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Biomarker panels must remain significantly associated with the outcomes across patient cohorts and add to pre-existing nomograms. We validated the prognostic ability of both IDC/C and CAPRA-S for BCR and time to BCR in over 600 patients (3 cohorts). CAPRA-S and IDC/C was significant in recurrence-free survival and an independent predictor of BCR. Background: Intraductal carcinoma and cr ibr iform (IDC/C) tumor features are well-established prognosticators of biochemical recurrence (BCR), metastasis, and prostate cancer (PCa)-specific mortality. However, approximately 70% of PCa patients undergoing a radical prostatectomy are IDC/C negative, yet up-to 20% of these patients progress and experience BCR. Thus, tumor histopathologic characteristics such as IDC/C alone are limited in their ability to predict disease progression. Conversely, several nomograms such as Cancer of the Prostate Risk Assessment-Surgery (CAPRA-S) have been developed to aid in the prognostication of BCR, but not yet widely applied in clinical settings. Materials and methods: In this study, we assessed the combined prognostic utility of IDC/C, and CAPRA-S for BCR in 3 PCa patient cohorts. Results: CAPRA-S + IDC/C improved the predictive accuracy of BCR in all 3 cohorts (P < .001). Specifically, among IDC/C negative cases, CAPRA-S improved the prognostication of BCR in low-risk (Cohort 1; P < .001, Cohort 2; P < .001, Cohort 3; P = .003), intermediate (Cohort 1; P < .001, Cohort 2; P = .006, Cohort 3; P = .03) and high-risk (Cohort 1-3; P < .001) patients. Conversely, IDC/C improved the prognostication of BCR among CAPRAS low-risk (Cohorts 1; P < .001 and Cohort 3; P = .003) patients. Conclusion: Our results suggest the investigation of histopathological IDC/C features in CAPRA-S low-risk patients and conversely, nomogram CAPRA-S among IDC/C negative patients improves the identification of patients likely to experience BCR, which would otherwise be missed through current assessment regimens. These patients can be offered more intensive monitoring and adjuvant therapies upfront to circumvent the development of recurrent cancer or overtreatment at the time of surgery. [ABSTRACT FROM AUTHOR]

Five monoclonal antibodies, mAb7.3, mAb29.3, mAb46.3, mAb12.3 and mAb36.3, raised to the LcrV virulence factor from Yersinia pestis were characterised for their Fab affinity against the purified protein and their Fc affinity to Protein A/G as a proxy for the FcγR receptor. Kinetic measurements were performed label-free in a localised particle plasmon array reader. The Fc-ProteinA/G complex first-order half-life was determined for each antibody and fell in the range of 0.8–3.8 h. The Fab first-order half-lives had ranged from 3.4 to 9.2 h although two antibodies, mAb12.3 and mAb36.3, showed low affinity interactions. Competitive binding studies of mixtures of the Fab-active antibodies were performed to measure the relative binding efficiency of one antibody in the presence of the other. A geometric relative positioning of the epitopes of mAb7.3, mAb29.3 and mAb46.3 was determined based on the footprint locus of the antibody and the percentage of competitive binding. The two known protective antibodies mAb7.3 and mAb29.3 showed greater interference, indicating epitopes close to one another compared to the non-protective mAb46.3 antibody. The Fab-Fc complex half-life screen and epitope mapping are potentially useful tools in the screening of therapeutic antibodies or vaccine candidates. [ABSTRACT FROM AUTHOR]

