A Rapid Human Lung Tissue Dissociation Protocol Maximizing Cell Yield and Minimizing Cellular Stress.

The human lung is a complex organ comprised of diverse populations of epithelial, mesenchymal, vascular and immune cells, which gains even greater complexity during disease states. To effectively study the lung at a single cell level, a dissociation protocol that achieves the highest yield of viable cells of interest with minimal dissociation-associated protein or transcription changes key. Here, we detail a rapid collagenase-based dissociation protocol (Col-Short), which provides a high-yield single cell suspension suitable for a variety of downstream applications. Diseased human lung explants were obtained and dissociated through the Col-Short protocol and compared to four other dissociation protocols. Resulting single cell suspensions were then assessed with flow cytometry, differential staining, and quantitative real-time PCR to identify major hematopoietic and non-hematopoietic cell populations, as well as their activation states. We observed that the Col-Short protocol provides the greatest number of cells per gram of lung tissue with no reduction in viability when compared to previously described dissociation protocols. Col-Short had no observable surface protein marker cleavage as well as lower expression of protein activation markers and stress-related transcripts compared to four other protocols. The Col-Short dissociation protocol can be used as a rapid strategy to generate single cells for respiratory cell biology research.

Dissociative Experiences Among Transgender Women: A Phenomenological Study.

There is little published research on dissociative experiences among transgender people, and none from an in-depth qualitative perspective. In-depth, open-ended interviews (N = 7, 6 trans women, 1 trans man) were conducted to explore how dissociation is experienced (particularly among trans women) and its possible relation to negative emotions. There were several similarities across the dissociative experiences described by participants: six felt themselves disconnected from their body as a whole (in contrast to feeling disconnected to a specific body part), and from the world around them and/or themselves. Four acted out different personalities, and five felt emotionally numb when they were dissociating. Six participants described that their dissociation lessened after they started hormone therapy. Respondents tended to distinguish between dysphoric and dissociative experiences: dysphoric phenomena were more clearly distressful while dissociative ones were more emotionally numb.

Investigating the applicability of novel hydrate dissociation inhibitors in oilwell cement through molecular simulations.

In the field of hydrate formation cementing, the method of developing the low hydration exothermic cement systems cannot effectively solve the problem of hydrate dissociation caused by the hydration heat release of cement. Therefore, we proposed a new approach to address this issue by employing cement additives that can effectively delay the dissociation of hydrate. In our previous work, we designed a novel hydrate dissociation inhibitor, PVCap/dmapma, however, its applicability with cement slurry remains unverified. In this study, we established a more realistic model of oilwell cement gel based on experimental data. Additionally, we investigated the potential effects of PVCap/dmapma on the microstructure and mechanical properties of cement gel through molecular simulations. The results suggest that PVCap/dmapma has no negative effect on the performance of cement slurry compared to Lecithin. By adding PVCap/dmapma to cement slurry, the problem of cementing in hydrate formations is expected to be solved.

Study of CO2 Adsorption Properties on the SrTiO3(001) Surface with Ambient Pressure XPS.

The adsorption properties of CO2 on the SrTiO3(001) surface were investigated using ambient pressure X-ray photoelectron spectroscopy under elevated pressure and temperature conditions. On the Nb-doped TiO2-enriched (1 × 1) SrTiO3 surface, CO2 adsorption, i.e., the formation of CO3 surface species, occurs first at the oxygen lattice site under 10-6 mbar CO2 at room temperature. The interaction of CO2 molecules with oxygen vacancies begins when the CO2 pressure increases to 0.25 mbar. The adsorbed CO3 species on the Nb-doped SrTiO3 surface increases continuously as the pressure increases but starts to leave the surface as the surface temperature increases, which occurs at approximately 373 K on the defect-free surface. On the undoped TiO2-enriched (1 × 1) SrTiO3 surface, CO2 adsorption also occurs first at the lattice oxygen sites. Both the doped and undoped SrTiO3 surfaces exhibit an enhancement of the CO3 species with the presence of oxygen vacancies, thus indicating the important role of oxygen vacancies in CO2 dissociation. When OH species are removed from the undoped SrTiO3 surface, the CO3 species begin to form under 10-6 mbar at 573 K, thus indicating the critical role of OH in preventing CO2 adsorption. The observed CO2 adsorption properties of the various SrTiO3 surfaces provide valuable information for designing SrTiO3-based CO2 catalysts.

