Semi-quantitative Analysis Research Articles

Preferences of Cancer Survivors for Follow-Up Care: A Systematic Review of Discrete Choice Experiments.

Cancer survivors frequently encounter multiple challenges, including physical, psychological, social, emotional, and financial difficulties. These challenges significantly impact their quality of life and recovery process. This systematic review intends to delineate and assess the evidence gathered from discrete choice experiments, aiming to unravel the preferences of cancer survivors towards their follow-up care. A systematic literature review was conducted across PubMed, Web of Science, Embase, CINAHL, PsycINFO, CNKI, Wanfang, and SinoMed databases up to July 2024. Two reviewers independently conducted the screening, data extraction, and quality appraisal of the studies. The collected data were synthesized through a narrative approach. Additionally, semi-quantitative analyses were used to evaluate the frequency, importance, and significance of each attribute. Eight studies conducted across four countries were analyzed, yielding 41 distinct attributes grouped into eight main categories and 14 subcategories, further refined by subject matter. Despite the differences in payment systems between countries, the type of provider topped the list of preferences for cancer survivors' follow-up care. Next in preference was the contact modality. Service quality (comprehensive plans, personalized service, detailed information) was also valued. However, opinions diverged on continuity, additional support, and service frequency. The attributes of follow-up care preferences integrated in this review demonstrate a diverse range among cancer survivors. Generally, they prioritize skilled professionals, flexible contact modality, and other high-quality follow-up care elements. Future studies should analyze cancer survivors' preferences for follow-up care from the perspectives of different stakeholders, and further consider internal and external factors to ensure authentic decision making.

Preliminary investigation of the effects of high-temperature thermal activation on the activity of graphite tailings and the mechanical properties of graphite tailings mortar

The disposal of graphite tailings, a byproduct of graphite mining, poses significant recycling challenges. This study explores eco-friendly cement mortar by partially replacing sand with activated graphite tailings, focusing on waste utilization. Activating the pozzolanic properties of graphite tailings at various thermal activation temperatures converts them into high-quality binder materials. Quantitative assessments of graphite tailings' activity, conducted through ion leaching tests and raw material diffraction analysis, produce an activity index Ka, evaluating their performance under different thermal activation temperatures. Variable testing of substitution rates and activation temperatures examines changes in mechanical properties, microstructure, and pore distribution of the graphite tailings mortar. Optimum results reveal a 30 % substitution rate and a 750°C activation temperature. Graphite tailings, as pozzolanic binder materials, enhance mortar strength by improving particle size distribution, reducing "micro-area bleeding" and "boundary effects". After thermal activation, reactive minerals on the tailings' surface undergo secondary hydration reactions with cementitious products, modifying chemical structures and micro-morphology at the interface. Semi-quantitative XRD analysis and FT-IR spectroscopy characterize the ensuing chemical reactions, unveiling molecular bond and functional group changes. The superior performance of thermally activated graphite tailings primarily stems from increased active SiO2 and Al2O3 post-activation. The active SiO2 and Al2O3 reacts with cementitious products to form C-(A)-S-H gels, filling micro-pores at the tailings-mortar interface, which is the key characteristic that enables graphite tailings to serve as high-quality pozzolanic binder aggregates. A predictive model for compressive strength, utilizing reference models from solid waste mortar concrete, quantifies the positive impact of chemical activation. The theoretical results obtained from this predictive model closely align with the experimental findings.

Using acoustic emission to understand the early-age reaction and cracking evolution of alkali-activated slag-brick powder paste

Real-time monitoring and accurately understanding the continuous occurring psychical-chemical characterization during the hardening process of alkali-activated materials present a formidable challenge. In this study, the early-age reaction process of alkali-activated slag-waste brick powder paste (ASWP) was in-situ and continuous monitored by using acoustic emission (AE) system and temperature. A novel machine learning method was employed to categorize the evolution of AE signals into three distinct stages: reaction acceleration, reaction stabilization and volume shrinkage stage.During reaction acceleration stage (0-2h), the waste brick powder (WBP) increased AE counts rate (4000-7300 counts/h), with RT values exceeding 200μs, indicating the settlement of WBP. Additionally, WBP reduced the high r-value AE activity in volume shrinkage stage, effectively mitigated the drastic volume shrinkage, and delayed its occurrence time by approximately 4 hours. Semi-quantitative analyses of FTIR and TG-DTG indicate that WBP reduced gel phase formation in the early-age stage, decreasing the relative areas in Q2 site by 16% before 36 hours. However, WBP positively impacted ASWP hydration at long ages, increasing Q2 site d areas by 60%-100%. AE technique offers a new perspective for in-depth understanding of the reaction mechanism and microstructural development of alkali-activated materials in situ and continuously.

