Biofilm formation under high shear stress increases resilience to chemical and mechanical challenges.

The effect that the hydrodynamic conditions under which biofilms are formed has on their persistence is still unknown. This study assessed the behaviour of Pseudomonas fluorescens biofilms, formed on stainless steel under different shear stress (τw) conditions (1, 2 and 4 Pa), to chemical (benzalkonium chloride - BAC, glutaraldehyde - GLUT and sodium hypochlorite - SHC) and mechanical (20 Pa) treatments (alone and combined). The biofilms formed under different τw  showed different structural characteristics. Those formed under a higher τw were invariably more tolerant to chemical and mechanical stresses. SHC was the biocide which caused the highest biofilm killing and removal, followed by BAC. The sequential exposure to biocides and mechanical stress was found to be insufficient for effective biofilm control. A basal layer containing biofilm cells mostly in a viable state remained on the surface of the cylinders, particularly for the 2 and 4 Pa-generated biofilms.

Accelerometer-based prediction of skeletal mechanical loading during walking in normal weight to severely obese subjects.

There is no objective way to monitor mechanical loading characteristics during exercise for bone health improvement. We developed accelerometry-based equations to predict ground reaction force (GRF) and loading rate (LR) in normal weight to severely obese subjects. Equations developed had a high and moderate accuracy for GRF and LR prediction, respectively, thereby representing an accessible way to determine mechanical loading characteristics in clinical settings. INTRODUCTION: There is no way to objectively prescribe and monitor exercise for bone health improvement in obese patients based on mechanical loading characteristics. We aimed to develop accelerometry-based equations to predict peak ground reaction forces (pGRFs) and peak loading rate (pLR) on normal weight to severely obese subjects. METHODS: Sixty-four subjects (45 females; 84.6 ± 21.7 kg) walked at different speeds (2-6 km·h-1) on a force plate-equipped treadmill while wearing accelerometers at lower back and hip. Regression equations were developed to predict pGRF and pLR from accelerometry data. Leave-one-out cross-validation was used to calculate prediction accuracy and Bland-Altman plots. Actual and predicted values at different speeds were compared by repeated measures ANOVA. RESULTS: Body mass and peak acceleration were included for pGRF prediction and body mass and peak acceleration transient rate for pLR prediction. All pGRF equation coefficients of determination were above 0.89, a good agreement between actual and predicted pGRFs, with a mean absolute percent error (MAPE) below 6.7%. No significant differences were observed between actual and predicted pGRFs at each walking speed. Accuracy indices from our equations were better than previously developed equations for normal weight subjects, namely a MAPE approximately 3 times smaller. All pLR prediction equations presented a lower accuracy compared to those developed to predict pGRF. CONCLUSION: Walking pGRF and pLR in normal weight to severely obese subjects can be predicted with moderate to high accuracy by accelerometry-based equations, representing an easy and accessible way to determine mechanical loading characteristics in clinical settings.

5'UTR +24T>C CR2 is not associated with nasopharyngeal carcinoma development in the North Region of Portugal.

OBJECTIVE: We have analysed the association of the +24T>C polymorphism (rs3813946) in CR2, the cellular receptor for Epstein-Barr virus (EBV), in the susceptibility for the development of nasopharyngeal carcinoma (NPC). METHODS: A retrospective case-control study was developed with peripheral blood samples from 111 individuals with NPC and 608 healthy individuals (controls) from the North region of Portugal. The genotyping analysis was performed by allelic discrimination real-time PCR using a TaqMan(®) SNP Genotyping Assay. RESULTS: The genotype distribution was 62.2% TT, 34.2% TC and 3.6% CC for NPC patients; and 65.0%, 30.6% and 4.4%, respectively, for controls. Our study showed no statistical association between the genotype distribution in controls and all types of NPC (P = 0.717); nevertheless, the analysis showed statistically significant differences (P = 0.038) regarding cases with well- or moderately differentiated types of NPC suggesting that +24CC/CT genotypes are associated with increased risk (OR = 4.16; 95% CI 1.28-15.7; P = 0.016). CONCLUSIONS: This is the first study in Western populations to characterize the association of the CR2 +24T>C polymorphism in NPC development, and our results suggest that more studies are required to clarify the impact on NPC susceptibility in different populations.

