingosine-1–phosphate receptor 2 in human mast cell activation,anaphylaxis, and pulmonary edema

Systemic exacerbation of allergic responses, in which mast cells play a critical role, results in life-threatening anaphylactic shock. Sphingosine-1–phosphate (S1P), a ligand for a family of G protein–coupled receptors, is a new addition to the repertoire of bioactive lipids secreted by activated mast cells. Yet little is known of its role in human mast cell functions and in anaphylaxis. We show that S1P₂ receptors play a critical role in regulating human mast cell functions, including degranulation and cytokine and chemokine release. Immunoglobulin E–triggered anaphylactic responses, including elevation of circulating histamine and associated pulmonary edema in mice, were significantly attenuated by the S1P₂ antagonist JTE-013 and in S1P₂-deficient mice, in contrast to anaphylaxis induced by administration of histamine or platelet-activating factor. Hence, S1P and S1P₂ on mast cells are determinants of systemic anaphylaxis and associated pulmonary edema and might be beneficial targets for anaphylaxis attenuation and prophylaxis.Allergic disease is endemic in developed nations, and none is more dramatic or deadly than systemic anaphylactic shock (Finkelman, 2007). Unfortunately, this collection of rapid changes to the cutaneous, vasculature, and pulmonary systems may also be the least understood of atopic diseases. A better understanding of the eliciting factors for systemic anaphylaxis is therefore critical for accurate diagnosis and therapeutic intervention. Mast cells, which reside in vascularized tissues and are strategically located at the interfaces of host and environment in skin and mucosal surfaces, are key effectors of IgE-mediated allergic disorders, including anaphylaxis, hay fever, eczema, and asthma. Mast cells express FcεRI (high-affinity receptors for IgE), which binds antigen (Ag)-specific IgE antibodies, and subsequent exposure to Ag initiates cross-linking and aggregation of FcεRI. This cascade of events triggers mast cell activation and secretion of a wide array of inflammatory mediators, such as histamine and other preformed mediators, bioactive lipids, including eicosanoids and platelet-activating factor (PAF), and numerous proinflammatory cytokines and chemokines (Kalesnikoff and Galli, 2008), which are essential to the pathogenesis of allergic diseases and anaphylaxis (Finkelman, 2007).A recent addition to the recognized repertoire of lipid mediators secreted by mast cells is the sphingolipid metabolite sphingosine-1–phosphate (S1P; Jolly et al., 2004; Mitra et al., 2006; Olivera et al., 2006), which has been implicated in initiation and maintenance of diverse aspects of immune cell activation and function (Rosen et al., 2008; for review see Schwab and Cyster, 2007; Rivera et al., 2008). Focus on S1P in immune responses has increased after the elucidation of its critical role in lymphocyte trafficking (Rosen et al., 2008; for review see Schwab and Cyster, 2007), and observations of local increases of S1P in inflammatory disorders such as asthma (Ammit et al., 2001) and rheumatoid arthritis (Kitano et al., 2006). S1P is a ligand for five G protein–coupled receptors, designated S1P_1-5, through which it exerts many of its actions (Spiegel and Milstien, 2003). Rodent mast cells express S1P₁ and S1P₂ receptors on their cell surface, which function in an autocrine manner to fine-tune mast cell functions (for review see Olivera, 2008; Price et al., 2008). Although S1P₁ is important for migration of mast cells toward Ag, S1P₂ contributes to the robustness of their degranulation (Jolly et al., 2004). FcεRI engagement on mast cells by allergens stimulates sphingosine kinase (SphK) 1 and 2, the two isoenzymes which produce S1P, and both contribute to mast cell functions (Jolly et al., 2004, 2005; Urtz et al., 2004; Olivera et al., 2006, 2007). SphK1, but not SphK2, was shown to be important for Ag-induced calcium mobilization, degranulation, and migration of human and rodent bone marrow–derived mast cells (Melendez and Khaw, 2002; Jolly et al., 2004, 2005; Oskeritzian et al., 2008). However, based on results with fetal liver–derived mast cells from SphK1 and SphK2 knockout mice, it was suggested that SphK2, but not SphK1, modulates calcium influx and downstream signaling, leading to degranulation and production of eicosanoids and cytokines (Olivera et al., 2007). Similarly, SphK2 was also necessary for secretion of both TNF and IL-6 by mature human mast cells (Oskeritzian et al., 2008). Remarkably, however, mice deficient in SphK1 displayed reduced levels of circulating S1P and were resistant to anaphylaxis, whereas anaphylactic responses in mice deficient in SphK2 were normal (Olivera et al., 2007). An intriguing finding of this study was the association between circulating concentrations of S1P and susceptibility to anaphylaxis, suggesting that S1P is a key factor that determines anaphylactic responses.Although much has been learned about the roles of the SphKs that produce S1P in mast cell functions and anaphylactic responses, little is known about how S1P executes these important functions in vivo and which of the S1P receptors is involved. To this end, we examined the roles of S1P receptors in developing and mature human mast cells and their importance in IgE-mediated passive systemic anaphylaxis (PSA) in mice. Our data provide direct pharmacological, genetic, and biological evidence for the importance of S1P and the S1P₂ receptor on mast cells in anaphylaxis and associated pulmonary edema. Hence, S1P and S1P₂ may be therapeutic targets for clinical intervention in this poorly understood disease.     

