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A study relating to research on the structure and properties of dendrimers was conducted to investigate the influence of solvents on the hydrodynamical volume of dendrimers. Specifically, a fluorophore-labeled dendrimer with a rubicene derivative as a core was covalently linked to the dendrons and fluoresence depolarization was used to derive the hydrodynamical volume of this labeled dendrimer. The experiment and its results are discussed.

Background: Methicillin-resistant Staphylococcus aureus (MRSA)-USA300 is notorious for its ability to cause community- and healthcare-acquired infections, which are even more difficult to treat when associated with a biofilm phenotype. We aimed to characterize the genetic determinants of biofilm formation in a USA300 skin abscess isolate (UAS391) that formed prolific biofilms. Methods: USA300 S. aureus strains, TCH1516 and FPR3757, were found to be closely related based on whole genome mapping (Argus? Optical Mapping System, Opgen Inc, Gaithersburg, USA) to UAS391 (96.3-99.1% similarity, P=0.0151), however differed markedly in biofilm formation (P=0.0001) on a dynamic assay (BioFlux 200, Fluxion Biosciences, USA). Comparison of whole genome sequences of these strains identified differences in a total of 18 genes. Corresponding Tn (bursa aurealis-bearing) knockout mutants in these target genes were obtained from a publicly available mutant library of the same clonal lineage (USA300-JE2) and were characterized phenotypically for biofilm formation. Tn mutants showing significant differences in biofilm formation were utilized for transduction into a plasmid-cured erythromycin-sensitive derivative of UAS391 and for complementation experiments. All strains were tested on the dynamic assay, and 17h-biofilms were stained (SYTO9, Life Technologies) and fluorescence intensity quantified by microscopy (Zeiss, ImageJ). Gene expression levels in Tn and transduced mutants were studied by quantitative reverse transcriptase PCR (StepOnePlusTM, Applied Biosystems®). Results: Comparison of the sequenced genomes of TCH1516, FPR3757 and UAS391 yielded a limited number of variant genes (n=18) that were hypothesized to account for the observed difference in biofilm-forming capacity. Screening of Tn mutants disrupted in these target genes identified one mutant (NE229) bearing a transposon insertion in SAUSA300_1119 (fakA), which exhibited increased biofilm formation similar to UAS391 (P=0.9320). Transduction experiments confirmed that fakA::Tn corresponded to 1.9- to 4.6-fold increase in biofilm formation depending on the USA300 strain background (P≤0.0007), while complementation of the TCH1516 wild-type fakA allele in UAS391 resulted in a 4.3-fold reduction in biofilm formation (P<0.0001). Conclusions: This sequential approach, consisting of strain typing, genome comparison and functional genomics, identified fakA, a recently described fatty acid kinase in S. aureus that is essential for phospholipid synthesis and also impacts the transcription of numerous virulence factors, as a negative regulator of biofilm formation in S. aureus USA300. [ABSTRACT FROM AUTHOR]

Objective: Postoperative cognitive dysfunction (PCD) is a subtle, prolonged deterioration in cognition after surgery. This complication has been frequently investigated, mainly after major (cardiac) surgery. However, the incidence after cochlear implantation is unknown. Therefore, the aim of the study was to investigate the incidence and possible risk factors of PCD in severely hearing-impaired older adults after cochlear implantation. Methods: In a prospective cohort study, 26 older participants (mean age: 70 (8) years), scheduled for cochlear implantation, were assessed prior to and one week after implantation by means of the Montreal Cognitive Assessment (MoCA). The incidence of PCD was calculated. In addition, the following possible risk factors were recorded: age, sex, education, duration of hearing impairment, preoperative signs of depression and anxiety, duration of anesthesia, anesthetic and surgical events and postoperative complications. Results: The incidence of PCD was 11.5%, defined by a Z-score of change in MoCA scores 1.96 (i.e. a decrease of 4 MoCA points, in the present study). The incidence of PCD was corrected for practice effects by incorporating data from a reference group. Besides an effect of age on the postoperative cognitive performance, no significant risk factors were identified. Conclusion: Given the considerably high incidence of PCD after cochlear implantation, routine cognitive screening before and after implantation is recommended to identify patients with PCD and to provide additional care for these patients. [ABSTRACT FROM AUTHOR]