JC polyomavirus nephropathy confirmed by using an in-house polymerase chain reaction method.

We report the case of an isolated JC virus (JCV) infection, without co-infection by polyoma BK virus (BKV), associated with nephropathy 4 years after kidney transplantation. Clinical suspicion followed the observation of a decrease in estimated glomerular filtration rate (eGFR) and a renal allograft biopsy revealing polyomavirus-associated tubulointerstitial nephritis and positivity for SV40. An in-house real-time polymerase chain reaction assay, targeting the presence of JCV and the absence of BKV in biopsy tissue, confirmed diagnosis. Thirteen months after diagnosis, and following therapeutic measures, eGFR remains stable.

Derepression of a baker's yeast strain for maltose utilization is associated with severe deregulation of HXT gene expression.

AIMS: We undertook to improve an industrial Saccharomyces cerevisiae strain by derepressing it for maltose utilization in the presence of high glucose concentrations. METHODS AND RESULTS: A mutant was obtained from an industrial S. cerevisiae strain following random UV mutagenesis and selection on maltose/5-thioglucose medium. The mutant acquired the ability to utilize glucose simultaneously with maltose and possibly also sucrose and galactose. Aerobic sugar metabolism was still largely fermentative, but an enhanced respirative metabolism resulted in a 31% higher biomass yield on glucose. Kinetic characterization of glucose transport in the mutant revealed the predominance of the high-affinity component. Northern blot analysis showed that the mutant strain expresses only the HXT6/7 gene irrespective of the glucose concentration in the medium, indicating a severe deregulation in the induction/repression pathways modulating HXT gene expression. Interestingly, maltose-grown cells of the mutant display inverse diauxy in a glucose/maltose mixture, preferring maltose to glucose. CONCLUSION: In the mutant here reported, the glucose transport step seems to be uncoupled from downstream regulation, because it seems to be unable to sense abundant glucose, via both repression and induction pathways. SIGNIFICANCE AND IMPACT OF THE STUDY: We report here the isolation of a S. cerevisiae mutant with a novel derepressed phenotype, potentially interesting for the industrial fermentation of mixed sugar substrates.

Numerical simulation of lateral and transforaminal lumbar interbody fusion, two minimally invasive surgical approaches.

The present study aims to compare spinal stability after two different minimally invasive techniques, the lateral lumbar interbody fusion (LLIF) and the transforaminal lumbar interbody fusion (TLIF) approaches. Two nonlinear three-dimensional finite element (FE) models of the L4-L5 functional spinal unit (FSU) were subjected to the loads that usually act on the lumbar spine. Findings show that the LLIF approach yields better results for torsion load case, due to the larger surface area of the implant. For extension, flexion and lateral bending loads, the TLIF approach presents smaller displacements probably due to the anterior placement of the cage and to the smaller damaged area of the annulus fibrosus.

Minimally invasive transforaminal and anterior lumbar interbody fusion surgery at level L5-S1.

This paper presents a finite element analysis to investigate the biomechanical changes caused by transforaminal (TLIF) and anterior lumbar interbody fusion (ALIF) at the L5-S1 level, applying two different implants: T_PAL (TLIF) and SynFix (ALIF). The main objective is to determine which one is more stable for patients. Numerical simulations of segmental motion show that, in the early postoperative phase, displacements and rotation angles obtained in ALIF are greater than the corresponding ones obtained in TLIF, as well as the principal stress values on the ligaments. So, TLIF performed with T_PAL is more stable than ALIF, especially during the recovery phase.

Low Golimumab Trough Levels at Week 6 Are Associated With Poor Clinical, Endoscopic and Histological Outcomes in Ulcerative Colitis Patients: Pharmacokinetic and Pharmacodynamic Sub-analysis of the Evolution Study.

BACKGROUND AND AIMS: Golimumab has an established exposure-response relationship in patients with ulcerative colitis [UC]. However, the association of serum golimumab trough levels [TL] with objective markers of disease activity, such as endoscopic and histological activity scores and concentrations of biomarkers, remains less understood. This report describes the relationship of serum golimumab TL at the end of the induction period [Week 6] with clinical, endoscopic, histological, and biomarker parameters. METHODS: This was an open-label, uncontrolled, prospective and interventional study. Moderate to severely active UC patients naïve to biologic therapy were treated with golimumab. Serum golimumab TL and faecal calprotectin levels were measured at baseline [Week 0 of induction] and Week 6. RESULTS: A total of 34 patients completed the induction phase [Week 6] and were included in this analysis. Overall, 47.1% and 14.7% of patients achieved clinical response and remission with significantly higher serum golimumab TL in patients with early response or remission [3.7 µg/mL vs 1.3 µg/mL, p = 0.0013; and 3.1 µg/mL vs 1.7 µg/mL, p = 0.0164, respectively]. In addition, golimumab TL were significantly higher in patients achieving histological remission [4.2 µg/mL vs 1.7 µg/mL, p = 0.0049]. Week 6 golimumab TL were inversely correlated with the total Mayo score [rs = -0.546; p = 0.0008], the Mayo endoscopic subscore [rs = -0.381; p = 0.0262], the Geboes histological activity score [rs = -0.464; p = 0.0057], and faecal calprotectin levels [rs = -0.497; p = 0.0044]. CONCLUSIONS: A higher early exposure to golimumab is associated with a better objective response in active UC patients and appears to drive the outcome at Week 6.

Strategies of leaf water uptake based on anatomical traits.

The ability of leaves to absorb fog water can positively contribute to the water and carbon balance of plants in montane ecosystems, especially in periods of soil water deficit. However, the ecophysiological traits and mechanisms responsible for variations in the speed and total water absorption capacity of leaves are still poorly known. This study investigated leaf anatomical attributes of seven species occurring in seasonal tropical high-altitude ecosystems (rocky outcrop and forest), which could explain differences in leaf water uptake (LWU) capacities. We tested the hypothesis that different sets of anatomical leaf attributes will be more marked in plant individuals living under these contrasting environmental conditions. Anatomical variations will affect the initial rate of water absorption and the total storage capacity, resulting in different strategies for using the water supplied by fog events. Water absorption by leaves was inferred indirectly, based on leaf anatomical structure and visual observation of the main access routes (using an apoplastic marker), the diffusion of water through the cuticle, and non-glandular or glandular trichomes in all species. The results suggest that three LWU strategies coexist in the species studied. The different anatomical patterns influenced the speed and maximum LWU capacity. The three LWU strategies can provide different adaptive advantages to adjust to temporal and spatial variations of water availability in these tropical high-altitude environments.

Correction to: Abstracts : 29th European Congress of Pathology.

In Poster Sessions, the first-author name was missing from the authorship group originally listed for Abstract PS-13-001 (page S176), entitled "Gastric carcinoma with lymphoid stroma: Analysis of microsatellite instability and Epstein-Barr virus status". The correct authorship group is shown above.

Phase diagram and re-entrant fermionic entanglement in a hybrid Ising-Hubbard ladder.

The degree of fermionic entanglement is examined in an exactly solvable Ising-Hubbard ladder, which involves interacting electrons on the ladder's rungs described by Hubbard dimers at half-filling on each rung, accounting for intrarung hopping and Coulomb terms. The coupling between neighboring Hubbard dimers is assumed to have an Ising-like nature. The ground-state phase diagram consists of four distinct regions corresponding to the saturated paramagnetic, the classical antiferromagnetic, the quantum antiferromagnetic, and the mixed classical-quantum phase. We have exactly computed the fermionic concurrence, which measures the degree of quantum entanglement between the pair of electrons on the ladder rungs. The effects of the hopping amplitude, the Coulomb term, temperature, and magnetic fields on the fermionic entanglement are explored in detail. It is shown that the fermionic concurrence displays a re-entrant behavior when quantum entanglement is being generated at moderate temperatures above the classical saturated paramagnetic ground state.

Pectin and cellulose cell wall composition enables different strategies to leaf water uptake in plants from tropical fog mountain.

Leaf water uptake (LWU) has been observed in plants of different ecosystems and this process is distinct among different species. Four plant species from the Brazilian fog mountain fields were evaluated in order to detect if leaf water uptake capacity is related to the cell wall composition of leaf epidermis. LWU measurements and their relation to anatomical and biochemical traits were analyzed. Cell wall composition was verified through immunocytochemistry using monoclonal antibodies recognizing pectin compounds, and histochemistry with calcofluor white to track cellulose. Differences in LWU among the four species were clearly revealed. Two species presented higher maximum leaf water content and the lowest values of water absorption speed. The other two species presented opposite behavior, namely, low leaf water uptake and the highest values of water absorption speed. The anatomical traits associated with the cell wall composition corroborated the data on the different LWU strategies. The species with abundant detection of cellulose in their epidermal cell walls absorbed more water, but more slowly, while those with abundant detection of pectins absorbed water at a higher speed. These results indicate that cell wall composition regarding pectin and cellulose are significant for water uptake by the leaf epidermis. Pectin provides greater porosity and absorption speed, while cellulose provides greater hydrophilicity and greater water uptake capacity. Current data indicate that the composition of epidermal cell walls is a relevant trait for leaf water uptake.

The performance of Remicade®-optimized quantification assays in the assessment of Flixabi® levels.

BACKGROUND: The advent of Remicade® biosimilars, Remsima®, Inflectra® and, more recently, Flixabi®, has brought along the potential to decrease the costs associated with this therapy, therefore increasing its access to a larger group of patients. However, and in order to assure a soft transition, one must make sure the assays and algorithms previously developed and optimized for Remicade perform equally well with its biosimilars. This study aimed to: (a) validate the utilization of Remicade-optimized therapeutic drug monitoring assays for the quantification of Flixabi; and (b) determine the existence of Remicade, Remsima and Flixabi cross-immunogenicity. METHODS: Healthy donors' sera spiked with Remicade, Remsima and Flixabi were quantified using three different Remicade-quantification assays, and the reactivity of anti-Remicade and anti-Remsima sera to Remicade and to its biosimilars was assessed. RESULTS: The results show that all tested Remicade-infliximab-optimized assays measure Flixabi as accurately as they measure Remicade and Remsima: the intraclass correlation coefficients between theoretical and measured concentrations varied from 0.920 to 0.990. Moreover, the interassay agreement values for the same compounds were high (intraclass correlation coefficients varied from 0.936 to 0.995). Finally, the anti-Remicade and anti-Remsima sera reacted to the different drugs in a similar fashion. CONCLUSIONS: The tested assays can be used to monitor Flixabi levels. Moreover, Remicade, Remsima and Flixabi were shown to have a high cross-immunogenicity, which supports their high similarity but prevents their switching in nonresponders with antidrug antibodies.

Postpartum follow up of gestational diabetes in a Tertiary Care Center.

BACKGROUND: Gestational diabetes is a risk factor for future development of type 2 diabetes. The primary aim of this study was to estimate the prevalence of postpartum glucose tolerance status evaluation in pregnancies complicated by gestational diabetes 6-12 weeks after delivery. The secondary one was to identify the factors that are implicated with postpartum glucose retesting. METHODS: This was a retrospective study performed with a cohort of women with gestational diabetes, with prenatal care and delivery at a tertiary care center, from January 2013 to April 2017. The diagnosis of gestational diabetes was based on IADPSG criteria (Fasting ≥ 92 mg/dl, 1 h ≥ 180 mg/dl and/or 2 h ≥ 153 mg/dl, respectively) and the diagnosis of type 2 diabetes and prediabetes were made using the 2016 ADA's criteria (fasting and 2 h after glucose load ≥ 126 mg/dl and/or ≥ 200 and 100-125 mg/dl and/or 140 and 199 mg/dl, respectively). All women had an appointment scheduled 6-12 weeks postpartum with the results of a 75-g oral glucose tolerance test (OGTT). RESULTS: Of the 152 evaluated women, 21 (13.8%) returned with the postpartum OGTT results. Of these, 9 (45.0%) had a diagnosis of prediabetes. The use of insulin during gestation was the only factor implicated in a higher adherence rate to postpartum testing OR 6.33 (p 0.002). No significance was found for other demographic and clinical variables (age, family income, years of study, parity, gestational age at first visit, smoking, family history of type 2 diabetes, diagnosis of gestational diabetes before the third trimester, pregestational body mass index, previous history of gestational diabetes and ethnicity). CONCLUSION: The majority of patients with gestational diabetes did not return postpartum to perform OGTT and in our study the only factor implicated in a higher postpartum return was the use of insulin during pregnancy. Considering that 45.0% were diagnosed with prediabetes, diabetes care teams should initially identify non-adherent patients.