Accessing the Low-Polar Molecular Composition of Boreal and Arctic Peat-Burning Organic Aerosol via Thermal Analysis and Ultrahigh-Resolution Mass Spectrometry: Structural Motifs and Their Formation.

Peatland fires emit organic carbon-rich particulate matter into the atmosphere. Boreal and Arctic peatlands are becoming more vulnerable to wildfires, resulting in a need for better understanding of the emissions of these special fires. Extractable, nonpolar, and low-polar organic aerosol species emitted from laboratory-based boreal and Arctic peat-burning experiments are analyzed by direct-infusion atmospheric pressure photoionization (APPI) ultrahigh-resolution mass spectrometry (UHRMS) and compared to time-resolved APPI UHRMS evolved gas analysis from the thermal analysis of peat under inert nitrogen (pyrolysis) and oxidative atmosphere. The chemical composition is characterized on a molecular level, revealing abundant aromatic compounds that partially contain oxygen, nitrogen, or sulfur and are formed at characteristic temperatures. Two main structural motifs are identified, single core and multicore, and their temperature-dependent formation is assigned to the thermal degradation of the lignocellulose building blocks and other parts of peat.

Retrograde and semantic amnesia in a case of post-treatment Lyme disease syndrome: did something lead to a psychogenic memory loss? A single-case study.

OBJECTIVE: To describe a case of Post-Treatment Lyme Disease Syndrome (PTLDS) with an atypical cognitive profile. METHOD: A 41-year-old PTLDS patient underwent comprehensive neuropsychological testing and psychological assessment. RESULTS: The patient exhibited impaired intensive attention but preserved selective attention. Executive functions were normal. Short-term and anterograde memory were intact, while retrograde and semantic memory were significantly impaired. The patient also experienced identity loss, specific phobias, dissociative symptoms, and depressed mood. CONCLUSIONS: Severe episodic-autobiographical and retrograde semantic amnesia was consistent with some reports of dissociative amnesia. Loss of identity and phobias were also highly suggestive of a psychogenic mechanism underlying amnesia.

The Mediating Role of Affect Dysregulation and Dissociation in the Relationship Between Attachment and Compulsive Online Shopping: A Path Analysis Model.

Objective: Online shopping has been described by the scientific literature both for its benefits and the potential risks when excessive usage is involved. Indeed, compulsive online shopping is commonly conceptualized as a potential behavioural addiction that substantially impacts the lives of individuals afflicted by it. In light of this, the present research aimed at exploring the association between possible risk and protective factors for compulsive online shopping, by specifically exploring the role of adult attachment, affect dysregulation, and dissociation. Method: 285 participants (75% females, 25% males; Mage = 31.57, SD = 11.379) were involved in the research and completed an online survey. The collected data was analysed implementing a path analysis model. Results: Results showed significant total effects in the relationship between secure/fearful attachment patterns and compulsive online shopping. Such associations were significantly mediated by the sequential effect of affect dysregulation and dissociation. Conclusions: The present study provides useful information to guide tailored interventions concerning both preventive activity and clinical practice.

Embedding Single Pd Atoms on NiGa Intermetallic Surfaces for Efficient and Selective Alkyne Semi-hydrogenation.

Catalytic removal of alkynes is essential in industry for producing polymer-grade alkenes from steam cracking processes. Non-noble Ni-based catalysts hold promise as effective alternatives to industrial Pd-based catalysts but suffer from low activity. Here we report embedding of single-atom Pd onto the NiGa intermetallic surface with replacing Ga atoms via a well-defined synthesis strategy to design Pd1-NiGa catalyst for alkyne semi-hydrogenation. The fabricated Pd1Ni2Ga1 ensemble sites deliver remarkably higher specific mass activity under superb alkene selectivity of >96% than the state-of-the-art catalysts under industry-relevant conditions. Integrated experimental and computational studies reveal that the single-atom Pd located synergizes with the neighbouring Ni sites to facilitate the σ-adsorption of alkyne and dissociation of hydrogen while suppress the alkene adsorption. Such synergistic effects confer the single-atom Pd on the NiGa intermetallic with a Midas touch for alkyne semi-hydrogenation, providing an effective strategy for stimulating low active Ni-based catalysts for other selective hydrogenations in industry.

