Ferric citrate for the treatment of hyperphosphatemia and iron deficiency anaemia in patients with NDD-CKD: a systematic review and meta-analysis.

Background: The application of ferric citrate therapy has yielded unexpected benefits in recent years for Chronic kidney disease patients suffering from hyperphosphatemia and iron deficiency -anaemia. Despite this, earlier research on the impact of ferric citrate on NDD-CKD has been contentious. Objective: The goal of the meta-analysis is to evaluate the evidence regarding the advantages and dangers of ferric citrate for the treatment of hyperphosphatemia and iron deficiency anaemia in NDD-CKD patients. Methods: Between the start of the study and June 2022, we searched PubMed, Embase, Cochrane, EBSCO, Scopus, Web of Science, Wan Fang Data, CNKI, and VIP databases for randomised controlled trials of iron citrate for hyperphosphatemia and anaemia in patients with NDD-CKD. For binary categorical data, risk ratios (OR) were employed, and for continuous variables, weighted mean differences The effect sizes for both count and measurement data were expressed using 95% confidence intervals Results: The meta-analysis includes eight trials with a total of 1281 NDD-CKD patients. The phosphorus-lowering effect of ferric citrate was greater compared to the control group (WMD, -0.55, 95% CI, -0.81 to -0.28; I2 = 86%, p < 0.001). Calcium (WMD, 0.092; 95% CI, -0.051 to 0.234; p > 0.05; I2 = 61.9%), PTH (WMD, -0.10; 95% CI, -0.44 to 0.23; I2 = 75%, p > 0.05) and iFGF23 (WMD, -7.62; 95% CI, -21.18 to 5.94; I2 = 20%, p > 0.05) levels were not statistically different after ferric citrate treatment compared to control treatment. Furthermore, ferric citrate increased iron reserves and haemoglobin. The ferric citrate group had considerably greater levels than the controls. Ferric citrate, on the other hand, may raise the risk of constipation, diarrhoea, and nausea. Conclusion: This meta-analysis found that ferric citrate had a beneficial effect in the treatment of NDD-CKD, particularly in reducing blood phosphorus levels when compared to a control intervention. It also shown that ferric citrate has a favourable effect on iron intake and anaemia management. In terms of safety, ferric citrate may increase the likelihood of gastrointestinal side effects.

Nondestructive ultrasound evaluation of microstructure-related material parameters of skeletal muscle: an in silico and in vitro study.

Direct and nondestructive assessment of material properties of skeletal muscle in vivo shall advance our understanding of intact muscle mechanics and facilitate personalized interventions. However, this is challenged by intricate hierarchical microstructure of the skeletal muscle. We have previously regarded the skeletal muscle as a composite of myofibers and extracellular matrix (ECM), formulated shear wave propagation in the undeformed muscle using the acoustoelastic theory, and preliminarily demonstrated that ultrasound-based shear wave elastography (SWE) could estimate microstructure-related material parameters (MRMPs): myofiber stiffness µf, ECM stiffness µm, and myofiber volume ratio Vf. The proposed method warrants further validation but is hampered by the lack of ground truth values of MRMPs. In this study, we presented analytical and experimental validations of the proposed method using finite-element (FE) simulations and 3D-printed hydrogel phantoms, respectively. Three combinations of different physiologically relevant MRMPs were used in the FE simulations where shear wave propagations in the corresponding composite media were simulated. Two 3D-printed hydrogel phantoms with the MRMPs close to those of a real skeletal muscle (i.e., µf=2.02kPa, µm=52.42kPa, and Vf=0.675,0.832) for ultrasound imaging were fabricated by an alginate-based hydrogel printing protocol that we modified and optimized from the freeform reversible embedding of suspended hydrogels (FRESH) method in literature. Average percent errors of (µf,µm,Vf) estimates were found to be (2.7%,7.3%,2.4%)in silico and (3.0%,8.0%,9.9%)in vitro. This quantitative study corroborated the potential of our proposed theoretical model along with ultrasound SWE for uncovering microstructural characteristics of the skeletal muscle in an entirely nondestructive way.