Butadiene rubbers (BRs) differing in isomeric content and cured with different amounts of dicumyl peroxide (DCP) have, according to TMA investigations, regions with two glass transition temperatures (low and high) and different thermal expansion properties. BR network has chemical and cluster type (physical localised) branching junctions. For rubbers of the same isomeric content, shares of such regions depend on concentration of DCP. This fact suggests cis–trans isomerization at curing, which increases the content of a high-temperature region with predominantly chemical branching junctions, because cluster type branching junctions start to relax at Tg″.It is supposed that an essential change in Tg, if DCP concentration in the rubber compound is varied, is determined by the different molecular mobility of the chains included in associated structures of cluster junctions of a network of cured rubber. Such mobility also increases if 1,2 (vinyl) isomer is present in the BR chain. The latter is probably distributed uniformly between the chains of both regions. In a low-temperature region it could act as an antiplasticizer, whereas in a high-temperature region as a plasticizer.The molecular weight distribution of the chains between the junctions of a low-temperature region is virtually bimodal for low DCP concentration in rubber compounds. This could result from some zones with different crosslinking degree within the network. It is believed that this region has branching junctions, which are of a low crosslink density of chemical types and a high crosslinking density of cluster type junctions.A wide spectrum of isomeric content in neat and cured BR when a medium content of DCP was used meant that a low-temperature region had unimodal and symmetric MWD between the junctions. In a high-temperature region the exact opposite phenomenon may be observed — separation of chain segment length giving a bimodal MWD. A possible reason for this is non-uniform distribution of 1,2 (vinyl) isomer within the chains of a high-temperature region. The shift in MWD between the junctions from bimodal to unimodal as DCP content is raised, which was identified using TMA, confirms our 2H NMR findings. The two splits in the 2H NMR spectra observed at lower DCP contents could result from regions with different crosslink density, as we have argued in a recently published paper. All these conclusions confirm the applicability of the adopted TMA for testing a complex structure of rubbers. [Copyright &y& Elsevier]

By using thermomechanical analysis (TMA) multiple relaxation transitions in the amorphous part of semi-crystalline polymers and their blends can be found. These result from differences in the interaction energies between segments of macromolecules, and as a result, in molecular mobility. TMA shows essential changes in the structure of low-density polyethylene (LDPE) resulting from the grafting of a maleic anhydride (LDPE-g-MAH) onto this semi-crystalline polymer. The grafting process did not suppress the ability of polyethylene to crystallize. Essential changes were found in molecular weight distribution and relaxation transitions of the ternary blends LDPE/PA6 (polyamide 6, PA6)/LDPE-g-MAH studied. For a concentration of PA6 up to 30 wt.%, a single relaxation transition is visible, which testifies that the components are miscible in the amorphous region. For blends with 40 and 50 wt.% of PA6, this structure was transformed and two relaxation transitions are visible. From the results obtained in this study it is concluded that the introduction 5 wt.% of grafted polyethylene is sufficient to produce a PA6/LDPE blend only for PA6 concentration up to 30 wt.% which is homogenous on “molecular” level. The transformation of the structure of the ternary polymer blend was explained by the interaction of the components during the melt mixing and changes in the structure of its amorphous regions. [Copyright &y& Elsevier]

Immune checkpoint inhibitors (ICI) therapies have demonstrated significant benefit in the treatment of many tumors including high grade urothelial cancer (HGUC) of the bladder. However, variability in patients' clinical responses highlights the need for biomarkers to aid patient stratification. ICI relies on an intact host immune response. In this context, we hypothesize that key players in the antitumor immune response such as markers of activated cytotoxic T lymphocytes (CD8, granzyme-B) and immune suppression (FOXP3) may help to identify patients who will derive the greatest therapeutic benefit from ICI. A major obstacle for deployment of such a strategy is the limited quantities of tumor-derived biopsy material. Therefore, in this technical study, we develop a multiplex biomarker with digital workflow. We explored the (1) concordance of conventional single stain results using digital image analysis, and (2) agreement between digital scoring versus manual analysis. (1) For concordance study of single and multiplex stains, triplicate core tissue microarrays of 207 muscle invasive, HGUC of bladder had sequential 4-micron sections cut and stained with CD8, FOXP3 and granzyme-B. An inhouse developed tri-chromogen multiplex immunohistochemistry (m-IHC) assay consisting of CD8 (green), granzyme B (brown), and FOXP3 (red) was used to stain the next sequential tissue section. (2) Agreement between manual and digital analysis was performed on 19 whole slide sections of HGUC cystectomy specimens. All slides were scanned using Aperio ScanScope AT Digital Scanner at 40X. Quantitative digital image analysis was performed using QuPath version 0.2.3 open-source software. Scores from triplicate cores were averaged for each HGUC specimen for each marker. Intraclass correlation coefficients were used to compare percent positive cells between the single- and multi-plex assays. Lin's concordance correlation coefficients were used for manual versus digital analysis. m-IHC offers significant advantages in characterizing the host immune microenvironment particularly in limited biopsy tissue material. Utilizing a digital image workflow resulted in significant concordance between m-IHC and individual single stains (p < 0.001 for all assessments). Moderate to good agreements were achieved between manual and digital scoring. Our technical work demonstrated potential uses of multiplex marker in assessing the host immune status and could be used in conjunction with PD-L1 as a predictor of response to ICI therapy. • High grade urothelial carcinoma has variable immune checkpoint inhibitors response. • PD-L1 and markers of immune activity may be useful predictors of treatment response. • Multiplex immunohistochemistry can be used to characterize tumor microenvironment. • Digital image analysis can be used effectively for quantifying multiplex stains. • Concordance was achieved between single stains and manual counts. [ABSTRACT FROM AUTHOR]

