Introducing an lsoprenaline Eluting Guidewire: Report on its Design and the Results of the Dose-Determining Pilot Study.

Introduction: Retrograde intrarenal surgery (RIRS) is associated with complications, many of which are related to the intrarenal pressure (IRP). We aim to describe the design of a novel isoprenaline-eluting guidewire ("IsoWire") and present the results from the first in vitro release studies and the first animal studies showing its effect on IRP. Materials and Methods: The IsoWire comprises a Nitinol core surrounded by a stainless-steel wire wound into a tight coil. The grooves created by this coil provided a reservoir for adding a hydrogel coating into which isoprenaline, a beta-agonist, was loaded. Animal studies were performed using a porcine model. For the control, IRP, heart rate (HR), and mean arterial pressure (MAP) were measured continuously for 6 minutes with a standard guidewire in place. For the experiment, the standard hydrophilic guidewire was removed, the IsoWire was inserted into the renal pelvis, and the same parameters were measured. Results: In vitro analysis of the isoprenaline release profile showed that most (63.9 ± 5.9%) of the loaded drug mass was released in the 1st minute, and almost all of the drug was released in the first 4 minutes exponentially. Porcine studies showed a 25.1% reduction in IRP in the IsoWire that released 10 µg in the 1st minute; however, there was a marked increase in HR. The average percentage reduction in IRP was 8.95% and 21.3% in the IsoWire that released 5 and 7.5 µg of isoprenaline, respectively, with no changes in HR or MAP. Conclusions: The IsoWire, which releases 5 and 7.5 µg of isoprenaline in the 1st minute, appears to be safe and effective in reducing the IRP. Further studies are needed to establish whether the isoprenaline-induced ureteral relaxation will render easier insertion of a ureteral access sheath, reduce IRP during sheathless RIRS, or even promote the practice of sheathless RIRS.

New CEACAM-targeting 2A3 single-domain antibody-based chimeric antigen receptor T-cells produce anticancer effects in vitro and in vivo.

Recently, a breakthrough immunotherapeutic strategy of chimeric antigen receptor (CAR) T-cells has been introduced to hematooncology. However, to apply this novel treatment in solid cancers, one must identify suitable molecular targets in the tumors of choice. CEACAM family proteins are involved in the progression of a range of malignancies, including pancreatic and breast cancers, and pose attractive targets for anticancer therapies. In this work, we used a new CEACAM-targeted 2A3 single-domain antibody-based chimeric antigen receptor T-cells to evaluate their antitumor properties in vitro and in animal models. Originally, 2A3 antibody was reported to target CEACAM6 molecule; however, our in vitro co-incubation experiments showed activation and high cytotoxicity of 2A3-CAR T-cells against CEACAM5 and/or CEACAM6 high human cell lines, suggesting cross-reactivity of this antibody. Moreover, 2A3-CAR T-cells tested in vivo in the BxPC-3 xenograft model demonstrated high efficacy against pancreatic cancer xenografts in both early and late intervention treatment regimens. Our results for the first time show an enhanced targeting toward CEACAM5 and CEACAM6 molecules by the new 2A3 sdAb-based CAR T-cells. The results strongly support the further development of 2A3-CAR T-cells as a potential treatment strategy against CEACAM5/6-overexpressing cancers.

Interoception, alexithymia, and anxiety among individuals with alcohol use disorder.