Applications of Surface Plasmon Resonance (SPR) to the Study of Diverse Protein-Ligand Interactions.

Functional characterization of enzymes/proteins requires determination of the binding affinity of small molecules or other biomolecules with the target proteins. Several available techniques, such as proteomics and drug discovery strategies, require a precise and high-throughput assay for rapid and reliable screening of potential candidates for further testing. Surface plasmon resonance (SPR), a well-established label-free technique, directly measures biomolecular affinities. SPR assays require immobilization of one interacting component (ligand) on a conductive metal (mostly gold or silver) and a continuous flow of solution containing potential binding partner (analyte) across the surface. The SPR phenomenon occurs when polarized light excites the electrons at the interface of the metal and the dielectric medium to generate electromagnetic waves that propagate parallel to the surface. Changes in the refractive index due to interaction between the ligand and analyte are measured by detecting the reflected light, providing real-time data on kinetics and specificity. A prominent use of SPR is identifying compounds in crude plant extracts that bind to specific molecules. Procedures that utilize SPR are becoming increasingly applicable outside the laboratory setting, and SPR imaging and localized SPR (LSPR) are cheaper and more portable alternative for in situ detection of plant or mammalian pathogens and drug discovery studies. LSPR, in particular, has the advantage of direct attachment to test tissues in live-plant studies. Here, we describe three protocols utilizing SPR-based assays for precise analysis of protein-ligand interactions. © 2024 Wiley Periodicals LLC. Basic Protocol 1: SPR comparison of binding affinities of viral reverse transcriptase polymorphisms Basic Protocol 2: SPR screening of crude plant extract for protein-binding agents Basic Protocol 3: Localized SPR-based antigen detection using antibody-conjugated gold nanoparticles.

Behavioural, autonomic, and neural responsivity in depersonalisation-derealisation disorder: A systematic review of experimental evidence.

Depersonalisation-derealisation disorder (DDD) is characterised by distressing experiences of separation from oneself and/or one's surroundings, potentially resulting from alterations in affective, cognitive, and physiological functions. This systematic review aimed to synthesise current experimental evidence of relevance to proposed mechanisms underlying DDD, to appraise existing theoretical models, and to inform future research and theoretical developments. Studies were included if they tested explicit hypotheses in DDD samples, with experimental manipulations of at least one independent variable, alongside behavioural, subjective, neurological, affective and/or physiological dependent variables. Some evidence for diminished subjective responsivity to aversive images and sounds, and hyperactivation in neurocircuits associated with emotional regulation when viewing aversive images emerged, corroborating neurobiological models of DDD. Inconsistencies were present regarding behavioural and autonomic responsivity to facial expressions, emotional memory, and self-referential processing. Common confounds included small sample sizes, medication, and comorbidities. Alterations in affective reactivity and regulation appear to be present in DDD; however, further research employing more rigorous research designs is required to provide stronger evidence for these possible mechanisms.

Electrochemically-assisted intensification of As(III) removal through integrated alkalization and oxidation process.

In response to the need for trace arsenic removal and detoxification, an electro-assisted self-alkalization and oxidant-free processes (ESOP) cell was developed and investigated. It was found that the ESOP removed 90.3 % of arsenic and reduced the As(III) concentration from 150 µg L-1 to less than 5 µg L-1 in its cathode chamber. The As removal involved migration of As(III) and As(V) from the cathode to the anode driven by electrical current. In the ESOP cathode, As(III) was dissociated to As(III) oxyanions via alkalization and then oxidized into As(V) by H2O2. Nearly 80 % of As(III) migration could be attributed to the oxidation by H2O2 and approximately 20 % dissociation by pH alkalization. The voltage-controlled conditions (1.2 -1.5 V) achieved a peak cumulative H2O2 concentration of 10.9 mg L-1. The ESOP demonstrated a high As(III) oxidation to As(V) conversion efficiency of 97.0 % as well as a low energy cost of 0.013 kWh m-3 at 1.2 V. The migrated arsenic was stabilized onto the anode electrode through in-situ electro-oxidation of As(III) and electrosorption of As(III, V); this would help with the post-treatment waste disposal. Those results have provided important insights into an electrochemical approach for highly efficient arsenic detoxification.