Auditory Receptive Field Net Based Automatic Snore Detection for Wearable Devices.

Although obstructive sleep apnea and hypopnea syndrome (OSAHS) is a common sleep disease, it is sometimes difficult to be detected in time because of the inconvenience of polysomnography (PSG) examination. Since snoring is one of the earliest symptoms of OSAHS, it can be used for early OSAHS prediction. With the recent development of wearable and IoT sensors, we proposed a deep learning-based accurate snore detection model for long-term home monitoring of snoring during sleep. To enhance the discriminability of features between snoring and non-snoring events, an auditory receptive field (ARF) net was proposed and integrated into the feature extraction network. Based on the feature maps derived by the feature extraction network, the detection model predicted a series of candidate boxes and corresponding confidence scores for each candidate box, which denoted whether the candidate box contained a snore event from the input sound waveforms. A snore detection dataset with a total duration of more than 4600 min was developed to evaluate the proposed model. The experimental results on this dataset revealed that the proposed model outperformed other traditional approaches and deep learning models.

Curcumin and berberine co-loaded liposomes for anti-hepatocellular carcinoma therapy by blocking the cross-talk between hepatic stellate cells and tumor cells.

Cancer-associated fibroblasts (CAFs) are a major component of the tumor microenvironment (TME). In hepatocellular carcinoma (HCC), quiescent hepatic stellate cells (HSCs) could be activated to become CAFs, which play a critical role in tumor progression and drug resistance. Therefore, recent efforts have been focused on combining anti-HSC and pro-apoptotic activities to improve anti-tumor efficacy of drugs. In this study, glycyrrhetinic acid and hyaluronic acid-modified liposomes (GA-HA-Lip) were prepared for co-delivery of curcumin (CUR) and berberine (BBR) for the treatment of HCC. Furthermore, we established the LX-2+BEL-7402 co-cultured cell model and implanted the m-HSCs+H22 cells into a mouse to evaluate the anti-tumor effect of CUR&BBR/GA-HA-Lip both in vitro and in vivo. The results showed that CUR&BBR/GA-HA-Lip could accumulate in tumor tissues and be taken up by HSCs and BEL-7402 cells simultaneously. Compared with free CUR, the combination therapy based on GA-HA-Lip exhibits stronger pro-apoptotic and anti-proliferation effect both in vitro and in vivo. The anti-tumor mechanistic study revealed that CUR&BBR/GA-HA-Lip could inhibit the activation of HSCs and restrain drug resistance of tumor cells. In summary, CUR&BBR/GA-HA-Lip could be a promising nano-sized formulation for anti-tumor therapy.

Therapeutic Response of Multifunctional Lipid and Micelle Formulation in Hepatocellular Carcinoma.

Hepatic stellate cells (HSCs), as an important part of the tumor microenvironment (TME), could be activated by tumor cells as cancer-associated fibroblasts (CAFs), thereby promoting the production of extracellular matrix (ECM) and favoring the development of tumors. Therefore, blocking the "CAFs-ECM" axis is a promising pathway to improve antitumor efficacy. Based on this, we developed a multifunctional nanosized delivery system composed of hyaluronic acid-modified pH-sensitive liposomes (CTHLs) and glycyrrheic acid-modified nanomicelles (DGNs), which combines the advantages of targeted delivery, pH-sensitivity, and deep drug penetration. To mimic actual TME, a novel HSCs+BEL-7402 cocultured cell model and a m-HSCs+H22 coimplanted mice model were established. As expected, CTHLs and DGNs could target CAFs and tumor cells, respectively, and promote the drug penetration and retention in tumor regions. Notably, CTHLs+DGNs not only exhibited a superior antitumor effect in three-level tumor-bearing mice but also presented excellent antimetastasis efficiency in lung-metastatic mice. The antitumor mechanism revealed that the lipid&micelle mixed formulations effectively inhibited the activation of CAFs, reduced the deposition of ECM, and reversed the epithelial-mesenchymal transition (EMT) of tumor cells. In brief, the nanosized delivery system composed of CTHLs and DGNs could effectively improve the therapeutic effect of liver cancer by blocking the "CAFs-ECM" axis, which has a good clinical application prospect.

pH-responsive Sulfated Hyaluronic Acid Nanoparticles Targeting Tumor Cells and CAFs for the Treatment of Breast Cancer.