The aim of this pilot study was to assess the role of immune markers in fast relapse (<2 years) of high-grade muscle invasive urothelial carcinomas of the bladder (HGUC) treated by cystectomy. A series of 40 such cases was investigated for immune protein (CD3, CD4, CD8, CD20, CD68, CD163, FOXP3 and PD-1) status by immunohistochemistry. Decreased expression of all immune cell markers was observed in tumors of patients who relapsed quickly. In Kaplan-Meier (log-rank test) analysis, low CD3, CD4 and CD8 expression was associated with fast relapse (P = 0.005, 0.028, 0.036 respectively). Additional evaluation of the immune transcriptome by NanoString Human PanCancer Immune Panel v.1.1 has identified 5 differentially expressed genes significantly associated with fast relapse. Among these, KLRB1 and HLA-DQA1 were also significant on Kaplan-Meier analysis (log-rank test P = 0.007 and 0.006, respectively). These findings strengthen the potential clinical utility and, hence, the need for further evaluation of immune markers in HGUC prognostication. [ABSTRACT FROM AUTHOR]

Iordanskii et al. BioPoCo (2018). • Morphology and diffusion characteristics of PHB-PLA electrospun fibers were determined. • For different fibre geometry affected by drug content, diffusion determines release profiles. • Water absorption, segmental dynamics and diffusivity are basic processes for drug delivery. An innovative remedies' implementation with a focus on the transition from custom macro- to nano-scaled therapeutic polymer vehicles relies upon the appropriate design of novel drug-polymer compositions, their functionalization at molecular and nano-scale levels, and addressed diffusion-kinetic processes that control drug distribution at any given time in both a proper polymer implant and surrounding biological media. In the framework of this paradigm, ultrathin fibers formed by blending poly(3-R-hydroxybutyrate) [PHB] with poly (l -lactic acid) [PLA] and loaded with modeling drug (dipyridamole [DPD]) were fabricated by solution electrospinning. PHB/PLA ratio and DPD content affected considerably the geometric shape, crystallinity of a single filament. As follows from the optical microscopy and SEM data, in the absence of the drug or at its low concentration (<3 wt.%), the PHB/PLA fibers appear as ellipsoid-like fragments alternating with cylindrical ones. At the higher content of the drug (3–5%), the abnormal ellipsoid-like structures disappear and all the fibers have cylindrical geometry. By the DSC technique it was shown that PHB/PLA ratio influenced on the crystallinity of fibers and modulated the general course of DSC thermograms. The ESR spectra of the TEMPO radical preliminary embedded into fiber volume represent a superposition of two elementary spectra belonging to the radical populations with different correlation times τ 1 and τ 2 that reflects heterogeneity of the PHB and PLA intercrystalline amorphous regions with slow and fast segmental dynamics respectively. The combination of structure-morphology characteristics of the ultrathin fibers determines the polymer dynamics and drug diffusivity as well as the corresponding profiles of drug release. A coherent model of drug release from the PHB fibrillar and ellipsoidal mats was advanced. [ABSTRACT FROM AUTHOR]

