Efficacy and safety of excimer light (308 nm) in the treatment of pityriasis lichenoides chronica.

INTRODUCTION: Pityriasis lichenoides chronica is the chronic end of the spectrum of pityriasis lichenoides which have several forms of papulosuamous conditions. Several treatments obtained complete clearance of the condition including phototherapy and specifically narrow band ultraviolet B. The Excimer light 308 is a monochromatic light that acts within the ultraviolet B wavelength and used as a targeted phototherapy in several skin conditions. METHODS: Thirty-four patients with histopathologically diagnosed pityriasis lichenoides chronica underwent treatment with biweekly sessions of excimer light 308 nm. Treatment continued until complete clearance was obtained or to a maximum of 48 sessions (24 weeks). RESULTS: Thirty-one patients obtained complete clearance with no recurrence till the end of the study period, two patients had partial response and only one patient showed poor response to treatment. CONCLUSION: Excimer light can be a safe and effective treatment of pityriasis lichinoides chronica in different ages and genders.

Energy Transfer and Radical-Pair Dynamics in Photosystem I with Different Red Chlorophyll a Pigments.

We establish a general kinetic scheme for the energy transfer and radical-pair dynamics in photosystem I (PSI) of Chlamydomonas reinhardtii, Synechocystis PCC6803, Thermosynechococcus elongatus and Spirulina platensis grown under white-light conditions. With the help of simultaneous target analysis of transient-absorption data sets measured with two selective excitations, we resolved the spectral and kinetic properties of the different species present in PSI. WL-PSI can be described as a Bulk Chl a in equilibrium with a higher-energy Chl a, one or two Red Chl a and a reaction-center compartment (WL-RC). Three radical pairs (RPs) have been resolved with very similar properties in the four model organisms. The charge separation is virtually irreversible with a rate of ≈900 ns-1. The second rate, of RP1 → RP2, ranges from 70-90 ns-1 and the third rate, of RP2 → RP3, is ≈30 ns-1. Since RP1 and the Red Chl a are simultaneously present, resolving the RP1 properties is challenging. In Chlamydomonas reinhardtii, the excited WL-RC and Bulk Chl a compartments equilibrate with a lifetime of ≈0.28 ps, whereas the Red and the Bulk Chl a compartments equilibrate with a lifetime of ≈2.65 ps. We present a description of the thermodynamic properties of the model organisms at room temperature.

Impact of Hydrodynamic Interactions on the Kinetic Pathway of Protein Folding.

Protein folding is a fundamental process critical to cellular function and human health, but it remains a grand challenge in biophysics. Hydrodynamic interaction (HI) plays a vital role in the self-organization of soft and biological materials, yet its role in protein folding is not fully understood despite folding occurring in a fluid environment. Here, we use the fluid particle dynamics method to investigate many-body hydrodynamic couplings between amino acid residues and fluid motion in the folding kinetics of a coarse-grained four-α-helices bundle protein. Our results reveal that HI helps select fast folding pathways to the native state without being kinetically trapped, significantly speeding up the folding kinetics compared to its absence. First, the directional flow along the protein backbone expedites protein collapse. Then, the incompressibility-induced squeezing flow effects retard the accumulation of non-native hydrophobic contacts, thus preventing the protein from being trapped in local energy minima during the conformational search of the native structure. We also find that the significance of HI in folding kinetics depends on temperature, with a pronounced effect under biologically relevant conditions. Our findings suggest that HI, particularly the short-range squeezing effect, may be crucial in avoiding protein misfolding.

Proceedings of the 2023 Viral Clearance Symposium, Session 4: Continuous Processing.

The continuous processing session at the 2023 Viral Clearance Symposium (VCS) focused on understanding how to effectively design viral clearance operations for use in continuous processes and methods to perform viral clearance studies. In this session, an approach to directly address control considerations with operating continuous-flow reactors for low pH viral inactivation was presented. Continuous-flow low pH incubation chamber design and implications for residence time determination were discussed. Additionally, viral clearance capability between batch operation and connected operation were demonstrated to be comparable for a connected bind-elute chromatography and flow-through chromatography step. Overall, this session provided additional scientific knowledge to support viral clearance strategies when implementing a continuous manufacturing process.

Proceedings of the 2023 Viral Clearance Symposium, Session 7: Up- and Downstream Virus Retentive Filtration.