Background: Interoception (i.e., the ability to recognize bodily signals), alexithymia (i.e., the inability to recognize emotional states) and negative affect (i.e., unpleasant feelings such as anxiety) have been associated with alcohol use disorder (AUD). Previous research suggests that interoception may underlie alexithymia, which in turn may be associated with negative affectivity. However, this remains to be empirically tested. This study investigates whether alexithymia mediates the association between interoception and anxiety and whether this association differs across individuals with AUD and a healthy control (HC) comparison group. Methods: The AUD group consisted of 99 participants enrolled in an 8-week abstinence-based inpatient treatment program. The HC group included 103 healthy individuals. The heartbeat counting task (HCT) was used to assess interoception (cardiac interoceptive accuracy). The Toronto Alexithymia Scale (TAS-20) was used to assess alexithymia. The Brief Symptom Inventory (BSI) was used to assess anxiety. Results: The moderated mediation model with interoception as the predictor, alexithymia as the mediator, and negative affect (i.e., state anxiety) as the dependent variable was tested. The analysis showed that the conditional indirect effect of interoception on anxiety via alexithymia was significant for individuals with AUD [ab = -0.300, bootstrap 95% CI = (-0.618, -0.088)], as well as for HCs [ab = -0.088, bootstrap 95% CI = (-0.195, -0.014)]; however, the conditional indirect effect significantly differed across HCs and individuals with AUD. Namely, the mediated effect was greater among individuals with AUD compared to the HC group. Conclusion: The results suggests that interoceptive impairment contributes to greater negative affect (i.e., state anxiety) via alexithymia especially for individuals with AUD. Improving emotion recognition via therapeutic methods focused on strengthening interoceptive abilities could improve outcomes for individuals receiving treatment for AUD.

Impact of an Aluminization Process on the Microstructure and Texture of Samples of Haynes 282 Nickel Alloy Produced Using the Direct Metal Laser Sintering (DMLS) Technique.

In this study, we examined the effects of an aluminization process on the microstructure and texture of Haynes 282 nickel samples fabricated using the direct metal laser sintering technique. The aluminization process involved the use of chemical vapor deposition with AlCl3 vapors in a hydrogen atmosphere at a temperature of 1040 °C for 8 h. Following the 3D printing and aluminization steps, we analyzed the microstructure of the Haynes 282 nickel alloy samples using light microscopy and scanning electron microscopy. Additionally, we investigated the texture using X-ray diffractometry. A texture analysis revealed that after the process of direct laser sintering of metals, the texture of the Haynes 282 nickel alloy samples developed a texture typical of cast materials. Then, in the aluminization process, the texture was transformed-from foundry-type components to a texture characteristic of recrystallization.

Distress tolerance and emotional regulation in individuals with alcohol use disorder.

Background: Previous research suggests that low distress tolerance may be associated with poor emotion regulation, contribute to drinking to cope motives, and predict alcohol-related problems in non-clinical populations. However, little is known about the ability to tolerate distress among individuals with alcohol use disorder (AUD) and its association with emotional dysregulation. The aim of this study was to examine the link between emotional dysregulation and a behavioral measure of distress tolerance among individuals with AUD. Methods: The sample consisted of 227 individuals with AUD enrolled in an 8-week abstinence-based inpatient treatment program. Behavioral distress tolerance was assessed using a test of ischemic pain tolerance and the Difficulties in Emotion Regulation Scale (DERS) was used to assess emotion dysregulation. Results: Distress tolerance was significantly associated with emotional dysregulation even when accounting for alexithymia, depressive symptomatology, age, and biological sex. Conclusion: The current study provides preliminary support for an association between low distress tolerance and emotion dysregulation in a clinical group of patients with AUD.

Abnormal behavioral and neural responses in the right dorsolateral prefrontal cortex during emotional interference for cognitive control in pedophilic sex offenders.

