Plate Adhesion Research Articles

ОПЫТ ИСПОЛЬЗОВАНИЯ ДИОДНОГО ЛАЗЕРА В КОМПЛЕКСНОМ ЛЕЧЕНИИ РАННИХ ПРОЯВЛЕНИЙ ХРОНИЧЕСКОГО ВОСПАЛЕНИЯ В ТКАНЯХ ПАРОДОНТА

The article presents data on the effect of a low-intensity diode laser and applications of adhesive plates based on biogel in the treatment of initial manifestations of chronic inflammation in periodontal tissues. Materials and methods. 3 equal randomized groups of 35 patients each with manifestations of chronic inflammation in periodontal tissues were created. In accordance with the protocol of local treatment, all patients underwent a complex of professional oral hygiene and motivation to conduct individual hygiene. In the main clinical subgroups, patients additionally received laser therapy and applications of adhesive plates on the gum, or treatment according to clinical protocols of StAR, 2001. The quantitative assessment of the periodontal condition was carried out using OHI-S indices – SBI, PMA, GI, PI. 
 The aim of the study is to evaluate the change in hygienic and periodontal indices in patients with early manifestations of chronic inflammation in periodontal tissues after using laser therapy in combination with applications of adhesive plates based on biogel and methods of local treatment according to the clinical protocols of Dental Association of Russia, 2001. Results. A comparative evaluation of the effectiveness of the effect of a diode laser and applications of adhesive plates based on biogel and local treatment according to the clinical protocols of Dental Association of Russia, 2001 was confirmed by dynamic observation and monitoring of changes in reversible and irreversible indices. Conclusions. The observed qualitative decrease in indices in patients against the background of laser therapy and applications of adhesive plates based on biogel from the initial indicators was 8.2, 7.3 and 3.4 times for OHI-S, 7.2, 6.8 and 1.9 times for PMA, 5.9, 3.9, 2.6 times for SBI, 6.2, 4.5, 0.98 times for GI times, for PI 6,3,1 and 0.57 times (p < 0.001). In clinical subgroups, against the background of local treatment, according to the clinical protocols of Dental Association of Russia, 2001, the decrease in the index of OHY-S from the initial indicators occurred 3.3, 4.4, 1.8 times, for the PMA index 2.3, 4.5 times, for the SBI index 3.0, 2.6 times, for the GI index on average 2.1 times, for the PI index 2.5, 1.4 times, while its increase was observed in the subgroup with clinically intact periodontal disease by 6 months of dynamic follow-up and a decrease by 1.4 times by the year of follow-up (p < 0.001).

Loss of connective tissue growth factor permits mesenchymal transition in ovarian cancer (260)

<b>Objectives:</b> Epithelial-mesenchymal transition (EMT) is a characteristic feature of ovarian cancer (OC) metastasis. Our study aimed to determine the role of Connective Tissue Growth Factor (CTGF) in EMT. <b>Methods:</b> R182 and R2615 are epithelial OC cell lines that are CK18+, b-catenin+, TWIST -, and vimentin+. Time-course experiments +/- Transforming Growth Factor-b (TGF-b) and a TGF-b inhibitor were performed to evaluate the timing of CTGF expression in EMT. R182 and R2615 CTGF knockouts (KO) were derived utilizing a Cas9/CRISPR-Cas9 lentivirus plasmid vector. Phenotypic characterization of wild types (WT) and CTGF KO cell lines were performed with the assays, including scratch wound, proliferation, and anoikis resistance assays. Scratch wound assays were treated with 50 ng/mL and 100 ng/mL concentrations of recombinant CTGF for 48 hours (hr). Cells were plated in triplicate, and confluence was measured. Proliferation was determined by cell count using Biotek Cytation over 72 hr. For anoikis resistance, cells were plated in triplicate in an ultra-low adhesive cell plate with cell growth media. Using Promega CellTiter assay, cell viability was quantified by absorbance at 490 nm at the following time points: 1hr, 24hr, 48hr, and 120hr. Western blots were performed to evaluate the expression of epithelial and mesenchymal markers. RNA sequence analysis was performed, and pathway analysis was evaluated using the iPathway guide. <b>Results:</b> TGF-b induced CTGF expression in both OC cell lines tested. CTGF was induced as early as 30 minutes post-treatment, continued to increase until 6hr, and expression was maintained until 24 hr. No CTGF expression was seen in the mesenchymal mR182 and mR2615 OC cell lines. CTGF KO was confirmed by western blot analysis and indel sequencing. CTGF KO had no effect on the expression of epithelial markers, b-catenin, CK18, and vimentin, but SNAIL, a mesenchymal marker, was induced. Loss of CTGF also had no effect on cell proliferation but promoted anoikis resistance. R182 CTGF KO cells displayed 80% viability when grown in detached conditions for 48hr compared to only 20% viability in WT cells. At 120hr, 30% of KO cells remain viable compared to <10% for WT cells. R2615 WT and KO cells demonstrated similar results. Finally, the scratch assay showed that CTGF KO cells decrease the time to confluence compared to WT. Administration of recombinant CTGF does not rescue the phenotype to WT levels.Fig. 1 <b>Conclusions:</b> CTGF is expressed early in epithelial OC cells during EMT and is regulated by TGF-b. Our data suggest that CTGF permits maintenance of an epithelial phenotype through inhibition of SNAIL expression, and loss of CTGF should be evaluated as a poor prognosis indicator in OC.