Session 7 of the 2023 Viral Clearance Symposium reviewed progresses in virus retentive filtrations applied to both upstream and downstream processing. Upstream topics included investigations and applications of media viral filtration for upstream cell culture viral risk mitigation. Downstream topics included evaluation of viral breakthrough in continuous processing using surrogate particles and demonstration of extensive viral filtration cycling with flow interruptions and long duration in connected process. Reuse of viral filters with proposed procedures was successfully demonstrated amid the supply chain challenge encountered during the pandemic. Discussions and additional considerations for the topics were also provided.

Proceedings of the 2023 Viral Clearance Symposium, Session 5: Viral Clearance Strategy and Process Understanding.

Session 5 of the 2023 Viral Clearance Symposium reviewed the strategy and process understanding of viral clearance testing. Topics included learnings from the past, leveraging surrogate-based methodologies, cleaning agents that inactivate enveloped baculoviruses, segregation, and retrovirus-like particles both in continuous process and in-use as spiking viruses. Overall, there were discussions over a wide array of viral clearance determinants.

Proceedings of the 2023 Viral Clearance Symposium, Session 1: Regulatory Updates.

At the time of the 2023 Viral Clearance Symposium in Vienna, the ongoing revision of ICH Guideline Q5A(R1) Viral Safety Evaluation of Biotechnology Products Derived from Cell Lines of Human or Animal Origin clearly was the dominant regulatory topic. At the symposium, the changes expected for Q5A(R2) to mirror advances of scientific knowledge, for example, the inclusion of new products, including viral-vector-derived ones, that can be subject to virus clearance, deliberations around continuous manufacturing processes, the use of prior knowledge to supplement or in part replace virus validation studies, and new molecular methods for detection of adventitious viruses, were discussed by a European and a US regulator as well as representatives from industry associations that had been involved with the drafting process.

Proceedings of the 2023 Viral Clearance Symposium: 2023 VCS Summary, Pending Questions, and Next Steps.

The 2023 Viral Clearance Symposium (VCS) was hosted by Takeda on 24 and 25 May 2023 in Vienna, Austria. The present conference extended the structure of the previous biennial symposia held between 2009 and 2019. As recapitulated in the introductory session, the genesis of the VCS, as described in the Proceedings of the 2009 VCS was "the worldwide regulatory and industry recognition that challenges, gaps, and opportunities exist, that it formally addressed could benefit the field as whole." This report provides a synopsis of the progress achieved at the conference resulting from detailed technical discussions and the pending questions that still require attention to address. The 2023 VCS was composed of nine individual sessions of short presentations followed by in-depth panel discussions from the presenters. Sessions included Regulatory Updates (with a focus on ICH Q5A(R2) efforts), including a summary of lessons learned from the 2019 VCS, and progress on these key areas mapped into 2023 VCS topics: Viral Clearance Strategy and Case Studies, New Modalities in Chromatography and Adsorptive Filters, Continuous Processing, Viral Clearance Strategy and Process Understanding, Virus Inactivation, Upstream and Downstream Virus Retentive Filtration and Cell Banks, and Advanced Technologies (advanced therapy medicinal products, next-generation sequencing).

Proceedings of the 2023 Viral Clearance Symposium, Session 3: 2023 VCS New Modalities in Chromatography and Adsorptive Filters.

The session provided an update on the application and mechanistic understanding of intensified unit operations (e.g., mixed mode depth filters, mixed mode AEX) since the last conference in 2019. One of the key gaps identified in the 2019 Viral Clearance Symposium session on the topic was for more investigation required to achieve a clear understanding of the molecular mechanisms of virus removal and the relevance of different moleculés interactions including resin, virus, and product. Further investigation into worst-case conditions for these unit operations is also warranted. One of the key outcomes from that 2019 discussion was also that multimodal anion exchangers can have robust and effective virus removal, depending on process and impurities-an observation that was recapitulated with more specific case studies and evidenced by broader application of these chromatographic resins in late-stage regulatory filings.

Estimation of Kinetics Using IMUs to Monitor and Aid in Clinical Decision-Making during ACL Rehabilitation: A Systematic Review.

