Addition of fibers derived from paper mill sludge in paper coatings: impact on microstructure, surface and optical properties.

Traditionally, cellulose nanofiber (CNF) production has primarily relied on virgin cellulose sources. Yet, the shift to using paper mill sludge (PMS) as a source for CNF underscores the significance of reusing and recycling industrial byproducts. PMS contains significant amounts of cellulose that can be extracted as a raw material. The purpose of present study is to provide a sustainable approach to PMS utilization as a paper coating additive in the cellulose nanofibrils (CNFPMS) form via simply scalable wire-wound rod coating method. The effect of CNFPMS additive amounts at two coating layers on microstructure and surface properties of coatings such as porosity, air permeability surface roughness and optical properties such as brightness, gloss and CIE L*a*b* is studied, which they can also provide insight for the eventual print performance. Results indicated that the obtained CNFPMS in paper coating shows 52% decrease in porosity, presenting significant improvement in the coating microstructure. The marginal increase in permeability coefficient and surface roughness, 54% and 10%, respectively, suggests improving color reproduction and preventing color density losses. Optical analysis showed slight decrease in brightness and gloss, as was expected. Notably, the lightness was improved, which also indicates increasing color gamut volume in printing applications. As a result, the current work offers a sustainable approach to manage PMS for use in paper coatings as a high-value-added material.

Pneumatic nano-sieve for CRISPR-based detection of drug-resistant bacteria.

The increasing prevalence of antibiotic-resistant bacterial infections, particularly methicillin-resistant Staphylococcus aureus (MRSA), presents a significant public health concern. Timely detection of MRSA is crucial to enable prompt medical intervention, limit its spread, and reduce antimicrobial resistance. Here, we introduce a miniaturized nano-sieve device featuring a pneumatically-regulated chamber for highly efficient MRSA purification from human plasma samples. By using packed magnetic beads as a filter and leveraging the deformability of the nano-sieve channel, we achieved an on-chip concentration factor of ∼15-fold for MRSA. We integrated this device with recombinase polymerase amplification (RPA) and clustered regularly interspaced short palindromic repeats (CRISPR)-Cas detection system, resulting in an on-chip limit of detection (LOD) of approximately 100 CFU mL-1. This developed approach provides a rapid, precise, and centrifuge-free solution suitable for point-of-care diagnostics, with the potential to significantly improve patient outcomes in resource-limited medical conditions.

Pneumatic Nano-Sieve for CRISPR-based Detection of Drug-resistant Bacteria.

The increasing prevalence of antibiotic-resistant bacterial infections, particularly methicillin-resistant Staphylococcus aureus (MRSA), presents a significant public health concern. Timely detection of MRSA is crucial to enable prompt medical intervention, limit its spread, and reduce antimicrobial resistance. Here, we introduce a miniaturized nano-sieve device featuring a pneumatically-regulated chamber for highly efficient MRSA purification from human plasma samples. By using packed magnetic beads as a filter and leveraging the deformability of the nano-sieve channel, we achieve an on-chip concentration factor of 15 for MRSA. We integrated this device with recombinase polymerase amplification (RPA) and clustered regularly interspaced short palindromic repeats (CRISPR)-Cas detection system, resulting in an on-chip limit of detection (LOD) of approximately 100 CFU/mL. This developed approach provides a rapid, precise, and centrifuge-free solution suitable for point-of-care diagnostics, with the potential to significantly improve patient outcomes in resource-limited medical conditions.

SEM Nano: An Electron Wave Optical Simulation for the Scanning Electron Microscope.

The simulation program "SEM Nano" is introduced to explain and visualize probe formation in field-emission scanning electron microscopes (SEMs). The program offers an easy and intuitive graphical user interface (GUI) to provide input in terms of understandable SEM parameters and visualization of the output. The simulations are based on wave optics treatment of the electron beam in the SEM column. Based on input parameters provided by the user, the spatial intensity distribution of electrons is calculated at the specimen by incorporating the effects of diffraction, aberrations, coherence, and noise. Given the specimen structure signal (So), the program has the capability to produce an image of the specimen using the electron probe intensity distribution. Finally, a feature is provided to reconstruct the electron probe intensity from the noisy image using a Wiener filter-based deconvolution.