Polyethylene liner dissociation in cementless total hip arthroplasty: Ensuring accurate diagnosis.

BACKGROUND: Uncemented acetabular components are widely used in modern total hip arthroplasty (THA). Modularity has numerous advantages including the ability to use supplementary screw fixation for the shell, and allow to switch from ceramic to polyethylene (PE) bearings and vice versa, and the use of lipped and face-changing liners. OBJECTIVE: Despite these advantages, a problem with modular PE liners is dissociation. This is a rare complication in modern implants. The rate of liner dissociation is reported to be very low between 0.17% and 0.8%. Typical symptoms are sudden onset of groin pain in a previously well-functioning hip joint, followed by grinding or clicking sensations during hip joint motions indicating, that the femoral head is articulating with the metal acetabular shell rather than with the PE liner. Any newly observed noise or squeaking from a THA should undergo radiographic investigation to exclude liner dissociation. CASE PRESENTATION: We present the case of an 88-yearold male patient who developed PE liner dissociation in a cementless THA with a Pinnacle acetabular component six years after the index operation. We recommended revision of the left hip, which was performed two days later. In the interim, the patient was advised to use a pair of crutches. During revision surgery, it was observed that the metal head made contact with the cementless cup shell without damaging it from a macroscopic standpoint. Consequently, a simple exchange of the PE liner was conducted, and a 36 mm metal head was implanted due to scratches on the original head. CONCLUSION: Early diagnosis facilitates a straightforward exchange of the liner along with the head, potentially preserving the osseous integrated cup shell and stem integrity. Routine radiologic follow-up allows to distinguish between PE liner dissociation and severe PE wear.

Mediator Role of Dissociative Experiences in the Effect of Childhood Traumas on Emotion Regulation Difficulty and Parental Child-Containing Function.

The aim of this study is to examine the mediating role of dissociation in emotional regulation and parental child-containing function skills of mothers exposed to childhood trauma. The research was conducted with 400 mothers between the ages of 20-60 who had childhood trauma and currently have children between the ages of 0-18. The relational screening model, one of the general screening models, was used in the research. The sample of the research was selected using the convenient sampling method and the simple random method. Research data were collected with the Sociodemographic Information Form, Childhood Trauma Scale (CTS), Parental Child-Containing Function Scale (PCCFS), Emotion Regulation Difficulty Scale (ERDS), and Dissociative Experiences Scale (DES). According to the results obtained in the study, physical abuse (ß = 0.197; 95% CI [0.124; 0.268]), physical neglect (ß = 0.232; 95% CI [0.161; 0.306]), emotional abuse (ß = 0.238; 95% CI [0.169; 0.309]), emotional neglect (ß = 0.210; 95% CI [0.150; 0.275]), and sexual abuse (ß = 0.139; 95% CI [0.058; 0.220]) were found to have a significant indirect effect on emotion regulation difficulties through dissociative experiences. In addition, physical abuse (ß = 0.122; 95% CI [0.071; 0.181]), physical neglect (ß = 0.151; 95% CI [0.084; 0.228]), emotional abuse (ß = 0.158; 95% CI [0.086; 0.238]), emotional neglect (ß = 0.159; 95% CI [0.093; 0.235]), and sexual abuse (ß = 0.086; 95% CI [0.039; 0.150] was found to have a significant indirect effect on parental child-containing function skills through dissociative experiences.

Immersive Virtual Reality Decreases Work Rate and Manipulates Attentional Focus During Self-Regulated Vigorous Exercise.

To determine the effect of immersive virtual reality use on finishing time of a vigorous-intensity self-regulated exercise task, and on relevant psychological variables. Healthy untrained adults (N = 21; 10 men/11 women; age = 22.9 ± 7.2 years; BMI = 24.0 ± 4.5 kg/m2) completed 1500-m exercise bouts on a rowing ergometer in a counterbalanced and randomized order, with and without use of a headset-delivered virtual reality fitness program. Heart rate, rating of perceived exertion, affective valence, and attentional focus were collected every 300 m, in addition to finishing time. Data were analyzed with repeated measures as appropriate. Intensity of both exercise bouts was considered vigorous according to heart rate results (>77% maximal heart rate). Finishing time was faster in the control condition (449.57 ± 82.39 s) than in the virtual reality condition (463.00 ± 91.78 s), p = .007. Compared to the control condition, the virtual reality condition was characterized by a more external attentional focus (52.38 ± 18.22 vs. 38.76 ± 17.81, p < .001). No differences were observed for remaining variables as a result of condition (p > .05 for all). When a headset-delivered VR program was used during a self-regulated vigorous-intensity exercise task, participants were 13.6 seconds (~3%) slower than in a control condition. Attentional focus was manipulated to be more external with VR use, which may have ultimately distracted from the exercise objective. Recommendations for selecting an appropriate virtual reality experience for a given exercise task are discussed.