After an ACL injury, rehabilitation consists of multiple phases, and progress between these phases is guided by subjective visual assessments of activities such as running, hopping, jump landing, etc. Estimation of objective kinetic measures like knee joint moments and GRF during assessment can help physiotherapists gain insights on knee loading and tailor rehabilitation protocols. Conventional methods deployed to estimate kinetics require complex, expensive systems and are limited to laboratory settings. Alternatively, multiple algorithms have been proposed in the literature to estimate kinetics from kinematics measured using only IMUs. However, the knowledge about their accuracy and generalizability for patient populations is still limited. Therefore, this article aims to identify the available algorithms for the estimation of kinetic parameters using kinematics measured only from IMUs and to evaluate their applicability in ACL rehabilitation through a comprehensive systematic review. The papers identified through the search were categorized based on the modelling techniques and kinetic parameters of interest, and subsequently compared based on the accuracies achieved and applicability for ACL patients during rehabilitation. IMUs have exhibited potential in estimating kinetic parameters with good accuracy, particularly for sagittal movements in healthy cohorts. However, several shortcomings were identified and future directions for improvement have been proposed, including extension of proposed algorithms to accommodate multiplanar movements and validation of the proposed techniques in diverse patient populations and in particular the ACL population.

Parameterization of Biomechanical Variables through Inertial Measurement Units (IMUs) in Occasional Healthy Runners.

Running is one of the most popular sports practiced today and biomechanical variables are fundamental to understanding it. The main objectives of this study are to describe kinetic, kinematic, and spatiotemporal variables measured using four inertial measurement units (IMUs) in runners during treadmill running, investigate the relationships between these variables, and describe differences associated with different data sampling and averaging strategies. A total of 22 healthy recreational runners (M age = 28 ± 5.57 yrs) participated in treadmill measurements, running at their preferred speed (M = 10.1 ± 1.9 km/h) with a set-up of four IMUs placed on tibias and the lumbar area. Raw data was processed and analysed over selections spanning 30 s, 30 steps and 1 step. Very strong positive associations were obtained between the same family variables in all selections. The temporal variables were inversely associated with the step rate variable in the selection of 30 s and 30 steps of data. There were moderate associations between kinetic (forces) and kinematic (displacement) variables. There were no significant differences between the biomechanics variables in any selection. Our results suggest that a 4-IMU set-up, as presented in this study, is a viable approach for parameterization of the biomechanical variables in running, and also that there are no significant differences in the biomechanical variables studied independently, if we select data from 30 s, 30 steps or 1 step for processing and analysis. These results can assist in the methodological aspects of protocol design in future running research.

Kinetics of Intermediate Release Enhances P450 11B2-Catalyzed Aldosterone Synthesis.

The mitochondrial enzyme cytochrome P450 11B2 (aldosterone synthase) catalyzes the 3 terminal transformations in the biosynthesis of aldosterone from 11-deoxycorticosterone (DOC): 11ß-hydroxylation to corticosterone, 18-hydroxylation, and 18-oxidation. Prior studies have shown that P450 11B2 produces more aldosterone from DOC than from the intermediate corticosterone and that the reaction sequence is processive, with intermediates remaining bound to the active site between oxygenation reactions. In contrast, P450 11B1 (11ß-hydroxylase), which catalyzes the terminal step in cortisol biosynthesis, shares a 93% amino acid sequence identity with P450 11B2, converts DOC to corticosterone, but cannot synthesize aldosterone from DOC. The biochemical and biophysical properties of P450 11B2, which enable its unique 18-oxygenation activity and processivity, yet are not also represented in P450 11B1, remain unknown. To understand the mechanism of aldosterone biosynthesis, we introduced point mutations at residue 320, which partially exchange the activities of P450 11B1 and P450 11B2 (V320A and A320V, respectively). We then investigated NADPH coupling efficiencies, binding kinetics and affinities, and product formation of purified P450 11B1 and P450 11B2, wild-type, and residue 320 mutations in phospholipid vesicles and nanodiscs. Coupling efficiencies for the 18-hydroxylase reaction with corticosterone as the substrate failed to correlate with aldosterone synthesis, ruling out uncoupling as a relevant mechanism. Conversely, corticosterone dissociation rates correlated inversely with aldosterone production. We conclude that intermediate dissociation kinetics, not coupling efficiency, enable P450 11B2 to synthesize aldosterone via a processive mechanism. Our kinetic data also suggest that the binding of DOC to P450 11B enzymes occurs in at least two distinct steps, favoring an induced-fit mechanism.

Deciphering the Roles of Extracellular Polymeric Substances (EPS) in Shaping Disinfection Kinetics through Permanent Removal via Genetic Disruption.

