Optimal Protocol Research Articles

Establishment of the complete life cycle of Calicophoron daubneyi under experimental conditions.

The complex life cycle of the rumen fluke Calicophoron daubneyi is similar to that of the liver fluke Fasciola hepatica. Interestingly, C. daubneyi and F. hepatica share the same intermediate host, Galba truncatula. However, in contrast to its relative, experimental production of metacercariae is a major challenge for C. daubneyi, hampering a detailed analysis of its life cycle, especially in the definitive host. G. truncatula snails collected from natural habitats were bred in glass Petri dishes and fed dried organic lettuce leaves. C. daubneyi eggs were obtained from feces of naturally infected cattle and incubated until miracidia were hatching. Subsequently, these miracidia were allowed to infect snails, which were kept under specific laboratory conditions to monitor the shedding of metacercariae. In total, 177 G. truncatula snails were exposed to C. daubneyi miracidia during eleven snail infection trials. Sixty-eight of these snails survived for longer than 30 days post-infection (p.i.). From day 35 p.i., seven snails from five trials started shedding an average number of 106 metacercariae (range: 38-186) per snail. Three ewe lambs (aged 7-10 months) were inoculated orally with 150 metacercariae each. A different batch of metacercariae (obtained from three different snail trials) was used for each lamb. Another two lambs served as controls. All animals were regularly examined clinically, hematologically and coproscopically, using sedimentation techniques for the detection of trematode eggs. Low numbers of C. daubneyi eggs were detected in fecal samples of two of the three inoculated lambs on day 86 post-inoculation (yielding ≤ 2 epg), but only one lamb continued to shed eggs (up to 6 epg) until the end of the experiment (day 104 post-inoculation). None of the animals showed any abnormal clinical findings or blood parameters throughout the course of the study. Production of C. daubneyi metacercariae under laboratory conditions is reported, followed by experimental infection of the definitive host, thus completing the full life cycle of this parasite under experimental conditions. However, neither the survival rate of the snails nor the amount of metacercariae produced were comparable to previously published experiments using F. hepatica, necessitating further optimization of the laboratory protocols. Nevertheless, the results can serve as a starting point for more in-depth studies of this increasingly important trematode.

Development of a 3-in-1 multifunctional cell processing system and optimization of cell type-dependent protocols for CPA addition/removal

Development of a 3-in-1 multifunctional cell processing system and optimization of cell type-dependent protocols for CPA addition/removal

Optimisation of low and ultra-low dose scanning protocols for ultra-extended field of view PET in a real-world clinical setting

True total-body and extended axial field-of-view (AFOV) PET/CT with 1m or more of body coverage are now commercially available and dramatically increase system sensitivity over conventional AFOV PET/CT. The Siemens Biograph Vision Quadra (Quadra), with an AFOV of 106cm, potentially allows use of significantly lower administered radiopharmaceuticals as well as reduced scan times. The aim of this study was to optimise acquisition protocols for routine clinical imaging with FDG on the Quadra the prioritisation of reduced activity given physical infrastructure constraints in our facility. Low-dose (1 MBq/kg) and ultra-low dose (0.5 MBq/g) cohorts, each of 20 patients were scanned in a single bed position for 10 and 15 min respectively with list-mode data acquisition. These data were then reconstructed simulating progressively shorter acquisition times down to 30 s and 1 min, respectively and then reviewed by 2 experienced PET readers who selected the shortest optimal and minimal acquisition durations based on personal preferences. Quantitative analysis was also performed of image noise to assess how this correlated with qualitative preferences. At the consensus minimum acquisition durations at both dosing levels, the coefficient of variance in the liver as a measure of image noise was 10% or less and there was minimal reduction in this measure between the optimal and longest acquisition durations. These data support the reduction in both administered activity and scan acquisition times for routine clinical FDG PET/CT on the Quadra providing efficient workflows and low radiation doses to staff and patients, while achieving high quality images.

Tailoring anaesthetic strategies for diabetes research: Acepromazine vs. medetomidine in Aachen minipigs.