Studies within the last decade have reported neural and behavioral differences in cognitive control between men with the pedophilic disorder who commit (CSO+) and do not commit (CSO-) child sexual abuse. Prior studies reported a higher number of errors in Go/Nogo task and lower activity of the prefrontal cortex in NoGo trials, in CSO+ compared with CSO-. Moreover, negative mood was reported as a risk factor for child sexual abuse in pedophilic men. We aimed to examine differences in brain function and behavior between CSO+ and CSO- patients regarding emotional interference on cognitive processes and inhibition. We recruited CSO+ (n = 11) and CSO- (n = 14) patients as well as matched healthy controls (HC) (n = 17). Participants performed the affective Go/NoGo task in a block-design functional magnetic resonance imaging experiment. The task comprised the following four conditions: Negative Go, including only Go stimuli and negatively valenced pictures; Negative NoGo, including 50% of Go and 50% of NoGo trials as well as negatively valenced pictures; and two corresponding conditions with neutral pictures. Brain analysis was restricted to the dorsolateral prefrontal (DLPFC), orbitofrontal, and anterior cingulate cortices. The HC and CSO- groups, but not the CSO+ group, showed significantly slower reactions in negative blocks compared with neutral blocks. Brain analysis revealed increased activation in the right DLPFC during emotional interference contrast (Negative > Neutral) in the HC and CSO- groups; however, there was decreased activation in the CSO+ group. In the CSO+ group, negative distractors did not increase cognitive control processes, which was observed in the CSO- and HC groups at the behavioral and neural levels. These results support previous reports indicating offender status is associated with cognitive and emotional impairments.

Investigations of Model Multilayer Ceramic Casting Molds in a Raw State by Nondestructive Methods.

This article presents the results of research on the use of modern nondestructive methods such as 3D scanning, thermography and computed tomography (CT) to assess the quality of multilayer ceramic molds. Tests were performed on spherical samples of multilayer ceramic molds in the raw state. Samples were made of molding sands composed of quartz and molochite powders, the alcoholic binder hydrolyzed ethyl silicate (ZKE) and an aqueous binder based on colloidal silica. Thickness measurements of spherical forms were made using a 3D scanner. Porosity measurements were made using CT. Additionally, thermography observations of the mold cooling process were made with controlled temperature and humidity. The results of temperature measurements of samples were compared with measurements of thickness and porosity. The practical goal was to determine the possibility of using thermography, 3D scanning and CT as a quick method for detecting mold defects by varying their thickness, porosity and cracks and for final verification of the ceramic molds' condition before casting.

Assessment of the Emission of Pollutants from Public Transport Based on the Example of Diesel Buses and Trolleybuses in Gdynia and Sopot.

The present study attempts to examine the research gap in terms of comparing the environmental impact of trolleybuses and diesel buses in the conditions of a country with an unfavourable energy mix. The analysed example concerns the trolleybus transport system in Gdynia, in northern Poland, which also partially serves the neighbouring city of Sopot. In the last few years, two bus lines have been electrified with trolleybuses in the In-Motion-Charging technology, which enables operation on sections without an overhead network. Using the actual operational data, a comparative analysis of the emissivity of diesel buses and trolleybuses used on the same lines in an identical operating regime was conducted. Moreover, an attempt was made to estimate the damage costs of the emission of air pollutants for the above-mentioned means of transport. Research has shown that trolleybuses significantly help to reduce emissions of nitrogen oxides, non-methane volatile organic compounds and particulate matter, while increasing sulphur dioxide emissions on the served lines. They also generate lower specific emissions of carbon dioxide compared to diesel buses. However, taking into account the differences in the number of seats in these vehicles, the length of routes resulting from a need to provide access to the necessary infrastructure and the total amount of kilometres covered on a given route, they may cause higher emissions per year and per the product life cycle than diesel buses. This is related to the unfavourable structure of energy production in Poland, which is dominated by coal sources. The research results clearly show that the use of trolleybuses in public transport contributes to a reduction of the damage costs of the emission of pollutants that amount to approximately EUR (€) 30,000-60,000 per year for the analysed lines.

Alcohol use and interoception - A narrative review.