XRCC1 protects transcription from toxic PARP1 activity during DNA base excision repair.

Genetic defects in the repair of DNA single-strand breaks (SSBs) can result in neurological disease triggered by toxic activity of the single-strand-break sensor protein PARP1. However, the mechanism(s) by which this toxic PARP1 activity triggers cellular dysfunction are unclear. Here we show that human cells lacking XRCC1 fail to rapidly recover transcription following DNA base damage, a phenotype also observed in patient-derived fibroblasts with XRCC1 mutations and Xrcc1-/- mouse neurons. This defect is caused by excessive/aberrant PARP1 activity during DNA base excision repair, resulting from the loss of PARP1 regulation by XRCC1. We show that aberrant PARP1 activity suppresses transcriptional recovery during base excision repair by promoting excessive recruitment and activity of the ubiquitin protease USP3, which as a result reduces the level of monoubiquitinated histones important for normal transcriptional regulation. Importantly, inhibition and/or deletion of PARP1 or USP3 restores transcriptional recovery in XRCC1-/- cells, highlighting PARP1 and USP3 as possible therapeutic targets in neurological disease.

XRCC1 prevents toxic PARP1 trapping during DNA base excision repair.

Mammalian DNA base excision repair (BER) is accelerated by poly(ADP-ribose) polymerases (PARPs) and the scaffold protein XRCC1. PARPs are sensors that detect single-strand break intermediates, but the critical role of XRCC1 during BER is unknown. Here, we show that protein complexes containing DNA polymerase ß and DNA ligase III that are assembled by XRCC1 prevent excessive engagement and activity of PARP1 during BER. As a result, PARP1 becomes "trapped" on BER intermediates in XRCC1-deficient cells in a manner similar to that induced by PARP inhibitors, including in patient fibroblasts from XRCC1-mutated disease. This excessive PARP1 engagement and trapping renders BER intermediates inaccessible to enzymes such as DNA polymerase ß and impedes their repair. Consequently, PARP1 deletion rescues BER and resistance to base damage in XRCC1-/- cells. These data reveal excessive PARP1 engagement during BER as a threat to genome integrity and identify XRCC1 as an "anti-trapper" that prevents toxic PARP1 activity.

Thermoplastic Blend Exhibiting Shape Memory-Assisted Self-Healing Functionality.

Here, we report on a polymer blend consisting of a soft thermoplastic polyurethane (TPU) elastomer and a low melting temperature thermoplastic healing agent (polycaprolactone, PCL). In this study, polymer blends containing up to 60 wt % PCL were prepared and the resulting mechanical, thermal, shape memory, and self-healing properties were studied. These immiscible polymers exhibit two well-separated transitions attributable to the melting of PCL and TPU hard segments. This viscoelastic behavior engendered shape memory capability at moderate processing temperatures (∼90 °C) and melt processability at elevated temperatures (>160 °C). The reversible plasticity shape memory (RPSM) effect was also characterized: when subjected to 125% strain at room temperature and subsequently heated to 90 °C, the samples nearly fully recovered to their original length. Moreover, upon heating to above PCL's melting temperature, the flow of PCL into an undeformed crack was shown to fill the crack void, thus promoting self-repair. Through the action of mild heating (90 °C/30 min), fracture surfaces are brought into intimate contact through the action of the RPSM effect and subsequently healed through the redistribution of molten PCL. The shape memory-assisted self-healing efficiency was evaluated by comparing the tensile force restoration after healing of a highly deformed, notched sample to its behavior prior to notching. It was observed that blends containing up to 30 wt % PCL showed nearly complete restoration of properties. In contrast, pure TPU showed only about 5% healing efficiency because of the absence of the PCL healing agent. Blends containing 50 and 60 wt % PCL likewise did not exhibit appreciable restoration of properties, and this was attributed to their propensity to neck during crack opening and poor mechanical properties at elevated temperatures. Blends may serve as a self-healing replacement for pure TPU in existing applications (e.g., automotive and sporting goods) or as a self-healing shape memory polymer in advanced products in soft robotic, biomedical, and microelectronic applications.

