Purinosomes spatially co-localize with mitochondrial transporters.

In this study, we advance our understanding of the spatial relationship between the purinosome, a liquid condensate consisting of six enzymes involved in de novo purine biosynthesis, and mitochondria. Previous research has shown that purinosomes move along tubulin toward mitochondria, suggesting a direct uptake of glycine from mitochondria. Here, we propose that the purinosome is located proximally to the mitochondrial transporters SLC25A13 and SLC25A38, facilitating the uptake of glycine, aspartate, and glutamate, essential factors for purine synthesis. We utilized the proximity ligation assay (PLA) and APEX proximity labeling to investigate the association between purinosome proteins and mitochondrial transporters. Our results indicate that purinosome assembly occurs close to the mitochondrial membrane under purine-deficient conditions, with the transporters migrating to be adjacent to the purinosome. Furthermore, both targeted and non-targeted analyses suggest that the SLC25A13-APEX2-V5 probe accurately reflects endogenous cellular status. These findings provide insights into the spatial organization of purine biosynthesis and lay the groundwork for further investigations into additional proteins involved in this pathway.

Impacts of coyote colonization on coastal mammalian predators.

Extreme ecosystem modification by humans has caused drastic reductions in populations and ranges of top mammalian predators, while simultaneously allowing synanthropic mesopredator species to expand. These conditions often result in inflated local densities of highly adaptable mesopredators that disrupt trophic dynamics and place unsustainable predation pressure on native prey populations. Colonization of a dominant predator may lead to top-down control of mesopredators and restore trophic balance. Coyotes are a novel colonizer of some coastal barrier islands of eastern North America, offering an opportunity to test how the addition of an apex predator impacts an established guild of mesopredators. To assess their trophic impact, we conducted 75,576 camera trapping hours over an 18-month study period, capturing > 1.5 million images across 108 coastal camera sites. Using two-species occupancy and habitat use models, we found sizeable effects of coyote habitat use on that of red foxes and free-ranging domestic cats, suggesting that coyotes function as apex predators in barrier island ecosystems. In fact, the only factor that determined the spatial pattern of highly ubiquitous red foxes was the sympatric habitat use of the largest carnivore in the food web-coyotes. That 'novel' apex predators can become established in coastal food webs illustrates the highly dynamic nature of conservation challenges for habitats and species at the edge of the sea.

Sino-Orbital IgG4 Related Disease- An Uncommon Entity.

IgG4 related disease (IgG4-RD) is an auto immune fibro-inflammatory condition, characterised by presence of IgG4 positive lymphoplasmacytic infiltrates and extensive fibrosis of the involved organ. It commonly affects pancreas, biliary tract and salivary glands. Sino-orbital involvement is a relatively rare presentation. There is extensive fibrosis of the involved organ. Biopsy is often diagnostic and it shows extensive necrosis and lymphoplasmocytic infiltrates. They show dramatic response to steroid therapy. Here we present three cases of IgG4-RD disease involving orbit and para nasal sinuses who were evaluated and treated in a tertiary care teaching hospital over a period of 2 years.

Granulomatosis With Polyangiitis Presenting As Orbital Apex Syndrome.

Granulomatosis with polyangiitis (GPA) is a rare small-vessel vasculitis that typically presents with a triad of sinonasal, pulmonary, and renal symptoms. Here, we present the case of a 43-year-old female with a history of substance use disorder who presented with vision changes and worsening left eye pain over five days. Previous evaluations raised concerns about GPA versus cocaine-induced vasculitis, but diagnostic confirmation was hindered by a lack of medical follow-up. Prompt multidisciplinary intervention led to significant improvement following steroid therapy and IV antibiotics, and the patient was ultimately diagnosed with a high GPA. This case highlights the complexities involved in diagnosing and managing GPA presenting as orbital apex syndrome, particularly in patients with comorbidities and non-adherence to medical follow-up.

[Analysis of curative effect of orbital apex syndrome caused by sinus disease].