Background: Methicillin-resistant Staphylococcus aureus (MRSA), a leading cause of chronic infections, forms prolific biofilms which afford an escape route from antibiotic treatment and host immunity. However, MRSA clones are genetically diverse, and mechanisms underlying biofilm formation remain under-studied. Such studies form the basis for developing targeted therapeutics. Here, we studied the temporal changes in the biofilm transcriptome of three pandemic MRSA clones: USA300, HEMRSA-15, and ST239., Methods: Biofilm formation was assessed using a static model with one representative strain per clone. Total RNA was extracted from biofilm and planktonic cultures after 24, 48, and 72 h of growth, followed by rRNA depletion and sequencing (Illumina Inc., San Diego, CA, United States, NextSeq500, v2, 1 × 75 bp). Differentially expressed gene (DEG) analysis between phenotypes and among early (24 h), intermediate (48 h), and late (72 h) stages of biofilms was performed together with in silico co-expression network construction and compared between clones. To understand the influence of SCC mec and ACME on biofilm formation, isogenic mutants containing deletions of the entire elements or of single genes therein were constructed in USA300., Results: Genes involved in primarily core genome-encoded KEGG pathways (transporters and others) were upregulated in 24-h biofilm culture compared to 24-h planktonic culture. However, the number of affected pathways in the ST239 24 h biofilm ( n = 11) was remarkably lower than that in USA300/EMRSA-15 biofilms (USA300: n = 27, HEMRSA-15: n = 58). The clfA gene, which encodes clumping factor A, was the single common DEG identified across the three clones in 24-h biofilm culture (2.2- to 2.66-fold). In intermediate (48 h) and late (72 h) stages of biofilms, decreased expression of central metabolic and fermentative pathways (glycolysis/gluconeogenesis, fatty acid biosynthesis), indicating a shift to anaerobic conditions, was already evident in USA300 and HEMRSA-15 in 48-h biofilm cultures; ST239 showed a similar profile at 72 h. Last, SCC mec +ACME deletion and opp3D disruption negatively affected USA300 biofilm formation., Conclusion: Our data show striking differences in gene expression during biofilm formation by three of the most important pandemic MRSA clones, USA300, HEMRSA-15, and ST239. The clfA gene was the only significantly upregulated gene across all three strains in 24-h biofilm cultures and exemplifies an important target to disrupt early biofilms. Furthermore, our data indicate a critical role for arginine catabolism pathways in early biofilm formation., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Vlaeminck, Lin, Xavier, De Backer, Berkell, De Greve, Hernalsteens, Kumar-Singh, Goossens and Malhotra-Kumar.)

