Higher-order mode filtering by a resistive layer.

A method for filtering higher-order acoustic modes using a resistive layer is proposed and applied to a two-dimensional rectangular waveguide with a quiescent fluid. An analogue of Cremer's criterion is discussed and used to obtain the optimal modal attenuation of the non-planar waves while the plane wave is preserved. Numerical validation of the concept is performed for a straight waveguide and an abrupt expansion in a waveguide.

Observation of Cytotoxicity of Phosphonium Derivatives Is Explained: Metabolism Inhibition and Adhesion Alteration.

The search for new antibiotics, substances that kill prokaryotic cells and do not kill eukaryotic cells, is an urgent need for modern medicine. Among the most promising are derivatives of triphenylphosphonium, which can protect the infected organs of mammals and heal damaged cells as mitochondria-targeted antioxidants. In addition to the antioxidant action, triphenylphosphonium derivatives exhibit antibacterial activity. It has recently been reported that triphenylphosphonium derivatives cause either cytotoxic effects or inhibition of cellular metabolism at submicromolar concentrations. In this work, we analyzed the MTT data using microscopy and compared them with data on changes in the luminescence of bacteria. We have shown that, at submicromolar concentrations, only metabolism is inhibited, while an increase in alkyltriphenylphosphonium (CnTPP) concentration leads to adhesion alteration. Thus, our data on eukaryotic and prokaryotic cells confirm a decrease in the metabolic activity of cells by CnTPPs but do not confirm a cytocidal effect of TPPs at submicromolar concentrations. This allows us to consider CnTPP as a non-toxic antibacterial drug at low concentrations and a relatively safe vector for delivering other antibacterial substances into bacterial cells.

Thrombin and Factor Xa Hydrolysis of Chromogenic Substrates in the Presence of Sulfated Derivatives of Galactomannan and Galactoglucomannan Natural Gels.

Polysaccharides are important structural components of all plant species. Gel-like polysaccharides have found wide application in various fields, including medicine, construction, and the food industry. In the present work, galactomannan and galactoglucomannan gel-like polysaccharides were modified with sulfate groups and their anticoagulant activity was studied. Sulfation with chlorosulfonic acid in pyridine and with sulfamic acid in pyridine and a sulfamic acid-urea deep eutectic solvent were used as synthesis routes. The resulting gel-like polysaccharide sulfates were studied by elemental analysis, Fourier-transform infrared spectroscopy, and gel permeation chromatography. It was established that the anticoagulant effect of sulfated galactoglucomannan (SGGM) and galactomannan (SGM-1 and SGM-2) is related to an independent antithrombin-independent decrease in the amidolytic activity of thrombin and factor Xa. It is shown that the inhibitory activity of SGGM and SGM-2 against the collagen-induced platelet aggregation can be an additional factor in selecting compounds that are most promising for modifying polymer surfaces to ensure resistance to blood clotting.

Auto-delineation framework for the focal liver reaction observed in post Stereotactic Body Radiation Therapy (SBRT) Primovist MRI scans.

Development of a novel auto-delineation methodology for observed hypointensity from focal liver reaction in hepatobiliary-specific contrast (Primovist) enhanced MRI acquired post Stereotactic Body Radiation Therapy (SBRT). Additionally, the methodology for the quantification of the threshold dose associated with the observed focal liver reaction was also established. An auto-delineation algorithm was created based on the correlation between intensity and radiation dose information. The error associated with the auto-delineation was quantified using virtual FLRs, as well as clinical patient scans. Patients underwent liver SBRT with a total dose prescription of 50 Gy in 5 fractions. An inherent correlation was established between the contrast-to-noise ratio (CNR) on MRI scans and expected performance of the algorithm using centre-of-mass (COM). Threshold dose associated with focal liver reaction was quantified for all ten patients and verified with associated most conformal isodose line. Based on the CNR vs COM error relationship, the expected median (range) auto-delineation COM error for ten patients was 0.5 (0 to 3.2) mm. The median threshold dose for ten clinical cases was 21.3 Gy based on the auto-delineation framework. This threshold dose was compared to the most conformal isodose line with the hypointensity; there was no significant difference observed (p = 0.6). We developed a framework for post-SBRT Primovist observed focal liver reaction localization. Furthermore, this study established an automated approach for the determination of the threshold dose associated with the hypointense region.