Objective:To compare the efficacy of endoscopic sinus surgery and conservative treatment for orbital apex syndrome caused by sinus lesions. Methods:The clinical data of 56 patients with orbital apex syndrome caused by sinus lesions who were admitted to the First Affiliated Hospital of Zhengzhou University from January 2018 to August 2023 were retrospectively analyzed and divided into a surgical group of 21 cases and a conservative group of 35 cases. The clinical features and prognosis of the two groups were compared. Results:Among the sinus lesions in the surgical group, 61.9% were fungal sinusitis, 28.6% were bacterial sinusitis, and 9.5% were sphenoid sinus tumors. In the conservative group, non-fungal sinusitis accounted for 65.7% and fungal sinusitis accounted for 34.3%. In addition to sinus lesions, patients had underlying diseases. In the surgical group, 71.4% had hypertension and 80.9% had diabetes; in the conservative group, 28.6% had hypertension and 42.9% had diabetes. After a follow-up of 1 month to 5 years, the symptom improvement rate in the surgical group was 85.7%, with 1 case of recurrence. No recurrence was found after reoperation, while the symptom improvement rate in the conservative group was 22.9%, and 6 cases recurred after symptom improvement, and were transferred to rhinology department. No recurrence was seen after surgery. Conclusion:Most of the sinus lesions in this study were fungal sinusitis. In addition, patients with underlying diseases such as diabetes, hypertension, nephrotic syndrome, etc. have reduced nasal immunity, which significantly increases the risk of disease. Since early nasal symptoms are not obvious, multidisciplinary cooperation in diagnosis and treatment is very necessary. Once imaging examination suggests orbital apex syndrome caused by sinus lesions, endoscopic sinus opening should be performed as soon as possible.

Clinicians' perspectives, inducements, preferences, and clinical experiences regarding the use of electronic apex locator and apex locator integrated engine-driven instrumentation: a cross-sectional study.

BACKGROUND: This study aimed to investigate the perspectives, preferences, and clinical experiences regarding using electronic apex locator and apex locator integrated instrumentation of dentists and endodontists. METHODS: A web-based questionnaire consisting of 3 parts and 23 closed-ended questions to achieve the objective of the study was carried out in ethical conditions between August and October 2023. The first part of survey included demographic information, while the second part was about evaluating electronic apex locator usage. In the last part, only participants' use of apex locator-integrated instrumentation was evaluated. Data were analyzed at a significance level of p < 0.05. RESULTS: A total of 297 clinicians, including 59 endodontists and 34 endodontic residents/Ph.D. students participated in the questionnaire. Endodontists and endodontic residents/Ph.D. students perform statistically significantly more root canal treatments per week on average (p = 0.001). For the working length determination method (multiple option question), 78.5% of participants use an electronic apex locator and 39.7% apex locator-integrated engines. However, the preference rate for electronic apex determination technique was generally 95.6%, with the full rate confirmation of endodontists and endodontic residents/Ph.D. students (100%). A total of 21 endodontists out of 59 prefer apex locator integrated engine-driven instrumentation. Although many of these specialized clinicians use this technique, they stated that they measure electronic working length passively for confirmation of the working length before (90.5%) and after the preparation (66.7%). CONCLUSIONS: Dentists, as well as endodontists, are skeptical about apex locator-integrated engine-driven instrumentation. Using this technique as a supporter rather than a primary way for preparation within safe limits may give safer results in terms of treatment outcomes.

The Management of Open Apex Using a Bioactive Material: A Case Report.

The management of teeth with open apices poses unique challenges in endodontics, requiring effective strategies to promote continued root development and maintain pulp vitality. This abstract explores the utilization of bioactive materials in the treatment of open apices, specifically focusing on their role in achieving optimal outcomes. Bioactive materials, such as Biodentine (Septodont, Saint-Maur-des-Fossés, France), have gained prominence for their favourable physiochemical properties, biocompatibility, and ability to stimulate dentinogenesis. The application of a bioactive material as an apical plug not only addresses immediate concerns but also contributes to long-term health and stability. This abstract reviews relevant literature, discusses clinical cases, and emphasizes the importance of tailoring treatment plans to the individual characteristics of open apex cases. The findings underscore the promising role of bioactive materials in reshaping the landscape of endodontic interventions for teeth with open apices, highlighting their potential to enhance both clinical and radiographic success.