Multifactor analysis on the effect of collagen concentration, cross-linking and fiber/pore orientation on chemical, microstructural, mechanical and biological properties of collagen type I scaffolds.

This work evaluates the effect of processing variables on some physicochemical and mechanical properties of multi- and unidirectional laminar collagen type I scaffolds. The processing variables considered in this study included microstructure orientation (uni- and multidirectional fiber/pore controlled by freeze-drying methodology), cross-linking (chemical - using genipin and glutaraldehyde, and physical - using a dehydrothermal method), and collagen concentration (2, 5 and 8mg/ml). The biocompatibility of the scaffolds obtained in each of the evaluated manufacturing processes was also assessed. Despite previous research on collagen-based platforms, the effects that these processing variables have on the properties of collagen scaffolds are still not completely understood. Unidirectional scaffolds presented higher resistance to failure under stress than multidirectional ones. The cross-linking degree was found to decrease when the concentration of collagen increased whilst using chemical cross-linkers, and to increase with the concentration of collagen for the dehydrothermal cross-linked scaffolds. Pore orientation indexes of both unidirectional and multidirectional scaffolds were not influenced by collagen concentration. Cross-linked scaffolds were more hydrophobic than non-cross-linked ones, and presented water vapor permeability adequate for use in low-to-moderate exuding wounds. Pore size ranges were compatible with cell in-growth, independently of the employed cross-linking and freezing methodologies. Moreover, scaffolds cross-linked with glutaraldehyde presented higher in-growth of primary oral mucosa fibroblasts than those cross-linked with genipin or with the dehydrothermal treatment. This multi-factor analysis is expected to contribute to the design of collagen type I platforms, which are usable on several potential soft tissue-engineering applications.

Toxicity of the bionematicide 1,4-naphthoquinone on non-target soil organisms.

The main goal of the present study was to evaluate the ecotoxicological effects of 1,4-naphthoquinone (1,4-NTQ), a natural-origin compound presenting nematicidal activity, that can be obtained from walnut husk, in plants and soil invertebrates, including non-target soil nematode communities. This research was part of an ongoing project that aims to develop environmentally-friendly nematicides obtained from agricultural residues. The battery of ISO tests included emergence and growth of corn (Zea mays) and rape (Brassica napus); avoidance with the earthworm Eisenia andrei and the collembolan Folsomia candida; and reproduction with the previous species plus the enchytraeid Enchytraeus crypticus. A novel soil nematode community assay was also performed. ISO tests and nematode assays were conducted using a natural uncontaminated soil that was spiked with a range of 1,4-NTQ concentrations. Toxicity of 1,4-NTQ was found for all test-species and the most sensitive were F. candida and E. andrei. After 7 days of exposure to 1,4-NTQ, nematode abundance decreased along the concentration gradient, and a partial recovery was observed after 14 days (1,4-NTQ <48 mg kg-1 soil). The number of nematode families consistently decreased in both periods. Overall, results indicate that a 1,4-NTQ concentration of <20 mg kg-1 could be environmentally safe but preliminary data suggest that it might be ineffective for the target-nematodes, root-knot nematodes, Meloidogyne spp., and root-lesion nematodes, Pratylenchus spp. In addition, if higher dosages of 1,4-NTQ bionematicide are necessary, the potential recovery of non-target organisms under real field scenarios also needs to be assessed.

Novel antiplatelet role for a protein disulfide isomerase-targeted peptide: evidence of covalent binding to the C-terminal CGHC redox motif.