The charge-assisted hydrogen-bonded organic framework (CAHOF) self-assembled from the conjugated acid of tetrakis(4-aminophenyl)methane and 2,6-naphthalenedisulfonate as a new class of recyclable Brønsted acid catalysts.

The acid-base neutralization reaction of commercially available disodium 2,6-naphthalenedisulfonate (NDS, 2 equivalents) and the tetrahydrochloride salt of tetrakis(4-aminophenyl)methane (TAPM, 1 equivalent) in water gave a novel three-dimensional charge-assisted hydrogen-bonded framework (CAHOF, F-1). The framework F-1 was characterized by X-ray diffraction, TGA, elemental analysis, and 1H NMR spectroscopy. The framework was supported by hydrogen bonds between the sulfonate anions and the ammonium cations of NDS and protonated TAPM moieties, respectively. The CAHOF material functioned as a new type of catalytically active Brønsted acid in a series of reactions, including the ring opening of epoxides by water and alcohols. A Diels-Alder reaction between cyclopentadiene and methyl vinyl ketone was also catalyzed by F-1 in heptane. Depending on the polarity of the solvent mixture, the CAHOF F-1 could function as a purely heterogeneous catalyst or partly dissociate, providing some dissolved F-1 as the real catalyst. In all cases, the catalyst could easily be recovered and recycled.

In Situ functionalization of Poly(hydroxyethyl methacrylate) Cryogels with Oligopeptides via ß-Cyclodextrin-Adamantane Complexation for Studying Cell-Instructive Peptide Environment.

Oligopeptides are versatile cell modulators resembling pleiotropic activities of ECM proteins and growth factors. Studying the role of cell-instructive peptide signals within 3D scaffolds, yet poorly known, requires effective approaches to introducing bioactive sequences into appropriate materials. We synthesized RGD and GHK motif based peptides 1 and 2 linked to the terminal adamantyl group (Ad) and their fluorescent derivatives 3 and 4. Poly(hydroxyethyl methacrylate) (pHEMA) cryogels with additional PEG/ß-cyclodextrin (CD) units were prepared as an inert macroporous scaffold capable to bind the adamantylated peptides via affinity CD-Ad complexation. According to toluidine blue staining, the CD moieties were effectively and stably incorporated in the pHEMA cryogels at nanomolar amounts per milligram of material. The CD component gradually increased the thickness and swelling ability of the polymer walls of cryogels, resulting in a noticeable decrease in macropore size and modulation of viscoelastic properties. The labeled peptides exhibited fast kinetics of specific binding to the CD-modified cryogels and were simultaneously immobilized by coincubation. The peptide loading approached ca. 0.31 mg per cm2 of cryogel sheet. A well-defined mitogenic effect of the immobilized peptides (2 < 1≪ 1 + 2) was revealed toward 3T3 and PC-12 cells. The synergistic action of RGD and GHK peptides induced a profound change in cell behavior/morphology attributed to a growth-factor-like activity of the composition. Altogether, our results provide an effective procedure for the preparation of CD-modified pHEMA cryogels and their uniform in situ functionalization with bioactive peptide(s) of interest and an informative study of cellular responses in the functionalized scaffolds.

Structure guided deformable image registration for treatment planning CT and post stereotactic body radiation therapy (SBRT) Primovist® (Gd-EOB-DTPA) enhanced MRI.