BACKGROUND: Tumor metastasis is a main cause of death in patients with breast cancer. The cross-talk between cancer-associated fibroblasts (CAFs) and tumor cells plays an important role in promoting tumor invasion and metastasis. It is important to develop a novel delivery system to inhibit tumor development by simultaneously targeting both CAFs and tumor cells. OBJECTIVES: The main objective of this research was to prepare nanoparticles to inhibit tumor proliferation and migration by blocking the cross-talk of tumor-CAFs. Additionally, a novel "MCF- 7+NIH/3T3" mixed cell model was established to mimic the tumor microenvironment (TME). METHODS: In this study, the pH-responsive nanoparticles (MIF/DOX-sul-HA NPs) based on sulfated hyaluronic acid (sul-HA) polymers were prepared for co-delivery of doxorubicin (DOX) and mifepristone (MIF). The effects of anti-proliferation and anti-metastasis of MIF/DOX-sul-HA NPs were investigated both in vitro and in vivo. RESULTS: The results showed that MIF/DOX-sul-HA NPs were nearly spherical in shape with narrow particle size distribution and pH-responsive drug release, and could be taken up by both MCF-7 and NIH/3T3 cells. Compared with MCF-7 cells alone, the anti-tumor effect of single DOX was weak in the "MCF-7+NIH/3T3" mixed cell model. MIF/DOX-sul-HA NPs exhibited strong effects of anti-proliferation and anti-metastasis than the free single drug. CONCLUSION: The sul-HA nanoparticles for co-delivery of DOX and MIF could be a promising combined therapy strategy for the treatment of breast cancer.

Noninvasive Assessment of In Vivo Passive Skeletal Muscle Mechanics as a Composite Material Using Biomedical Ultrasound.

OBJECTIVE: This study develops a biomedical ultrasound imaging method to infer microstructural information (i.e., tissue level) from imaging mechanical behavior of skeletal muscle (i.e., organ level). METHODS: We first reviewed the constitutive model of skeletal muscle by regarding it as a transversely isotropic (TI) hyperelastic composite material, for which a theoretical formula was established among shear wave speed, deformation, and material parameters (MPs) using the acoustoelasticity theory. The formula was evaluated by finite element (FE) simulations and experimentally examined using ultrasound shear wave imaging (SWI) and strain imaging (SI) on in vivo passive biceps brachii muscles of two healthy volunteers. The imaging sequence included 1) generation of SW in multiple propagation directions while resting the muscle at an elbow angle of 90°; 2) generation of SW propagating along the myofiber direction during continuous uniaxial muscle extension by passively changing the elbow angle from 90° to 120°. Ultrasound-quantified SW speeds and muscle deformations were fitted by the theoretical formula to estimate MPs of in vivo passive muscle. RESULTS: Estimated myofiber stiffness, stiffness ratio of myofiber to extracellular matrix (ECM), and ECM volume ratio all agreed with literature findings. CONCLUSION: The proposed mathematical formula together with our in-house ultrasound imaging method enabled assessment of microstructural material properties of in vivo passive skeletal muscle from organ-level mechanical behavior in an entirely noninvasive way. SIGNIFICANCE: Noninvasive assessment of both micro and macro properties of in vivo skeletal muscle will advance our understanding of complex muscle dynamics and facilitate treatment and rehabilitation planning.

Spontaneous extension wave for in vivo assessment of arterial wall anisotropy.