Pre-established anaesthetic protocols in animal models might unexpectedly interfere with the main outcome of scientific projects and therefore they need to account for the specific research goals. We aimed to optimize the anaesthetic protocol and animal handling strategies in a diabetes-related-study exemplifying how the anaesthetic approach must be adjusted for individual research targets. Aachen minipigs were used as a model to test long-lasting skin glucose sensors for diabetic human patients. A total of 6 animals participated in two or three rounds of experiments. Each round lasted 2 months, with a maximum of 2 rounds per year. In each round, animals were anaesthetised 4 times: for glucose sensors insertion, twice for glucagon stress tests (GST), and a last time for removal of sensors. Acepromazine (ACE) was compared to medetomidine (MED) in association with butorphanol (BUT) and Ketamine (KET) and 4 parameters were analysed to define the optimum anaesthetic protocol including: sedation level, anaesthesia duration, effects on blood glucose and safety. ACE-BUT demonstrated a weaker sedative effect but reduced overall experimental time, minimized anaesthetic risk and minimally interfered with the glucose metabolism. The improvement obtained by animal conditioning and handling strategies applied in this study were not objectively estimated, although the aversion behavior was completely abolished. Based on the analysed parameters, the use of acepromazine is proposed to be superior when Aachen Minipigs are used specifically as a model for diabetes-related studies, albeit the recommendations for the anaesthesia of minipigs suggest otherwise.

Enhancing conservation and genetic resource management through cross-transferability exploration and optimization of DNA extraction protocol for Buchanania cochinchinensis (Lour.) M. R. Almeida

Enhancing conservation and genetic resource management through cross-transferability exploration and optimization of DNA extraction protocol for Buchanania cochinchinensis (Lour.) M. R. Almeida

P-839. Clinical Diagnostic Performance of Droplet Digital PCR for Pathogen Detection in Patients with Escherichia coli Bloodstream Infections

Abstract Background Rapid diagnosis and early administration of appropriate antimicrobials are crucial to improve the prognosis and decrease the mortality of patients with bacterial bloodstream infections (BSIs). Droplet digital PCR (ddPCR) is a new tool for pathogen detection in BSIs. This study aimed to examine the sensitivity of ddPCR and explore the association between bacterial DNA load in whole blood and time-to-positivity (TTP) of blood cultures (BCs) in patients with Escherichia coli BSIs. Relationship between E. coli DNA load in whole blood and time-to-positivity of blood cultures Methods This prospective study included 59 patients with E. coli BSIs confirmed using BCs enrolled at the Hiroshima University Hospital between June, 2023 and April, 2024. The E. coli DNA load in whole blood, which was simultaneously obtained from two sets of BCs, was evaluated using ddPCR. For each patient, only the TTP of the first positive BC obtained using the BacT/ALERT Virtuo System was used for analysis. Results The median TTP was 9.0 (IQR: 8.3–10.2) h, and 15.2% (9/59) of the patients exhibited septic shock during BC sampling. In 83.1% (49/59) of the whole blood samples, E. coli DNA was detected via ddPCR. Patients with positive ddPCR results exhibited significantly shorter TTP than those with negative ddPCR results (median TTP: 8.8 vs. 10.5 h; p < 0.001). The rate of positive results for both sets of BCs was significantly higher in patients with positive ddPCR results than in those with negative ddPCR results (87.8 vs. 40.0%, p = 0.003). The E. coli DNA load was 75–200,475 copies/mL. Moreover, ddPCR revealed a significant correlation between the E. coli DNA load in whole blood and TTP (r: –0.58; 95% confidence interval: –0.74 to –0.36; p < 0.001). All patients with septic shock exhibited positive ddPCR results and significantly higher E. coli DNA loads in whole blood than those without septic shock (median E. coli DNA load: 6,375 vs. 113 copies/mL; p < 0.001). Conclusion Overall, ddPCR effectively detected E. coli DNA in the whole blood of patients with septic shock with a relatively short TTP. Notably, E. coli DNA load was associated with the infection severity of the patient. Further optimization of the bacterial DNA extraction protocol from blood and reaction systems and conditions can enhance the sensitivity of ddPCR. Disclosures All Authors: No reported disclosures