Interoception, defined as the ability to perceive and interpret body signals, may play an important role in alcohol use disorder (AUD). Earlier studies suggested an association between interoception impairment and known risk factors for AUD (e.g., alexithymia, emotion dysregulation, impulsivity, pain). Neurobiological studies show that the neurotoxicity of alcohol affects various elements of the interoceptive system (especially the insula) at structural and functional levels, with differential short/long term impacts. Conversely, primary interoceptive impairments may promote alcohol consumption and foster the evolution towards addiction. Despite convincing evidence demonstrating that interoception impairment may be an important contributor to the development and course of AUD, only a few studies directly evaluated interoceptive abilities in AUD. The research shows that interoceptive accuracy, the objective component of interoception, is lower in AUD individuals, and is correlated with craving and emotion dysregulation. Interoceptive sensibility is in turn higher in AUD individuals compared to healthy controls. Moreover, there is evidence that therapy focused on improving the ability to sense signals from the body in addiction treatment is effective. However, important methodological limitations in interoceptive measures persist, and it is therefore necessary to further investigate the associations between interoception and AUD.

Polymer Binders of Ceramic Nanoparticles for Precision Casting of Nickel-Based Superalloys.

This study presents the general characteristics of binders used in precision casting of Nickel-based superalloys. Three groups of binders were described: resins, organic compounds, and materials containing nanoparticles in alcohol or aqueous systems. This study also includes literature reports on materials commonly used and those recently replaced by water-soluble binders, i.e., ethyl silicate (ES) and hydrolysed ethyl silicate (HES). The appearance of new and interesting solutions containing nano-alumina is described, as well as other solutions at the initial stage of scientific research, such as those containing biopolymers, biodegradable polycaprolactone (PCL), or modified starch. Special attention is paid to four binders containing nano-SiO2 intended for the first layers (Ludox AM, Ludox SK) and structural layers (EHT, Remasol) of shell moulds. Their morphology, viscosity, density, reactions, and electrokinetic potential were investigated. The binders were characterized by a high solid-phase content (>28%), viscosity, and density close to that of water (1-2 mPa·s) and good electrokinetic stability. The nanoparticles contained in the binders were approximately spherically shaped with an average particle size of 16-25 nm.

Pain Sensitivity, Negative Affect, and Alcohol Use Disorder Status: A Moderated Mediation Study of Emotion Dysregulation.

Previous work suggests that the association between pain and emotional processes among individuals with alcohol use disorder (AUD) may differ from healthy controls. This study investigates whether pain sensitivity mediates the association between negative affect and emotional dysregulation and whether this association differs across AUD status using moderated mediation. The sample included 165 individuals diagnosed with AUD and 110 healthy controls. Of interest was pain sensitivity, as assessed with the Pain Sensitivity Questionnaire, negative affect, as assessed with the Beck Depression Inventory, and emotional dysregulation, as assessed with the Difficulties in Emotional regulation Scale. Age, biological sex, and current pain severity were included as covariates. The results support a moderated partial mediation model that explained 44% of the variance in emotional dysregulation. The findings indicate that negative affect is related to higher pain sensitivity across groups. Moreover, pain sensitivity partially mediated the association between negative affect and emotional dysregulation, but in opposite directions depending on AUD status. Among healthy controls, greater pain sensitivity was related to better emotional regulation, while greater pain sensitivity led to greater emotional dysregulation among individuals with AUD. The potential parallels in the underlying neurobiological mechanisms of emotionality, pain, and AUD suggest that interventions targeting pain may improve adaptive affect regulation skills, which in turn could reduce negative affect and its effect on pain sensitivity among individuals with AUD.

The synergistic effect between interoceptive accuracy and alcohol use disorder status on pain sensitivity.

BACKGROUND: Interoceptive accuracy and pain sensitivity are both risk factors in the development of alcohol use disorder (AUD). However, the synergistic association between these two factors has not been investigated in an AUD sample. Therefore, the aim of the current study was to investigate whether the association between interoceptive accuracy and sensitivity to pain differed across AUD status. METHODS: The study group included 165 individuals diagnosed with AUD (88.1% men) and 110 healthy controls (HCs; 74.5% men). Interoceptive accuracy was assessed with the Schandry Task. The Pain Sensitivity Questionnaire was utilized to measure sensitivity to pain. Anxiety, biological sex, and age were included as covariates in a model examining the role of AUD status as a moderator in the association between interoceptive accuracy and pain sensitivity. RESULTS: A significant interaction was found between interoceptive accuracy and AUD status (b = -4.580, 95% CI = [-8.137, -1.022], p = 0.012, ΔR2 = 0.032). Findings indicate that interoceptive accuracy was negatively associated with pain sensitivity among individuals with AUD, while there was a trend for an opposite association among healthy controls. CONCLUSION: We hypothesize that persistent alcohol drinking may contribute to disruption of the normative association between interoception and pain. Future studies should be conducted to develop knowledge on this association and to investigate its possible therapeutic significance and implications.