Histogram Analysis Parameters Derived from Conventional T1- and T2-Weighted Images Can Predict Different Histopathological Features Including Expression of Ki67, EGFR,VEGF, HIF-1α, and p53 and Cell Count in Head and Neck Squamous Cell Carcinoma

Molecular Imaging and Biology - To analyze associations between histogram analysis parameters derived from conventional magnetic resonance imaging (MRI) and different histopathological features in...     

The Vα14 invariant natural killer T cell TCR forces microbial glycolipids and CD1d into a conserved binding mode

Invariant natural killer T cells (iNKT cells) rapidly produce effector cytokines. In this study, we report the first crystal structures of the iNKT cell T cell receptor (TCR) bound to two natural, microbial glycolipids presented by CD1d. Binding of the TCR induced CDR3-α-dependent structural changes in the F' roof of CD1d; these changes resemble those occurring in the absence of TCR engagement when the highly potent synthetic antigen α-galactosylceramide (α-GalCer) binds CD1d. Furthermore, in the Borrelia burgdorferi α-galactosyl diacylglycerol-CD1d complex, TCR binding caused a marked repositioning of the galactose sugar into an orientation that closely resembles α-GalCer. The TCR-dependent reorientation of the sugar, together with the induced CD1d fit, may explain the weaker potency of the microbial antigens compared with α-GalCer. We propose that the TCR of iNKT cells binds with a conserved footprint onto CD1d, regardless of the bound glycolipid antigen, and that for microbial antigens this unique binding mode requires TCR-initiated conformational changes.     

Genetic and biochemical characterization of a novel metallo-β-lactamase, TMB-1, from an Achromobacter xylosoxidans strain isolated in Tripoli, Libya

An Achromobacter xylosoxidans strain from the Tripoli central hospital produced a unique metallo-β-lactamase, designated TMB-1, which is related to DIM-1 (62%) and GIM-1 (51%). bla(TMB-1) was embedded in a class 1 integron and located on the chromosome. The TMB-1 β-lactamase has lower k(cat) values than both DIM-1 and GIM-1 with cephalosporins and carbapenems. The K(m) values were more similar to those of GIM-1 than those of DIM-1, with the overall k(cat)/K(m) values being lower than those for GIM-1 and DIM-1.     

Histogram Analysis of T1-Weighted,T2-Weighted,and Postcontrast T1-Weighted Images in Primary CNS Lymphoma: Correlations with Histopathological Findings—a Preliminary Study

Purpose

Previously, some reports mentioned that magnetic resonance imaging (MRI) can predict histopathological features in primary CNS lymphoma (PCNSL). The reported data analyzed diffusion-weighted imaging findings. The aim of this study was to investigate possible associations between histopathological findings, such as tumor cellularity, nucleic areas and proliferation index Ki-67, and signal intensity on T1-weighted and T2-weighted images in PCNSL.

Procedures

For this study, 18 patients with PCNSL were retrospectively investigated by histogram analysis on precontrast and postcontrast T1-weighted and fluid-attenuated inversion recovery (FLAIR) images. For every patient, histopathology parameters, nucleic count, total nucleic area, and average nucleic area, as well as Ki-67 index, were estimated.

Results

Correlation analysis identified several statistically significant associations. Skewness derived from precontrast T1-weighted images correlated with Ki-67 index (p = ? 0.55, P = 0.028). Furthermore, entropy derived from precontrast T1-weighted images correlated with average nucleic area (p = 0.53, P = 0.04). Several parameters from postcontrast T1-weighted images correlated with nucleic count: maximum signal intensity (p = 0.59, P = 0.017), P75 (p = 0.56, P = 0.02), and P90 (p = 0.52, P = 0.04) as well as SD (p = 0.58, P = 0.02). Maximum signal intensity derived from FLAIR sequence correlated with nucleic count (p = 0.50, P = 0.03).