Pathogenic ARH3 mutations result in ADP-ribose chromatin scars during DNA strand break repair.

Neurodegeneration is a common hallmark of individuals with hereditary defects in DNA single-strand break repair; a process regulated by poly(ADP-ribose) metabolism. Recently, mutations in the ARH3 (ADPRHL2) hydrolase that removes ADP-ribose from proteins have been associated with neurodegenerative disease. Here, we show that ARH3-mutated patient cells accumulate mono(ADP-ribose) scars on core histones that are a molecular memory of recently repaired DNA single-strand breaks. We demonstrate that the ADP-ribose chromatin scars result in reduced endogenous levels of important chromatin modifications such as H3K9 acetylation, and that ARH3 patient cells exhibit measurable levels of deregulated transcription. Moreover, we show that the mono(ADP-ribose) scars are lost from the chromatin of ARH3-defective cells in the prolonged presence of PARP inhibition, and concomitantly that chromatin acetylation is restored to normal. Collectively, these data indicate that ARH3 can act as an eraser of ADP-ribose chromatin scars at sites of PARP activity during DNA single-strand break repair.

Pathological mutations in PNKP trigger defects in DNA single-strand break repair but not DNA double-strand break repair.

Hereditary mutations in polynucleotide kinase-phosphatase (PNKP) result in a spectrum of neurological pathologies ranging from neurodevelopmental dysfunction in microcephaly with early onset seizures (MCSZ) to neurodegeneration in ataxia oculomotor apraxia-4 (AOA4) and Charcot-Marie-Tooth disease (CMT2B2). Consistent with this, PNKP is implicated in the repair of both DNA single-strand breaks (SSBs) and DNA double-strand breaks (DSBs); lesions that can trigger neurodegeneration and neurodevelopmental dysfunction, respectively. Surprisingly, however, we did not detect a significant defect in DSB repair (DSBR) in primary fibroblasts from PNKP patients spanning the spectrum of PNKP-mutated pathologies. In contrast, the rate of SSB repair (SSBR) is markedly reduced. Moreover, we show that the restoration of SSBR in patient fibroblasts collectively requires both the DNA kinase and DNA phosphatase activities of PNKP, and the fork-head associated (FHA) domain that interacts with the SSBR protein, XRCC1. Notably, however, the two enzymatic activities of PNKP appear to affect different aspects of disease pathology, with reduced DNA phosphatase activity correlating with neurodevelopmental dysfunction and reduced DNA kinase activity correlating with neurodegeneration. In summary, these data implicate reduced rates of SSBR, not DSBR, as the source of both neurodevelopmental and neurodegenerative pathology in PNKP-mutated disease, and the extent and nature of this reduction as the primary determinant of disease severity.

Exploring the Parameter Space of Point Spread Function Determination for the Scanning Electron Microscope-Part II: Effect on Image Restoration Quality.

Point spread function (PSF) deconvolution is an attractive software-based technique for resolution improvement in the scanning electron microscope (SEM) because it can restore information which has been blurred by challenging operating conditions. In Part 1, we studied a modern PSF determination method for SEM and explored how various parameters affected the method's ability to accurately estimate the PSF. In Part 2, we extend this exploration to PSF deconvolution for image restoration. The parameters include reference particle size, PSF smoothing (K), background correction, and restoration denoising (λ). Image quality was assessed by visual inspection and Fourier analysis. Overall, PSF deconvolution improved image quality. Low λ enhanced image sharpness at the cost of noise, while high λ created smoother restorations with less detail. λ should be chosen to balance feature preservation and denoising based on the application. Reference particle size within ±0.9 nm and K within a reasonable range had little effect on restoration quality. Restorations using background-corrected PSFs had superior quality compared with using no background correction, but if the correction was too high, the PSF was cut off causing blurrier restorations. Future efforts to automatically determine parameters would remove user guesswork, improve this method's consistency, and maximize interpretability of outputs.

Exploring the Parameter Space of Point Spread Function Determination for the Scanning Electron Microscope-Part I: Effect on the Point Spread Function.