Extracellular polymeric substances (EPS) ubiquitously encapsulate microbes and play crucial roles in various environmental processes. However, understanding their complex interactions with dynamic bacterial behaviors, especially during the disinfection process, remains very limited. In this work, we investigated the impact of EPS on bacterial disinfection kinetics by developing a permanent EPS removal strategy. We genetically disrupted the synthesis of exopolysaccharides, the structural components of EPS, in Pseudomonas aeruginosa, a well-known EPS-producing opportunistic pathogen found in diverse environments, creating an EPS-deficient strain. This method ensured a lasting absence of EPS while maintaining bacterial integrity and viability, allowing for real-time in situ investigations of the roles of EPS in disinfection. Our findings indicate that removing EPS from bacteria substantially lowered their susceptibility threshold to disinfectants such as ozone, chloramine B, and free chlorine. This removal also substantially accelerated disinfection kinetics, shortened the resistance time, and increased disinfection efficiency, thereby enhancing the overall bactericidal effect. The absence of EPS was found to enhance bacterial motility and increase bacterial cell vulnerability to disinfectants, resulting in greater membrane damage and intensified reactive oxygen species (ROS) production upon exposure to disinfectants. These insights highlight the central role of EPS in bacterial defenses and offer promising implications for developing more effective disinfection strategies.

Construction of a direct Z-scheme Cs3Bi2Cl9/g-C3N4 heterojunction composite for efficient photocatalytic degradation of various pollutants in water: Performance, kinetics and degradation mechanism.

The use of emerging composite materials has been booming to remove environmental pollutants. The aim of this research is to develop a new composite based on Cs3Bi2Cl9 perovskite and graphitic carbon nitride (g-C3N4) to investigate the photocatalytic performance under visible light irradiation. To achieve this, we produce the Cs3Bi2Cl9/g-C3N4 heterojunctions through a simple self-assembly synthesis. The as-synthesized composites are characterized using XRD, FTIR, FESEM, TEM, BET and EDX techniques. The photocatalytic performance of Cs3Bi2Cl9/g-C3N4 is examined in the degradation of various water contaminants, including 4-nitrophenol (4-NP), tetracycline antibiotic (TC), methylene blue (MB) and methyl orange (MO). The experimental results indicate the superior photocatalytic performance of the composites in the degradation of pollutants compared to pure Cs3Bi2Cl9 and g-C3N4. The 10% Cs3Bi2Cl9/g-C3N4 composite achieves the optimal degradation efficiency of 100, 92, 98.7, and 85.1% of 4-NP, TC, MB, and MO, respectively. This superior photocatalytic activity attributes to improved optical and electrochemical properties, including enhanced absorption ability, narrowing band gap, promoted separation efficiency of photogenerated carriers, and a high redox potential, which is confirmed by UV-vis DRS, PL, EIS, and CV analyses. The 10% Cs3Bi2Cl9/g-C3N4 composite also demonstrates high photocatalytic stability after four consecutive cycles. Radical trapping tests show that superoxide radicals (•O2-), holes (h+), and hydroxyl radicals (•OH) contribute to the photocatalytic process. Based on the obtained data, a direct Z-scheme heterojunction mechanism is proposed. Overall, this research offers a new stable photocatalyst with excellent prospect for photocatalytic applications.

Thiol-functionalized cellulose for mercury polluted water remediation: Synthesis and study of the adsorption properties.

Mercury pollution poses a global health threat due to its high toxicity, especially in seafood where it accumulates through various pathways. Developing effective and affordable technologies for mercury removal from water is crucial. Adsorption stands out as a promising method, but creating low-cost materials with high selectivity and capacity for mercury adsorption is challenging. Here we show a sustainable method to synthesize low-cost sulfhydrylated cellulose with ethylene sulfide functionalities bonded glucose units. Thiol-functionalized cellulose exhibits exceptional adsorption capacity (1325 mg g-1) and selectivity for Hg(II) over other heavy metals (Co, Cu, Zn, Pb) and common cations (Ca++, Mg++) found in natural waters. It performs efficiently across a wide pH range and different aqueous matrices, including wastewater, and can be regenerated and reused multiple times without significant loss of performance. This approach offers a promising solution for addressing mercury contamination in water sources.

Monocarboxylate Transporters 1 and 2 Are Responsible for L-Lactate Uptake in Differentiated Human Neuroblastoma SH-SY5Y Cells.