Simple Determination of Affinity Constants of Antibodies by Competitive Immunoassays.

The affinity constant, also known as the equilibrium constant, binding constant, equilibrium association constant, or the reciprocal value, the equilibrium dissociation constant (Kd), can be considered as one of the most important characteristics for any antibody-antigen pair. Many methods based on different technologies have been proposed and used to determine this value. However, since a very large number of publications and commercial datasheets do not include this information, significant obstacles in performing such measurements seem to exist. In other cases where such data are reported, the results have often proved to be unreliable. This situation may indicate that most of the technologies available today require a high level of expertise and effort that does not seem to be available in many laboratories. In this paper, we present a simple approach based on standard immunoassay technology that is easy and quick to perform. It relies on the effect that the molar IC50 approaches the Kd value in the case of infinitely small concentrations of the reagent concentrations. A two-dimensional dilution of the reagents leads to an asymptotic convergence to Kd. The approach has some similarity to the well-known checkerboard titration used for the optimization of immunoassays. A well-known antibody against the FLAG peptide, clone M2, was used as a model system and the results were compared with other methods. This approach could be used in any case where a competitive assay is available or can be developed. The determination of an affinity constant should belong to the crucial parameters in any quality control of antibody-related products and assays and should be mandatory in papers using immunochemical protocols.

Three prime repair exonuclease 1 preferentially degrades the integration-incompetent HIV-1 DNA through favorable kinetics, thermodynamic, structural, and conformational properties.

HIV-1 integration into the human genome is dependent on 3'-processing of the viral DNA. Recently, we reported that the cellular Three Prime Repair Exonuclease 1 (TREX1) enhances HIV-1 integration by degrading the unprocessed viral DNA, while the integration-competent 3'-processed DNA remained resistant. Here, we describe the mechanism by which the 3'-processed HIV-1 DNA resists TREX1-mediated degradation. Our kinetic studies revealed that the rate of cleavage (kcat) of the 3'-processed DNA was significantly lower (approximately 2-2.5-fold) than the unprocessed HIV-1 DNA by TREX1. The kcat values of human TREX1 for the processed U5 and U3 DNA substrates were 3.8 s-1 and 4.5 s-1, respectively. In contrast, the unprocessed U5 and U3 substrates were cleaved at 10.2 s-1 and 9.8 s-1, respectively. The efficiency of degradation (kcat/Km) of the 3'-processed DNA (U5-70.2 and U3-28.05 pM-1s-1) was also significantly lower than the unprocessed DNA (U5-103.1 and U3-65.3 pM-1s-1). Furthermore, the binding affinity (Kd) of TREX1 was markedly lower (∼2-fold) for the 3'-processed DNA than the unprocessed DNA. Molecular docking and dynamics studies revealed distinct conformational binding modes of TREX1 with the 3'-processed and unprocessed HIV-1 DNA. Particularly, the unprocessed DNA was favorably positioned in the active site with polar interactions with the catalytic residues of TREX1. Additionally, a stable complex was formed between TREX1 and the unprocessed DNA compared the 3'-processed DNA. These results pinpoint the mechanism by which TREX1 preferentially degrades the integration-incompetent HIV-1 DNA and reveal the unique structural and conformational properties of the integration-competent 3'-processed HIV-1 DNA.

Electron Capture vs Transfer Dissociation for Site Determination of Tryptic Peptide Tyrosine Sulfation: Direct Detection of Fibrinogen Sulfation Sites and Identification of Novel Isobaric Interferences.