Fusarium is one of the most important fungal genera of plant pathogens that affect the cultivation of a wide range of crops. Agricultural losses caused by Fusariumoxysporumf.sp.cubense ( Foc ) directly affect the income, subsistence, and nourishment of thousands of farmers worldwide. For Viet Nam, predictions on the impact of Foc for the future are dramatic, with an estimated loss in the banana production area of 8% within the next five years and up to 71% within the next 25 years. In the current study, we applied a combined morphological-molecular approach to assess the taxonomic identity and phylogenetic position of the different Foc isolates collected in northern Viet Nam. In addition, we aimed to estimate the proportion of the different Fusarium races infecting bananas in northern Viet Nam. The morphology of the isolates was investigated by growing the collected Fusarium isolates on four distinct nutritious media (PDA, SNA, CLA, and OMA). Molecular phylogenetic relationships were inferred by sequencing partial rpb1 , rpb2 , and tef1a genes and adding the obtained sequences into a phylogenetic framework. Molecular characterization shows that c. 74% of the Fusarium isolates obtained from infected banana pseudostem tissue belong to F.tardichlamydosporum . Compared to F.tardichlamydosporum , F.odoratissimum accounts for c.10% of the Fusarium wilt in northern Viet Nam, demonstrating that Foc TR4 is not yet a dominant strain in the region. Fusariumcugenangense - considered to cause Race 2 infections among bananas - is only found in c. 10% of the tissue material that was obtained from infected Vietnamese bananas. Additionally, one of the isolates cultured from diseased bananas was phylogenetically not positioned within the F.oxysporum species complex (FOSC), but in contrast, fell within the Fusariumfujikuroi species complex (FFSC). As a result, a possible new pathogen for bananas may have been found. Besides being present on several ABB 'Tay banana', F.tardichlamydosporum was also derived from infected tissue of a wild Musalutea , showing the importance of wild bananas as a possible sink for Foc ., (Loan Le Thi, Arne Mertens, Dang Toan Vu, Tuong Dang Vu, Pham Le Anh Minh, Huy Nguyen Duc, Sander de Backer, Rony Swennen, Filip Vandelook, Bart Panis, Mario Amalfi, Cony Decock, Sofia I. F. Gomes, Vincent S. F. T. Merckx, Steven B. Janssens.)

The antibiotic growth promoters (AGPs) flavophospholipol and virginiamycin have been widely used for decades in food animal production. AGP activity is believed to be partly modulated by gut microbial composition although exact AGP-induced changes remain unclear. In a controlled intervention study, we studied the effect of flavophospholipol and virginiamycin on the broiler chicken ileal microbiota spanning from birth to 39 days. Using 16S rRNA gene profiling and prediction of metabolic activity, we show that both AGPs result in dynamic microbial shifts that potentially increase anti-inflammatory mechanisms and bioavailability of several essential nutrients by decreasing degradation (flavophospholipol) or increasing biosynthesis (virginiamycin). Further, virginiamycin-supplemented broilers showed increased colonization with potentially pathogenic bacteria, Clostridium perfringens, Campylobacter, and Escherichia/ Shigella spp. Overall, we show that both AGPs induce microbial changes potentially beneficial for growth. However, the increase in (foodborne) pathogens shown here with virginiamycin use could impact not only broiler mortality but also human health. IMPORTANCE Antibiotic growth promoters (AGPs) are commonly used within poultry farming to increase muscle growth. Microbial composition in the gut is known to be influenced by AGP use although exact AGP-induced changes remain unclear. Utilizing 16S rRNA gene profiling, this study provides a first head-to-head comparison of the effect of the two most commonly used AGPs, flavophospholipol and virginiamycin, on the broiler chicken ileum microbiota over time. We found that supplementation with both AGPs altered ileal microbial composition, thereby increasing potential bioavailability of essential nutrients and weight gain. Flavophospholipol showed a slight benefit over virginiamycin as the latter resulted in more extensive microbial perturbations including increased colonization by enteropathogens, which could impact broiler mortality.