The purpose of this study was to assess the performance of structure-guided deformable image registration (SG-DIR) relative to rigid registration and DIR using TG-132 recommendations. This assessment was performed for image registration of treatment planning computed tomography (CT) and magnetic resonance imaging (MRI) scans with Primovist® contrast agent acquired post stereotactic body radiation therapy (SBRT). SBRT treatment planning CT scans and posttreatment Primovist® MRI scans were obtained for 14 patients. The liver was delineated on both sets of images and matching anatomical landmarks were chosen by a radiation oncologist. Rigid registration, DIR, and two types of SG-DIR (using liver contours only; and using liver structures along with anatomical landmarks) were performed for each set of scans. TG-132 recommended metrics were estimated which included Dice Similarity Coefficient (DSC), Mean Distance to Agreement (MDA), Target Registration Error (TRE), and Jacobian determinant. Statistical analysis was performed using Wilcoxon Signed Rank test. The median (range) DSC for rigid registration was 0.88 (0.77-0.89), 0.89 (0.81-0.93) for DIR, and 0.90 (0.86-0.94) for both types of SG-DIR tested in this study. The median MDA was 4.8 mm (3.7-6.8 mm) for rigid registration, 3.4 mm (2.4-8.7 mm) for DIR, 3.2 mm (2.0-5.2 mm) for SG-DIR where liver structures were used to guide the registration, and 2.8 mm (2.1-4.2 mm) for the SG-DIR where liver structures and anatomical landmarks were used to guide the registration. The median TRE for rigid registration was 7.2 mm (0.5-23 mm), 6.8 mm (0.7-30.7 mm) for DIR, 6.1 mm (1.1-20.5 mm) for the SG-DIR guided by only the liver structures, and 4.1 mm (0.8-19.7 mm) for SG-DIR guided by liver contours and anatomical landmarks. The SG-DIR shows higher liver conformality as per TG-132 metrics and lowest TRE compared to rigid registration and DIR in Velocity AI software for the purpose of registering treatment planning CT and post-SBRT MRI for the liver region. It was found that TRE decreases when liver contours and corresponding anatomical landmarks guide SG-DIR.

Dosimetric and radiobiological impact of abdominal compression on adjacent gastro-intestinal critical structures for patients treated with upper and mid-abdominal stereotactic body radiotherapy.

OBJECTIVES: We evaluated the dosimetric and radiobiological impact of abdominal compression (AC) on nearby gastrointestinal critical structures (GI-CS) and reported toxicities of patients treated with non-hepatic abdominal stereotactic body radiotherapy (SBRT). METHODS: Two sets of CT scans, planning scans with AC and pre-treatment diagnostic scans without AC (non-AC) were compared for patients treated with a prescription dose to planning target volume (PTV) ≥25 Gy/5-fractions at a single institution. Target volumes were delineated on both sets of scans and PTV was expanded isotropically by 2 cm (PTV+2) and 4 cm (PTV+4). All GI-CS were summated to create a composite CS (GI-lumen). Rigid registration of AC and non-AC scans was done using Velocity AI (Varian Medical Systems) to obtain dose distribution information. Lymann-Probit and logit models were used for radiobiological calculations. Toxicity scores were obtained from prospectively collected clinical data. RESULTS: A total of 12 patients were included. Mean PTV volumes were 190.3cc and 196.4cc with AC and non-AC (p=0.95). Significant improvement in V30 of GI-lumen was seen with AC (0.11cc vs. 4.97cc, p=0.04). There were no differences in the normal tissue complication probabilities of the individual GI-CS or the summary indices except a notable trend towards better NTCP for small bowel late effects with AC (0.21% vs. 2.45%; p=0.055). Three patients had acute grade-1 anorexia, one patient had acute grade-2 gastritis. There was no grade ≥3 GI toxicity. At a median follow-up of 2.6 years, total of 8/12 (66.7%) patients developed local recurrence of whom 4 (33.3%) had isolated local recurrence. CONCLUSION: Use of AC did not result in any dosimetric or radiobiological inferiority for GI-CS. The current cohort completed their treatment with minimal toxicity.

Checklist to the Elatostema (Urticaceae) of Vietnam including 19 new records, ten new combinations, two new names and four new synonyms.