L-Lactate transport via monocarboxylate transporters (MCTs) in the central nervous system, represented by the astrocyte-neuron lactate shuttle (ANLS), is crucial for the maintenance of brain functions, including memory formation. Previously, we have reported that MCT1 contributes to L-lactate transport in normal human astrocytes. Therefore, in this study, we aimed to identify transporters that contribute to L-lactate transport in human neurons. SH-SY5Y cells, which are used as a model for human neurons, were differentiated using all-trans-retinoic acid. L-Lactate uptake was measured using radiolabeled L-lactate, and the expression of MCT proteins was confirmed Western blotting. L-Lactate transport was pH-dependent and saturated at high concentrations. Kinetic analysis suggested that L-lactate uptake was biphasic. Furthermore, MCT1, 2 selective inhibitors inhibited L-lactate transport. In addition, the expression of MCT1 and 2 proteins, but not MCT4, was confirmed. In this study, we demonstrated that MCT1 and 2 are major contributors to L-lactate transport in differentiated human neuroblastoma SH-SY5Y cells from the viewpoint of kinetic analysis. These results lead to a better understanding of ANLS in humans, and further exploration of the factors that can promote MCT1 and 2 functions is required.

Measurement of KRAS-GTPase Activity.

Oncogenic mutations in KRAS typically impact the GAP-mediated and intrinsic GTP hydrolysis activity resulting in elevated levels of cellular KRAS-GTP. The development of biochemical assays for GTPase activity provides an opportunity to quantitatively measure the impact of these mutations on GTP hydrolysis. Here we describe a biochemical assay that measures the release of free phosphate upon hydrolysis of the GTP nucleotide and allows the measurement of intrinsic or GAP-stimulated GTP hydrolysis by KRAS. This assay can be used to measure GTPase activity under single turnover conditions.

A new insight into the surface adsorption in the solution phase: A modification of the Langmuir isotherm.

The Langmuir isotherm, originally developed to study the adsorption of gases, has been modified in this research to investigate the adsorption of solutes in the solution phase. The modification considers the adsorption of solvent molecules and the interactions between adsorbed particles and the species in the solution. Three equations have been obtained to calculate the contribution of these additional effects on the Gibbs free energy, enthalpy, and entropy of solute adsorption based on the new isotherm. The study evaluated the efficiency of the new isotherm in the adsorption of some metal ions in an aqueous solution and found that it is more accurate than the Langmuir isotherm and provides a deeper insight into the adsorption process in the solution phase. PRACTITIONER POINTS: Modification of the Langmuir isotherm for adsorption in solution. Comparison of the efficiency of the Langmuir and modified Langmuir isotherms. Accurate determination of ∆ H ads o $$ \Delta {H}_{ads}^o $$ and ∆ S ads o $$ \Delta {S}_{ads}^o $$ for Pb(II), Cd(II), and Ni(II) adsorption.

Mechanism of DNA origami folding elucidated by mesoscopic simulations.

Many experimental and computational efforts have sought to understand DNA origami folding, but the time and length scales of this process pose significant challenges. Here, we present a mesoscopic model that uses a switchable force field to capture the behavior of single- and double-stranded DNA motifs and transitions between them, allowing us to simulate the folding of DNA origami up to several kilobases in size. Brownian dynamics simulations of small structures reveal a hierarchical folding process involving zipping into a partially folded precursor followed by crystallization into the final structure. We elucidate the effects of various design choices on folding order and kinetics. Larger structures are found to exhibit heterogeneous staple incorporation kinetics and frequent trapping in metastable states, as opposed to more accessible structures which exhibit first-order kinetics and virtually defect-free folding. This model opens an avenue to better understand and design DNA nanostructures for improved yield and folding performance.

Engineering an artificial catch bond using mechanical anisotropy.

Catch bonds are a rare class of protein-protein interactions where the bond lifetime increases under an external pulling force. Here, we report how modification of anchor geometry generates catch bonding behavior for the mechanostable Dockerin G:Cohesin E (DocG:CohE) adhesion complex found on human gut bacteria. Using AFM single-molecule force spectroscopy in combination with bioorthogonal click chemistry, we mechanically dissociate the complex using five precisely controlled anchor geometries. When tension is applied between residue #13 on CohE and the N-terminus of DocG, the complex behaves as a two-state catch bond, while in all other tested pulling geometries, including the native configuration, it behaves as a slip bond. We use a kinetic Monte Carlo model with experimentally derived parameters to simulate rupture force and lifetime distributions, achieving strong agreement with experiments. Single-molecule FRET measurements further demonstrate that the complex does not exhibit dual binding mode behavior at equilibrium but unbinds along multiple pathways under force. Together, these results show how mechanical anisotropy and anchor point selection can be used to engineer artificial catch bonds.