The effects of the carbon backbone chain length on the EPR spectra of linear perfluoro-n-alkanes (PFAs) γ-irradiated at 77 K was studied for the short chain n-C6F14, n-C8F16, n-C12F26, and n-C16F34 molecules as well as the polymer polytetrafluoroethylene (PTFE). The experimental data show that the processes occurring during radiolysis of perfluoro-n-alkanes and polytetrafluoroethylene are very similar. EPR spectra of irradiated perfluoro-n-alkanes at low radiation dose show superimposed signals from three radicals: –F2CC᛫FCF2–, –CF2C᛫F2 and F3C᛫. The signal intensity decreases with perfluoro-n-alkanes chain length. At doses above 2.0 MGy, a constant increase in concentration of the radicals –F2CC᛫FCF2– and –CF2C᛫F2 is observed with decreasing chain length. The concentration of these radicals formed during radiolysis of PFA is described by the ratio: [–CF2C᛫F2]/[–F2CC᛫FCF2–] ≈3/(n − 2), where n is the number of carbon atoms in the linear perfluoroalkanes. Density functional theory was used to calculate the structures of the radicals and C–F bond energies in model perfluoro-n-alkanes as well as the EPR spectra of the associated radicals. This data is used to provide further insight into the radiation stability of PTFE. Four topographical structures of polytetrafluoroethylene, one amorphous and three crystalline, were identified by thermomechanical analysis. In the crystal phase, γ-irradiation results in their transformation to the amorphous form. The helical structure of individual perfluroalkanes readily distorts on removal of a fluorine and this will have an impact on the overall structure of the material. Such structural reorganization can lead to loss of the mechanical stability of polytetrafluoroethylene. [Copyright &y& Elsevier]

Abstract: The structure of the plate specimens obtained from the molten PA6 that was cooled at rates between 2 and 2000°C/min have been studied. The cooling rate of 2000°C/min did not ensure a complete amorphization of the specimens. The amorphous phase created by supercooling is unstable and at room temperature undergoes a noticeable cold crystallization. The access of water to the specimen from the surrounding air accelerates this process. Variations in the cooling rate of the melt reflect in rearrangement of the amorphous phase of PA6. Owing to the effect of interaction with the crystalline phase, the amorphous regions undergo changes in molecular weight and MWD of the chain segments between junctions in topological regions, temperatures of the glass transition and β-relaxation, compaction, etc. [Copyright &y& Elsevier]

Mid-infrared photodissociation spectra of NH3+–Arn (n=1–6) complexes in the electronic ground state have been recorded in the vicinity of the N–H stretch vibrations of the ammonia cation. The rovibrational analysis of the transitions in the spectrum of the NH3+–Ar dimer (n=1) is consistent with a planar, proton-bound equilibrium structure with C2v symmetry. The three N–H stretching fundamentals occur at ν1(a1)=3177.4±1 cm−1, ν3(a1)=3336.0±1 cm−1, and ν3(b2)=3396.26±0.13 cm−1, and the combination band of ν1 with the intermolecular stretching vibration is observed at ν1+νs(a1)=3305.5±2 cm−1. The relatively long lifetime with respect to predissociation (τ>250 ps) and modest complexation-induced frequency shifts (|Δν1,3|<70 cm−1) of the N–H stretch fundamentals imply weak coupling between the intramolecular and intermolecular degrees of freedom. The linear intermolecular proton bond in the ground vibrational state is characterized by an interatomic H–Ar separation of 2.27 A˚ and a harmonic stretching force constant of &ap;12 N/m. Observed tunneling splittings in the ν3(b2) band are attributed to hindered internal rotation through potential barriers separating the three equivalent H-bound global minima. By comparison with theoretical data, the frequency of the infrared forbidden ν1 fundamental of free NH3+ is estimated from the NH3+–Ar spectrum as 3234±15 cm−1, the currently most accurate value based upon experimental measurements. The vibrational spectra of the larger NH3+–Arn complexes (n=2–6) display distinct frequency shifts and splittings of the N–H stretching vibrations as a function of cluster size. The spectra are consistent with cluster geometries in which the first three Ar ligands fill a primary solvation subshell by forming equivalent intermolecular proton bonds (n=1–3) leading to planar structures with either C2v or D3h symmetry. The next two Ar ligands fill a second subshell by forming equivalent p-bonds to the two lobes of the 2pz orbital of the central N atom leading to cluster structures with C3v (n=4) and D3h symmetry (n=5). The first Ar solvation shell around the interior NH3+ ion is closed at n=5 and the 6th Ar ligand occupies a position in the second solvation shell. The dissociation energies of the H-bonds and p-bonds are estimated from photofragmentation branching ratios as D0(H)&ap;950±150 cm−1 and D0(p)&ap;800±300 cm−1, respectively. In general, the intermolecular H-bonds significantly weaken the intramolecular N–H bonds, whereas the p-bonds slightly strengthen them. Properties of the intermolecular bonds and the cluster growth in NH3+–Arn are compared to related AHk+–Arn cluster systems. [Copyright &y& Elsevier]