Another type of natural wave, traced from longitudinal wall motion and propagation along the artery, is observed in our in vivo human carotid artery experiments. We coin it as extension wave (EW) and hypothesize that EW velocity (EWV) is associated with arterial longitudinal stiffness. The EW is thus assumed to complement the pulse wave (PW), whose velocity (PWV) is tracked from the radial wall displacement and linked to arterial circumferential stiffness through the Moens-Korteweg equation, as indicators for arterial mechanical anisotropy quantification by noninvasive high-frame-rate ultrasound. The relationship between directional arterial stiffnesses and the two natural wave speeds was investigated in wave theory, finite-element simulations based on isotropic and anisotropic arterial models, and in vivo human common carotid artery (n = 10) experiments. Excellent agreement between the theory and simulations showed that EWV was 2.57 and 1.03 times higher than PWV in an isotropic and an anisotropic carotid artery model, respectively, whereas in vivo EWV was consistently lower than PWV in all 10 healthy human subjects. A strong linear correlation was substantiated in vivo between EWV and arterial longitudinal stiffness quantified by a well-validated vascular-guided wave imaging technique (VGWI). We thereby proposed a novel index calculated as EWV2/PWV2 as an alternative to assess arterial mechanical anisotropy. Simulations and in vivo results corroborated the effect of mechanical anisotropy on the propagation of spontaneous waves along the arterial wall. The proposed anisotropy index demonstrated the feasibility of the concurrent EW and PW imaged by high frame-rate ultrasound in grading of arterial wall anisotropy.NEW & NOTEWORTHY An extension wave formed by longitudinal wall displacements was observed by high-frame-rate ultrasound in the human common carotid artery in vivo. A strong correlation between extension wave velocity and arterial longitudinal stiffness complements the well-established pulse wave, which is linked to circumferential stiffness, to noninvasively assess direction-dependent wall elasticity of the major artery. The proposed anisotropy index, which directly reflects arterial wall microstructure and function, might be a potential risk factor for screening (sub-) clinical cardiovascular diseases.

Walled vessel-mimicking phantom for ultrasound imaging using 3D printing with a water-soluble filament: design principle, fluid-structure interaction (FSI) simulation, and experimental validation.

The geometry and stiffness of a vessel are pertinent to blood dynamics and vessel wall mechanical behavior and may alter in diseased conditions. Ultrasound-based ultrafast Doppler (uDoppler) imaging and shear wave imaging (SWI) techniques have been extensively exploited for the assessment of vascular hemodynamics and mechanics. Their performance is conventionally validated on vessel-mimicking phantoms (VMPs) prior to their clinical use. Compared with commercial ones, customized VMPs are favored for research use because of their wider range of material properties, more complex lumen geometries, or wall structures. Fused deposition modeling (FDM) 3D printing technique with plastic filaments is a promising method for making VMPs with a complex vessel lumen. However, it may require a toxic solvent or a long dissolution time currently. In this paper, we present a safe, efficient and geometrically flexible method where FDM 3D printing with a water-soluble polyvinyl alcohol (PVA) filament is exploited to fabricate a walled three-branch VMP (VMP-I). As a key step in fabrication, to avoid dissolution of the PVA-printed vessel core by the solution of the tissue-mimicking material, paraffin wax was used for isolation. Paraffin wax is easy to coat (i.e. without any special equipment), of satisfactory thickness (∼0.1 mm), chemically stable, and easy to remove after fabrication, thus making the proposed method practicable for ultrasound imaging studies. VMP-I was examined by B-mode imaging and power Doppler imaging (PDI) to verify complete dissolution of PVA-printed vessel core in its lumen, confirming good fabrication quality. The flow velocities in VMP-I were estimated by uDoppler imaging with a -0.8% difference, and the shear wave propagation speeds for the same phantom were estimated by SWI with a -6.03% difference when compared with fluid-structure interaction (FSI) simulation results. A wall-less VMP of a scaled and simplified coronary arterial network (VMP-II) was additionally fabricated and examined to test the capability of the proposed method for a complex lumen geometry. The proposed fabrication method for customized VMPs is foreseen to facilitate the development of ultrasound imaging techniques for blood vessels.

Characterization of a new hemihydrate rebaudioside B crystal having lower aqueous solubility.