Physical activity for the elderly: systematic review and synthesis of digital health interventions

Abstract Background Physical activity (PA) is key for the well-being of the elderly. Despite its proven efficacy, many older adults remain sedentary. Digital health interventions offer promising results. The study aimed to appraise scientific literature evidence regarding digital interventions to improve physical activity levels among community-dwelling adults aged 60 years or older. Methods A systematic review and semiquantitative analysis were performed. Digital interventions were included if using eHealth/mHealth tools, app-based technologies and text messaging. An intervention control was required. Main outcomes consisted in physical activity levels and related measures. A semiquantitative approach was chosen for evidence synthesis, using a harvest plot to visually facilitate the weighing of results with sample size and quality assessment. Results A total of 10 studies (13 publications) were included, from which five main intervention types were identified (2 studies per each intervention type). A semiquantitative evidence synthesis via a harvest plot was performed, to base the studies evaluation on the simultaneous consideration of sample size, study quality by type and effectiveness of intervention. Interventions adopting dynamically-adjusted exercise plans via wearables and app-based digital peer-support resulted the most successful, whereas PA digital coaching interventions resulted the least recommendable for adoption. Motivating interventions through digital platforms and digital diaries showed inconclusive results. Conclusions Personalized interventions using dynamically-adjusted wearable-based technology and app-based digital peer-support ones resulted as the most promising. Their ability to provide real-time, personalized feedback and foster social support is key for health promotion. It is crucial to leverage these promising tools for enhancing population health, integrating them into comprehensive, equitable, sustainable and engaging public health interventions. Key messages • Interventions using personalized and dynamically-adjusted plans result as the most effective to increase physical activity amond the elderly. • It is key to integrate effective digital health tools into comprehensive, equitable, sustainable and engaging public health interventions to enhance population health.

Calcareous nannoplankton fluctuation within the Albian-Cenomanian Boundary Event of the Tethyan Himalaya

A hemipelagic succession 29m thick, situated in South Tibet within the Tethyan Himalaya tectonic unit, has been investigated for its calcareous nannofossil content. A total of 17 samples were subject to qualitative and semi-quantitative analysis. The studied interval belongs to the upper Albian-lowermost Cenomanian and extends into the UC0 nannofossil zone; based on the last occurrence of Hayesites albiensis, the UC0a and UC0b-c subzones were recognized. The most abundant nannofossil of the Youxia section is Watznaueria barnesiae. Other common taxa are Eiffellithus turriseiffelii, Eprolithus floralis, Rhagodiscus spp., and Zeugrhabdotus spp. In the lowermost part of the studied section, below the beginning of the Albian-Cenomanian Boundary Event (ACBE), i.e., prior to the δ13C positive excursion related to OAE1d, the nannofossils confined to high paleolatitudes, namely Repagulum parvidentatum, Seribiscutum primitivum, and Sollasites horticus, are present with a low abundance. This occurrence is believed to be evidence of a short episode of cooler surface waters linked to a transgressive event. The nannofossil abundance and diversity, along with the fluctuation patterns of the nutrient and temperature indices throughout the section, reflects a primary signal of mesotrophic to eutrophic conditions from the base of the succession up to the two oldest δ13C peaks of ACBE, both late Albian in age and within the OAE1d. By contrast, the dominance of Watznaueria barnesiae, representing more than 80% of the total assemblages, along with the significant drop in abundance and diversity shown by nannofossils within late phases of ACBE, are interpreted as a diagenetic signal. Mesotrophic to eutrophic conditions returned towards the top of the studied succession, where Biscutum constans and Zeugrhabdotus erectus again show a higher abundance.

Multiconfigurational Excitonic Couplings in Homo- and Heterodimer Stacks of Azobenzene-Derived Dyes.

Molecular excitons play a major role within dye aggregates and hold significant potential for (opto)electronic and photovoltaic applications. Numerous studies have documented alterations in the spectral properties of dye homoaggregates, but only limited work has been reported for heteroaggregates. In this article, dimeric dye stacks were constructed from azobenzene-like dyes with identical or distinct structures, and their excitonic features were computationally investigated. Our results show that strong exciton coupling is not limited to identical chromophores, as often assumed, based on a recently made available Frenkel Exciton Hamiltonian and multiconfigurational plus second-order perturbation theory energetics methodology. Heteroaggregate stacks were found to exhibit different absorption features from the corresponding interacting monomers, indicating considerable coupling interactions between units. We analyzed how such coupling may vary according to various aspects, such as the relative positions of the interacting monomers or the differences in their energetics. Such qualitative and semiquantitative analyses allow the evaluation of the excitonic behavior of these dye aggregates to encourage further efforts toward a deeper understanding of the excitonic properties of tailored dye heteroaggregate systems.