Essentials Inhibitors of protein disulfide isomerase (PDI) have been considered a new antithrombotic class. CxxC is a PDI-targeted peptide that has been previously shown to inhibit its reductase activity. CxxC binds to surface PDI and inhibits ADP- and thrombin-evoked platelet activation and aggregation. CxxC binds to Cys400 on CGHC redox motif of PDI a' domain, a site for PDI prothrombotic activity. SUMMARY: Background Protein disulfide isomerase (PDI) plays a major role in platelet aggregation, and its inhibitors have emerged as novel antithrombotic drugs. In previous work, we designed a peptide based on a PDI redox motif (CGHC) that inhibited both PDI reductase activity and PDI-modulated superoxide generation by neutrophil Nox2. Thus, we hypothesized that this peptide would also inhibit platelet aggregation by association with surface PDI. Methods Three peptides were used: CxxC, containing the PDI redox motif; Scr, presenting a scrambled sequence of the same residues and AxxA, with cysteines replaced by alanine. These peptides were tested under platelet aggregation and flow cytometry protocols to identify their possible antiplatelet activity. We labeled membrane free thiol and electrospray ionization liquid chromatography tandem mass spectrometry to test for an interaction. Results CxxC decreased platelet aggregation in a dose-dependent manner, being more potent at lower agonist concentrations, whereas neither AxxA nor Scr peptides exerted any effect. CxxC decreased aIIbb3 activation, but had no effect on the other markers. CxxC also decreased cell surface PDI pulldown without interfering with the total thiol protein content. Finally, we detected the addition of one CxxC molecule to reduced PDI through binding to Cys400 through mass spectrometry. Interestingly, CxxC did not react with oxidized PDI. Discussion CxxC has consistently shown its antiplatelet effects, both in PRP and washed platelets, corroborated by decreased aIIbb3 activation. The probable mechanism of action is through a mixed dissulphide bond with Cys400 of PDI, which has been shown to be essential for PDI's actions. Conclusion In summary, our data support antiplatelet activity for CxxC through binding to Cys400 in the PDI a0 domain, which can be further exploited as a model for sitedriven antithrombotic agent development.

Proactive therapeutic drug monitoring of infliximab: a comparative study of a new point-of-care quantitative test with two established ELISA assays.

BACKGROUND: Therapeutic drug monitoring is a powerful strategy known to improve the clinical outcomes and to optimise the healthcare resources in the treatment of autoimmune diseases. Currently, most of the methods commercially available for the quantification of infliximab (IFX) are ELISA-based, with a turnaround time of approximately 8 h, and delaying the target dosage adjustment to the following infusion. AIM: To validate the first point-of-care IFX quantification device available in the market - the Quantum Blue Infliximab assay (Buhlmann, Schonenbuch, Switzerland) - by comparing it with two well-established methods. METHODS: The three methods were used to assay the IFX concentration of spiked samples and of the serum of 299 inflammatory bowel diseases (IBD) patients undergoing IFX therapy. RESULTS: The point-of-care assay had an average IFX recovery of 92%, being the most precise among the tested methods. The Intraclass Correlation Coefficients of the point-of-care IFX assay vs. the two ELISA-based established methods were 0.889 and 0.939. Moreover, the accuracy of the point-of-care IFX compared with each of the two reference methods was 77% and 83%, and the kappa statistics revealed a substantial agreement (0.648 and 0.738). CONCLUSIONS: The Quantum Blue IFX assay can successfully replace the commonly used ELISA-based IFX quantification kits. This point-of-care IFX assay is able to deliver the results within 15 min makes it ideal for an immediate target concentration adjusted dosing. Moreover, it is a user-friendly desktop device that does not require specific laboratory facilities or highly specialised personnel.

Astra blue and basic fuchsin double staining of plant materials.

Methods for double staining plant materials using astra blue and basic fuchsin are described here. These methods can be applied to free hand and microtome sections embedded in paraffin, paraplast or historesin. Also, they can be used to study isolated epidermal peels and pollen preparations. Temporary, semipermanent and permanent preparations were studied. Astra blue stained polysaccharides of the cell wall such as cellulose and pectins. Basic fuchsin showed an affinity for lignified, suberized or cutinized walls. The easy preparation of the reagents, excellent color contrast of the histological preparations, and brief staining times of some methods makes them useful for both routine research and didactic purposes. Also, excellent color or black and white photomicrography can be obtained after the double staining described here.