Tyrosine sulfation, an understudied but crucial post-translational modification, cannot be directly detected in conventional nanoflow liquid chromatography-tandem mass spectrometry (nanoLC-MS/MS) due to the extreme sulfate lability. Here, we report the detection of sulfate-retaining fragments from LC-electron capture dissociation (ECD) and nanoLC-electron transfer higher energy collision dissociation (EThcD). Sulfopeptide candidates were identified by Proteome Discoverer and MSFragger analysis of nanoLC-HCD MS/MS data and added to inclusion lists for LC-ECD or nanoLC-EThcD MS/MS. When this approach failed, targeted LC-ECD with fixed m/z isolation windows was performed. For the plasma protein fibrinogen, the known pyroglutamylated sulfopeptide QFPTDYDEGQDDRPK from the beta chain N-terminus was identified despite a complete lack of sulfate-containing fragment ions. The peptide QVGVEHHVEIEYD from the gamma-B chain C-terminus was also identified as sulfated or phosphorylated. This sulfopeptide is not annotated in Uniprot but was previously reported. MSFragger further identified a cysteine-containing peptide from the middle of the gamma chain as sulfated and deamidated. NanoLC-EThcD and LC-ECD MS/MS confirmed the two former sulfopeptides via sulfate-retaining fragment ions, whereas an unexpected fragmentation pattern was observed for the third sulfopeptide candidate. Manual interpretation of the LC-ECD spectrum revealed two additional isobaric identifications: a trisulfide-linked cysteinyl-glycine or a carbamidomethyl-dithiothreiotol covalent adduct. Synthesis of such adducts confirmed the latter identity.

Elsberg Syndrome With Albuminocytologic Dissociation - A Guillain-Barré Syndrome Mimic or Guillain-Barré Syndrome Variant?

Introduction: Elsberg Syndrome is a presumed infectious lumbosacral radiculitis, with or without accompanying lumbar myelitis, that is often attributed to herpes simplex virus type 2 (HSV-2). Case: A 58-year-old man presented with lower extremity anesthesia, ataxic gait, radiological evidence of radiculitis, and CSF albuminocytologic dissociation. Polymerase chain reaction testing of CSF confirmed HSV-2 infection. Conclusion: A variety of presentations are reported within the scope of Elsberg Syndrome, potentially with distinct disease mechanisms. Delayed onset of neurological symptoms after resolution of rash and absence of pleocytosis raises the possibility that some patients meeting criteria for Elsberg Syndrome have a post-infectious immune-mediated neuropathy. We advise a lower threshold for PCR testing of herpes viruses in patients with acute neuropathy and albuminocytologic dissociation, particularly in cases with early sacral involvement.

Constructing a Built-In Electric Field To Accelerate Water Dissociation for Efficient Alkaline Hydrogen Evolution.

The alkaline hydrogen evolution reaction (HER) is intricately linked to the water dissociation kinetics. The quest for new strategies to accelerate this step is a pivotal aspect of enhancing the HER performance. Herein, we designed and synthesized a heterogeneous nickel phosphide/cobalt phosphide nanowire array grown on nickel foam (Ni2P/CoP/NF) to form a p-n junction structure. The built-in electric field (BEF) in the p-n junction optimizes the binding ability of hydrogen and hydroxyl intermediates, efficiently promoting water dissociation for the alkaline HER. Consequently, Ni2P/CoP/NF exhibits a lower overpotential of 58 and 118 mV at 30 and 100 mA cm-2, respectively, and high stability over 40 h at 300 mA cm-2 for the HER in 1 M KOH. Computational calculations combined with experiment results testify that the BEF presence in the p-n junction of Ni2P/CoP/NF effectively promotes water dissociation, regulates intermediate adsorption/desorption, and boosts electron transport. This study presents a rational design approach for high-performance heterogeneous electrocatalysts.

Unraveling the Layers: Dissociative Identity Disorder as a Response to Trauma.

Dissociative identity disorder (DID), previously recognized as multiple personality disorder, impacts approximately 1.5% of the population. The Diagnostic and Statistical Manual of Mental Disorders, Fifth Edition, Text Revision (DSM-5-TR), outlines various dissociative disorders (DDs), including depersonalization/derealization disorder, dissociative amnesia, DID, unspecified DD, and other specified DDs. Among these, DID stands out as the most severe, characterized by persistent depersonalization/derealization and dissociative amnesia. This case report explores the symptomatology of DID, available treatments, and the role of trauma.