The degenerated glenohumeral joint.

This study aimed to define the histopathology of degenerated humeral head cartilage and synovial inflammation of the glenohumeral joint in patients with omarthrosis (OmA) and cuff tear arthropathy (CTA). Additionally, the potential of immunohistochemical tissue biomarkers in reflecting the degeneration status of humeral head cartilage was evaluated. Specimens of the humeral head and synovial tissue from 12 patients with OmA, seven patients with CTA, and four body donors were processed histologically for examination using different histopathological scores. Osteochondral sections were immunohistochemically stained for collagen type I, collagen type II, collagen neoepitope C1,2C, collagen type X, and osteocalcin, prior to semiquantitative analysis. Matrix metalloproteinase (MMP)-1, MMP-3, and MMP-13 levels were analyzed in synovial fluid using enzyme-linked immunosorbent assay (ELISA). Cartilage degeneration of the humeral head was associated with the histological presentation of: 1) pannus overgrowing the cartilage surface; 2) pores in the subchondral bone plate; and 3) chondrocyte clusters in OmA patients. In contrast, hyperplasia of the synovial lining layer was revealed as a significant indicator of inflammatory processes predominantly in CTA. The abundancy of collagen I, collagen II, and the C1,2C neoepitope correlated significantly with the histopathological degeneration of humeral head cartilage. No evidence for differences in MMP levels between OmA and CTA patients was found. This study provides a comprehensive histological characterization of humeral cartilage and synovial tissue within the glenohumeral joint, both in normal and diseased states. It highlights synovitis and pannus formation as histopathological hallmarks of OmA and CTA, indicating their roles as drivers of joint inflammation and cartilage degradation, and as targets for therapeutic strategies such as rotator cuff reconstruction and synovectomy.

Innovative strategy for the treatment of oily wastewater by in-situ synthesis of nitrogen-doped biochar supported FeS for activation of peroxymonosulfate

ABSTRACT Disposing of oily wastewater poses a significant challenge in treating oilfield-produced wastewater treatment. This study developed a FeSNC-9/PMS system for the effective degradation of total petroleum hydrocarbons (TPHs) in oily wastewater, while increasing the value of excess sludge, thus achieving the dual purpose of waste treatment. This work involved the in-situ preparation of a porous nitrogen-doped biochar-supported iron sulphide catalyst material using surplus sludge from SBR. Compared to undoped FeS (NC-9), FeSNC-9 exhibited excellent pore structure and abundant functional groups. Fe-Nx served as an effective connecting site between FeS species and the graphite network of biochar. The FeSNC-9/PMS system significantly degraded 74.21% of TPHs within 300 min. The FeSNC-9/PMS system demonstrated remarkable TPHs degradation efficiency across a wide temperature range and under both weak acidity and near-neutral conditions The dominant reactive oxygen species were identified as SO4 •− and •OH, with O2 •− and 1O2 also confirmed as active species. Gas chromatography semi-quantitative analysis showed that the long-chain alkanes of C20-C30 in total petroleum hydrocarbons were significantly degraded into short-chain alkanes or completely mineralized. This work provides new insights for the low-cost and high-efficiency treatment of TPHs in oilfield-produced water, and delves into the activation mechanism of PMS and the degradation pathways of TPHs.

PD-L1 expression in testicular germ cell tumors undergoing spontaneous regression.

Spontaneous regression of testicular germ cell tumors is a well-known phenomenon; however, the precise mechanisms of spontaneous regression are still unknown. Our study aimed to investigate programmed death-ligand 1 (PD-L1) expression in spontaneously regressed testicular germ cell tumors, exploring the link between the immune response and spontaneous regression. From a sample of 356 testicular germ cell tumors, we singled out 5 completely regressed and 6 partially regressed tumors. In four out of six cases with partial regression, a residual seminoma component was found, while in the remaining two cases, an embryonal carcinoma component was found. Comparisons were made with 20 pure seminomas and 20 mixed germ cell tumors (MGCTs). A semiquantitative immunohistochemical analysis of PD-L1 expression in tumor cells and intra/peritumoral lymphocytes was performed. There was no PD-L1 expression in tumors with complete regression. All partially regressed tumors showed expression in intra/peritumoral lymphocytes within the tumor remnants. Expression was significantly more frequent in pure seminomas compared to MGCTs (P = 0.004). A positive correlation was demonstrated between the seminoma component and the proportion of PD-L1 positive lymphocytes, with a Kendall's Tau-b coefficient of 0.626 (P < 0.001). Tumor cells showed PD-L1 expression in three MGCTs within the embryonal carcinoma component. Our results support an immunological mechanism of spontaneous tumor regression, with the strongest potential in testicular tumors containing seminoma components. However, further research is necessary to determine the role of PD-L1 ligand more precisely in the microenvironment of spontaneously regressed tumors.