Norepinephrine promotes anoikis resistance in fallopian tube secretory cells through beta adrenergic receptor signaling (247)

<b>Objectives:</b> High-grade serous ovarian cancer (HGSOC) remains the most lethal gynecologic malignancy with a 5-year survival rate approaching 40%. Unfortunately, most women present at an advanced stage due to subtle symptomatology at disease onset and inadequate screening protocols. Thus, understanding the cellular events that lead to the development and malignant transformation of fallopian tube (FT) precursor cells into metastatic disease may lead to actionable targets to improve outcomes. Fallopian tube secretory epithelium is considered the cell of origin of HGSOC, where <i>TP53</i> and other genetic defects accrue to permit exfoliation and seeding of the ovary and peritoneum, leading to widespread metastasis. Follicular fluid bathes the fallopian tube monthly during the reproductive years and contains several factors contributing to carcinogenesis. Recently, neurotransmitters such as norepinephrine (NE) have been implicated in ovarian cancer growth and metastasis through multiple mechanisms and are also present within the follicular fluid. Thus, the objective of the current study was to evaluate the effects of NE on FT epithelial cell function under 3D conditions. <b>Methods:</b> FT cell lines (FT189, FT190, FT194, FT237, FT240, FT246, FT282), previously immortalized through several mechanisms, were plated into round bottom ultralow adhesion plates with 10µM NE or vehicle control (VHC), in the presence or absence of adrenergic antagonist propranolol (10µM). Spheroids were serially imaged over 24h. For FT237 and FT246, live/dead cell staining was performed using ReadyProbe cell viability assay, and apoptotic activity was assessed via annexin staining analyzed by flow cytometry. qPCR and western blotting were used to assess receptor expression. <b>Results:</b> All FT lines examined expressed <i>ADRB1</i> and <i>ADRB2</i> by both western blot and qPCR. Treatment with NE resulted in significant compaction of spheroid formation by 33% (+/- 5%, p <0.001) at 24h compared to VHC. This response was completely abrogated by preexposure to propranolol (p <0.05). NE nor propranolol had a significant impact on cell viability under immunofluorescence. However, annexin staining revealed a significant 5% increase in cell survival with a concomitant decrease in early and late apoptotic cell populations exposed to NE in 3D culture (p<0.05), compared to VHC. FT exposure to NE resulted in significantly increased expression of EMT genes, <i>SNAI1</i> and <i>SNAI2</i>, which was completely reversed by pretreatment with propranolol (p<0.05). <b>Conclusions:</b> Taken together, these findings demonstrate that NE exposure promotes FT cell spheroid formation and EMT gene transcription through activation of ADRB-mediated signaling. Thus, NE may potentiate metastasis of early ovarian cancer precursor lesions through stimulation of EMT genes and enhancing anoikis resistance, two factors that are critical to the coelomic spread of ovarian cancer. Future studies examining NE-induced carcinogenesis in FT cells are warranted.

Effect of cyclic aging on mode I fracture energy of dissimilar metal/composite DCB adhesive joints

In some industrial applications, adhesive joints are cyclically exposed to a moist environment, where cyclic moisture absorption and desorption can significantly alter the fracture energy of the bonded joints. Most previous studies are based on monotonic aging conditions, while the performance of bonded joints under cyclic aging is not well explored The aim of the current study is to investigate the effect of cyclic aging on mode I fracture energy of dissimilar DCB (double cantilever beam) adhesive joints. Accordingly, bulk adhesive plates were manufactured and exposed to 4 aging cycles. After the aging process, at different exposure times, the aged adhesive plates were used to bond dissimilar Al/GFRP substrates using a secondary adhesive. Then the prepared DCBs were tested and subsequently the mode I fracture energy of the adhesive was determined. Meanwhile, using gravimetrical tests and numerical simulation, moisture diffusion of the adhesive layer in different exposure times was analysed. Using experimental and numerical results, the variation of fracture energy as a function of moisture uptake was studied. In addition, glass transition temperature (Tg) and chemical bonding of the aged adhesive were analysed in different aging cycles. The results showed that by increasing the number of aging cycles, the reduction rate of mode I fracture energy between the aging cycles decreases.