Background: Highly sensitive and specific urinary biomarkers for the early detection of bladder cancer (BC) to improve the performance of urinary cytology are needed.Objective: To investigate the usefulness of methylation markers in voided urine to identify BC presence and grade.Design, Settings, and Participants: Using genome-wide methylation strategies in Toronto, Canada and Liège, Belgium, we have identified differentially methylated genes (TWIST1, RUNX3, GATA4, NID2, and FOXE1) in low-grade vs. high-grade BC tissue and urine. We accrued urine samples from 313 patients using a 2:1 ratio in a case-control setting from Toronto, Canada, Halifax, Canada, and Zurich, Switzerland. We studied the usefulness of these 5 methylated genes to identify BC and discriminate cancer grade in voided urine specimens. Urinary cell sediment DNA was evaluated using qPCR-based MethyLight assay. Multivariable logistic regression prediction models were created.Results and Limitations: We included 211 BC patients (180 nonmuscle invasive) and 102 controls. In univariate analyses, all methylated genes significantly predicted BC vs. no BC, and high grade vs. low grade (all P < 0.05). In multivariable analysis, NID2, TWIST1, and age were independent predictors of BC (all P < 0.05). Sensitivity of NID2 and TWIST1 to predict BC and BC grade was 76.2% and 77.6%, respectively, whereas specificity was 83.3% and 61.1%, respectively. Multivariable models predicting BC overall and discriminating between high-grade and low-grade BC reached area under the receiver operating characteristics curves of 0.89 and 0.78, respectively.Conclusions: This multi-centric study in a real life scenario (different countries, techniques, and pathologists) supports the promise of epigenetic urinary markers in noninvasively detecting BC. With sensitivities and specificities in the range of 80%, the overall performance characteristics of this panel of methylated genes probably does not allow such signature to significantly alter clinical care at this stage but is worth further studying for instance in BC surveillance or screening in high-risk populations. [ABSTRACT FROM AUTHOR]

Purpose: Patients with clinically localized, high-risk prostate cancer are often treated with surgery, but exhibit variable prognosis requiring long-term monitoring. An ongoing challenge for such patients is developing optimal strategies and biomarkers capable of differentiating between men at risk of early recurrence (<3 years) that will benefit from adjuvant therapies and men at risk of late recurrence (>5 years) who will benefit from long-term monitoring and/or salvage therapies.Patients and Methods: DNA methylation changes for 12 genes associated with disease progression were analyzed in 453 prostate tumors. A 4-gene prognostic model (4-G model) for biochemical recurrence (BCR) was derived utilizing LASSO from Cohort 1 (n = 254) and validated in Cohort 2 (n = 199). Subsequently, the 4-G model was evaluated for its association with salvage radiotherapy (RT) and/or hormone therapy, and the additive potential to CAPRA-S to develop an integrative gene model was assessed.Results: The 4-G model was significantly associated with BCR in both cohorts (chi-squared analysis P≤ 0.004) and specifically, with late recurrence at 5+ years (P < 0.001, Cohort 1; P= 0.028, Cohort 2). Multivariable Cox proportional regression analysis identified the 4-G model as significantly associated with salvage RT or hormone therapy in Cohort 1 (hazard ratio (HR) 1.64, 95% confidence interval (CI) 1.29-2.10, P< 0.001) and further validated in Cohort 2 (HR 1.63, 95% CI 1.18-2.25, P< 0.001). The integrative model outperformed prostate-specific antigen and the 4-G model alone for predicting BCR and was associated with patients who received hormone therapy 3+ years postsurgery.Conclusions: We have identified and validated a novel integrative gene model as an independent prognosticator of BCR and demonstrated its association with late BCR. These patients require more long-term postsurgical monitoring and could be spared the comorbidities of adjuvant therapies. [ABSTRACT FROM AUTHOR]