In methicillin-sensitive Staphylococcus aureus (MSSA), the tricarboxylic acid (TCA) cycle is known to negatively regulate production of the major biofilm-matrix exopolysaccharide, PIA/PNAG. However, methicillin-resistant S. aureus (MRSA) produce a primarily proteinaceous biofilm matrix, and contribution of the TCA-cycle therein remains unclear. Utilizing USA300-JE2 Tn-mutants (NARSA) in genes encoding TCA- and urea cycle enzymes for transduction into a prolific biofilm-forming USA300 strain (UAS391-Ery s ), we studied the contribution of the TCA- and urea cycle and of proteins, eDNA and PIA/PNAG, to the matrix. Genes targeted in the urea cycle encoded argininosuccinate lyase and arginase ( argH ::Tn and rocF ::Tn), and in the TCA-cycle encoded succinyl-CoA synthetase, succinate dehydrogenase, aconitase, isocitrate dehydrogenase, fumarate hydratase class II, and citrate synthase II ( sucC ::Tn, sdhA/B ::Tn, acnA ::Tn, icd ::Tn, fumC ::Tn and gltA ::Tn). Biofilm formation was significantly decreased under no flow and flow conditions by argH:: Tn, fumC:: Tn , and sdhA/B:: Tn (range OD 492 0.374-0.667; integrated densities 2.065-4.875) compared to UAS391-Ery S (OD 492 0.814; integrated density 10.676) ( p ≤ 0.008). Cellular and matrix stains, enzymatic treatment (Proteinase K, DNase I), and reverse-transcriptase PCR-based gene-expression analysis of fibronectin-binding proteins (fnbA/B) and the staphylococcal accessory regulator (sarA) on pre-formed UAS391-Ery s and Tn-mutant biofilms showed: (i) < 1% PIA/PNAG in the proteinaceous/eDNA matrix; (ii) increased proteins under no flow and flow in the matrix of Tn mutant biofilms (on average 50 and 51 (±11)%) compared to UAS391-Ery s (on average 22 and 25 (±4)%) ( p < 0.001); and (iii) down- and up-regulation of fnbA/B and sarA , respectively, in Tn-mutants compared to UAS391-Ery S (0.62-, 0.57-, and 2.23-fold on average). In conclusion, we show that the biofilm matrix of MRSA-USA300 and the corresponding Tn mutants is PIA/PNAG-independent and are mainly composed of proteins and eDNA. The primary impact of TCA-cycle inactivation was on the protein component of the biofilm matrix of MRSA-USA300.

Isolation profoundly influences social behavior in all animals. In humans, isolation has serious effects on health. Drosophila melanogaster is a powerful model to study small-scale, temporally-transient social behavior. However, longer-term analysis of large groups of flies is hampered by the lack of effective and reliable tools. We built a new imaging arena and improved the existing tracking algorithm to reliably follow a large number of flies simultaneously. Next, based on the automatic classification of touch and graph-based social network analysis, we designed an algorithm to quantify changes in the social network in response to prior social isolation. We observed that isolation significantly and swiftly enhanced individual and local social network parameters depicting near-neighbor relationships. We explored the genome-wide molecular correlates of these behavioral changes and found that whereas behavior changed throughout the six days of isolation, gene expression alterations occurred largely on day one. These changes occurred mostly in metabolic genes, and we verified the metabolic changes by showing an increase of lipid content in isolated flies. In summary, we describe a highly reliable tracking and analysis pipeline for large groups of flies that we use to unravel the behavioral, molecular and physiological impact of isolation on social network dynamics in Drosophila., Competing Interests: I have read the journal’s policy and the authors of this manuscript declare the following competing interests: Authors Hans Nicasy and Manu Sneyders work for Peira, a for profit company. Authors Steve De Backer and, Barbara Weyn worked for DCI Labs, a for profit company.

Background: Alzheimer's disease (AD) is a progressive neurodegenerative disorder and the most common cause of dementia in the elderly population. In this study, we used the APP/PS1 transgenic mouse model to explore the feasibility of using diffusion kurtosis imaging (DKI) as a tool for the early detection of microstructural changes in the brain due to amyloid-β (Aβ) plaque deposition., Methods: We longitudinally acquired DKI data of wild-type (WT) and APP/PS1 mice at 2, 4, 6 and 8 months of age, after which these mice were sacrificed for histological examination. Three additional cohorts of mice were also included at 2, 4 and 6 months of age to allow voxel-based co-registration between diffusion tensor and diffusion kurtosis  metrics and immunohistochemistry., Results: Changes were observed in diffusion tensor (DT) and diffusion kurtosis (DK) metrics in many of the 23 regions of interest that were analysed. Mean and axial kurtosis were greatly increased owing to Aβ-induced pathological changes in the motor cortex of APP/PS1 mice at 4, 6 and 8 months of age. Additionally, fractional anisotropy (FA) was decreased in APP/PS1 mice at these respective ages. Linear discriminant analysis of the motor cortex data indicated that combining diffusion tensor and diffusion kurtosis metrics permits improved separation of WT from APP/PS1 mice compared with either diffusion tensor or diffusion kurtosis metrics alone. We observed that mean kurtosis and FA are the critical metrics for a correct genotype classification. Furthermore, using a newly developed platform to co-register the in vivo diffusion-weighted magnetic resonance imaging with multiple 3D histological stacks, we found high correlations between DK metrics and anti-Aβ (clone 4G8) antibody, glial fibrillary acidic protein, ionised calcium-binding adapter molecule 1 and myelin basic protein immunohistochemistry. Finally, we observed reduced FA in the septal nuclei of APP/PS1 mice at all ages investigated. The latter was at least partially also observed by voxel-based statistical parametric mapping, which showed significantly reduced FA in the septal nuclei, as well as in the corpus callosum, of 8-month-old APP/PS1 mice compared with WT mice., Conclusions: Our results indicate that DKI metrics hold tremendous potential for the early detection and longitudinal follow-up of Aβ-induced pathology.