A Rare Tick Tale: A Novel Case of the Australian Paralysis Tick Causing Multiple Cranial Neuropathies.

The Australian paralysis tick (Ixodes holocyclus) is found along the east coast of Australia. Tick bites may result in paralysis ranging from muscular weakness to ascending paralysis requiring respiratory support. Ocular complications and facial nerve involvement are rare. We present a rare occurrence of tick-bite-associated visual loss, proptosis, and multiple cranial neuropathies not previously reported in the literature. The tick was removed, and the patient's symptoms improved following treatment with steroids and oral doxycycline. The vision and sensory changes are not explained by the Ixodes toxin; thus, we hypothesize this is related to orbital apex inflammation.

KAT6A Condensates Impair PARP1 Trapping of PARP Inhibitors in Ovarian Cancer.

Most clinical PARP inhibitors (PARPis) trap PARP1 in a chromatin-bound state, leading to PARPi-mediated cytotoxicity. PARPi resistance impedes the treatment of ovarian cancer in clinical practice. However, the mechanism by which cancer cells overcome PARP1 trapping to develop PARPi resistance remains unclear. Here, it is shown that high levels of KAT6A promote PARPi resistance in ovarian cancer, regardless of its catalytic activity. Mechanistically, the liquid-liquid phase separation (LLPS) of KAT6A, facilitated by APEX1, inhibits the cytotoxic effects of PARP1 trapping during PARPi treatment. The stable KAT6A-PARP1-APEX1 complex reduces the amount of PARP1 trapped at the DNA break sites. In addition, inhibition of KAT6A LLPS, rather than its catalytic activity, impairs DNA damage repair and restores PARPi sensitivity in ovarian cancer both in vivo and in vitro. In conclusion, the findings demonstrate the role of KAT6A LLPS in fostering PARPi resistance and suggest that repressing KAT6A LLPS can be a potential therapeutic strategy for PARPi-resistant ovarian cancer.

One year "ADAPT system" use for proximal femoral fracture osteosynthesis with intramedullary nail . A case control study.

Introduction: Proximal femoral fractures (PFF) are a significant health concern among the elderly, often leading to complications and high mortality rates. Intramedullary nailing is widely considered the most effective treatment for lateral proximal femoral fractures (LPFF), with the Tip Apex Distance (TAD) being a crucial predictor of surgical success. This study aimed to compare outcomes between patients treated with and without the ADAPT (ADAptive Positioning Technology) system, which aids in the precise placement of the cephalic screw. Materials and Methods: A retrospective analysis was conducted on 97 patients with intertrochanteric fractures treated in 2022. Patients were divided into two groups: those treated with the ADAPT system (group I, n=34) and those treated without it (group II, n=63). Fractures were classified according to AO/OTA classification. The primary outcomes measured were operative time, cephalic screw angle, TAD, and incidence of lag screw cut-out. Statistical analyses included chi-square tests and t-tests, with significance set at P < 0.05. Results: The ADAPT system did not significantly reduce TAD (18.21 mm in the ADAPT group vs. 19.94 mm in the control group, p=0.149). Operative times were similar between the groups. The incidence of lag screw cut-out was low in both groups, with no significant differences. The study confirmed a strong correlation between higher TAD and increased risk of screw cut-out, underscoring the importance of precise screw placement. Discussion: Computer-assisted surgery, such as the ADAPT system, aims to enhance the accuracy of cephalic screw placement. In this study, the ADAPT system didn't demonstrate a statistically significant advantage in reducing TAD or preventing screw cut-out. Nevertheless, the critical role of TAD in preventing fixation failure was reaffirmed, emphasising the need for precise surgical techniques. Conclusion: While the ADAPT system did not show a significant advantage in reducing TAD or preventing screw cut-out in this study, the importance of achieving optimal TAD in cephalomedullary nailing was reinforced. Future research should continue to explore the role of computer-assisted systems in enhancing surgical accuracy and improving outcomes for patients with LPFF.