In vitro high-throughput screening of the antimicrobial activity of different compounds against Xylella fastidiosa subsp. pauca

The worldwide distribution and the host range of the phytopathogenic bacterium Xylella fastidiosa (Xf) have significantly changed in the last decade with numerous outbreaks reported in the Old Continent. Among the different European isolates, those of the subspecies pauca have been ranked as highly pathogenic, being the causal agents of the olive quick decline decimating olive trees in southern Italy. Significant research investments have been devoted towards finding therapeutic approaches to mitigate the impact of the infections in highly susceptible host species. This study aimed to evaluate in vitro efficacy against Xylella fastidiosa subsp. pauca (Xfp) of different classes of products, including metal ions, micronutrients, antibiotics, and phenolic compounds. The slow and fastidious growth of the bacterium requires optimization of specific protocols to assess antibacterial activities and the effect on biofilm formation. The results showed a dose–response effect against Xf for most products. Notably, among micronutrients and phenolic compounds, CuSO4·5H2O, Dentamet®, pyrocatechol and 4-methylcatechol showed the highest bactericidal and antibiofilm activity. At the same time, antibiotics demonstrated substantial bacteriostatic activity effectively inhibiting biofilm formation. For metal ions, such as CoCl2, K2B4O7·4H2O and MnSO4·H2O, significant effects on bacterial cell viability were recorded but were not able to completely kill the bacterium. Regarding the antibiofilm activity, some of them were able to inhibit biofilm formation, while others increased its formation. Ca(NO3)2·4H2O and Na2MoO4 were found to promote the growth of Xf. The methodologies described proved to be useful for profiling the antimicrobial activity of a large panel of compounds and the data collected provide evidence of their in vitro effectiveness.Graphical

Automated estimation of individualized organ-specific dose and noise from clinical CT scans

Objective. Radiation dose and diagnostic image quality are opposing constraints in x-ray computed tomography (CT). Conventional methods do not fully account for organ-level radiation dose and noise when considering radiation risk and clinical task. In this work, we develop a pipeline to generate individualized organ-specific dose and noise at desired dose levels from clinical CT scans.Approach. To estimate organ-specific dose and noise, we compute dose maps, noise maps at desired dose levels and organ segmentations. In our pipeline, dose maps are generated using Monte Carlo simulation. The noise map is obtained by scaling the inserted noise in synthetic low-dose emulation in order to avoid anatomical structures, where the scaling coefficients are empirically calibrated. Organ segmentations are generated by a deep learning-based method (TotalSegmentator). The proposed noise model is evaluated on a clinical dataset of 12 CT scans, a phantom dataset of 3 uniform phantom scans, and a cross-site dataset of 26 scans. The accuracy of deep learning-based segmentations for organ-level dose and noise estimates was tested using a dataset of 41 cases with expert segmentations of six organs: lungs, liver, kidneys, bladder, spleen, and pancreas.Main results. The empirical noise model performs well, with an average RMSE approximately 1.5 HU and an average relative RMSE approximately 5% across different dose levels. The segmentation from TotalSegmentator yielded a mean Dice score of 0.8597 across the six organs (max = 0.9315 in liver, min = 0.6855 in pancreas). The resulting error in organ-level dose and noise estimation was less than 2% for most organs.Significance. The proposed pipeline can output individualized organ-specific dose and noise estimates accurately for personalized protocol evaluation and optimization. It is fully automated and can be scalable to large clinical datasets. This pipeline can be used to optimize image quality for specific organs and thus clinical tasks, without adversely affecting overall radiation dose.

Studying Cationic Liposomes for Quick, Simple, and Effective Nucleic Acid Preconcentration and Isolation.

To ensure high quality of food and water, the identification of traces of pathogens is mandatory. Rapid nucleic acid-based tests shorten traditional detection times while maintaining low detection limits. Challenging is the loss of nucleic acids during necessary purification processes, since elution off solid surfaces is not efficient. We therefore propose the development of a vanishing surface strategy in which cationic liposomes efficiently capture nucleic acids. A lipoplex is formed that can be easily centrifuged down and washed if needed. