Preparation Protocol Research Articles

Navigating the combinations of platelet-rich fibrin with biomaterials used in maxillofacial surgery.

Platelet-rich fibrin (PRF) is a protein matrix with growth factors and immune cells extracted from venous blood via centrifugation. Previous studies proved it a beneficial biomaterial for bone and soft tissue regeneration in dental surgeries. Researchers have combined PRF with a wide range of biomaterials for composite preparation as it is biocompatible and easily acquirable. The results of the studies are difficult to compare due to varied research methods and the fact that researchers focus more on the PRF preparation protocol and less on the interaction of PRF with the chosen material. Here, the literature from 2013 to 2024 is reviewed to help surgeons and researchers navigate the field of commonly used biomaterials in maxillofacial surgeries (calcium phosphate bone grafts, polymers, metal nanoparticles, and novel composites) and their combinations with PRF. The aim is to help the readers select a composite that suits their planned research or medical case. Overall, PRF combined with bone graft materials shows potential for enhancing bone regeneration both in vivo and in vitro. Still, results vary across studies, necessitating standardized protocols and extensive clinical trials. Overviewed methods showed that the biological and mechanical properties of the PRF and material composites can be altered depending on the PRF preparation and incorporation process.

Gas Chromatography Tandem Mass Spectrometry (GC-MS/MS) for High-Throughput Screening of Pesticides in Rice Samples Collected in Bangladesh and Saudi Arabia

Gas Chromatography Tandem Mass Spectrometry (GC-MS/MS) with modified QuEChERS sample preparation has been applied to the high-throughput screening of pesticide residuals in rice collected from Bangladesh and Saudi Arabia markets. Both countries consume high volumes of rice, which is a fundamental food for their populations. We report optimized sample preparation and mass spectrometry analysis protocols, which can be rapidly deployed in analytical laboratories. The screening of four groups (organophosphorus, synthetic pyrethroid, organonitrogen, and organochlorine) of a total of 115 pesticides can be performed within ~10 min using a matrix-matched calibration. For most compounds, the limits of detection and quantification (LOD/LOQ) are well below the maximum residue levels (MRLs) of the main regulators. The method generally demonstrates acceptable recovery values (91 compounds 75–125% and 10 compounds 30–75%). Out of 55 rice samples analyzed, 16 samples (29%) contained pesticide residues above LOQ. Four samples contained chlorpyrifos with concentrations ranging from 21.3 to 71.9 µg/kg, ten samples contained tebuconazole (34.7–69.0 µg/kg), and three samples contained pirimiphos methyl (10.7–20.7 µg/kg). The concentrations of the pesticide residues detected in these samples are well below MRL of FAO/WHO (chlorpyrifos, 500 µg/kg; tebuconazole, 1500 µg/kg; pirimiphos methyl, 7000 µg/kg).

Standardization of Protocol for Production of Synthetic Seed of Brahmi (Bacopa monnieri) and Assessment of their Longevity under Differential Storage Condition

Brahmi [Bacopa monnieri (L) Pennel] is an important medicinal small succulent, medicinal herb belongs to the family ‘Scrophulariaceae’ which is commonly known as Jala Brahmi or Nira-Brahmi. National Medicinal Plants Board (NMPB), estimated an annual demand of Bacopa in 2004-2005 was 6621.8 tonnes, with a 7% annual growth rate. Micropropagation techniques and production of synthetic seeds can be a breakthrough to overcome the low availability of Brahmi propagating materials. So, the present study was conducted to standardize the protocol for preparation of synthetic seeds of Brahmi using nodal segment. The experiment was conducted with five treatments by using combinations of different concentrations of sodium alginate and calcium chloride and preparing two types of synthetic seeds (single layer and double layer). Further the longevity of both single layer and double layered synthetic seeds was studied under two different storage temperatures at 25°C (ambient condition) and 4°C (refrigerator) up to 40 days by inoculating them in two different types of media. The result showed no significant effect of concentrations of gelling agent and complexing agent on the germination percentage of synthetic seeds. The number of shoots per synthetic seed varied from 22.00 to 29.35 with 100% germination and maximum shoots/synthetic seed and shoot length was obtained by using T5 (single layered with MS media, 3% sodium alginate and 2.5% calcium chloride). Maximum number of days of storage can be observed in the single layer synthetic seeds when stored at 4°C along with media followed by double layer synthetic seeds when stored at 4°C along with media. However, seeds stored on vermiculite got dried up in the initial days of storage. Germination was found to be higher in seeds stored at 4°C when compared to seeds stored at 25°C. The study as a whole elucidated the opportunity for the use of synthetic seed as an alternative to traditional propagation system for Brahmi.