Quantitative assessment of poly- and perfluoroalkyl substances (PFASs) in aqueous film forming foam (AFFF)-impacted soils: a comparison of analytical protocols.

Quantitatively assessing all per- and poly fluoroalkyl substances (PFASs) in an environmental sample, particularly soils impacted by aqueous film forming foams (AFFFs), has proven to be a challenge. To make such an assessment, a comprehensive sample processing procedure and analytical tool must be used. However, doubts remain whether current analytical tools such as high-resolution mass spectrometry (HRMS) with targeted quantitation and semi-quantitative analysis of suspects (Semi-Q HRMS) or total organic fluorine (TOF) are capable of accurately quantifying all non-polymeric PFASs in a sample. Further, current comprehensive soil PFAS HRMS methods are incompatible with TOF, preventing direct comparisons of the approaches. To enable direct comparisons, a soil sample processing procedure that is comprehensive as well as compatible with multiple analytical tools is needed. In this study, we assessed the performance of a previously developed soil PFAS method, EPA Method 1633, and a hybrid solid phase extraction (SPE)-based method for characterizing AFFF-impacted soil composites while maintaining compatibility with multiple analytical tools (i.e., Semi-Q HRMS and TOF). Comparative results for AFFF-impacted soil composites indicate analysis via EPA Method 1633 (as compared to the novel hybrid method) results in maybeup to 75% of the PFAS mass being missed by only analyzing for compounds listed in EPA Method 1633. Simply expanding the EPA Method 1633 analyte list was insufficient to account for the missing mass: up to 69% of the PFAS mass was still missed because of EPA Method 1633's extraction and cleanup bias. Additionally, the novel method developed offers a more comprehensive analysis with minimal reductions to sensitivity when compared to those reported in EPA Method 1633, with limits of quantification ranging from 0.12 to 2.4 ng/g as compared to 0.16-4.0 ng/g, respectively. For these reasons, an alternative hybrid SPE-based method is proposed for comprehensive evaluation of PFASs in AFFF-impacted soils.

Neodymium-Facilitated Visualization of Extreme Phosphate Accumulation in Fibroblast Filopodia: Implications for Intercellular and Cell–Matrix Interactions

A comprehensive understanding of intercellular and cell–matrix interactions is essential for advancing our knowledge of cell biology. Existing techniques, such as fluorescence microscopy and electron microscopy, face limitations in resolution and sample preparation. Supravital lanthanoid staining provides new opportunities for detailed visualization of cellular metabolism and intercellular interactions. This study aims to describe the structure, elemental chemical, and probable origin of zones of extreme lanthanoid (neodymium) accumulation that form during preparation for scanning electron microscopy (SEM) analysis in corneal fibroblasts filopodia. The results identified three morphological patterns of neodymium staining in fibroblast filopodia, each exhibiting asymmetric staining within a thin, sharp, and extremely bright barrier zone, located perpendicular to the filopodia axis. Semi-quantitative chemical analyses showed neodymium-labeled non-linear phosphorus distribution within filopodia, potentially indicating varying phosphate anion concentrations and extreme phosphate accumulation at a physical or physicochemical barrier. Phosphorus zones labeled with neodymium did not correspond to mitochondrial clusters. During apoptosis, the number of filopodia with extreme and asymmetric phosphorus accumulation increases. Supravital lanthanoid staining coupled with SEM allows detailed visualization of intercellular and cell–matrix interactions with high contrast and resolution. These results enhance our understanding of phosphate anion accumulation and transfer mechanisms in cells under normal conditions and during apoptosis.

Computer Vision-Assisted Robotized Sampling of Volatile Organic Compounds.

In conventional chemical analysis, samples are homogenized, extracted, purified, and injected into an analytical instrument manually or with a certain degree of automation. Such complex methods can provide superior performance in terms of sensitivity or selectivity. However, in some cases, it would be advantageous to possess a method that circumvents those preparatory steps, which require much attention. Here, we present a facile analytical approach to sampling volatile organic compounds (VOCs). Solid specimens emitting VOCs can be dropped onto the drop-off zone at a random position without any special alignment. A computer vision system recognizes specimen position, and a robotic arm places a sampling probe in the proximity of the specimen. The probe aspirates the VOCs─emitted by the specimen─with the aid of a suction force. A portion of the gaseous extract is transferred to the tritium-based ion source of a drift-tube ion mobility spectrometer. The ion mobility spectrum is immediately displayed in the customized graphical user interface (GUI). The sampling system also features a function for flushing extract ducts with hot nitrogen gas. Multiple specimens can be dropped for analysis at the same time. In one embodiment, the system can distinguish fresh meat from spoiled meat. When two meat specimens are placed on the drop-off zone, they are immediately sampled by the robotic arm, analyzed, and labeled on the digital image displayed on the GUI. Thus, the developed autosampling platform provides a hassle-free way of qualitative or semiquantitative analysis of raw specimens.