Conclusion

Histogram-derived parameters of conventional MRI sequences can reflect different histopathological features in PSNCL.

     

Upregulation of Tim-3 and PD-1 expression is associated with tumor antigen–specific CD8+ T cell dysfunction in melanoma patients

The paradoxical coexistence of spontaneous tumor antigen–specific immune responses with progressive disease in cancer patients furthers the need to dissect the molecular pathways involved in tumor-induced T cell dysfunction. In patients with advanced melanoma, we have previously shown that the cancer-germline antigen NY-ESO-1 stimulates spontaneous NY-ESO-1–specific CD8⁺ T cells that up-regulate PD-1 expression. We also observed that PD-1 regulates NY-ESO-1–specific CD8⁺ T cell expansion upon chronic antigen stimulation. In the present study, we show that a fraction of PD-1⁺ NY-ESO-1–specific CD8⁺ T cells in patients with advanced melanoma up-regulates Tim-3 expression and that Tim-3⁺PD-1⁺ NY-ESO-1–specific CD8⁺ T cells are more dysfunctional than Tim-3⁻PD-1⁺ and Tim-3⁻PD-1⁻ NY-ESO-1–specific CD8⁺ T cells, producing less IFN-γ, TNF, and IL-2. Tim-3–Tim-3L blockade enhanced cytokine production by NY-ESO-1–specific CD8⁺ T cells upon short ex vivo stimulation with cognate peptide, thus enhancing their functional capacity. In addition, Tim-3–Tim-3L blockade enhanced cytokine production and proliferation of NY-ESO-1–specific CD8⁺ T cells upon prolonged antigen stimulation and acted in synergy with PD-1–PD-L1 blockade. Collectively, our findings support the use of Tim-3–Tim-3L blockade together with PD-1–PD-L1 blockade to reverse tumor-induced T cell exhaustion/dysfunction in patients with advanced melanoma.There is ample evidence that patients with melanoma can develop immune responses directed against antigens expressed by their own tumor (Boon et al., 2006). Among these antigens, cancer-germline antigens (CGAs) are expressed by tumors of many different histological types, including melanoma, but not by normal tissues, except testis. Because germ cells in testis do not express HLA molecules on their surface (Haas et al., 1988), CGAs represent strictly tumor-specific T cell targets (Boon et al., 2006). Among CGAs, NY-ESO-1 has been shown to stimulate spontaneous cellular and humoral responses that are detectable only in patients with advanced NY-ESO-1–expressing cancer (Stockert et al., 1998; Jäger et al., 2000; Mandic et al., 2005; Fourcade et al., 2008). Understanding the failure of spontaneous NY-ESO-1–specific T cell responses to promote regression of NY-ESO-1⁺ tumors is therefore critical for the design of novel therapeutic interventions aimed at overcoming tumor-induced immune escape.We have previously shown that the large majority of spontaneous NY-ESO-1–specific CD8⁺ T cells up-regulates programmed death 1 (PD-1) expression (Fourcade et al., 2009), which appears to be associated with T cell exhaustion/dysfunction in chronic viral infections in animals and humans (Barber et al., 2006; Day et al., 2006; Petrovas et al., 2006; Trautmann et al., 2006). We observed that PD-1 up-regulation on spontaneous NY-ESO-1–specific CD8⁺ T cells occurs along with T cell activation and is not directly associated with an inability to produce cytokines ex vivo upon stimulation with cognate antigen. Blockade of the PD-1–programmed death ligand 1 (PD-L1) pathway in combination with prolonged antigen stimulation with PD-L1⁺ APCs or melanoma cells augmented the frequencies of cytokine-producing, proliferating, and total NY-ESO-1–specific CD8⁺ T cells. Our findings are in line with previous studies of PD-1 expression by HIV- and SIV-specific CD8⁺ T cells, demonstrating that PD-1 is a regulator of antigen-specific CD8⁺ T cell expansion in the context of chronic antigen exposure, although it does not exhibit a major impact upon their functionality on a cell-per-cell basis (Petrovas et al., 2006, 2007). To further determine whether other molecular pathways are involved in tumor antigen–specific T cell dysfunction, we studied T cell immunoglobulin and mucin-domain–containing molecule 3 (Tim-3) expression on spontaneous NY-ESO-1–specific CD8⁺ T cells from patients with advanced melanoma and investigated whether Tim-3 up-regulation defines a subgroup of dysfunctional tumor antigen–specific CD8⁺ T cells. Tim-3 is a transmembrane protein constitutively expressed on Th1/Tc1 cells in mice and humans (Monney et al., 2002). Several lines of evidence support the role of Tim-3 as an inhibitory molecule that down-regulates effector Th1/Tc1 cell responses. In mice, blocking the Tim-3–Tim-3L pathway resulted in hyperproliferation of Th1-type cells and abrogated the induction of peripheral and transplantation tolerance (Sabatos et al., 2003; Sánchez-Fueyo et al., 2003). Tim-3 interacts with its ligand galectin-9 to induce cell death in Th1 cells (Zhu et al., 2005). In humans, Tim-3 expression is defective in CD4⁺ T cells producing high levels of IFN-γ, as well as those isolated from cerebrospinal fluid of patients with multiple sclerosis (Koguchi et al., 2006). Recently, Tim-3 up-regulation has been reported in HIV-specific and HCV-specific CD8⁺ T cells in patients with progressive HIV infection and chronic hepatitis C, respectively (Jones et al., 2008; Golden-Mason et al., 2009). Tim-3⁺ HIV- and HCV-specific CD8⁺ T cells were distinct from the PD-1⁺ CD8⁺ T cells and exhibited T cell dysfunction. However, it is unknown whether tumor antigen–specific CD8⁺ T cells in patients with advanced cancers express Tim-3.In this study, we show that a fraction of PD-1⁺ NY-ESO-1–specific CD8⁺ T cells, which represents the large majority of circulating NY-ESO-1–specific CD8⁺ T cells in patients with advanced melanoma, up-regulates Tim-3 expression. Tim-3⁺PD-1⁺ NY-ESO-1–specific CD8⁺ T cells are highly dysfunctional compared with Tim-3⁻PD-1⁺ and Tim-3⁻PD-1⁻ NY-ESO-1–specific CD8⁺ T cells. Tim-3–Tim-3L pathway blockade alone or in combination with PD-1–PD-L1 pathway blockade enhanced NY-ESO-1–specific CD8⁺ T cell numbers and functions. Collectively, our findings support the use of Tim-3–Tim-3L blockade in association with PD-1–PD-L1 blockade to reverse tumor-induced T cell exhaustion/dysfunction in patients with advanced melanoma.     