Platelet-rich fibrin in the management of oral mucosal lesions: a scoping review

ObjectivesOral mucosal lesions are prevalent and often cause pain, thus impacting patients' quality of life. Platelet-rich fibrin (PRF) has emerged as a promising autologous biomaterial for wound healing, yet comprehensive evidence regarding its efficacy in treating oral mucosal lesions is limited. This study aims to update the current evidence on the effectiveness of PRF in treating various types of oral mucosal lesions.Materials and methodsWe conducted a literature search in PubMed, Scopus, Embase, and Web of Science databases until April 2024. The search included studies that investigated the use of PRF in treating oral mucosal lesions. Twelve studies met the inclusion criteria, comprising three case reports, three randomized controlled trials, two animal studies, three split-mouth trials, and one retrospective study. We performed data extraction according to a predefined form.ResultsPRF was applied in two forms—membranes and injectable gels—to treat a range of oral mucosal lesions, including ulcerative, red and white, pigmented, and potentially malignant or malignant lesions. Compared to control groups or conventional treatments, PRF generally demonstrated superior outcomes regarding faster healing, lesion size reduction, symptom relief, and lower recurrence rates. Histological and molecular analyses from some studies also indicated PRF's regenerative and anti-inflammatory effects.ConclusionPRF shows promise as an effective and safe alternative to current treatments for oral mucosal lesions due to its autologous nature, ease of preparation, and wound-healing capabilities. However, further research is needed to standardize PRF preparation protocols and confirm its long-term efficacy across different lesion types.

12209 Validation Of A Novel Placenta Explant Protocol For Studying Circadian Rhythms In The Mouse Placenta In Early, Mid And Late Pregnancy

Abstract Disclosure: J.S. Lee: None. A.M. Yaw: None. H.M. Hoffmann: None. Introduction: The placenta serves as an interface facilitating the exchange of essential nutrients and oxygen between the mother and fetus. Preeclampsia, a complication of pregnancy driven by placental dysfunction, is one of the leading causes of maternal, fetal, and neonatal morbidity and mortality. While deregulation of the genes driving circadian rhythm has been associated with preeclampsia, the role of circadian rhythms in the placenta largely remains unknown. To address this gap of knowledge, our goal was to establish and validate an organotypic ex vivo preparation to study placenta circadian rhythms at gestation day (GD) 10, GD14, and GD18 in the mouse. Methods: To establish an organotypic placenta culture GD10, GD14, and GD18 placental explants, we sectioned the placenta into 350μm sections using a vibratome. The explants were prepared from the validated circadian Per2::Luciferase reporter mouse, allowing automated circadian rhythms recording of the explants in a LumiCycle. We compared explant success rate and rhythm quality in 2x2mm section vs whole slice explants at GD14. Using hematoxylin and eosin (H&E) staining we validated the explants to be decidua which is strictly of maternal origin, labyrinth which is strictly of fetal origin, and the junctional zone which consists of binucleated cells fused of maternal and fetal cells. Results: Through H&E staining, we confirmed the decidua is enriched with neutrophils, the junctional zone contains spongiotrophoblasts and glycogen trophoblasts, and the labyrinth has red blood cells. Both explant types were enriched with >80% of cells from the targeted region. We next established a protocol to record Per2::Luciferase rhythms for <6 days. The fragility of the GD10 placentas required the whole explant protocol, whereas the GD18 2x2mm sectioned explants produced strong rhythms with a high success rate. At GD14, the whole explants resulted in stronger rhythms with higher amplitude and higher goodness of fit compared to the 2x2mm explants. Conclusion: We found that depending on GD recording success differs. Specifically, the whole placental explant provides a more robust and reliable model for studying circadian rhythms in GD10 and GD14, whereas 2x2mm explants are successful at GD18. This novel placental preparation protocol will be valuable in studying disease- and drug-induced changes to circadian rhythms in a placenta layer-specific manner in the mouse. Presentation: 6/1/2024