Poor aqueous solubility of some minor steviol glycosides (SGs) has prevented their potential widespread usage as non-nutritional high intensity sweeteners in beverage industry. Rebaudioside B (reb B) is one of the minor SGs found in stevia leaf, and has a better taste quality than many of the major SGs. However, reb B suffers from poor aqueous solubility and low dissolution rate, which greatly limits its application, especially in beverages. In our effort to enhance its solubility by using natural means, we discovered that under certain conditions reb B forms hemihydrate crystal, which has much lower solubility and dissolution rate than commercial powder reb B product. The crystal was characterized by Fourier Transform Infrared spectroscopy (FT-IR), Scanning Electron Microscopy (SEM), and X-ray Diffraction (XRD). This may offer more insight into the interaction of SGs with water at molecular level, and therefore provide new guidance on current efforts to enhance the solubility of SGs.

[Simultaneous quantitative analysis of four lignanoids in Schisandra chinensis by quantitative analysis of multi-components by single marker].

The aim of the study is to establish a new method of quality evaluation and validate its feasibilities by the simultaneous quantitative assay of four lignanoids in Schisandra chinensis. A new quality evaluation method, quantitative analysis of multi-components by single marker (QAMS), was established and validated with Schisandra chinensis. Four main lignanoids, schisandrin, schisantherin A, deoxyschizandrin and gamma-schizandrin, were selected as analytes and schisandrin as internal reference substance to evaluate the quality. Their contents in 13 different batches of samples, collected from different bathes, were determined by both external standard method and QAMS. The method was evaluated by comparison of the quantitative results between external standard method and QAMS. No significant differences were found in the quantitative results of four lignanoids in 13 batches of S. chinensis determined by external standard method and QAMS. QAMS is feasible for determination of four lignanoids simultaneously when some authentic standard substances were unavailable, and the developed method can be used for quality control of S. chinensis.

Spectroscopic and calorimetric evaluation of chemically induced protein denaturation in HuH-7 liver cancer cells and impact on cell survival.

Solid tumors such as hepatocellular carcinoma are very often not amenable to chemotherapy and radiotherapy. Local ablation methods, including chemical ablation with absolute ethanol, are therefore an option for treatment but lack of information about the mechanism of devitalization leading to cell death is a hindrance to further adoption. Systemic toxicity also has limited the amount of ethanol that can be used in a single treatment session. Therefore we evaluated the mechanism of urea, a denaturant with little or no systemic toxicity, for potential use in chemical ablation. In this study we report on the use of three methods to analyze the effects in cell culture with a view towards eventual clinical application. Human hepatoma HuH-7 cells were analyzed at several time points after treatment using FTIR, DSC, and Raman microspectroscopy based on MTT and PI-exclusion viability assays. Time course fractional denaturation data plotted against viability show that a 50% viability drop occurs after only a 10-20% drop in overall protein denaturation. Other methods of cell death such as apoptosis may also be operative, but this result implies that protein denaturation is one of the major mechanisms of cell death. This is in line with what has been previously suggested for purely thermal methods, and opens the way to mechanism-based improvements in chemical ablation of solid tumors.

The polypyrimidine/polypurine motif in the mouse mu opioid receptor gene promoter is a supercoiling-regulatory element.

The mu opioid receptor (MOR) is the principle molecular target of opioid analgesics. The polypyrimidine/polypurine (PPy/u) motif enhances the activity of the MOR gene promoter by adopting a non-B DNA conformation. Here, we report that the PPy/u motif regulates the processivity of torsional stress, which is important for endogenous MOR gene expression. Analysis by topoisomerase assays, S1 nuclease digests, and atomic force microscopy showed that, unlike homologous PPy/u motifs, the position- and orientation-induced structural strains to the mouse PPy/u element affect its ability to perturb the relaxation activity of topoisomerase, resulting in polypurine strand-nicked and catenated DNA conformations. Raman spectrum microscopy confirmed that mouse PPy/u containing-plasmid DNA molecules under the different structural strains have a different configuration of ring bases as well as altered Hoogsteen hydrogen bonds. The mouse MOR PPy/u motif drives reporter gene expression fortyfold more effectively in the sense orientation than in the antisense orientation. Furthermore, mouse neuronal cells activate MOR gene expression in response to the perturbations of topology by topoisomerase inhibitors, whereas human cells do not. These results suggest that, interestingly among homologous PPy/u motifs, the mouse MOR PPy/u motif dynamically responds to torsional stress and consequently regulates MOR gene expression in vivo.