Evaluation of different methodologies for primary human dermal fibroblast spheroid formation: automation through 3D bioprinting technology

Cell spheroids have recently emerged as an effective tool to recapitulate native microenvironments of living organisms in an in vitro scenario, increasing the reliability of the results obtained and broadening their applications in regenerative medicine, cancer research, disease modeling and drug screening. In this study the generation of spheroids containing primary human dermal fibroblasts was approached using the two-widely employed methods: hanging-drop and U-shape low adhesion plate (LA-plate). Moreover, extrusion-based three-dimensional (3D) bioprinting was introduced to achieve a standardized and scalable production of cell spheroids, decreasing considerably the possibilities of human error. This was ensured when U-shape LA-plates were used, showing an 85% formation efficiency, increasing up to a 98% when it was automatized using the 3D bioprinting technologies. However, sedimentation effect within the cartridge led to a reduction of 20% in size of the spheroid during the printing process. Hyaluronic acid (HA) was chosen as viscosity enhancer to supplement the bioink and overcome cell sedimentation within the cartridge due to the high viability values exhibited by the cells—around 80%—at the used conditions. Finally, (ANCOVA) of spheroid size over time for different printing conditions stand out HA 0.4% (w/v) 60 kDa as the viscosity-improved bioink that exhibit the highest cell viability and spheroid formation percentages. Besides, not only did it ensure cell spheroid homogeneity over time, reducing cell sedimentation effects, but also wider spheroid diameters over time with less variability, outperforming significantly manual loading.

Interaction of printing speed and temperature settings on the build plate adhesion of 3D-printed polylactide acid parts

ABSTRACT Fused filament fabrication, also called material extrusion, is a widespread used 3D-printing process which allows to manufacture parts with a high freedom of design at low costs. A common issue is the deformation of parts during the printing process caused by thermal stress and an insufficient adhesion. Despite the widespread usage of material extrusion only few publications dealing with this topic and it is yet nearly unknown, which process parameters have an influence onto the part adhesion nor how they interact among each other. This publication investigates how the adhesion between part and build surface is affected by the most important process parameters. The influence of the first layer’s height, printing speed, nozzle temperature, and build surface temperature was determined for three different build surface materials. Furthermore, the interactions of these parameters among each other were investigated. It was found that the first layer’s height has a strong influence onto the adhesion, which can be increased up to 0.56 MPa. Also, a strong dependence of printing speed, build surface temperature and nozzle temperature among each other was found.

Abstract 374: Small intestinal organoids to study the effects of tight junction modulators that may improve the absorption of orally-administered, anti-neoplastic drugs

Abstract Although oral dosing is the most convenient route of drug administration, most anti-neoplastic drugs are administered intravenously due to poor bioavailability because of the epithelial barriers. A major organ involved in oral drug absorption is the small intestine where enterocytes are linked together by tight junctions (TJs). TJs regulate paracellular transport by creating a selectively-permeable barrier to small molecules, ions and water, thereby restricting the uptake of drugs. Transient modulation of the TJs is considered a potential strategy to improve drug delivery. Human small intestinal (SI) organoids are 3D in vitro models of the epithelium which recapitulate the structure and function of the small intestine. However, their geometry prevents access to the apical compartment of the epithelium (the outer, or luminal, surface being located within the organoid with the basal surface on the outside), making them unsuitable for the study of orally-administered drugs. Here we describe methods to generate transwell cell monolayers and inverted or ‘apical-out’ organoids, both derived from normal human small intestine, that allow direct access to the apical surface. We demonstrate that both formats provide an in vivo-like cell composition, comprising of all the intestinal cellular types, including stem cells, Paneth cells, goblet cells, enteroendocrine cells and enterocytes, and respond to treatment with ‘tool drug’ TJ modulators such as sodium caprate, chitosan, EGTA and various cytokines. Briefly, for the generation of transwell-grown cell monolayers, SI organoids were dissociated into single cells and plated in matrix-coated transwell inserts. The monolayer formation was followed by live imaging and Trans-Epithelial Electrical Resistance (TEER) measurements. Treatments with TJ modulators were applied after TEER reached a plateau and effects on TJs were evaluated by TEER reduction, FITC-Dextran permeability increase and analysis of TJ proteins by immunofluorescence. For the generation of apical-out organoids, SI organoids were mechanically broken into fragments and seeded in low adhesion plates until the organoids switched polarity. Treatments with TJ modulators were applied and effects on TJs were evaluated by immunofluorescence. In conclusion, our models provide an innovative in vitro solution to study the effect of TJ modulators on increasing paracellular transport of orally-administered, anti-neoplastic drugs. Furthermore, as anti-neoplastic drugs can also have toxic effects on the intestinal epithelium by affecting TJs, thereby creating a leaky barrier, the models are useful to predict such effects and stratify drug candidates prior to in vivo studies. Citation Format: Valentina Ubertini, Stephanie French, James Wilson, Frida Ponthan. Small intestinal organoids to study the effects of tight junction modulators that may improve the absorption of orally-administered, anti-neoplastic drugs [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 374.

Connective tissue growth factor expression maintains the epithelial phenotype of ovarian cancer in early epithelial to mesenchymal transition.

e17544 Background: Epithelial to Mesenchymal Transition (EMT) is essential to the process of metastasis in ovarian cancer (OC). Early expressions of proteins and pathways during EMT essential for understanding mechanism and potential therapeutic targets in OC. The aim of our study is to evaluate the role and mechanism of Connective Tissue Growth Factor (CTGF) in OC. Methods: R182 and R2615 are well-characterized epithelial OC cell lines. CTGF expression +/- Transforming Growth Factor -b (TGF-b) was determined via Western blot. R182 and R2615 CTGF knock out (KO) were derived utilizing a Cas9/CRISPR-Cas9 lentivirus plasmid vector and confirmed via PCR. Anoikis resistance and invasion assays were performed to characterize phenotypes of R182 and R2615 wild type (WT) and KO cells. For anoikis resistance, cells were plated in triplicate in an ultra-low adhesive (ULA) cell plate Promega CellTiter assay which measured cell viability quantified by absorbance at 450 nm at 0, 24, 48, and 72 hrs. For invasion assay, 3000 cells were suspended in 50% reduced growth factor Basement Membrane Extract. Human recombinant CTGF was added at 50 and 100 ng/mL concentrations. Cells were plated in tissue culture plate and placed in Cytation 5/Biospa and imaged at 4-hour interval for up to 6 days. Western blot evaluated expression of mesenchymal markers. Chemo-sensitivity was measured by half maximal inhibitory concentration (IC50) for Cisplatin between the WT and KO cells. RNA sequence analysis was performed for both R182 WT and KO cells and analyzed utilizing iPathway Guide. Results: CTGF is constitutively expressed in R182 and R2615 OC cells line as early as 6 hrs of culture and after 30 min of TGF-b treatment. Loss of CTGF promoted anoikis resistance. At 72 hr, R182 CTGF WT cells displayed 75% viability while R182 KO only have 10% viability. Further, we demonstrated that loss of CTGF increases invasion. Administration of exogeneous CTGF in KO cells suppresses invasion in a dose dependent manner demonstrated. Along with epithelial markers CK-18 and beta-catenin, the KO cells expressed the mesenchymal marker SNAIL. No difference in chemo-sensitivity to Cisplatin was seen between WT and KO in either cell line. RNA sequence analysis identified 1106 out of 14054 differentially expressed genes between the WT and KO cells. PI3KAkt, cell adhesion molecules and ECM receptor interaction were pathway that were most significantly regulated. Conclusions: CTGF expression maintains the epithelial phenotype of OC cells during EMT. Loss of CTGF allows anoikis resistance and invasion which are vital characteristics in the metastatic nature of OC. We suggest that loss of CTGF expression in OC cells could be a biomarker for more aggressive OC.

Hinge point emergence in mammalian spinal neurulation

Neurulation is the process in early vertebrate embryonic development during which the neural plate folds to form the neural tube. Spinal neural tube folding in the posterior neuropore changes over time, first showing a median hinge point, then both the median hinge point and dorsolateral hinge points, followed by dorsolateral hinge points only. The biomechanical mechanism of hinge point formation in the mammalian neural tube is poorly understood. Here we employ a mechanical finite element model to study neural tube formation. The computational model mimics the mammalian neural tube using microscopy data from mouse and human embryos. While intrinsic curvature at the neural plate midline has been hypothesized to drive neural tube folding, intrinsic curvature was not sufficient for tube closure in our simulations. We achieved neural tube closure with an alternative model combining mesoderm expansion, nonneural ectoderm expansion, and neural plate adhesion to the notochord. Dorsolateral hinge points emerged in simulations with low mesoderm expansion and zippering. We propose that zippering provides the biomechanical force for dorsolateral hinge point formation in settings where the neural plate lateral sides extend above the mesoderm. Together, these results provide a perspective on the biomechanical and molecular mechanism of mammalian spinal neurulation.