Elatostema (Urticaceae) comprises several hundred herbaceous species distributed in tropical and subtropical Africa, Asia, Australia and Oceania. The greatest species richness occurs on limestone karst in Southeast Asia. Taxonomic revisions of Elatostema are largely out of date and contradict each other with respect to the delimitation of Elatostema and Pellionia. Most herbaria in SE Asia and worldwide contain significant amounts of unidentified material. As part of a broader revision of Elatostema in SE Asia, we present an updated checklist for Vietnam based on field visits, a review of specimens in herbaria worldwide, a review of type material and nomenclature. We recognize 77 taxa (75 species and two infraspecific taxa) of Elatostema in Vietnam, 23 of which were previously ascribed to Pellionia. Nineteen of these are new records for the country, i.e., E. attenuatoides, E. austrosinense, E. backeri, E. brunneinerve, E. crassiusculum, E. crenatum, E. fengshanense, E. glochidioides, E. malacotrichum, E. nanchuanense, E. oblongifolium, E. obtusum, E. oppositum, E. pergameneum, E. prunifolium, E. pseudolongipes, E. pycnodontum, E. salvinioides and E. xichouense. We place E. baviensis in synonymy of E. platyphyllum, E. colaniae in synonymy of E. myrtillus, P. macroceras in synonymy of E. hookerianum, and P. tetramera in synonymy of E. dissectum for the first time. Fourteen taxa (18% of all the recognized taxa) are endemic to Vietnam, which makes Elatostema one of the richest genera for endemic species in this country; this level of endemism is comparable to levels observed in Orchidaceae. Our checklist suggests that the highest diversity and endemism of Elatostema occurs in northern Vietnam, and that there is the greatest floristic similarity of northern Vietnam to SW China. The relationship among floristic regions is also investigated. We could find no records of Elatostema for 33 out of 63 provincial units of Vietnam, including all the southernmost provinces. We propose that further studies on the diversity of Elatostema in central and southern Vietnam are severely needed.

Characterization of nanoDot optically stimulated luminescence detectors and high-sensitivity MCP-N thermoluminescent detectors in the 40-300 kVp energy range.

PURPOSE: To investigate empirically the energy dependence of the detector response of two in vivo luminescence detectors, LiF:Mg,Cu,P (MCP-N) high-sensitivity TLDs and Al2 O3 :C OSLDs, in the 40-300-kVp energy range in the context of in vivo surface dose measurement. As these detectors become more prevalent in clinical and preclinical in vivo measurements, knowledge of the variation in the empirical dependence of the measured response of these detectors across a wide spectrum of beam qualities is important. METHOD: We characterized a large range of beam qualities of three different kilovoltage x-ray units: an Xstrahl 300 Orthovoltage unit, a Precision x-Ray X-RAD 320ix biological irradiator, and a Varian On-Board Imaging x-ray unit. The dose to water was measured in air according to the AAPM's Task Group 61 protocol. The OSLDs and TLDs were irradiated under reference conditions on the surface of a water phantom to provide full backscatter conditions. To assess the change in sensitivity in the long term, we separated the in vivo dosimeters of each type into an experimental and a reference group. The experimental dosimeters were irradiated using the kilovoltage x-ray units at each beam quality used in this investigation, while the reference group received a constant 10 cGy irradiation at 6 MV from a Varian clinical linear accelerator. The individual calibration of each detector was verified in cycles where both groups received a 10 cGy irradiation at 6 MV. RESULTS: The nanoDot OSLDs were highly reproducible, with ±1.5% variation in response following >40 measurement cycles. The TLDs lost ~20% of their signal sensitivity over the course of the study. The relative light output per unit dose to water of the MCP-N TLDs did not vary with beam quality for beam qualities with effective energies <50 keV (~150 kVp/6 mm Al). At higher energies, they showed a reduced (~75-85%) light output per unit dose relative to 6 MV x rays. The nanoDot OSLDs exhibited a very strong (120-408%) dependency of the light output relative to 6 MV x rays. Variations up to 15% between different x-ray units with equivalent effective energies were also observed. CONCLUSIONS: While convenient for clinical use, nanoDot OSLDs exhibit a strong variation in their measured light output per unit dose relative to 6 MV in the 40-300 kV x-ray range. This variability differs unit-to-unit, limiting their effective use for in vivo dosimetry applications in the kilovoltage x-ray energy range. MCP-N TLDs offer a much more stable response, but suffer from variations in sensitivity over time dependent on radiation history, which requires careful experimental handling.