Applications of Platelet Concentrates (PCs) in Regenerative Onco-Urology: A Systematic Review of Literature

Objective: To assess the effectiveness of Platelet Concentrates (PCs) in the contest of Hemorrhagic, Actinic, and Radiation Cystitis, plus Urethral Obstruction or Stenosis. Eligibility criteria: Open article in English or Italian regarding in situ applications of PCs for the selected pathologies. Information sources: MEDLINE, Cochrane Library, and ELSEVIER. Risk of bias: High (and discussed). Methods for synthesis of results: Selection of relevant contents, resumed by digital tools, checked by authors and used throughout the manuscript. Included studies: 13 screened articles + 7 personal sources + 37 “extra” articles. Synthesis of results: Pre-clinical and clinical studies demonstrated substantial symptom relief, mucosal restoration, and improved growth factor levels, reducing recurrence rates and complications. However, preparation protocols and results varied among studies. Limitations of evidence: Frequent low-quality studies with mall sample size, plus heterogeneous experimental setups and nomenclature/preparations. Interpretation: PCs demonstrate promise due to their bioactive components, enhancing tissue repair and reducing inflammation with no significant adverse events. Despite positive outcomes in pre-clinical and clinical studies, variability in preparation protocols and small sample sizes, together with inconsistent results, highlight the need for high-quality research to validate PCs’ clinical efficacy and cost-effectiveness.

Electrodeposition of graphene oxide on titanium foil. Application field: Rechargeable batteries

The method of electrodepositing graphene oxide on titanium utilizes a thin grade 1 titanium foil with a thickness of 0.2 mm, a width of 10 mm, and a rectangular graphite plate with a parallelepiped shape with a thickness of 15 mm, a width of 40 mm, and a length of 100 mm. The titanium foil has a cut on one side, forming an isosceles triangle with a slightly inclined tip, creating an angle where the distance between the tip and the vertical symmetry axis of the foil is 3 mm. The titanium foil is positioned parallel to the graphite plate, with the tip as the only electrical contact. The two electrodes, titanium and graphite, assembled in this way, are immersed in an electrolytic solution composed of water and sulfuric acid. When a direct current voltage is applied between the two electrodes, a strong electric field is generated at the contact point, resulting in high temperature and the production of ultraviolet rays. Following a treatment described later in the so-called “preparation protocol,” reduced graphene oxide (rGO) is produced in suspension in the solution at the contact point between the tip of the titanium foil and the graphite plate, which, due to the electric field, tends to deposit on the titanium surface. A possible application found experimentally refers to rechargeable batteries.

Evaluating Bacterial Viability in Faecal Microbiota Transplantation: A Comparative Analysis of InVitro Cultivation and Membrane Integrity Methods.