In-vitro Evaluation of the Accuracy of Electronic Apex Locators and Cone Beam Computed Tomography in the Detection of Oblique Root Fractures.

OBJECTIVES: This study aimed to compare the accuracy of cone-beam computed tomography (CBCT) scanning and three different electronic apex locators (EALs) in the detection of simulated oblique root fractures (ORF) in different localizations. METHODS: The study utilized a total of eighty human maxillary incisors, which were categorized into two groups based on the location of the ORF (apical and middle third of the root) formed on the buccal side of the root surface. The measurement of the distance between the incisal edge and the intersection of the ORF with the root canal was conducted using a stereomicroscope. This measurement is referred to as the actual working length (AWL). Additionally, three EALs-Dentaport ZX, EndoRadar Pro, and Propex II-were utilized to determine the electronic working length (EWL). Furthermore, CBCT images were employed to assess the distance, known as the CBCT working length (CWL). The differences were determined by subtracting AWL from EWL and CWL. RESULTS: Based on the accuracy of the devices, there were no significant differences observed among Dentaport ZX, EndoRadar, Propex II, and CBCT measures in both the apical and middle third HRF groups, within the acceptable range of 0.5 mm and 1 mm. CONCLUSION: The accuracy of Dentaport ZX, Propex II, and CBCT was higher in the middle third ORF group compared to the apical third ORF group, with a tolerance of 0.5 mm. However, there were no significant differences seen among the devices.

The usefulness of ultrasonography as a dynamic measurement system for visualizing root canal working length: an in vivo study.

OBJECTIVE: Although apex locators are generally effective tools for determining root canal working length, they may produce inaccurate results in some cases. The present study aimed to evaluate the efficacy of ultrasonography as an alternative method for measuring root canal length. MATERIALS AND METHODS: Forty-seven anterior teeth with apical lesions were selected for the study. Initially, an electronic apex locator was used to measure the working length. Subsequently, ultrasonography was employed to visualize the root apex and determine the working length. During ultrasound imaging, a K-file No. 15 was inserted into the root canal until its tip was visible on the ultrasound monitor. Measurements obtained from both methods were compared using an independent sample t-test. Correlations were assessed with the Pearson correlation coefficient, and agreement was determined using the Bland‒Altman plot. RESULTS: The mean working canal length was 19.9 mm for the apex locator and 20.6 mm for the ultrasonography-guided method. No significant differences were observed between the data obtained using the apex locator method and the data obtained using the ultrasonography guidance method. Furthermore, a high level of agreement was identified between the two techniques. CONCLUSION: Ultrasonography can be used to visualize the apex effectively and determine canal length, especially when canal length determination is uncertain for various reasons.

Comparison of endoscopic multiport approaches to the petrous apex: contralateral transmaxillary versus contralateral medial transorbital corridor.