Pharmacokinetics and pharmacodynamics of TBR-652, a novel CCR5 antagonist, in HIV-1-infected, antiretroviral treatment-experienced, CCR5 antagonist-naïve patients

TBR-652 is a novel CCR5 antagonist with potent in vitro anti-HIV activity. The objective of this study was to determine the pharmacokinetics (PK) and pharmacodynamics (PD) of TBR-652 in HIV-1-infected, antiretroviral treatment-experienced, CCR5 antagonist-naïve patients. A double-blind, placebo-controlled, randomized, dose-escalating study of TBR-652 monotherapy given once daily orally for 10 days was performed, followed by a 40-day follow-up period. Approximately 10 patients/dose level received 25, 50, 75, 100, and 150 mg TBR-652 or placebo (4:1). Blood was collected at different intervals for PK and HIV-1 RNA assessments. PK analysis of TBR-652 was performed using noncompartmental methods. PK/PD was modeled using a maximum inhibitory effect model (E_max) and 50% inhibitory concentrations (IC₅₀). TBR-652 was well absorbed in the systemic circulation. TBR-652 concentration levels declined slowly, with mean elimination half-lives ranging from 22.5 to 47.62 h across dose levels. TBR-652 treatment resulted in potent, dose-dependent decreases in viral load, with statistically significant decreases in nadir HIV-1 RNA compared to baseline for all dose levels. Suppression of HIV-1 RNA persisted over the 40-day follow-up period. A steep exposure-effect relationship was observed, with an E_max of −1.43 log₁₀ copies/ml and IC₅₀ of 13.1 ng/ml. TBR-652 was generally safe and well tolerated at all dose levels studied. Short-term monotherapy treatments of TBR-652 in HIV-1-infected patients resulted in promising PK and PD results, with a clear exposure-response relationship at the current dose levels studied. Data from this study support further development of TBR-652 in HIV-infected patients.     