Molding Binder Influence on the Porosity and Gas Permeability of Ceramic Casting Molds.

The investment casting process is widely used in the aerospace industry to produce complex engine parts. The article determines the properties of quartz powders, nanosilica dioxide binders, and multilayer samples of ceramic casting molds. The properties of spherical molds obtained using an alcohol-water system derived from hydrolyzed ethyl silicate (ZKE) and LUDOX PX-30 (type Q1) were compared with those obtained in water systems derived from Remasol Plus and Remasol Premium binders (type Q2). The spherical samples are composed of seven layers made with the use of an immersion-sprinkling method. To assess the properties of the molds, X-ray microscopy (µCT), scanning electron microscopy (SEM), and gas permeability analysis over a temperature range of 20-950 °C were utilized. The binder type is proven to affect the properties of the casting mold samples. The material obtained in the water system, Q2, has advantageous properties such as a high porosity and gas permeability.

Electrocardiographic identification of the culprit coronary artery in acute non-ST-elevation myocardial infarction: predictive value of N-wave and T-wave precordial instability.

BACKGROUND: Recently, novel ischemic electrocardiographic changes have been described, which may be clinically significant in the identification of the culprit coronary vessel in patients with non-ST-elevation myocardial infarction (NSTEMI). We sought to determine the predictive value of N-wave, T-wave precordial instability, de-Winter ST/T-wave complex, and inferolateral myocardial infarction in the identification of the culprit artery in patients with NSTEMI referred for early invasive (<24 h) treatment. METHODS: A total of 148 patients with NSTEMI, aged 40-91 years, were enrolled from a cohort of 510 consecutive NSTEMI subjects, hospitalized in our center in 2015-2017. RESULTS: Of the evaluated ischemic ECG changes, the most common finding in patients with culprit left circumflex (LCx)/obtuse marginal artery or right coronary artery was T-wave precordial instability (28.3 and 13.5%, respectively), whereas in individuals with culprit left anterior descending/diagonal artery, T-wave precordial instability and N-wave in leads II, III or aVF occurred equally often (16.0%). A significant relationship was found between the occurrence of N-wave in inferolateral leads and culprit LCx/obtuse marginal. In multivariable analysis, N-wave in lead aVL [odds ratio (OR) 2.10; 95% confidence interval (CI), 1.15-3.81], and T-wave precordial instability (OR 1.56; 95% CI, 1.02-2.41) were independent predictors of culprit LCx/obtuse marginal. The accuracy of N-wave in lead aVL in predicting the culprit LCx/obtuse marginal was 73.9% and was higher than the accuracy of T-wave precordial instability, which was 69.1%. CONCLUSIONS: In patients with NSTEMI referred for early invasive treatment, the presence of N-wave or T-wave precordial instability may be of greater clinical importance in the prediction of culprit LCx/obtuse marginal than classic ischemic changes.

Analysis of Microstructure and Properties of a Ti-AlN Composite Produced by Selective Laser Melting.