OBJECTIVE: Accessing the petrous apex (PA) via an endoscopic endonasal approach (EEA) is challenging due to its posterior and lateral anatomical relationship with the paraclival carotid artery. Typically, the EEA requires the mobilization or compression of the vessel and the use of angled-lens endoscopes and instruments. A sublabial contralateral transmaxillary (CTM) corridor has been used to overcome these challenges. Still, it requires extensive osteo-meatal disruption and drilling of the medial pterygoid process, which risks the vidian nerve and increases nasal morbidity. Furthermore, the CTM corridor positions the endoscope in the same horizontal plane as the instruments passing through the nostrils, leading to fencing. The authors propose a novel minimally invasive route to the PA, the precaruncular contralateral medial transorbital (cMTO) corridor, to address these issues. This anatomical study compares the EEA+CTM and EEA+cMTO corridors in accessing the PA. METHODS: The authors dissected 14 fresh, preinjected cadaveric specimens (28 sides) using neuronavigation to complete EEA, cMTO, and CTM on each side. In addition to qualitative analysis, they measured and compared the working distance between the entry point (nose, orbit, maxilla) and the petrosal process of the sphenoid bone (PPSB), superomedial PA, and foramen lacerum (FL); angle of attack (AoA); area of surgical freedom; endoscope-instrument fencing angle; and visual angle for each approach. RESULTS: The cMTO corridor provided the shortest working distance to the petroclival region (PA = 67.4 ± 4.47 mm, PPSB = 67.57 ± 4.33 mm, and FL = 66.30 ± 4.77 mm) compared to the CTM (PA = 75.85 ± 3.63 mm, PPSB = 76 ± 3.96 mm, and FL = 74.52 ± 4.26 mm) and to the EEA (PA = 85.16 ± 3.16 mm, PPSB = 84.55 ± 3.02 mm, and FL = 83.42 ± 3.21 mm, p < 0.001). Both CTM and cMTO corridors had a similar visual angle to the PA (20.72° ± 2.16° and 21.63° ± 1.84°, respectively), offering a similar but significantly better visualization than EEA alone (44.71° ± 3.24°, p < 0.001). The cMTO corridor provided better instrument maneuverability than the CTM, as evidenced by a significantly greater fencing angle (30.9° ± 4.9°) than with the CTM (21.7° ± 4.02°, p < 0.001). The vertical AoAs for the EEA, cMTO, and CTM corridors were 9.79° ± 1.75°, 10.65° ± 0.82°, and 9.82° ± 1.43°, respectively (p = 0.009), whereas in the horizontal plane, these were 9.29° ± 1.51°, 9.10° ± 0.73°, and 10.49° ± 1.43° (p < 0.001), respectively. Both the CTM and cMTO corridors offered similar areas of surgical freedom (678.06 ± 99.5 mm2 and 673.59 ± 104.8 mm2, p = 0.986), but they were more significant than that provided by the EEA 487.29 ± 112.9 mm2 (p < 0.001). CONCLUSIONS: The EEA+cMTO multiport technique may be a better alternative than the EEA+CTM multiport approach for targeting the petroclival region. However, clinical validation is required to confirm these laboratory findings.

Microsurgical Clipping of a Ruptured Basilar Apex Aneurysm: Contending with a Formidable Clinical Scenario.

Basilar apex aneurysms (BAAs) are challenging and mortal vascular pathologies. In this operative video, we present the case of a 58-year-old female who was admitted with a severe headache and loss of consciousness. She was diagnosed with a ruptured BAA (Fisher grade 4) and merited microsurgical management after careful consideration of aneurysmal features, available clinical setting resources, and discussion with her family. Anatomical nuances for approaching these lesions stepwise are demonstrated in the attached video. Unfortunately, despite adequate surgical treatment, she developed postoperative complications such as cerebral vasospasm and pneumonia, which led to her death. Overall, this technical report depicts the formidable nature of BAA and prompts the need for optimized protocols during the postoperative stage.

Heterozygous Apex1 deficiency exacerbates lipopolysaccharide-induced systemic inflammation in a murine model.

The biological role of apurinic/apyrimidinic endonuclease 1/redox factor-1 (Apex1) in modulating systemic inflammation remains unclear. This study aimed to assess the impact of Apex1 deficiency on systemic inflammation triggered by lipopolysaccharide (LPS) in a murine model. The methods involved transcriptomic analysis and assessments of inflammatory responses in age-matched 8-week-old Apex1+/- and wild-type Apex1+/+ mice, generated using the CRISPR/Cas9 system. Apex1+/- mice displayed no overt changes in body weight, however, Apex1 protein expressions in tissues were significantly reduced compared to wild-type mice. Furthermore, in Apex1+/- mice transcriptomic analysis showed that genes associated with antioxidant pathways were downregulated, and levels of superoxide production, 8-hydroxy-2'-deoxyguanosine (8-OHdG), and malondialdehyde (MDA) were increased. Moreover, hematological analysis showed increased neutrophil levels and a twofold increase in the count of splenic lymphocyte antigen 6 family member G+ (Ly6G+) neutrophils in the Apex1+/- mice compared to those in Apex1+/+ mice. Furthermore, following LPS treatment, the levels of cytokines and chemokines, including interleukin-1ß, interleukin-10, tumor necrosis factor-α, and monocyte chemoattractant protein 1, increased in the Apex1+/- mice. The Kaplan-Meier curve showed a significant reduction in the survival rates of Apex1+/- mice treated with LPS compared to those of Apex1+/+ mice. The hepatic and lung injury scores and Ly6G+ neutrophil infiltration levels also increased in Apex1+/- mice after LPS treatment. These results showed that Apex1 deficiency exacerbated the LPS-induced tissue damage in the lung and liver. These findings illustrate that in vivo Apex1 deficiency exacerbates LPS-induced systemic inflammation, tissue damage, and mortality in a murine model, highlighting the crucial role of Apex1 in mitigating inflammatory responses and maintaining a holistic physiological equilibrium.

Dynamic changes in the proximitome of neutral sphingomyelinase-2 (nSMase2) in TNFα stimulated Jurkat cells.

Ceramides generated by the activity of the neutral sphingomyelinase 2 (nSMase2) play a pivotal role in stress responses in mammalian cells. Dysregulation of sphingolipid metabolism has been implicated in numerous inflammation-related pathologies. However, its influence on inflammatory cytokine-induced signaling is yet incompletely understood. Here, we used proximity labeling to explore the plasma membrane proximal protein network of nSMase2 and TNFα-induced changes thereof. We established Jurkat cells stably expressing nSMase2 C-terminally fused to the engineered ascorbate peroxidase 2 (APEX2). Removal of excess biotin phenol substantially improved streptavidin-based affinity purification of biotinylated proteins. Using our optimized protocol, we determined nSMase2-proximal biotinylated proteins and their changes within the first 5 min of TNFα stimulation by quantitative mass spectrometry. We observed significant dynamic changes in the nSMase2 microenvironment in response to TNFα stimulation consistent with rapid remodeling of protein networks. Our data confirmed known nSMase2 interactors and revealed that the recruitment of most proteins depended on nSMase2 enzymatic activity. We measured significant enrichment of proteins related to vesicle-mediated transport, including proteins of recycling endosomes, trans-Golgi network, and exocytic vesicles in the proximitome of enzymatically active nSMase2 within the first minutes of TNFα stimulation. Hence, the nSMase2 proximal network and its TNFα-induced changes provide a valuable resource for further investigations into the involvement of nSMase2 in the early signaling pathways triggered by TNFα.

Comparison of femoral tunnel position and knee function in anterior cruciate ligament reconstruction: a retrospective cohort study using measuring-fluoroscopy method versus bony marker method.