Faecal microbiota transplantation (FMT) is a developing therapy for disorders related to gut dysbiosis. Despite its growing application, standardised protocols for FMT filtrate preparation and quality assessment remain undeveloped. The viability of bacteria in the filtrate is crucial for FMT's efficacy and for validating protocol execution. We compared two methods-in vitro cultivation and membrane integrity assessment-for their accuracy, reproducibility and clinical applicability in measuring bacterial viability in frozen FMT stool filtrate. Bacterial viability in stool filtrate was evaluated using (i) membrane integrity through fluorescent DNA staining with SYTO9 and propidium iodide, followed by flow cytometry and (ii) culturable bacteria counts (colony-forming units, CFU) under aerobic or anaerobic conditions. Using different types of samples (pure bacterial culture, stool of germ-free and conventionally bred mice, native and heat-treated human stool), we refined the bacterial DNA staining protocol integrated with flow cytometry for assessment of bacterial viability in frozen human stool samples. Both the membrane integrity-based and cultivation-based methods exhibited significant variability in bacterial viability across different FMT filtrates, without correlation. The cultivation-based method showed a mean coefficient of variance of 30.3%, ranging from 7.4% to 60.1%. Conversely, the membrane integrity approach yielded more reproducible results, with a mean coefficient of variance for viable cells of 6.4% ranging from 0.2% to 18.2%. Bacterial viability assessment in stool filtrate using the membrane integrity method offers robust and precise data, making it a suitable option for faecal material evaluation in FMT. In contrast, the cultivation-dependent methods produce inconsistent outcomes.

Clinical Outcomes of Full Arch Immediate Fixed Prostheses Supported By a Novel Implant System with Low Speed Site Preparation A 5 Years Retrospective Case Series

Background: The aim of this case series was to evaluate the 5-years clinical and radiographic outcomes of immediate full-arch fixed prosthesis supported by a novel implant system characterized by a low-speed site preparation protocol. Methods: Six medically compromised patients (3 females and 3 males) were consecutively enrolled and treated. Each patient received a fixed full-arch rehabilitations supported by two axial and two tilted implants. A total of 24 implants (N1 System, Nobel Biocare AB, Göteborg, Sweden) were placed in three mandibles and three maxillae using a low-speed drilling protocol with no irrigation. Prosthetic loading was applied within 2 hours of surgery, while then definitive restorations were placed 4 months later. Patients were scheduled for follow-up every 6 months up to 2 years and then annually. Radiographic evaluation of marginal bone level change was performed on regular intervals. Results: Between February 2019 and July 2019, six patients received a full-arch fixed prosthesis on four implants. All implants reached a minimum insertion torque of 30 Newton for immediate loading. After 5 years of observation (range 62-67 months), no implants failed and all definitive prostheses were stable and functional, resulting in a cumulative survival rate of 100%. After 5 years, the marginal bone loss was 1.46 ± 0.16 mm for the mandible and 1.85 ± 0.18 mm for the maxilla. Two implants (one axial and 1 tilted) in 1 patient treated in the maxillary arch reported more then 2 mm of marginal bone loss after 5 years. Conclusions: Within the limitations of this study, the successful medium-term results seem to confirm that immediate fixed full-arch rehabilitation, supported by two anterior axial and two posterior tilted implants, could be an effective and biologically beneficial treatment for complete arch edentulism. The reduced bone remodeling evidenced after 5 years of function confirm the biologically friendly concept behind this novel implant system.

Optimizing renal transporter immunodetection: consequences of freeze-thaw during sample preparation.