Sponge-like Scaffolds for Colorectal Cancer 3D Models: Substrate-Driven Difference in Micro-Tumors Morphology.

Macroporous scaffolds (cryogels) for the 3D cell culturing of colorectal cancer micro-tumors have been fabricated by cross-linking chitosan and carboxymethyl chitosan (CMC) with 1,4-butandiol diglycidyl ether (BDDGE) under subzero temperature. Due to the different intrinsic properties and reactivity of CMC and chitosan under the same cross-linking conditions, Young′s moduli and swelling of the permeable for HCT 116 cells cryogels varied in the broad range 3–41 kPa and 3500–6000%, respectively. We have demonstrated that the morphology of micro-tumors can be controlled via selection of the polymer for the scaffold fabrication. Although both types of the cryogels had low cytotoxicity and supported fast cell proliferation, round-shaped tightly packed HCT 116 spheroids with an average size of 104 ± 30 µm were formed in CMC cryogels (Young′s moduli 3–6 kPa), while epithelia-like continuous sheets with thickness up to 150 µm grew in chitosan cryogel (Young′s modulus 41 kPa). There was an explicit similarity between HCT 116 micro-tumor morphology in soft (CMC cryogel) or stiff (chitosan cryogel) and in ultra-low attachment or adhesive culture plates, respectively, but cryogels provided the better control of the micro-tumor’s size distribution and the possibility to perform long-term investigations of drug–response, cell–cell and cell–matrix interactions in vitro.

MP59-10 EVALUATING EX VIVO INNATE IMMUNE TRAINING AS A PREDICTIVE MEASURE OF RESPONSIVENESS TO BCG IMMUNOTHERAPY IN PATIENTS WITH NON-MUSCLE INVASIVE BLADDER CANCER

You have accessJournal of UrologyCME1 May 2022MP59-10 EVALUATING EX VIVO INNATE IMMUNE TRAINING AS A PREDICTIVE MEASURE OF RESPONSIVENESS TO BCG IMMUNOTHERAPY IN PATIENTS WITH NON-MUSCLE INVASIVE BLADDER CANCER Song Jiang, Gil Redelman-Sidi, Eugene J. Pietzak, Bernard H. Bochner, and Michael Glickman Song JiangSong Jiang More articles by this author , Gil Redelman-SidiGil Redelman-Sidi More articles by this author , Eugene J. PietzakEugene J. Pietzak More articles by this author , Bernard H. BochnerBernard H. Bochner More articles by this author , and Michael GlickmanMichael Glickman More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000002642.10AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Intravesical instillation of the live attenuated mycobacterium Bacillus Calmette-Guerin (BCG) remains the most effective adjuvant therapy for non-muscle invasive bladder cancer. An increasing body of evidence suggests that the innate immune system has characteristics that involve a heterologous memory of past insults through a process involving epigenetic reprogramming, termed trained immunity. We seek to investigate the potential for innate immune training as a pre-treatment predictor of BCG-response. METHODS: A prospective biospecimen collection was performed on venous blood from non-muscle invasive bladder cancer patients prior to receiving intravesical BCG therapy. To investigate the role of trained immunity in primary monocytes, cells were isolated from peripheral blood mononuclear cells (PBMC) by plate adhesion. In vitro training was performed via stimulation with 10:1 MOI BCG. Cells were then washed in media and allowed to rest 6 days. Subsequently, cells were stimulated with lipopolysaccharide (a TLR4 agonist). Cell free supernatant was measured for TNF-alpha production by enzyme-linked immunosorbent assay. RESULTS: In vitro training of primary monocytes induced an increase in proinflammatory cytokine production upon re-stimulation. This phenotype was dose dependent on stimulation with LPS and confined to the monocyte population. In our in vitro training experiments, there was demonstrable interpatient variability seen pre-BCG exposure in a cohort of NMIBC patients. These results will be correlated with future long term BCG response in this prospective cohort. CONCLUSIONS: We have developed a reliable assay to measure ex vivo innate immune training potential in primary monocytes collected from patient samples. These findings will be correlated with prospective clinical data on BCG responsiveness and recurrence free survival. Such insight will have potential clinical impact on the development of biomarkers and clinical assays that could predict immune responsiveness to BCG a priori, as well as identify possible immunological adjuvants that could enhance the effect of intravesical BCG. Source of Funding: NCI Specialized Programs of Research Excellence (SPORE) in Bladder Cancer P50 CA221745 © 2022 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 207Issue Supplement 5May 2022Page: e1005 Advertisement Copyright & Permissions© 2022 by American Urological Association Education and Research, Inc.MetricsAuthor Information Song Jiang More articles by this author Gil Redelman-Sidi More articles by this author Eugene J. Pietzak More articles by this author Bernard H. Bochner More articles by this author Michael Glickman More articles by this author Expand All Advertisement PDF DownloadLoading ...

Peel-off behavior of a double-plate adhesion system

Peeling and adhesion mechanics of slender structures is of great significance in engineering and science. For a complex adhesion system with soft and slender structures, predicting its deformation can be highly difficult due to the nonlinear nature of the associated governing equations. Additionally, due to the unknown parameters in the highly nonlinear governing equations, it is impossible to solve the governing equations by the standard numerical method directly. In this paper, we perform an elaborate study on the peel-off behavior of two circular adhesive plates in the displacement-controlled cases from the theoretical and numerical perspectives. The governing equations of the system are derived using the energy method based on the finite deformation theory. Since there are many unknown parameters in the governing equations, we adopt an optimization-based approach to determine the peel-off radius as well as the motion parameters at the interface between the peel-off region and the adhesion region. The numerical examples in this work reveal that the peel-off radius becomes larger with the increase of the applied deflection ratio between two plates. We also observe that the infinitesimal deformation theory always underestimates the peel-off radius and this underestimation can lead to as high as 10% error in the predicted peel-off radius. Overall, this work proposes a novel optimization-based method to study peeling mechanics of a double-plate adhesion system and can be generalized to more sophisticated cases. The results of this work can also provide some insights for us to understand the peeling phenomena of an adhesion system.

Defining innate immune training potential as a predictor of Bacillus Calmette-Guérin immunotherapy response in nonmuscle-invasive bladder cancer.

522 Background: Intravesical instillation of the live attenuated mycobacterium Bacillus Calmette-Guerin (BCG) remains the most effective therapy for the treatment of non-muscle invasive bladder cancer. The precise mechanism of BCG is thought to involve phagocytosis, induction of inflammation, activation of the innate immune apparatus, and an effective cell-mediated T cell response. An increasing body of evidence suggests that the innate immune system has characteristics that involve a heterologous memory of past insults through a process thought to involve epigenetic reprogramming, termed trained immunity. We seek to investigate the potential for innate immune training as a predictor of the anti-tumor effects of BCG. Methods: A prospective biospecimen collection was performed on venous blood from non-muscle invasive bladder cancer patients prior to receiving intravesical BCG therapy. To investigate the role of trained immunity in primary monocytes, cells were isolated from peripheral blood mononuclear cells (PBMC) by plate adhesion. 2-5 x 105 cells/well were assayed in 96-well format, cultured in RPMI 1640 supplemented with 10% Fetal Bovine Serum and 2mM L-glutamine and stimulated with 10:1 MOI BCG for 24 hours. Cells were then washed in media and allowed to rest 6 days. Media was changed on day 3 of rest. Subsequently, cells were stimulated with 25 ng/ml lipopolysaccharide (a TLR4 agonist) for 24 hours. Cell free supernatant was measured for TNF-alpha production by enzyme-linked immunosorbent assay. Results: In vitro training of primary monocytes induced an increase in proinflammatory cytokine production upon re-stimulation. This phenotype was dose dependent on stimulation with LPS, however, the relative change between trained and untrained cells was diminished at higher concentrations of agonist. In our in vitro training experiments, there was demonstrable interpatient variability seen pre-BCG exposure in a cohort of NMIBC patients. These results will be correlated with future long term BCG response in this prospective cohort. Conclusions: We present data on a trained innate immune phenotype on primary monocytes collected from patient samples. These data serve as proof of principle for the ability to induce a trained phenotype in vitro. These findings will be correlated with prospective clinical data on BCG responsiveness and recurrence free survival to allow us to garner further insight into the role of trained immunity in BCG immunotherapy. Such insight will have potential clinical impact on the development of biomarkers and clinical assays that could predict immune responsiveness to BCG a priori, as well as identify possible immunological adjuvants that could enhance the effect of intravesical BCG.