Associations Between [18F]FDG-PET and Complex Histopathological Parameters Including Tumor Cell Count and Expression of KI 67, EGFR,VEGF, HIF-1α, and p53 in Head and Neck Squamous Cell Carcinoma

Molecular Imaging and Biology - Head and neck squamous cell carcinoma (HNSCC) is one of common cancers worldwide. Positron emission tomography (PET) with 2-deoxy-2-[18F]fluoro-d-glucose ([18F]FDG)...     

The Autism 1–2–3 Project,a short-duration and low-intensity intervention that targets basic social and communication skills and parent involvement,shows promise and is worthy of future research

This review provides a summary and appraisal commentary on the treatment review by Wong, V. C. N., & Kwan, Q. K. (2010). Randomized controlled trial for early intervention for autism: A pilot study of the Autism 1–2–3 Project. Journal of Autism and Developmental Disorders, 40, 677–688.

     

Effects of a Small Dose of Ethanol and Calcium Carbimide-Induced Acetaldehyde Intoxication on Human Platelet Aggregation,Associated Thromboxane Formation and Urinary Excretion of 2,3-dinor-6-Keto Prostaglandin F1α

Abstract

We studied ADP-induced platelet aggregation, associated thromboxane B₂ (TXB₂) formation, urinary excretion of the prostacyclin metabolite 2,3-dinor-6-keto prostaglandin F_1α (2,3-dinor-6-keto PGF_1α) and formation of malondialdehyde in 10 healthy volunteers after ingestion of a small dose of ethanol (0.25?g per kg of body weight) and calcium carbimide (50?mg). Platelet aggregation in platelet-rich plasma (PRP) was suppressed (p≥0.05) by ethanol, but no change occurred in platelet TXB₂ formation. Ingestion of calcium carbimide caused significant elevations in blood acetaldehyde (p≥0.001) and ethanol (p≥0.05) levels, but acetaldehyde did not influence platelet aggregability or the aggregation-associated TXB₂ formation. However, calcium carbimide per se significantly (p≥0.05) elevated TXB₂ formation. No effects were found on plasma malondialdehyde levels and urinary excretion of 2,3-dinor-6-keto PGF_1α. These observations indicate that a small dose of ethanol attenuates platelet aggregation without any significant effect on aggregation-associated TXB₂ formation. By contrast, ingestion of calcium carbimide per se may enhance TXB₂ formation.     

Action of Phα1β, a Peptide From the Venom of the Spider Phoneutria nigriventer,on the Analgesic and Adverse Effects Caused by Morphine in Mice

Opioids are standard therapy for the treatment of pain; however, adverse effects limit their use. Voltage-gated calcium channel blockers may be used to increase opioid analgesia, but their effect on opioid-induced side effects is little known. Thus, the goal of this study was to evaluate the action of the peptide Phα1β, a voltage-gated calcium channel blocker, on the antinociceptive and adverse effects produced by morphine in mice. A single administration of morphine (3–10 mg/kg) was able to reduce heat nociception as well as decrease gastrointestinal transit. The antinociception caused by a single injection of morphine was slightly increased by an intrathecal injection of Phα1β (30 pmol/site). Repeated treatment with morphine caused tolerance, hyperalgesia, withdrawal syndrome, and constipation, and the Phα1β (.1–30 pmol/site, intrathecal) was able to reverse these effects. Finally, the effects produced by the native form of Phα1β were fully mimicked by a recombinant version of this peptide. Taken together, these data show that Phα1β was effective in potentiating the analgesia caused by a single dose of morphine as well as in reducing tolerance and the adverse effects induced by repeated administration of morphine, indicating its potential use as an adjuvant drug in combination with opioids.PerspectiveThis article presents preclinical evidence for a useful adjuvant drug in opioid treatment. Phα1β, a peptide calcium channel blocker, could be used not only to potentiate morphine analgesia but also to reduce the adverse effects caused by repeated administration of morphine.     

‘Normal’ toddlers’ comprehension of the word UNDER in Polish seems facilitated when training about understanding its meaning takes place in situations with familiar toys,and might be enhanced for transfer of meaning to situations with novel toys when training includes contrasts with the earlier developing prepositions of IN or UNDER1

This review provides a summary and appraisal commentary on the treatment review by Al Otaiba, S., Puranik, C. S., Ziolkowski, R. A., &; Montgomery, T. M. (2009). Effectiveness of early phonological awareness interventions for students with speech or language impairments. The Journal of Special Education, 43, 107–128.

Source of funding and disclosure of interest: This study was supported by a grant from National Institute of Child Health and Human Development and the Institute of Education Sciences, and the original authors of this research report no conflicts of interest.