Renewable biodegradable films of poly(3-hydroxybutyrate) (PHB) were produced via thermo-compression and casting method (specimens 1 and 2). On specimens 1, gas permeability coefficients with CO 2 , O 2 , N 2 , and C 2 H 4 gases were measured at 277 K, 288 K and 296 K. The physicochemical characteristics of PHB based on calorimetric, mechanical, and thermal examination were performed for both specimens. SEM and probe ESR technique showed the dramatic differences in their surface morphology and segmental motilities in polymer bulk. In the selected temperature range, the gas permeability ranking (P i ) for the thermo-compressed films is as follows: P(CO 2 ) > P(O 2 ) > P(N 2 ) ≈ P(C 2 H 4 ). The gas diffusion coefficients were considered to be dependent on the gas critical volume, in accordance with the semi-logarithmic function, consistent with a free volume model. The activation energies of permeability, diffusivity and spin probe rotation, as well as the apparent heat of sorption equilibrium, forecast the values of PHB barrier characteristics in the range of foodstuffs stored temperatures. [ABSTRACT FROM AUTHOR]

Abstract: K95 experimental lubricant being a product of fluorine reaction with a blend of mutually soluble poly(fluorine alcohols) and poly(fluorine esters) with molecular weight 240–900g/mol was studied as an additive for rubber compounds based on blend of NR and BR. It was thermally stable till 270–300°C. For comparison, it was tested simultaneously with homogenizing agent, Struktol MS40. Lubricant K95 added in a quantity of 0.5wt% reduced the viscosity of rubber compound; it also improved compound flow in the mold. Mechanical properties of cured rubber not decrease while resistance to abrasion and fatigue increased. K95 participated in forming strong physical junctions (lower molecular weight between junctions of thermally stable network) while Struktol MS40 reduced the networking degree of rubber. As a result, it acts a multifunctional additive for NR/BR rubber. [Copyright &y& Elsevier]

Purpose: Prostate cancer (CaP) patients with low-grade tumors are enrolled in active surveillance (AS) programs and monitored with digital rectal exams (DREs), prostate-specific antigen (PSA) tests, and periodic invasive biopsies. Patients are "reclassified" with higher-risk disease if they show signs of disease progression. However, AS patients who will reclassify cannot be easily identified upfront and suffer morbidities associated with biopsy. Biomarkers derived from noninvasively obtained specimens such as serum or urine samples are promising alternatives to monitor patients with clinically insignificant cancer. Previously, we have characterized and validated a urinary DNA methylation panel and a serum miRNA panel for the prediction of patient reclassification in 2 independent AS cohorts. In this exploratory study, we have investigated cell-free miRNAs in the urinary supernatant combined with urinary DNA methylation markers to form an integrative panel for prediction of AS patient reclassification.Methods: Post-DRE urine was collected from 103 CaP patients on active surveillance. Urinary sediment DNA methylation levels of selected genes were previously analyzed using qPCR-based MethyLight assay. Using qRT-PCR, we analyzed the urinary supernatants for relative quantities of 10 miRNAs previously shown to be associated with AS reclassification. Logistic regression and Receiver Operating Characteristics curve analyses were performed to assess the predictive ability of miRNAs and DNA methylation biomarkers.Results: We identified a 3-marker panel, consisting of miR-24, miR-30c and CRIP3 methylation, that was significant for prediction of patient reclassification (Odds ratio = 2.166, 95% confidence interval = 1.22-3.847) with a negative predictive value of 90.9%. Our 3-marker panel also demonstrated additive value to PSA for prediction of patient reclassification (c-statistic = 0.717, ROC bootstrapped 1000 iteration P = 0.041).Conclusion: A urinary integrated panel of methylation and miRNA markers is a promising approach to identify AS patients at risk for reclassification. Our 3-marker panel, with its high negative predictive value, would be beneficial to identify and preclude AS patients with truly indolent cancer and to personalize monitoring strategies for AS patients. [ABSTRACT FROM AUTHOR]