BACKGROUND: Previous studies have shown that surgical technique errors especially the wrong bone tunnel position are the primary reason for the failure of anterior cruciate ligament (ACL) reconstruction. In this study, we aimed to compare the femoral tunnel position and impact on knee function during the ACL reconstruction using measuring combined with fluoroscopy method and bony marker method for femoral tunnel localization. METHODS: A retrospective cohort study of patients undergoing ACL reconstruction using the bony marker method or measuring combined with fluoroscopy for femoral tunnel localization was conducted between January 2015 and January 2020. A second arthroscopic exploration was performed more than 1 year after surgery. Data regarding patient demographics, the femoral tunnel position, results of the Lysholm score, the International Knee Documentation Committee (IKDC) score, KT-1000 side-to-side difference, pivot shift grade, and Lachman grade of the knee were collected. RESULTS: A total of 119 patients were included in the final cohort. Of these, 42 cases were in the traditional method group, and 77 cases were in the measuring method group. The good tunnel position rate was 26.2% in the traditional method group and 81.8% in the measuring method group (p < 0.001). At the final follow-up, the Lysholm and IKDC scores were significantly greater in the measuring method group than the traditional method group (IKDC: 84.9 ± 8.4 vs. 79.6 ± 6.4, p = 0.0005; Lysholm: 88.8 ± 6.4 vs. 81.6 ± 6.4, p < 0.001). Lachman and pivot shift grades were significantly greater in the measuring method group (p = 0.01, p = 0008). The results of KT-1000 side-to-side differences were significantly better in the measuring method group compared with those in the traditional method group (p < 0.001). CONCLUSIONS: The combination of the measuring method and intraoperative fluoroscopy resulted in a concentrated tunnel position on the femoral side, a high rate of functional success, improved knee stability, and a low risk of tunnel deviation. This approach is particularly suitable for surgeons new to ACL reconstructive surgery.

Endoscopic-assisted transorbital extended orbital exenteration: A multi-institutional preclinical study.

BACKGROUND: Sinonasal malignancies with orbital invasion have dismal prognosis even when treated with orbital exenteration (OE). Sugawara et al. developed a surgical strategy called "extended-OE (EOE)," showing encouraging outcomes. We hypothesized that a similar resection is achievable under endoscopic guidance through the exenterated orbit (endoscopic-EOE). METHODS: The study was conducted in three institutions: University of Vienna; Mayo Clinic; University of Insubria; 48 orbital dissections were performed. A questionnaire was developed to evaluate feasibility and safety of each step, scoring from 1 to 10, ("impossible" to "easy," and "high risk" to "low risk," respectively), most likely complication(s) were hypothesized. RESULTS: The step-by-step technique is thoroughly described. The questionnaire was answered by 25 anterior skull base surgeons from six countries. Mean, median, range, and interquartile range of both feasibility and safety scores are reported. CONCLUSIONS: Endoscopic-EOE is a challenging but feasible procedure. Clinical validation is required to assess real-life outcomes.

A proximity proteomics pipeline with improved reproducibility and throughput.

Proximity labeling (PL) via biotinylation coupled with mass spectrometry (MS) captures spatial proteomes in cells. Large-scale processing requires a workflow minimizing hands-on time and enhancing quantitative reproducibility. We introduced a scalable PL pipeline integrating automated enrichment of biotinylated proteins in a 96-well plate format. Combining this with optimized quantitative MS based on data-independent acquisition (DIA), we increased sample throughput and improved protein identification and quantification reproducibility. We applied this pipeline to delineate subcellular proteomes across various compartments. Using the 5HT2A serotonin receptor as a model, we studied temporal changes of proximal interaction networks induced by receptor activation. In addition, we modified the pipeline for reduced sample input to accommodate CRISPR-based gene knockout, assessing dynamics of the 5HT2A network in response to perturbation of selected interactors. This PL approach is universally applicable to PL proteomics using biotinylation-based PL enzymes, enhancing throughput and reproducibility of standard protocols.

Research on species categorical authentication of accelerants based on flame characteristics analysis.

Species categorical authentication of accelerants has traditionally relied on fire debris analysis. To explore a novel method for identifying the accelerants species, four commonly used accelerants for arson were loaded onto different substrates and ignited at different locations. The entire combustion process was recorded and flame characteristics were analyzed. The results showed that the probability density function (PDF) of flame apex angle counts within a certain period after ignition can be used to distinguish accelerant species, and this method is not affected by accelerant loading amount, ignition location, and substrate, demonstrating strong stability and universality, while the temporal variation of flame area and the value obtained by dividing half of the flame width by the flame height (tangent of flame cone angle) can effectively differentiate gasoline and diesel. The utilization of flame characteristics for identifying accelerants species holds significant implications for arson investigation.