Phytoplankton, zooplankton, submerged vegetation and main nutrients have been monitored in 48 eutrophic ponds from the Brussels Capital Region (Belgium) between 2005 and 2008. Nine ponds have been biomanipulated in order to improve their ecological quality and prevent the occurrence of noxious cyanobacterial blooms. The 4-year study of a large number of ponds allowed identification of the factors having the strongest influence on phytoplankton growth. Continuous monitoring of the biomanipulated ponds allowed the significance of changes caused by biomanipulation to be tested as well as the main reasons of biomanipulation successes and failures to be elucidated. The main factors controlling phytoplankton in the ponds studied appeared to be grazing by large cladocerans and inhibition of phytoplankton growth by submerged vegetation. Biomanipulation resulted in a significant decrease in phytoplankton biomass in general and biomass of bloom-forming cyanobacteria in particular that were associated with a significant increase in large Cladocera density and size. In six out of nine ponds biomanipulation resulted in the restoration of submerged vegetation. The maintenance of the restored clearwater state in the biomanipulated ponds was strongly dependent on fish recolonisation and nutrient level. In the absence of fish, the clearwater state could be maintained by submerged vegetation or large zooplankton grazing alone. In case of fish recolonisation, restoration of extensive submerged vegetation could buffer, to a considerable degree, the effect of fish except for ponds with high nutrient levels.

Voxel-based analysis (VBA) methods are increasingly being used to compare diffusion tensor image (DTI) properties across different populations of subjects. Although VBA has many advantages, its results are highly dependent on several parameter settings, such as those from the coregistration technique applied to align the data, the smoothing kernel, the statistics, and the post-hoc analyses. In particular, to increase the signal-to-noise ratio and to mitigate the adverse effect of residual image misalignments, DTI data are often smoothed before VBA with an isotropic Gaussian kernel with a full width half maximum up to 16 x 16 x 16 mm(3). However, using isotropic smoothing kernels can significantly partial volume or voxel averaging artifacts, adversely affecting the true diffusion properties of the underlying fiber tissue. In this work, we compared VBA results between the isotropic and an anisotropic Gaussian filtering method using a simulated framework. Our results clearly demonstrate an increased sensitivity and specificity of detecting a predefined simulated pathology when the anisotropic smoothing kernel was used., (2009 Wiley-Liss, Inc.)

In this paper, a nonrigid coregistration algorithm based on a viscous fluid model is proposed that has been optimized for diffusion tensor images (DTI), in which image correspondence is measured by the mutual information criterion. Several coregistration strategies are introduced and evaluated both on simulated data and on brain intersubject DTI data. Two tensor reorientation methods have been incorporated and quantitatively evaluated. Simulation as well as experimental results show that the proposed viscous fluid model can provide a high coregistration accuracy, although the tensor reorientation was observed to be highly sensitive to the local deformation field. Nevertheless, this coregistration method has demonstrated to significantly improve spatial alignment compared to affine image matching.