Methods and Protocols最新文献

This study evaluated the concurrent validity of the Vitruve linear encoder compared to the T-Force device for measuring mean propulsive velocity (MPV) and peak velocity (PV) during the free-weight bench press exercise. Thirteen resistance-trained men participated in three sessions, during which MPV and PV were recorded simultaneously by both devices. The data were analysed using one-way ANOVA, Pearson's correlation, Bland-Altman analysis, and effect size calculations, with statistical significance set at p ≤ 0.05. The results showed discrepancies between the Vitruve and T-Force devices across different intensity levels. Specifically, the Vitruve device generally reported higher MPV and lower PV values, particularly at moderate and low intensities. Vitruve was deemed useful for MPV measurements, especially at velocities below 0.65 m/s during free-weight bench press exercises. In conclusion, the Vitruve device overestimated MPV and underestimated PV at moderate and low loads (>0.65 m·s^-1), with the discrepancies increasing as velocity rose. It can provide valuable data for monitoring and assessing resistance training programs focused on MPV at heavier loads (<0.65 m·s^-1). Researchers and practitioners should take these findings into account when incorporating the Vitruve into velocity-based strength training protocols.

Implant-supported prosthetic rehabilitation for patients with severely atrophic jaws is challenging due to complex anatomical considerations and the limitations of conventional augmentation techniques. This study explores the potential of subperiosteal (juxta-osseous) implants as an alternative solution, using finite element analysis (FEA) to evaluate mechanical performance. Realistic jaw models, developed from radiographic data, are utilized to simulate various implant configurations and load scenarios. Results indicate that different screw placements, implant designs, and structural modifications can significantly influence stress distribution and biomechanical behavior. Upper and lower jaw models were assessed under multiple load conditions to determine optimal configurations. Findings suggest that strategic adjustments, such as adding posterior screws or altering implant connections, can enhance load distribution and reduce stress concentration, particularly in critical areas. Tensile loads in critical bone areas near cortical fixing screws exceeded 50 MPa under anterior loading, while configurations with larger load distributions reduced stress on both implant and bone. The study provides evidence-based insights into optimizing subperiosteal implant design to improve stability, longevity, and patient outcomes.

Procyanidins are widely distributed in plant-derived foods, and consist of flavanol oligomers and polymers. Recently, the crown procyanidin sub-family, characterised by a unique macrocyclic structure, has been identified in grapes and wine. This study reports the development and validation of a rapid and quantitative analytical method measuring crown procyanidin concentration in red and white wines using ultra-high-performance liquid chromatography (UHPLC) coupled with a Q-TOF mass spectrometer. Validation followed international standards, demonstrating high sensitivity (LOQ = 0.033 mg/L), accuracy (recovery = 88.21% to 107.64%), repeatability (RSD = 1.99% to 11.03%), and intermediate reproducibility (RSD = 2.51% to 19.05%). Minimal matrix effects were observed, ensuring reliable and precise quantification across both wine types. The applicability of the method was confirmed through the successful analysis of wine samples, leading to the first quantification of crown procyanidins in white wine. Concentrations ranged from 0.81 mg/L to 15.88 mg/L in the different analysed wines. This validated method provides a valuable tool for the study of crown procyanidin profiles in various wine matrices and establishes a foundation for future research into the role of crown procyanidins in wines and other food and beverage matrices where these compounds may be present.

Background: Advanced airway management is of fundamental importance in almost all areas of anesthesiology, emergency medicine, and critical care. Securing the airway is of the utmost importance, as this is a prerequisite for the oxygenation of the human organism. The clinical relevance of airway management is particularly evident in the fact that the primary cause of significant anesthesia-related complications can be attributed to this field. The need for the systematic recording of these procedures and their complications, as well as structured training in airway management and the evaluation of outcome parameters, is, therefore, evident.

Methods: The HAPA-registry is a prospective and monocentric observational trial at the Department of Anesthesiology, Medical Faculty Heidelberg, University of Heidelberg, Germany. All patients requiring general anesthesia with consecutive advanced airway management during a surgical procedure were included. We, therefore, planned to include approximately 9000 patients in the first period. Following successful airway management, the anesthetist completed a case report form (CRF) in person. The intention was to record airway management cases on an annual basis for a period of several months, thus ensuring that the register remains up-to-date and that airway management procedures are continuously recorded.

Discussion: The aim of this study was to establish an airway registry that enables the systematic recording and evaluation of different methods of airway management. The registry can be used to monitor and evaluate the implementation of current guidelines and recommendations for action, as well as the rates of success and complications.

Multi-drug delivery systems have gained increasing interest from the pharmaceutical industry. Alongside this is the interest in amorphous solid dispersions as an approach to achieve effective oral delivery of compounds with solubility-limited bioavailability. Despite this, there is limited information regarding predicting the behavior of two or more drugs (in amorphous forms) in a polymeric carrier and whether molecular interactions between the compounds, between each compound, and if the polymer have any effect on the physical properties of the system. This work studies the interaction between model drug combinations (two of ibuprofen, malonic acid, flurbiprofen, or naproxen) dispersed in a polymeric matrix of hypromellose acetate succinate (HPMCAS) using a solvent evaporation technique. Hildebrand and Hansen calculations were used to predict the miscibility of compounds as long as the difference in their solubility parameter values was not greater than 7 MPa^1/2. It was observed that the selected APIs (malonic acid, ibuprofen, naproxen, and flurbiprofen) were miscible within the formed polymeric matrix. Adding the API caused depression in the Tg of the polymer to certain concentrations (17%, 23%, 13%) for polymeric matrices loaded with malonic acid, ibuprofen, and naproxen, respectively. Above this, large crystals started to form, and phase separation was seen. Adding two APIs to the same matrix resulted in reducing the saturation concentration of one of the APIs. A trend was observed and linked to Hildebrand and Hansen solubility parameters (HSP).