Sample Preparation and Determination of Trace Elements in High-Entropy Alloys by Total Reflection X-Ray Fluorescence (TXRF)

High-entropy alloys (HEAs) have attracted significant attention in recent years owing to their exceptional material properties; however, trace impurities in these alloys can significantly affect their strength, toughness, and radiation resistance. This study employs total reflection X-ray fluorescence (TXRF) to determine the trace impurities in three common HEA powders prepared via suspension techniques. High-Z (Ca and Mn) and low-Z (Mg and Al) elements were quantified using the internal standard and calibration curve methods, respectively. The levels of high-Z elements are consistent with those determined using inductively coupled plasma – optical emission spectrometry (ICP-OES). Although the physical mechanisms exerted various effects on low-Z elements, qualitative and semi-quantitative analyses remained feasible, demonstrating the effectiveness of TXRF for impurities in HEAs. Additionally, the combination of suspension sample preparation and TXRF affords a nondestructive, rapid, and cost-effective methodology that requires minimal sample quantities. Consequently, TXRF is well suited for the rapid determination of impurities in HEAs.

7559 TRIM21 Regulates Activation of ERK1/2 to Promote Growth and Drug Resistance in Pituitary Neuroendocrine Tumors

Abstract Disclosure: Y. Liu: None. F. Liu: None. C. Li: None. T. Zhang: None. L. Xue: None. Z. Wu: None. Objective: To investigate the role of the TRIM family genes in pituitary neuroendocrine tumors (PitNETs) and provide new targets and strategies for the clinical treatment of these tumors. Methods: A CRISPR screening system targeting the TRIM family was constructed and validated using next-generation sequencing. Stable cell lines overexpressing or knocking out TRIM21 were generated, and functional validation was performed using assays such as CCK-8, colony formation, and xenograft tumor growth in nude mice. RNA-seq, mass spectrometry, immunohistochemistry, and immunoblotting were employed to explore the signaling pathways and mechanisms underlying TRIM21's actions. Immunoprecipitation, immunofluorescence, GST-pulldown, NanoBiT assays, and ubiquitination experiments were conducted to investigate the effects of TRIM21 on ERK1/2. Immunohistochemistry and immunoblotting were used to explore the role of TRIM21 in drug- resistant prolactinomas and the correlation between TRIM21 and p-ERK. Results: Through screening the proliferative and drug-resistant effects of the TRIM family in MMQ and GH3 cells, we found that knocking out TRIM21 may inhibit cell growth and enhance drug sensitivity. Subsequently, stable cell lines overexpressing or knocking out TRIM21 were generated in MMQ and GH3 cells, revealing that TRIM21 overexpression promotes pituitary tumor cell growth and resistance to drug treatment, while TRIM21 knockout inhibits cell growth and enhances drug sensitivity. Cell-derived xenograft models showed that TRIM21 overexpression promoted growth of GH3 cells in vivo and resistance to cabergoline (CAB), while TRIM21 knockout suppressed growth of MMQ cells in vivo. Further exploration of the signaling pathways influenced by TRIM21 revealed that TRIM21 knockout downregulates the MAPK signaling pathway, while TRIM21 overexpression upregulates it. TRIM21 overexpression upregulates p-ERK1/2 in the MAPK pathway, while TRIM21 knockout decreases p-ERK. However, there is no significant influence on ERK1/2, JNK, p-JNK, p38, or p-p38. Mechanically, TRIM21 interacted with ERK1/2, enhancing the K27-linked ubiquitination of ERK1/2 and promoting their interaction with MEK1/2. In addition, the TRIM21 with D-docking mutant or ERK1/2 with CD-domain mutant resulted in the loss of their interaction. Furthermore, TRIM21 and p-ERK levels were significantly elevated in drug-resistant prolactinomas and immunohistochemical semi-quantitative analysis demonstrated a positive correlation between TRIM21 and p-ERK. Conclusion: The ubiquitin ligase TRIM21 catalyzes K27-linked ubiquitination of ERK1/2, enhancing its interaction with MEK1/2 and activation, thereby promoting growth and drug resistance in pituitary neuroendocrine tumors. TRIM21 may serve as a potential target for the treatment of pituitary neuroendocrine tumors. Presentation: 6/3/2024

Screening and Functional Evaluation of Four Larix kaempferi Promoters.