Investigation of suitable material and adhesion promoter combinations for fused filament fabrication on flexible silicone build plates

PurposeThe extension of the vacuum-assisted multipoint moulding (VAMM) technology to a broader field of geometries makes it necessary to extend it with attachments to the enhanced vacuum-assisted multipoint moulding with additive attachments (EMMA) technology. Therefore, it is necessary to build additive manufactured attachments on a curved silicone surface by fused filament fabrication (FFF). The main challenge of FFF on a silicone-made build plate is the adhesion of the part on the build plate. Hence, the purpose of this paper is to find suitable and reliably manufacturable material and adhesion promoter combinations for the use of the FFF on silicone build plates.Design/methodology/approachThe combinations of seven different filaments and four adhesion promoters were investigated with an experimental study. Therefore, four different specimen geometries were built with the different combinations and tested in a tensile test, and some of the specimens were analysed with a confocal laser scanning microscope (CLSM).FindingsThis study proves that the FFF on unheated silicone building plates is possible for several material combinations. As a filament material, polylactide can reliably be manufactured with all of the investigated adhesion promoters on the silicone build plate. Thereby, the highest adhesion strengths were achieved with an adhesive foil as an adhesion promoter, whereas the glue stick is the most appropriate solution. The investigations with the CLSM showed that there are large differences in the manifestation of the first layer depending on the adhesion promoter used.Originality/valueTo the best of the authors’ knowledge, this study is the first to demonstrate the manufacturing of FFF-made attachments on silicone build plates for the EMMA process. This paper provides measurement data on the build plate adhesion of the attachments on silicone-made build plates.

Universal Dental Adhesives: Cost-Effectiveness and Duration of Use

The purpose of this study was to conduct a cost-effectiveness analysis (CEA) of different brands of universal dental adhesives used for composite restorations. Four adhesive brands were included: Single-Bond Universal (SB), Tetric N-Bond Universal VivaPen (TN), OptiBond All-In-One (OB), and G-Premio Bond (GP). Adhesives were applied 5 times daily in a standardized class II cavity onto a plastic tooth. A precision-analysis scale was used to measure all of the following parameters before and after use: adhesive bottle, applicator, dosing plate, and plastic tooth. CEA was done by measuring the amount of material utilized/day, waste/day, efficacy, efficiency, average cost-effectiveness ratio (ACER), and incremental cost-effectiveness ratio (ICER). Data were analyzed using Kruskal–Wallis and Dunn’s tests with Bonferroni correction at 0.05 significance level. CEA tested parameters were significantly different between groups (p &lt; 0.001) except for ICER (p = 0.112). GP was the least effective (median = 0.062), and SB was the least efficient (median = 0.366). The highest and lowest ACER values were associated with TN (median cost ≈ USD 317) and SB (median cost ≈ USD 317), respectively. ICER analysis reported an incremental cost for extra material utilized per milliliter of ≈USD 208 for TN, USD 3.8 for GP, and USD −38 for OB, compared to SB. TN seems to be the most efficient and cost-effective dental adhesive.

Populações de insetos-praga: diversidade e similaridade em cultura agrícola

The anthropic actions expressed by conventional-modern agriculture greatly alter the environment, interfering with the balance of natural processes and making its recovery mechanisms precarious. Thus, the interest in the use of microorganisms in agricultural practices has increased significantly in recent years, both in promoting plant growth and in the biological control of plant pests and diseases. The objective of the present work was to identify the insect pest population in the cucumber crop after foliar application of effective microorganisms (EM). The experiment was carried out in a rural family farm located in the city of São Sebastião-AL. Yellow (4) and blue (4) adhesive plates were used in a usable area of ​​128.8 m2. EM was diluted at a concentration of 4.7% and the applied dose was 50 mL/plant. Six collections were carried out, from December 2020 to February 2021. The individuals sampled were screened and identified at the level of class, order, family and when identified as a pest, at the level of gender. A total of 1301 invertebrate individuals of the Insecta and Arachnida classes were collected. Of these, 55.11% were identified as pest insects and 44.8% as visiting arthropods. The taxon that presented the highest relative frequency was the order Diptera (30.6%) followed by the orders Hemiptera (25.4%), Hymenoptera (24%), Coleoptera (13%), Lepidoptera (5.7%), Araneae (0.8%) and Odonata (0.5%). The pests were representatives of the genera Diabrotica spp, Anastrepha spp, Myzus spp, Bemisia spp, Atta spp and Diaphania spp. 24.7% of the visiting arthropods were predators. The arthropod population increase occurred from the 40th day after planting, with growth peak at the 57th day.