Quinine is known for treating malaria, muscle cramps, and, more recently, has been used as an additive in tonic water due to its bitter taste. However, it was shown that excessive consumption of quinine can have severe side effects on health. In this work, we utilized fluorescence spectroscopy to measure the concentration of quinine in commercial tonic water samples. An external standard method was used to calculate the concentrations of quinine in two commercially available tonic water brands, namely Canada Dry and Schweppes, and compare them to the maximum allowable concentration of quinine in beverages. Upon analysis of the data collected by five different groups, the levels of quinine were found to be above the average concentration in most commercial tonic water samples, but below the maximum permitted concentration. Moreover, the five replicate sets of data demonstrated high reproducibility of the method employed in this study. The simple yet instructive protocol that we developed can be adapted to determine the concentration of other fluorescent compounds in foods and beverages. Further, the presented method and detailed protocol can be easily adopted for undergraduate labs and in chemical education.

The micronucleus test is one of the most popular genotoxicity assays. In order to avoid underestimation of micronuclei frequencies by counting non-replicating cells, the cytokinesis-blocked micronucleus test has been developed. In this technique, only binucleated cells are scored. One underestimated problem is the potential difficulty in discriminating binucleated from mononucleated cells when using DAPI staining, i.e., the possibility that two neighboring mononucleated cells could be mistaken for a binucleated one. The new protocol presented here comprises the addition of acridine orange in order to stain the cytoplasm (in addition to DAPI to stain nuclei and micronuclei). This new technique can increase the sensitivity of the cytokinesis-blocked micronucleus test and avoid underestimation of micronuclei frequencies, an important issue when high doses are employed.

Cobalt is a trace element, crucial for red blood cell formation and neurological function. Cobalt toxicity is often only diagnosed after severe manifestations, including visual impairment. We aimed to investigate whether optical coherence tomography (OCT) and magnetic resonance imaging (MRI) can effectively detect cobalt-induced ocular toxicity in a murine model. Five wild-type mice (WT, C57Bl6) received daily intraperitoneal cobalt chloride injections for 28 days with a dosage of 12.5 mg/kg. Another 5 WT mice served as controls. After 28 days, all mice underwent manganese contrast-enhanced MRI and OCT examinations. Macroscopic and histological analysis of the enucleated eyes were performed. MRI revealed an increased signal in the optic nerves of injected mice. Anterion OCT provided in vivo visualization of the entire eye, demonstrating incipient cataract formation in the cobalt-injected mice. Both Spectralis domain OCT and Anterion, followed by histological analyses, confirmed preserved retinal structure with decreased thickness in the cobalt-injected group, with only minor neuronal damage and cell loss. Optic nerve analysis demonstrated myelin loss and increased inflammation with high levels of reactive gliosis. This study demonstrates optic neuropathy induced by cobalt toxicity, as shown by increased optic nerve signal on MRI without significant retinopathy. Anterion OCT showed incipient cataracts in the anterior segment.

This differential extraction protocol details the steps for isolating DNA from sample pads used in lateral flow immunochromatographic (LFI) tests, particularly for cases involving mixed biological samples such as semen and menstrual blood, or other evidence related to sexual assault. This procedure utilizes a differential extraction technique applied to sample pads from immunochromatographic tests, where the sample pads serve as the substrate. The method involves two sequential lysis steps to effectively separate non-sperm and sperm fractions, enabling the targeted isolation of distinct cell types for downstream DNA analysis. The efficiency of this procedure is demonstrated by the results within this paper, which highlights the successful recovery of both male autosomal and Y-STR profiles, even in mixed samples with a high female presence. Overall, this protocol demonstrates the effective recovery of DNA from sample pads, which is beneficial for forensic practitioners dealing with limited sample quantities, underscoring the value of using these pads in forensic analysis.

Culex pipiens is a major vector of pathogens, including West Nile and Usutu viruses, that poses a significant public health risk. Monitoring pyrethroid resistance in mosquito populations is essential for effective vector control. This study aims to evaluate four DNA extraction protocols-QIAsymphony, DNAzol^® Direct reagent, PrepMan^® Ultra Sample Preparation Reagent (USPR), and Chelex^® 100-to identify an optimal method to extract DNA from individual Culex pipiens, as part of a high-throughput surveillance of pyrethroid resistance using Real-Time Genotyping PCR. The target is the L1014F mutation in the voltage-sensitive sodium channel (VSSC) gene, which confers knockdown (kdr) resistance to pyrethroids. Mosquitoes were collected from wintering and summer habitats in Lazio and Tuscany, Italy, and DNA was extracted using the four methods. The quality, quantity, extraction time, and cost of the DNA were compared among the various methods. The PrepMan^® USPR protocol was the most efficient, providing high-quality DNA with a 260/280 purity ratio within the optimal range at the lowest cost and in a short time. This method also demonstrated the highest amplification success rate (77%) in subsequent real-time PCR assays, making it the preferred protocol for large-scale genotyping studies.