Renal transporters (cotransporters, channels, and claudins) mediate homeostasis of fluids and electrolytes and are targets of hormonal and therapeutic regulators. Assessing renal transporter abundance with antibody probes by immunoblotting is an essential tool for mechanistic studies. Although journals require authors to demonstrate antibody specificity, there are no consensus guidelines for kidney sample preparation leading to lab-to-lab variability in immunoblot results. In this study, we determined the impact of sample preparation, specifically freeze-thawed (Frozen) versus freshly processed (Fresh) kidneys (female and male rats and mice) on immunoblot signal detection of 15 renal transporters and the impact of protease inhibitors during homogenization. In female Sprague-Dawley rat kidneys homogenized with aprotinin, Na2EDTA, PMSF, and phosphatase inhibitors, immunodetection signals were ∼50% lower in Frozen versus Fresh samples for most transporters. Inclusion of additional inhibitors (Roche cOmplete Protease Inhibitor, "+") only partially increased transporter immunoblot signals to near Fresh levels. In male Sprague-Dawley rats, immunoblot signal density was lower in Frozen+ versus Fresh+ despite additional inhibitors. In C57BL/6 male mice, immunoblot signals from proximal tubule transporters were lower in Frozen versus Fresh by ∼25-50% and greater in Frozen+. In contrast, immunodetection signal was equivalent in female Frozen+ versus female Fresh+ for claudin 2, villin, AQP1, NKCC2, NCC, ENaCβ, ENaCɣ, claudin 7, AQP2, NKAα1, and NKAβ1. Thus, kidney sample preparation variables, including freeze-thaw and protease inhibition, have substantial transporter-specific effects on quantification of renal transporter abundance by immunoblot. These findings underscore the critical importance of assessing and reporting the impact of sample preparation protocols on transporter recovery to ensure robust rigor and reproducibility. NEW & NOTEWORTHY Freeze-thawing kidneys before homogenization is widely accepted in renal research. This study demonstrates that if kidneys are freeze-thawed just once before homogenization, immunoblot signals are reduced in a transporter-specific manner in rats and mice dependent on sex and that immunoblot signals can be partially recovered by adding additional protease inhibitors. These findings underscore the critical importance of assessing the impact of sample preparation, including freeze-thaw versus fresh, to ensure robust rigor and reproducibility.

Investigating the Predictive Biomethods. of Hypertension (HTN) in Human Foetal Astrocytes (HFAs) using Modified Proteomic Methods

Background: Hypertension (HTN) is a major risk factor for Cerebrovascular and cardiovas-cular diseases, causing premature deaths. Hence, there is an urgent need for predictive bi-omarkers to detect HTN before its onset. Methods • Cell Culture Technique: Growth of HFAs, followed by conversion of normal (A2) HFAs intoreactive (A1) type, by using ATP solution. • Confocal Microscopy: Quantification of Glial Fibrillary Acidic Protein (GFAP) in A1 and A2 HFAs, by using antibody ab 7260 with AF 488. Proteomics Methos • Extraction of proteins from A1 and A2 HFAs, using Single-pot, Sol-id-phase-enhanced Sample Preparation (SP3) protocol. • Liquid Chromatography- Mass Spectrometry techniques for obtaining the lev-els of calcium-gated proteins in A1 and A2 cell-solutions. Results • Micrographs obtained by Leica Microsystem indicated thicker filaments in A1 HFAs compared to A2 HFAs. • Confocal results showed higher levels of GFAP in A1 HFAs (P> 0.05). • Proteomics data analysed by PEAKS software revealed that proteins, such as GFAP were higher in reactive HFAs. Conclusions: The results showed higher levels of GFAP in A1 HFAs. Therefore, early detection of GFAP in human body fluids could be a reliable predictive biomarker for HTN

Mouse Wound Models and Preparation of Single-Cell Suspensions.

The management of acute full-thickness skin injuries presents a considerable challenge in clinical practice. The complexity of wound healing, involving diverse cell populations and the intricate wound microenvironment, complicates the assessment of treatment effects and their interactions using traditional experimental approaches. Single-cell transcriptome sequencing technology has revolutionized the understanding of cellular functions and mechanisms during wound healing. However, the variability in methodologies for creating mouse wound models introduces confounding factors that can impact experimental results. Furthermore, the process of dissociating mouse skin tissues presents significant technical hurdles, highlighting the critical need for high-quality single-cell suspensions for accurate transcriptome sequencing. Therefore, this study aims to optimize the construction of mouse wound models to accurately assess changes in wound closure while eliminating variables that could influence the subsequent sequencing result. Additionally, the preparation of single-cell suspensions was performed using both enzyme preparation protocols and test kit protocols to optimize cost and effectiveness.

Updated annotation and meta-analysis of Brugia malayi transcriptomics data reveals consistent transcriptional profiles across time and space with some study-specific differences in adult female worm transcriptional profiles.