Spatial patterns and cell surface clusters in perineuronal nets.

Perineuronal nets (PNN) ensheath GABAergic and glutamatergic synapses on neuronal cell surface in the central nervous system (CNS), have neuroprotective effect in animal models of Alzheimer disease and regulate synaptic plasticity during development and regeneration. Crucial insights were obtained recently concerning molecular composition and physiological importance of PNN but the microstructure of the network remains largely unstudied. Here we used histochemistry, fluorescent microscopy and quantitative image analysis to study the PNN structure in adult mouse and rat neurons from layers IV and VI of the somatosensory cortex. Vast majority of meshes have quadrangle, pentagon or hexagon shape with mean mesh area of 1.29µm(2) in mouse and 1.44µm(2) in rat neurons. We demonstrate two distinct patterns of chondroitin sulfate distribution within a single mesh - with uniform (nonpolar) and node-enriched (polar) distribution of the Wisteria floribunda agglutinin-positive signal. Vertices of the node-enriched pattern match better with local maxima of chondroitin sulfate density as compared to the uniform pattern. PNN is organized into clusters of meshes with distinct morphologies on the neuronal cell surface. Our findings suggest the role for the PNN microstructure in the synaptic transduction and plasticity.

Supramolecular architecture of betulin diacetate complexes with arabinogalactan from Larix sibirica.

Supramolecular ensembles of arabinogalactan (AG) and its complexes with betulin diacetate (BDA) were studied in water and dimethyl sulfoxide (DMSO) using ablation, induced by submillimeter radiation from the free electron laser. Solutions of 1wt% AG resulted in formation of aerosol particles with a maximum size of 60-70nm. In contrast, with DMSO as the solvent, the majority of particles were significantly smaller. Nevertheless, the addition of water shifted the particle size distribution to a larger size, suggesting the cross-linking of AG chains due to hydrogen bonding through water molecules. The ensembles of molecules were larger in solutions of the AG-BDA complex as compared to pure AG aqueous solution, and the distribution was narrow. The role of side chain interactions in the formation of AG-BDA complexes in aqueous solutions was confirmed by NMR.

Antibacterial effects of quaternary bis-phosphonium and ammonium salts of pyridoxine on Staphylococcus aureus cells: A single base hitting two distinct targets?

We studied the effects of quaternary bis-phosphonium and bis-ammonium salts of pyridoxine with lipophilic substituents on the survival and morphology of Staphylococcus aureus cells. We found that, while originating from the same base, they exhibit considerably different antimicrobial mechanisms. In the presence of Ca(2+) ions the MIC and MBC values of ammonium salt increased 100-fold, suggesting that Ca(2+) ions can successfully impede the membrane Ca(2+) ions exchange required for ammonium salt incorporation. In contrast, in the presence of quaternary phosphonium salt, the artificial capsular-like material was formed around the cells and the filamentous and chain-like growth of the cells was observed suggesting the disruption of the cell division mechanisms. Altogether, both pyridoxine derivatives successfully inhibited the growth of gram-positive bacteria (Staphylococcus aureus, Staphylococcus epidermidis, Bacillus subtilis) and Escherichia coli considerably, while demonstrated nearly no effect against Klebsiella pneumoniae and Pseudomonas aeruginosa. We suggest that due to their effects on distinct and likely complementary targets the derivatives of pyridoxine represent potentially perspective antibacterials with complicated adaptation and thus with lower risk of drug resistance development.

Water: Promising Opportunities For Tunable All-dielectric Electromagnetic Metamaterials.

We reveal an outstanding potential of water as an inexpensive, abundant and bio-friendly high-refractive-index material for creating tunable all-dielectric photonic structures and metamaterials. Specifically, we demonstrate thermal, mechanical and gravitational tunability of magnetic and electric resonances in a metamaterial consisting of periodically positioned water-filled reservoirs. The proposed water-based metamaterials can find applications not only as cheap and ecological microwave devices, but also in optical and terahertz metamaterials prototyping and educational lab equipment.

Integration of Quercetin-Iron Complexes into Phosphatidylcholine or Phosphatidylethanolamine Liposomes.

It is well known that flavonoids can chelate transition metals. Flavonoid-metal complexes exhibit a high antioxidative and therapeutic potential. However, the complexes are frequently hydrophobic ones and low soluble in water, which restricts their medical applications. Integration of these complexes into liposomes may increase their bioavailability and therapeutic effect. Here, we studied the interaction of quercetin-iron complexes with dimyristoylphosphatidylcholine (DMPC) or palmitoyl-oleoyl phosphatidylethanolamine (POPE) multilamellar liposomes. Differential scanning calorimetry (DSC) and freeze-fracture electron microscopy revealed that quercetin-iron complexes did not interact with liposomes. Quercetin however could penetrate lipid bilayers, when added to liposomes at a temperature above lipid melting. Iron cations added later penetrated into the lipid bilayers and produced complexes with quercetin in the liposomes. The quercetin-iron entry in POPE liposomes was improved when the suspension was heated above the temperature of the bilayer-hexagonal HII phase transition of the lipid. The approach proposed facilitates the integration of quercetin-iron complexes into liposomes and may promote their use in medicine.

Assessment of human immune responses to H7 avian influenza virus of pandemic potential: results from a placebo-controlled, randomized double-blind phase I study of live attenuated H7N3 influenza vaccine.

INTRODUCTION: Live attenuated influenza vaccines (LAIVs) are being developed to protect humans against future epidemics and pandemics. This study describes the results of a double-blinded randomized placebo-controlled phase I clinical trial of cold-adapted and temperature sensitive H7N3 live attenuated influenza vaccine candidate in healthy seronegative adults. OBJECTIVE: The goal of the study was to evaluate the safety, tolerability, immunogenicity and potential shedding and transmission of H7N3 LAIV against H7 avian influenza virus of pandemic potential. METHODS AND FINDINGS: Two doses of H7N3 LAIV or placebo were administered to 40 randomly divided subjects (30 received vaccine and 10 placebo). The presence of influenza A virus RNA in nasal swabs was detected in 60.0% and 51.7% of subjects after the first and second vaccination, respectively. In addition, vaccine virus was not detected among placebo recipients demonstrating the absence of person-to-person transmission. The H7N3 live attenuated influenza vaccine demonstrated a good safety profile and was well tolerated. The two-dose immunization resulted in measurable serum and local antibody production and in generation of antigen-specific CD4⁺ and CD8⁺ memory T cells. Composite analysis of the immune response which included hemagglutinin inhibition assay, microneutralization tests, and measures of IgG and IgA and virus-specific T cells showed that the majority (86.2%) of vaccine recipients developed serum and/or local antibodies responses and generated CD4⁺ and CD8⁺ memory T cells. CONCLUSIONS: The H7N3 LAIV was safe and well tolerated, immunogenic in healthy seronegative adults and elicited production of antibodies broadly reactive against the newly emerged H7N9 avian influenza virus. TRIAL REGISTRATION: ClinicalTrials.gov NCT01511419.

DNA-aptamer targeting vimentin for tumor therapy in vivo.

In recent years, new prospects for the use of nucleic acids as anticancer drugs have been discovered. Aptamers for intracellular targets can regulate cellular functions and cause cell death or proliferation. However, intracellular aptamers have limited use for therapeutic applications due to their low bioavailability. In this work, we selected DNA aptamers to cell organelles and nucleus of cancer cells, and showed that an aptamer NAS-24 binds to vimentin and causes apoptosis of mouse ascites adenocarcinoma cells in vitro and in vivo. To deliver the aptamer NAS-24 inside cells, natural polysaccharide arabinogalactan was used as a carrier reagent. The mixture of arabinogalactan and NAS-24 was injected intraperitonealy for 5 days into mice with adenocarcinoma and inhibited adenocarcinoma growth more effectively than free arabinogalactan or the aptamer alone. The use of aptamers to intracellular targets together with arabinogalactan becomes a promising approach for anticancer therapy.

Lipophilicity of flavonoid complexes with iron(II) and their interaction with liposomes.