Promoters are powerful tools for breeding new varieties using transgenic technology. However, the low and unstable expression of target genes is still a limiting factor in Larix kaempferi (Lamb.) Carr (Japanese larch) genetic transformation. In this study, we analyzed L. kaempferi transcriptome data, screened out highly expressed genes, cloned their promoters, and constructed plant expression vectors containing the β-glucuronidase (GUS) reporter gene driven by these promoters. Recombinant vectors were introduced into the L. kaempferi embryogenic callus by means of the Agrobacterium-mediated transient or stable genetic transformation method, and the promoter activity was then determined by measuring GUS expression and its enzyme activity in the transformed materials. Four highly expressed genes were identified: L. kaempferi Zhang Chen Yi-1 (LaZCY-1), Zhang Chen Yi-2 (LaZCY-2), Translationally Controlled Tumor Protein (LaTCTP), and ubiquitin (LaUBQ). The 2000 bp fragments upstream of ATG in these sequences were cloned as promoters and named pLaZCY-1, pLaZCY-2, pLaTCTP, and pLaUBQ. Semi-quantitative and quantitative RT-PCR analyses of transient genetic transformation materials showed that all four promoters could drive GUS expression, indicating that they have promoter activities. Semi-quantitative and quantitative RT-PCR analyses and the histochemical staining of stable genetic transformation materials showed that the pLaUBQ promoter had higher activity than the other three L. kaempferi promoters and the CaMV35S promoter. Thus, the pLaUBQ promoter was suggested to be used in larch genetic transformation.

B-281 Performance Characteristics Evaluation of the ARK Diagnostics Immunoassay for Xylazine/4-HydroxyXylazine in Urine

Abstract Background Xylazine exposure is currently common in some US cities, but routine laboratory testing is not available. We evaluated the ARK Diagnostics immunoassay for qualitative detection of xylazine/4-hydroxy Xylazine in urine, prior to its anticipated release as a Forensic Use Only assay. Evaluation of performance characteristics of the ARK assay were conducted at two university hospitals, using either the semi-quantitative application of the assay, as performed on the Roche Cobas 503 analyzer (Institution A), or the qualitative application of the assay, as performed on the Beckman Coulter AU480 analyzer (Institution B). Methods Xylazine (X) and 4-hydroxy Xylazine (4-OHX) were obtained from Cayman Chemical (Ann Arbor, MI). Pooled xylazine-free urine was spiked either with X or 4-OHX in the range of 0-2000 ng/mL. De-identified, to-be-discarded patient urine samples were obtained from among those submitted for routine drugs-of-abuse panel testing. X measurements on all samples were performed by LC-MS/MS; samples were segregated into X-NEGATIVE (X&amp;lt;10 ng/mL) and X-POSITIVE (X&amp;gt;=10 ng/mL). The ARK assay was conducted according to manufacturer’s instructions, either as a semi-quantitative assay (Institution A, Roche analyzer) or as a qualitative assay (Institution B, Beckman Coulter analyzer), with 10 ng/mL as the manufacturer-specified cutoff for ARK POSITIVE, using manufacturer-supplied calibration standards ranging from 0-500 ng/mL X. Results The ARK semi-quantitative analysis (Roche analyzer) showed essentially equivalent curvilinear response curves for either X or 4-OHX at concentrations below 1000 ng/mL; for the semi-quantitative ARK assay, we use “&amp;gt;1200 ng/mL” for X-ARK &amp;gt;1200 ng/mL. By mixing studies (Roche), there were neither positive nor negative interferences observed as assessed for common drugs of abuse or for common therapeutic drugs. The presence of hemoglobin, bilirubin or moderate lipemia were devoid of assay interference. Precision studies for manufacturer-supplied controls were as follows: intra-assay CVs were 6.9% for the NEGATIVE control (5.1±0.35 ng/mL) and 5.9% for the POSITIVE control (15.7±0.53 ng/mL) (n=20); interassay CVs were 7.8% for the NEGATIVE control (5.1±0.40 ng/mL) and 5.7% for the POSITIVE control (15.9±0.90 ng/mL) (n=20). For the qualitative ARK assay as performed on the Beckman analyzer, POSITIVE and NEGATIVE controls ran invariably according to their designations. For the semi-quantitative assay application (Roche analyzer): among 78 X-NEGATIVE samples by LC-MS/MS, there were 0% ARK-POSITIVE results (false-positive rate=0%); among 74 X-POSITIVE samples by LC-MS/MS, there was one ARK-NEGATIVE result (false-negative rate=1.4%); this sample (X=65 ng/mL) was found to have a high amorphous crystalline content, and was unable to be reanalyzed. For the qualitative assay application (Beckman Coulter analyzer): among 78 X-NEGATIVE samples there was one ARK-POSITIVE sample (false-positive rate=1.3%) for a sample with xylazine=8.8 ng/mL by LC-MS/MS, with undetectable 4-OHX (&amp;lt;10 ng/mL); among 74 X-POSITIVE samples, there were no ARK-NEGATIVE results (false-negative rate=0%). Conclusions The ARK xylazine immunoassay demonstrated acceptable analytical performance with respect to detection of xylazine in urine at 10 ng/mL. With additional consideration of the assay’s cross reactivity with the major xylazine metabolite, 4-OH-xylazine, the assay was judged to be highly suitable for routine use as a screening test for detection of xylazine exposure via urine testing.