Genomics, transcriptomics, and proteomics have significantly advanced our understanding of obligately host-associated microbes, where interrogation of the biology is often limited by the complexity of the biological system and limited tools. This includes the causative agents of many neglected tropical diseases, including filarial nematodes. Therefore, numerous transcriptomics studies have been undertaken on filarial nematodes. Most of these transcriptomics studies focus on Brugia malayi, which causes lymphatic filariasis and is a laboratory model for human filarial disease. Here, we undertook a meta-analysis of the publicly available B. malayi transcriptomics data enabling the direct cross comparison of samples from almost a dozen studies. This reanalysis highlights the consistency of transcriptomics results across many different studies and experimental designs from across the globe for over a decade of research, across many different generations of a sequencing technology, library preparation protocols, and differential expression tools. Males and microfilariae across samples had similar expression profiles. However, female samples were clustered into two differential expression patterns that were significantly different from one another. Largely, we confirm previous results for all studies reanalyzed including tissue-specific gene expression and anti-Wolbachia doxycycline treatment of microfilaria. However, we did not detect previously reported differential expression upon in vitro or in vivo treatment with ivermectin, albendazole, and DEC, instead identifying a consistent lack of transcriptomic change upon exposure to these anthelminthic drugs. Updated annotation has been provided that denotes poorly supported genes including those overlapping rRNAs.

L'angioplastie du Tronc commun gauche en ambulatoire : Comment s'y prendre ?

The steady development of same-day discharge (SDD) PCIs in recent years means that, by 2024, we will be able to tackle increasingly complex lesions, including those of the left main artery. This strategy is supported by the literature, but requires an appropriate patient selection (based on medical and social criteria) and precise assessment of the anatomical complexity of the lesion and the foreseeable risks of the PCI. Strict adherence to pre-established service protocols for patient preparation and follow-up, as well as the establishment of clinical and paraclinical criteria for discharge to a conventional hospital unit, are essential to the success of an SDD PCI program.

An initial game-theoretic assessment of enhanced tissue preparation and imaging protocols for improved deep learning inference of spatial transcriptomics from tissue morphology.

The application of deep learning to spatial transcriptomics (ST) can reveal relationships between gene expression and tissue architecture. Prior work has demonstrated that inferring gene expression from tissue histomorphology can discern these spatial molecular markers to enable population scale studies, reducing the fiscal barriers associated with large-scale spatial profiling. However, while most improvements in algorithmic performance have focused on improving model architectures, little is known about how the quality of tissue preparation and imaging can affect deep learning model training for spatial inference from morphology and its potential for widespread clinical adoption. Prior studies for ST inference from histology typically utilize manually stained frozen sections with imaging on non-clinical grade scanners. Training such models on ST cohorts is also costly. We hypothesize that adopting tissue processing and imaging practices that mirror standards for clinical implementation (permanent sections, automated tissue staining, and clinical grade scanning) can significantly improve model performance. An enhanced specimen processing and imaging protocol was developed for deep learning-based ST inference from morphology. This protocol featured the Visium CytAssist assay to permit automated hematoxylin and eosin staining (e.g. Leica Bond), 40×-resolution imaging, and joining of multiple patients' tissue sections per capture area prior to ST profiling. Using a cohort of 13 pathologic T Stage-III stage colorectal cancer patients, we compared the performance of models trained on slide prepared using enhanced versus traditional (i.e. manual staining and low-resolution imaging) protocols. Leveraging Inceptionv3 neural networks, we predicted gene expression across serial, histologically-matched tissue sections using whole slide images (WSI) from both protocols. The data Shapley was used to quantify and compare marginal performance gains on a patient-by-patient basis attributed to using the enhanced protocol versus the actual costs of spatial profiling. Findings indicate that training and validating on WSI acquired through the enhanced protocol as opposed to the traditional method resulted in improved performance at lower fiscal cost. In the realm of ST, the enhancement of deep learning architectures frequently captures the spotlight; however, the significance of specimen processing and imaging is often understated. This research, informed through a game-theoretic lens, underscores the substantial impact that specimen preparation/imaging can have on spatial transcriptomic inference from morphology. It is essential to integrate such optimized processing protocols to facilitate the identification of prognostic markers at a larger scale.