We studied complex formation of flavonoids quercetin and taxifolin with iron(II) and the complex influence on phase transitions of phospholipid bilayer. UV-Vis spectroscopy revealed that the stoichiometry of flavonoid/iron complexes was equal to 3:2 and 2:1. Molecular modeling and experimental measurements demonstrated the increase of flavonoids lipophilicity after the complex formation. A considerable influence of quercetin-iron complex on Palmitoyl-Oleoyl-Phosphatidylethanolamine transitions from bilayer to hexagonal HII phase was detected by differential scanning calorimetry. The obtained data are related to flavonoid/iron complexes bioavailability, their influence on cell membrane functioning, and should be considered in designing liposomal vehicles for drug and gene delivery.

Endogenous thrombospondin-1 regulates leukocyte recruitment and activation and accelerates death from systemic candidiasis.

Disseminated Candida albicans infection results in high morbidity and mortality despite treatment with existing antifungal drugs. Recent studies suggest that modulating the host immune response can improve survival, but specific host targets for accomplishing this goal remain to be identified. The extracellular matrix protein thrombospondin-1 is released at sites of tissue injury and modulates several immune functions, but its role in C. albicans pathogenesis has not been investigated. Here, we show that mice lacking thrombospondin-1 have an advantage in surviving disseminated candidiasis and more efficiently clear the initial colonization from kidneys despite exhibiting fewer infiltrating leukocytes. By examining local and systemic cytokine responses to C. albicans and other standard inflammatory stimuli, we identify a crucial function of phagocytes in this enhanced resistance. Subcutaneous air pouch and systemic candidiasis models demonstrated that endogenous thrombospondin-1 enhances the early innate immune response against C. albicans and promotes activation of inflammatory macrophages (inducible nitric oxide synthase⁺, IL-6(high), TNF-α(high), IL-10(low)), release of the chemokines MIP-2, JE, MIP-1α, and RANTES, and CXCR2-driven polymorphonuclear leukocytes recruitment. However, thrombospondin-1 inhibited the phagocytic capacity of inflammatory leukocytes in vivo and in vitro, resulting in increased fungal burden in the kidney and increased mortality in wild type mice. Thus, thrombospondin-1 enhances the pathogenesis of disseminated candidiasis by creating an imbalance in the host immune response that ultimately leads to reduced phagocytic function, impaired fungal clearance, and increased mortality. Conversely, inhibitors of thrombospondin-1 may be useful drugs to improve patient recovery from disseminated candidiasis.

sFRP-1 binds via its netrin-related motif to the N-module of thrombospondin-1 and blocks thrombospondin-1 stimulation of MDA-MB-231 breast carcinoma cell adhesion and migration.

Secreted frizzled-related protein (sFRP)-1 is a Wnt antagonist that inhibits breast carcinoma cell motility, whereas the secreted glycoprotein thrombospondin-1 stimulates adhesion and motility of the same cells. We examined whether thrombospondin-1 and sFRP-1 interact directly or indirectly to modulate cell behavior. Thrombospondin-1 bound sFRP-1 with an apparent K(d)=48nM and the related sFRP-2 with a K(d)=95nM. Thrombospondin-1 did not bind to the more distantly related sFRP-3. The association of thrombospondin-1 and sFRP-1 is primarily mediated by the amino-terminal N-module of thrombospondin-1 and the netrin domain of sFRP-1. sFRP-1 inhibited α3ß1 integrin-mediated adhesion of MDA-MB-231 breast carcinoma cells to a surface coated with thrombospondin-1 or recombinant N-module, but not adhesion of the cells on immobilized fibronectin or type I collagen. sFRP-1 also inhibited thrombospondin-1-mediated migration of MDA-MB-231 and MDA-MB-468 breast carcinoma cells. Although sFRP-2 binds similarly to thrombospondin-1, it did not inhibit thrombospondin-1-stimulated adhesion. Thus, sFRP-1 binds to thrombospondin-1 and antagonizes stimulatory effects of thrombospondin-1 on breast carcinoma cell adhesion and motility. These results demonstrate that sFRP-1 can modulate breast cancer cell responses by interacting with thrombospondin-1 in addition to its known effects on Wnt signaling.