Debonding failure analysis of FRP-plated RC beams via an inter-element cohesive fracture approach

Debonding mechanisms in the externally reinforced structural elements have been widely studied by the scientific community since they lead to premature failure in concrete structures. Different numerical models, based on simplified strength approaches, have been proposed in the literature to simulate such debonding phenomena, but often are unable to accurately predict the development of diffuse crack patterns which are typical for reinforced concrete (RC) structures. In this context, the present work aims to propose a cohesive modeling technique, based on a diffuse interface fracture approach, to in-depth investigate the plate-end and mid-span debonding failures in FRP-strengthened RC structural elements. This model, already presented by some of the authors, is used in combination with bond-slip elements, to simulate the stress transfer between the rebars and the surrounding concrete, and with additional cohesive elements along the adhesive/concrete (AC) and adhesive/plate (AP) material interfaces. Numerical simulations have been performed by using the present fracture approach for predicting the load-carrying capacity and the related failure modes of real-scale retrofitted RC elements. The reliability and the effectiveness of the proposed fracture approach have been demonstrated through suitable comparisons with available experimental results. In particular, this model results to be an innovative and versatile numerical tool able to analyze the debonding mechanisms in FRP-strengthened RC elements as well as to provide more accurate crack patterns, including multiple crack branching and coalescence, than commonly used existing models.

High-temperature strength properties of epoxy-based adhesive joints and the effects of hightemperature aging

Lap joints were prepared using epoxy adhesive and aluminum plates, and the properties of high-temperature strength were studied. Tensile test and aging test were performed at high temperature. The effects of test temperature were systematically investigated. The obtained conclusions were following. When the specimen was exposed at temperature of 135℃, the strength decreased significantly. The adhesive changed its chemical structure by oxidation. The relationship between test temperature and tensile strength could be expressed using the Arrhenius equation.

MONITORING AND CONTROL OF THE PEST TUTA ABSOLUTA (MEYRICK) IN TOMATO CROPS UNDER HIGH PLASTIC TUNNELS

Tuta absoluta – tomato leafminer (Meyrick) (Lepidoptera: Gelechiidae) is a major pest of tomato crops in protected areas, which attacks both foliage and fruit, in all phases of plant growth. Its invasion has led to yield and quality decrease, increased costs to control and high dependence on chemical insecticides. With pheromone traps it is possible to monitor and reduce the density of the pest population. The experience was established in 2020, at a farmer in Dobreni village (Giurgiu county). In the first and second crop cycle, the population of the pest Tuta absoluta was monitored, by placing Delta pheromone traps in tomato crops. The Sahmat F1 tomato hybrid was planted in both crop cycles. Pest monitoring was performed weekly, by changing the adhesive plates with pheromones from Delta traps, on the dates: 03.06, 10.06, 17.06, 24.06, 01.07, 08.07, 15.07, 22.07 in the first cycle of crop and on the dates: 29.07, 05.08, 12.08, 19.08, 02.09 , 09.09, 16.09, 23.09 in the second cycle. The appearance and evolution of the attack were recorded in the both crop cycles and its influence on production was established.

Changes in the filtration function of the spleen after surgery following traumatic organ injuries

The incidence of splenic injuries among all closed injuries of the abdominal cavity is from 15.5 to 30.0 %, and a mortality rate is between 7 % and 26 %. The tactics in the treatment of splenic injuries is determined by the degree of traumatic injury, the patient's condition during the operation, and a concomitant pathology. Studies of tissue regeneration of the operated spleen allow identifying two main options for its regeneration, associated with the nature of blood circulation in the preserved part. Objective — to study the regeneration and filtration function of the spleen after various types of surgery for the management of traumatic spleen injuries. Materials and methods. The results of treatment of 85 patients in the period 2015 — 2020 were analyzed. To investigate the changes in the filtration function of the spleen, a study of the peripheral blood (general blood test) was performed to assess the shape of erythrocytes. In order to determine the size of the residual splenic parenchyma, its structure and regenerative processes were evaluated and sonographic examination was carried out. Results. The average increase in the size of the residual splenic tissue after subtotal resection of the organ with the formation of couplings of the parenchyma averages 40.2 ± 3.4 % one year after surgery, and in patients who underwent subtotal resection of the spleen with covering the cut plane with adhesive hemostatic plate 70 % — 49.14 ± 6.77 %. The study of changes in the filtration function of the spleen in patients, who underwent subtotal resection of the spleen, showed the appearance of target cells, acanthocytes and halocytes in the peripheral blood, but their number was insignificant and did not exceed normal (not more than 3 %). Any destroyed and pathologically altered erythrocytes were not visualized in patients, who underwent atypical resection of the spleen (mass deficit less than 30 %). Conclusions. Spleen regeneration is determined both by the nature of blood supply to the residual splenic tissue and its size, and does not depend on the nature of the pathological process. Organ‑preserving surgical interventions on the spleen allow maintaining the filtration function of the spleen.