Differential gene expression activity among species-specific polypyrimidine/polypurine motifs in mu opioid receptor gene promoters.

The mu opioid receptor (MOR) is the principle molecular target of opioid analgesics. An appropriate understanding of MOR gene expression across species is critical for understanding its analgesic functions in humans. Here, we undertake a cross-species analysis of the polymorphic polypyrimidine/polypurine (PPy/u) motif, a key enhancer of MOR gene expression. The mouse PPy/u motif is highly homologous to those of rat (67%) and human (83%), but drives reporter gene expression tenfold and fivefold more effectively than those of rat and human, respectively. Circular dichroism profiles of PPy/u oligonucleotides from different species showed that they are primarily different in structure. Conformational studies of reporter plasmids using confocal Raman spectra, S1 nuclease and restriction enzymes demonstrated that the structural difference is the result of changes in the phosphodiester backbone. Furthermore, these conformational disparities produce differences in torsional stress, as shown by topoisomerase II relaxation and activation of different levels of gene expression under hypertonic conditions. This study demonstrates that homologous PPy/u motifs adopt unique species-specific conformations with different mechanisms and activities for gene expression. We further discuss how structural aspects of transcription regulatory elements, rather than the sequence itself, are significant when studying functional gene expression regulatory elements.

Cellular uptake and nanoscale localization of gold nanoparticles in cancer using label-free confocal Raman microscopy.

This work demonstrates the use of confocal Raman microscopy (CRM) to measure the dynamics of cellular uptake and localization of gold nanoparticles (GNP) with nanoscale resolution. This is important as nanoparticle cellular interactions are increasingly under investigation to support applications as diverse as drug delivery, gene transfection and a variety of heat and radiation based therapeutics. At the heart of these applications is a need to know the dynamics of nanoparticle cellular uptake and localization (i.e., cell membrane, cytoplasm or nucleus). This process can change dramatically based on size, charge, shape and ligand attached to the nanoparticle. While electron microscopy, atomic emission spectroscopy and histology can be used to assess cellular uptake, they are labor intensive and post-mortem and can miss critical dynamics of the process. For this reason investigators are increasingly turning to optically active nanoparticles that allow direct microscopic interrogation of uptake. Here we show that CRM adds to this evolving armamentarium as a fast, noninvasive, and label-free technique to dynamically study cellular uptake of GNPs with subcellular detail in cancer. Raman laser interaction with GNPs inside cells shows unique spectroscopic features corresponding to the intracellular localization of GNPs over 2 to 24 h at the membrane, cytoplasm or nucleus that are separately verified by histology (silver staining) and electron microscopy. These results show that CRM has the potential to facilitate high-throughput study of the dynamics and localization of a variety of GNPs in multiple cell types.

Spatial distribution of the state of water in frozen mammalian cells.

We describe direct determination of the state of intracellular water, measurement of the intercellular concentration of a cryoprotectant agent (dimethylsulfoxide), and the distribution of organic material in frozen mammalian cells. Confocal Raman microspectroscopy was utilized at cryogenic temperatures with single live cells to conduct high spatial resolution measurements (350 × 350 × 700 nm), which yielded two, we believe, novel observations: 1), intracellular ice formation during fast cooling (50°C/min) causes more pronounced intracellular dehydration than slow cooling (1°C/min); and 2), intracellular dimethylsulfoxide concentration is lower (by as much as 50%) during fast cooling, decreasing the propensity for intracellular vitrification. These observations have a very significant impact for developing successful biopreservation protocols for cells used for therapeutic purposes and for cellular biofluids.

Effect of the coating morphology on the drug release from engineered drug-polymer nanocomposites.

The ElectroNanospray process (Nanocopoeia, Inc) transforms drugs and polymers into many nanoscale material states including powders, liquids, encapsulated particles, and coatings. This enabling technology platform allows application of polymers and drugs to the surface of medical devices such as coronary stents in a single-stage process. Modification of ElectroNanospray process parameters resulted in surface coatings with rich morphologies ranging in appearance from smooth and heterogeneous to highly porous and rough (open matrix). The traditional approach of measuring percent release over time by HPLC shows that the drug release profiles change significantly with coating morphology. In this study, we employed high resolution imaging techniques such as SEM, Atomic Force Microscopy (AFM) and Confocal Raman Microscopy to elucidate the drug release process on these coatings in situ, indicating a correlation of release kinetics with coating morphology.

Real-time confocal Raman imaging of a drug delivery system on cardiac leads.

Drug delivery systems incorporated onto the end of cardiac leads are used to reduce inflammation and fibrosis at the lead-tissue interface and enable optimal lead performance. In this research, confocal Raman microscopy was used to capture chemical images of the drug delivery system on pacemaker leads in different elution media in real-time. Raman images in ambient air showed that drug was dispersed in the polymer matrix as discrete particles with size ranging from 1 to 3 microm. Upon immersion into an aggressive elution medium, drug near the surface dissolved immediately and solvent started to penetrate into the polymer matrix through channels from which drug was eluted. The drug depletion depth was a function of time, which was consistent with the drug release profiles obtained by HPLC. Comparing the drug elution in aggressive solvent and biorelevant solvent, a mechanism of drug release is proposed.

Modifications of surfactant distributions and surface morphologies in latex films due to moisture exposure.

Migration of surfactants in water-based, pressure-sensitive adhesive (PSA) films exposed to static and cyclic relative humidity conditions was investigated using confocal Raman microscopy (CRM) and atomic force microscopy (AFM). Studied PSA films contain monomers n-butyl acrylate, vinyl acetate, and methacrylic acid and an equal mass mixture of anionic and nonionic nonylphenol ethoxylate emulsifiers. A leveling of surfactant concentration distributions is observed via CRM after films stored at 50% relative humidity (RH) are exposed to a 100% RH for an extended time period, while relatively small increases in surface enrichment occur when films are stored at 0% RH. Use of CRM for binary mixtures containing anionic or nonionic surfactant and latex that has undergone dialysis to remove nonpolymeric components indicates that surfactant-polymer compatibility governs to a great extent surface enrichment, but not changes observed with humidity variations. AFM images show that upon drying latex coatings, surfactant and other additives collect in large aggregation regions, which protrude from film surfaces. These structures are absent at high humidity, which appears to result from lateral spreading across the polymer surface. When humidity is reduced, aggregation regions reform but appear to be smaller and more evenly dispersed, and by cycling humidity between 0 and 100% RH, interfacial enrichment can be seen to diminish. Presented results provide greater insights into the distribution behavior of surfactants in latex films and potential mechanisms for observed issues arising for these systems.

Freezing-induced phase separation and spatial microheterogeneity in protein solutions.

Amid decades of research, the basic mechanisms of lyo-/cryostabilization of proteins and more complex organisms have not yet been fully established. One major bottleneck is the inability to probe into and control the molecular level interactions. The molecular interactions are responsible for the significant differences in the outcome of the preservation processes. (1) In this communication, we have utilized confocal Raman microspectroscopy to quantify the freezing-induced microheterogeneity and phase separation (solid and liquid) in a frozen solution composed of a model protein (lysozyme) and a lyo-/cryoprotectant (trehalose), which experienced different degrees of supercooling. Detailed quantitative spectral analysis was performed across the ice, the freeze-concentrated liquid (FCL) phases, and the interface region between them. It was established that the characteristics of the microstructures observed after freezing depended not only on the concentration of trehalose in the solution but also on the degree of supercooling. It was shown that, when samples were frozen after high supercooling, small amounts of lysozyme and trehalose were occluded in the ice phase. Lysozyme preserved its native-like secondary structure in the FCL region but was denatured in the ice phase. Also, it was observed that induction of freezing after a high degree of supercooling of high trehalose concentrations resulted in aggregation of the sugar and the protein.