The point spread function (PSF) of the scanning electron microscope (SEM) can be determined using a recently developed nanoparticle calibration method. Many parameters are involved in PSF determination and introduce a previously unstudied amount of uncertainty into the PSF size and shape. Signal type, support material thickness, reference particle size, PSF smoothing (K), and background correction were investigated regarding their effect on the PSF. Experimental data were complemented by CASINO simulations. Differences in detector position between the observed particles and the method's simulated reference particles caused shifting between secondary electron PSFs and backscattered electron PSFs. Support material thickness did not have a practical effect on the PSF at the tested voltages. Uncertainty in reference particle size varied the PSF full width at half maximum (FWHM) within ±0.7 nm at 2σ, with virtually no uncertainty in some cases. K and background correction within a reasonable range of values resulted in PSF FWHM differences within ±0.9 nm, except at 2 kV for K with an upper bound of ±1.9 nm due to increased noise. Tailoring K and background correction case-by-case would result in smaller differences. The interconnection of these parameters may help in future efforts to calculate their best selection.

Efficient Single-Strand Break Repair Requires Binding to Both Poly(ADP-Ribose) and DNA by the Central BRCT Domain of XRCC1.

XRCC1 accelerates repair of DNA single-strand breaks by acting as a scaffold protein for the recruitment of Polß, LigIIIα, and end-processing factors, such as PNKP and APTX. XRCC1 itself is recruited to DNA damage through interaction of its central BRCT domain with poly(ADP-ribose) chains generated by PARP1 or PARP2. XRCC1 is believed to interact directly with DNA at sites of damage, but the molecular basis for this interaction within XRCC1 remains unclear. We now show that the central BRCT domain simultaneously mediates interaction of XRCC1 with poly(ADP-ribose) and DNA, through separate and non-overlapping binding sites on opposite faces of the domain. Mutation of residues within the DNA binding site, which includes the site of a common disease-associated human polymorphism, affects DNA binding of this XRCC1 domain in vitro and impairs XRCC1 recruitment and retention at DNA damage and repair of single-strand breaks in vivo.

The Determination and Application of the Point Spread Function in the Scanning Electron Microscope.

A method is presented to determine the spatial distribution of electrons in the focused beam of a scanning electron microscope (SEM). Knowledge of the electron distribution is valuable for characterizing and monitoring SEM performance, as well as for modeling and simulation in computational scanning electron microscopy. Specifically, it can be used to characterize astigmatism as well as study the relationship between beam energy, beam current, working distance, and beam shape and size. In addition, knowledge of the distribution of electrons in the beam can be utilized with deconvolution methods to improve the resolution and quality of backscattered, secondary, and transmitted electron images obtained with thermionic, FEG, or Schottky source instruments. The proposed method represents an improvement over previous methods for determining the spatial distribution of electrons in an SEM beam. Several practical applications are presented.

Custom cerium oxide nanoparticles protect against a free radical mediated autoimmune degenerative disease in the brain.

Cerium oxide nanoparticles are potent antioxidants, based on their ability to either donate or receive electrons as they alternate between the +3 and +4 valence states. The dual oxidation state of ceria has made it an ideal catalyst in industrial applications, and more recently, nanoceria's efficacy in neutralizing biologically generated free radicals has been explored in biological applications. Here, we report the in vivo characteristics of custom-synthesized cerium oxide nanoparticles (CeNPs) in an animal model of immunological and free-radical mediated oxidative injury leading to neurodegenerative disease. The CeNPs are 2.9 nm in diameter, monodispersed and have a -23.5 mV zeta potential when stabilized with citrate/EDTA. This stabilizer coating resists being 'washed' off in physiological salt solutions, and the CeNPs remain monodispersed for long durations in high ionic strength saline. The plasma half-life of the CeNPs is ∼4.0 h, far longer than previously described, stabilized ceria nanoparticles. When administered intravenously to mice, the CeNPs were well tolerated and taken up by the liver and spleen much less than previous nanoceria formulations. The CeNPs were also able to penetrate the brain, reduce reactive oxygen species levels, and alleviate clinical symptoms and motor deficits in mice with a murine model of multiple sclerosis. Thus, CeNPs may be useful in mitigating tissue damage arising from free radical accumulation in biological systems.