Selective Laser Melting (SLM) is a manufacturing technique that is currently used for the production of functional parts that are difficult to form by the traditional methods such as casting or CNC (Computer Numerical Control) cutting from a wide range of metallic materials. In our study, a mixture of commercially pure titanium (Ti) and 15% at. aluminum nitride (AlN) was Selective Laser Melted to form three-dimensional objects. The obtained 4 mm edge cubes with an energy density that varied from 70 to 140 J/mm3 were examined in terms of their microstructure, chemical and phase composition, porosity, and Vickers microhardness. Scanning Electron Microscopy (SEM) observations of the etched samples showed inhomogeneities in the form of pores and unmelted and partly melted AlN particles in the fine-grained dendritic matrix, which is typical for titanium nitrides and titanium aluminum nitrides. The AlN particles remained unmelted in samples, but no porosity was observed in the interface area between them and the dendritic matrix. Additionally, samples fabricated with the presintering step had zones with different sizes of dendrites, suggesting a differing chemical composition of the matrix and the possibility of the formation of the phases forming an Ti-Al-N ternary system. The chemical composition in the microareas of the samples was determined using Energy Dispersive X-Ray Spectroscopy (EDS) and revealed differences in the homogeneity of the samples depending on the SLM process parameters and the additional presintering step. The phase composition, examined using X-ray Diffraction analysis (XRD), showed that samples were formed from Ti, TiN, and AlN phases. Porosity tests carried out using a computer microtomography revealed porosities in a range from 7% to 17.5%. The formed material was characterized by a relatively high hardness exceeding 700 HV0.2 over the entire cross-section, which depended on the manufacturing conditions.

Predictive value of electrocardiographic ST­segment elevation myocardial infarction equivalents for detecting acute coronary artery occlusion in patients with non-ST­segment elevation myocardial infarction.

BACKGROUND: The sensitivity and accuracy of 12-lead ECG for the detection of acute total occlusion (TO) of the culprit coronary artery in non-ST-elevation myocardial infarction (NSTEMI) is still suboptimal, particularly for posterolateral circulation.   Aims: We evaluated the prevalence and predictive value of electrocardiographic STEMI-equivalents (i.e. de-Winter ST/T-wave complex, N-wave, T-wave precordial instability, and posterior myocardial infarction) for detecting acute coronary artery occlusion in NSTEMI patients referred for early invasive treatment. METHODS: A total of 165 NSTEMI patients were enrolled. The patients were grouped according to the coronary angiography findings into those with TO (TIMI 0) in the culprit artery (n=43) and those with preserved flow in this vessel (TIMI 1-3) (n=122). RESULTS: The main findings of this study were as follows: 1) 31.5% of patients had at least one STEMI-equivalent, mostly N-wave in lead II, III or aVF; 2) the most common STEMI-equivalent in subjects with acute TO was T-wave precordial instability; 3) there was a significant relationship between the prevalence of STEMI-equivalents and acute coronary artery occlusion; 4) among all evaluated ECG parameters, only ST-segment depression in leads I, aVL, V6 was an independent predictor of acute TO in multivariate analysis; 5) ST-segment depression in leads I, aVL, V6 had higher specificity, positive and negative predictive values as well as accuracy in predicting acute TO of the culprit vessel, as compared to STEMI-equivalents. CONCLUSIONS: STEMI-equivalents do not seem to have a relevant advantage over classic ischaemic ECG changes in the prediction of acute coronary artery occlusion in NSTEMI patients.

Structural and electronic properties of Fe monolayer on BaTiO3(0 0 1).

The structural, electronic, and phonon properties of the BaTiO3(0 0 1) surface and the Fe/BaTiO3(0 0 1) interface have been studied within the density functional theory. Attention is paid to the lattice instabilities (soft phonon modes) that induce ferroelectric distortions in the surface and the interface. A phonon-induced monoclinic (Cm) thin-film counterpart of the low-temperature rhombohedral (R3m) ferroelectric bulk BaTiO3 phase is found. The changes in crystal structure, electronic density of states, atomic charges, and magnetic moments associated with the ferroelectric distortions are discussed comparing the results of the standard GGA and the hybrid DFT calculations. The magnetoelectric coupling at the Fe/BaTiO3(0 0 1) interface is investigated by the analysis of changes in magnetic moments on Fe and Ti atoms induced by the atomic displacements perpendicular and parallel to the surface.