The usefulness of evaluating PTEN expression for accurate grading of phyllodes tumors

Phyllodes tumors (PTs) are classified as benign, borderline, or malignant based on histologic characteristics. However, because histological criteria are subjective and diagnosis requires integrating multiple findings, discrepancies often occur between observers. Therefore, it is necessary to discover biomarkers based on the molecular characteristics of PTs. This study aimed to identify dysregulated microRNAs (miRNAs) in PTs through miRNA profiling and determine whether expression of their target genes could be useful as PT biomarkers. MiRNA profiling was performed on 13 PTs (three malignant, three borderline, seven benign) and six fibroadenomas, and predicted target genes of dysregulated miRNAs were selected using three computation algorithms. The expression of two miRNAs, miR-155 and miR-200c, was 1.69-fold and 1.61-fold higher, respectively, in borderline and malignant PT groups than in the benign PT group (p < 0.05). KEGG pathway analysis revealed that the 374 target genes of these two miRNAs (miR-155 and miR200c) participated in several signaling pathways, adherens junction, cell cycle, and pathway in cancer. Immunohistochemical staining for PTEN, one of candidate target genes of miR-200c, was performed on whole slides of PT tissue classified as malignant (n = 9), borderline (n = 12), or benign (n = 21). Stromal tumor cells of high-grade PTs (borderline and malignant) had significantly lower PTEN expression than those of low-grade PTs (benign) (p-value ≤0.001). Semiquantitative analysis of PTEN expression, score 0–8, revealed that it correlated with histologic findings. Low PTEN expression (s-score of 6 or less) was used as a diagnostic criterion for high-grade PTs, it showed 100 % sensitivity and 95.2 % specificity in 42 cases of PTs. Currently, PT grading based solely on subjective histologic findings is challenging, but semiquantitative PTEN expression analysis could provide a more accurate and objective way to grade PTs.

Influences of Ipomoea batatas Anti-Cancer Peptide on Tomato Defense Genes.

This study investigates the impact of IbACP (Ipomoea batatas anti-cancer peptide) on defense-related gene expression in tomato leaves, focusing on its role in plant defense mechanisms. Previously, IbACP was isolated from sweet potato leaves, and it was identified as a peptide capable of inducing an alkalinization response in tomato suspension culture media. Additionally, IbACP was found to regulate the proliferation of human pancreatic adenocarcinoma cells. Elucidate IbACP's molecular influence on defense-related gene expression in tomato leaves using next-generation sequencing analysis. To assess the impact of IbACP on defense-related gene expression, transcriptome data were analyzed, encompassing various functional categories such as photosynthesis, metabolic processes, and plant defense. Semi-quantitative reverse-transcription polymerase chain reaction analysis was employed to verify transcription levels of defense-related genes in tomato leaves treated with IbACP for durations ranging from 0 h (control) to 24 h. IbACP induced jasmonic acid-related genes (LoxD and AOS) at 2 h, with a significant up-regulation of salicylic acid-dependent gene NPR1 at 24 h. This suggested a temporal antagonistic effect between jasmonic acid and salicylic acid during the early hours of IbACP treatment. Downstream ethylene-responsive regulator genes (ACO1, ETR4, and ERF1) were consistently down-regulated by IbACP at all times. Additionally, IbACP significantly up-regulated the gene expressions of suberization-associated anionic peroxidases (TMP1 and TAP2) at all time points, indicating enhanced suberization of the plant cell wall to prevent pathogen invasion. IbACP enhances the synthesis of defense hormones and up-regulates downstream defense genes, improving the plant's resistance to biotic stresses.