The current state of tolerance induction in vascularized composite allotransplantation.

Significant advancements have been made in the field of vascularized composite allotransplantation (VCA); however, like solid organ transplantation, bypassing the recipient's immune response remains a significant obstacle to long-term allograft survival. Therefore, strategies to overcome acute and chronic rejection and minimize immunosuppressive therapy are crucial for the future of VCA. This review highlights recent attempts to induce tolerance in VCA and discusses key findings through a clinical lens. Promising VCA tolerance protocols are being investigated, with five recent studies illustrating various successes. These preclinical approaches demonstrate a correlation between the presence of donor-derived T cells and VCA tolerance, the importance of using clinically available reagents within preclinical protocols, and the ability to induce sustained tolerance through nonmyeloablative methods. Furthermore, environmental factors, such as NB-UVB light are being investigated for their immunomodulation properties and may influence VCA graft rejection. To widen the scope of VCA, minimization of immunosuppression is needed. Overall, tolerance induction protocols should have a low-toxicity level, minimally invasive induction therapies, and utilize short-term immunosuppressive medications. By examining the milestones of recent studies, researchers can gain new technical approaches to immune modulation and make data-driven amendments to tolerance protocols in preparation for clinical translation.

Electrochemical Analysis of Key Anticancer Herbal Drugs: Progress and Innovations in Emodin, Rutin, Berberine, Shikonin, and Sophoridine

Background: Anticancer herbal drugs have gained significant attention in pharmaceutical research due to their complex chemical profiles and multifaceted therapeutic effects. Electrochemical analysis has emerged as a powerful tool for studying these compounds, offering unique insights into their behavior and properties. Methods: This review examines recent advances in the electrochemical analysis of five key anticancer herbal drugs: emodin, rutin, berberine, shikonin, and sophoridine. Various electrochemical techniques, including cyclic voltammetry, differential pulse voltammetry, and square wave voltammetry, are discussed in relation to their application in detecting and characterizing these compounds. Results: Significant progress has been made in developing highly sensitive and selective electrochemical sensors for these herbal drugs. Nanomaterial-modified electrodes have consistently improved detection limits and expanded linear ranges. Compound-specific innovations in electrode modifications and measurement techniques have been tailored to the unique electrochemical properties of each drug. Conclusion: Electrochemical analysis of anticancer herbal drugs has advanced substantially, offering powerful tools for studying and utilizing these compounds in cancer research and treatment. Future directions include the development of multi-analyte sensors, integration with microfluidic technologies, and application of artificial intelligence for data analysis. Challenges remain in improving the stability of modified electrodes and standardizing protocols for sample preparation and analysis.