Development and Characterization of a Camelid Single Domain Antibody-Urease Conjugate That Targets Vascular Endothelial Growth Factor Receptor 2.

Angiogenesis is the process of new blood vessel formation and is essential for a tumor to grow beyond a certain size. Tumors secrete the pro-angiogenic factor vascular endothelial growth factor, which acts upon local endothelial cells by binding to vascular endothelial growth factor receptors (VEGFRs). In this study, we describe the development and characterization of V21-DOS47, an immunoconjugate that targets VEGFR2. V21-DOS47 is composed of a camelid single domain anti-VEGFR2 antibody (V21) and the enzyme urease. The conjugate specifically binds to VEGFR2 and urease converts endogenous urea into ammonia, which is toxic to tumor cells. Previously, we developed a similar antibody-urease conjugate, L-DOS47, which is currently in clinical trials for non-small cell lung cancer. Although V21-DOS47 was designed from parameters learned from the generation of L-DOS47, additional optimization was required to produce V21-DOS47. In this study, we describe the expression and purification of two versions of the V21 antibody: V21H1 and V21H4. Each was conjugated to urease using a different chemical cross-linker. The conjugates were characterized by a panel of analytical techniques, including SDS-PAGE, size exclusion chromatography, Western blotting, and LC-MSE peptide mapping. Binding characteristics were determined by ELISA and flow cytometry assays. To improve the stability of the conjugates at physiologic pH, the pIs of the V21 antibodies were adjusted by adding several amino acid residues to the C-terminus. For V21H4, a terminal cysteine was also added for use in the conjugation chemistry. The modified V21 antibodies were expressed in the E. coli BL21 (DE3) pT7 system. V21H1 was conjugated to urease using the heterobifunctional cross-linker succinimidyl-[(N-maleimidopropionamido)-diethyleneglycol] ester (SM(PEG)2), which targets lysine resides in the antibody. V21H4 was conjugated to urease using the homobifunctional cross-linker, 1,8-bis(maleimido)diethylene glycol (BM(PEG)2), which targets the cysteine added to the antibody C-terminus. V21H4-DOS47 was determined to be the superior conjugate as the antibody is easily produced and purified at high levels, and the conjugate can be efficiently generated and purified using methods easily transferrable for cGMP production. In addition, V21H4-DOS47 retains higher binding activity than V21H1-DOS47, as the native lysine residues are unmodified.

Non-apoptotic Fas signaling regulates invasiveness of glioma cells and modulates MMP-2 activity via NFkappaB-TIMP-2 pathway.

Fas (CD95/APO-1) is a cell surface "death receptor" that mediates apoptosis upon engagement by its ligand, FasL. Paradoxically, Fas/FasL can also promote cell invasion among non-apoptotic cells; here, we show that Fas/FasL signaling can promote tumor invasion when apoptosis is compromised. We have developed a recombinant FasL Interfering Protein (FIP) to interfere with Fas signaling in C6 glioma cells expressing both Fas receptor and its ligand. FIP administration did not affect cell viability but impaired cell motility and invasiveness of glioma cells. Blockade of Fas signaling reduced MMP-2 activity in glioma cells, that was associated with down-regulation of MAPK signaling, and AP-1 and NFkappaB-driven transcription. FIP treatment did not affect mmp-2 and mt1-mmp expression but significantly attenuated timp-2 expression and TIMP-2 amount in the culture medium. Studies with pharmacological inhibitors of JNK/c-Jun (SP600125) and NFkappaB (BAY11-7082) signaling pathways demonstrated that timp-2 expression is regulated by NFkappaB transcription factor. Our findings show that non-apoptotic Fas signaling activated in the autocrine manner or through microenvironment derived factors can regulate invasiveness of glioma cells via modulation of MMP-2 activation, likely by controlling TIMP-2 expression.