Expression of Wnt signaling proteins in rare congenital bladder disorders

Introduction and AimsCongenital bladder anomalies are rare and are a leading cause of end stage renal failure in children. The Wnt signaling pathway, important during embryonic development, has been implicated in the pathogenesis of these conditions through regulation of gene expression, including essential transcription factors. We investigated the expression of four Wnt transcriptional targets, namely, Pygopus 1 (Pygo1), FRA1, TCF7L1 and Connexin 43 (Cx43) in three rare congenital bladder disorders: bladder exstrophy (BE), neurogenic bladder (NGB) and posterior urethral valves (PUV). MethodsBladder tissue samples were collected from patients at the Great Ormond Street Hospital for Sick Children, London, UK, with control (normally-functioning bladder, N=9), BE (N=15), NGB (N=6) and PUV (N=5). Histological analysis was performed using the van Gieson stain to differentiate smooth muscle (SM) and connective tissue (CT) compartments. An unbiased, automated, semi-quantitative immunofluorescence analysis was performed to measure the labelling intensity of four Wnt-related proteins in tissue from these four groups. Results and DiscussionThere was a significant (p<0.05) increase in the expression of Pygo1 in the smooth muscle of all anomalies examined and also in the connective tissue in PUV compared to control. Cx43 also showed overexpression in the smooth muscle across all conditions; however, there was a reduced expression in NGB and an increase in PUV in connective tissue. TCF7L1 showed a significant decrease in both tissue compartments for NGB, whereas FRA1 expression remained unchanged across all anomalies. We also measured colocalization of Wnt-related proteins. TCF7L1 exhibited increased colocalization with Pygo1 and FRA1 in exstrophy compared to control. These results suggest a complex dysregulation of the Wnt pathway in congenital bladder disorders. ConclusionWnt signaling-related proteins show dysregulation in congenital bladder disorders compared to control tissue. Understanding these mechanisms should help towards non-invasive early diagnosis, drug target discovery and development of treatment strategies for these conditions.Summary tableSummary of Wnt-related protein expression and colocalization in bladder exstrophy (BE), neurogenic bladder (NGB) and posterior urethral valves (PUV) disorder compared to normally-functioning control bladder. The table shows the levels of expression as significantly up (↑) down (↓) or no change (-) of Pygopus 1 (Pygo1), Connexin 43 (Cx43), FRA1 and TCF7L1 expression in smooth muscle and connective tissue compartments in BE, NGB and PUV relative to control bladder tissue samples. The table also shows colocalization of six protein pairs of Pygo1 and FRA1; Pygo1 and Cx43; Pygo1 and TCF7L1; FRA1 and Cx43; FRA1 and TCF7L1 and Cx43 and TCF7L1 in bladder disorders compared to control.Summary tableProtein Expression (Smooth muscle)Pygo1Cx43FRA1TCF7L1BE↑↑--NGB↑↑-↓PUV↑↑--Protein Expression (Connective tissue)BE----NGB-↓-↓PUV↑↑--ColocalizationPygo1/FRA1Pygo1/Cx43Pygo1/TCF7L1FRA1/Cx43FRA1/TCF7L1Cx43/TCF7L1Exstrophy--↑-↑-NGB------PUV------

Healing sequelae following tooth extraction and dental implant placement in an aged, ovariectomy model.

At present, a lack of consensus exists regarding the clinical impact of osteoporosis on alveolar bone metabolism during implant osseointegration. While limited preclinical and clinical evidence demonstrates a negative influence of osteoporosis on dental extraction socket healing, no preclinical studies offer data on the results of implant placement in 6-mo-old, ovariectomized (OVX) Sprague-Dawley rats. This study aimed to investigate the outcomes of dental tooth extraction socket healing and implant placement in a rodent model of osteoporosis following daily vehicle (VEH) or abaloparatide (ABL) administration. Micro-CT and histologic analysis demonstrated signs of delayed wound healing, consistent with alveolar osteitis in extraction sockets following 42d of healing in both the VEH and ABL groups. In a semiquantitative histological analysis, the OVX-ABL group demonstrated a tendency for improved socket regeneration with a 3-fold greater rate for moderate socket healing when compared to the OVX-VEH group (43% vs 14%), however, this finding was not statistically significant (p=.11). No significant differences were observed between vehicle and test groups in terms of implant outcomes (BMD and bone volume/total volume) at 14- and 21-d post-implant placement. Abaloparatide (ABL) significantly increased BMD of the femoral shaft and intact maxillary alveolar bone sites in OVX animals, demonstrating the therapeutic potential for oral hard tissue regeneration. The present model involving estrogen-deficiency-induced bone loss demonstrated an impaired healing response to dental extraction and implant installation.