OPTIMIZING ULTRASONIC gDNA FRAGMENTATION FOR NGS SEQUENCING

The fragmentation of genomic DNA (gDNA) is a crucial step in many Illumina library preparation protocols, including the TSO 500 assay, aimed at achieving fragment sizes ranging from 90 to 250 base pairs (bp) for next-generation sequencing (NGS). However, challenges arise in practice due to variations in fragmentation times, which are influenced by different extraction methods and sample sources. To optimize ultrasonic gDNA fragmentation across various extraction methods for NGS sequencing applications. Shearing of gDNA was performed using the Covaris S220 system, employing microtubes containing a total volume of 130 microliters of diluted Tris EDTA (pH 8.0) and 100 ng of gDNA, following the manufacturer's protocol. gDNA samples were extracted using three different kits: AllPrep DNA/RNA FFPE (Qiagen) from FFPE blocks, ReliaPrep FFPE gDNA Miniprep system (Promega) from glass slides, and Gentra Puregene (Qiagen) from cell lines. The shearing times varied depending on the extraction method and sample source. gDNA extracted using the AllPrep DNA/RNA FFPE kit from FFPE blocks required shearing times ranging from 300 to 400 seconds. In contrast, gDNA extracted using the ReliaPrep FFPE gDNA Miniprep system from glass slides exhibited shearing times of approximately 480 to 500 seconds. Lastly, gDNA extracted using the Gentra Puregene kit from cell lines showed shearing times ranging from 150 to 180 seconds. Our study demonstrates the influence of gDNA extraction methods and sample sources on ultrasonic fragmentation efficiency for NGS sequencing. The observed differences in shearing times highlight the need for method optimization to achieve consistent and optimal fragment sizes across diverse sample types. These findings contribute to the development of standardized protocols for gDNA fragmentation, facilitating reliable and reproducible NGS data generation in various research and clinical settings. Further investigations into the underlying factors affecting fragmentation efficiency are warranted to advance our understanding and improve NGS library preparation methodologies.

Functionalized Amorphous Carbon Materials via Reactive Molecular Dynamics Simulations.

We derive a database of atomistic structural models of amorphous carbon materials endowed with exohedral functional groups. We start from phases previously derived using the DynReaxMas method for reactive molecular dynamics simulations, which exhibit atomistic and medium-length-scale features in excellent agreement with available experimental data. Given a generic input structure/phase, we develop postprocessing simulation algorithms mimicking experimental preparation protocols aimed at: (1) "curing" the phase to decrease the defect concentration; (2) automatically selecting the most reactive carbon atoms via interaction with a probe molecular species, and (3) stabilizing the phase by saturating the valence of carbon atoms with single-bond functional groups. Although we focus on oxygen-bearing functionalities, they can be replaced with other monovalent groups, such as -H, -COOH, -CHO, so that the protocol is quite general. We finally classify reactive sites in terms of their location within the structural framework and coordination environment (edges, tunnels, rings, aromatic carbons becoming aliphatic) and try to single out descriptors that correlate with tendency to functionalization.

Application of linaclotide in bowel preparation for colonoscopy in patients with constipation: A prospective randomized controlled study.

Colonoscopy plays a crucial role in the early diagnosis and treatment of colorectal cancer. Adequate bowel preparation is essential for clear visualization of the colonic mucosa and lesion detection. However, inadequate bowel preparation is common in patients with constipation, and there is no standardized preparation protocol for these patients. This study aimed to explore the effectiveness and tolerability of a pre-colonoscopy combination regimen of linaclotide and polyethylene glycol (PEG). In this prospective, single-center, randomized controlled trial, 322 participants were divided into two groups: a 3-L PEG+870-μg linaclotide group (administered as a single dose for 3days) and a 4-L PEG group. The primary endpoints were the Boston Bowel Preparation Scale (BBPS) score and the rate of adequate and excellent bowel preparation. Secondary endpoints were the rates of detection of colonic adenomas and polyps, cecal intubation rates, colonoscopy time, adverse reactions, patient satisfaction, and physician satisfaction. The study included 319 patients. The 3-L PEG+linaclotide group showed significantly higher rates of adequate and excellent bowel preparation than the 4-L PEG group (89.4% vs 73.6% and 37.5% vs 25.3%, respectively; P<0.05). The mean BBPS score for the right colon in the 3-L PEG+linaclotide group was significantly higher than that in the 4-L PEG group. There were no significant between-group differences regarding the detection rates of colonic polyps and adenomas (44.4% vs 37.7% and 23.1% vs 20.1%, respectively; P>0.05). There were no significant between-group differences regarding cecal intubation rates, colonoscopy operation, and withdrawal times. However, patient tolerance and sleep quality were better in the 3-L PEG+linaclotide group. The combination of 3-L PEG and 870-μg linaclotide, because of its lower volume of intake, can be considered as an alternative bowel preparation regimen for constipated patients undergoing colonoscopy, especially for the elderly.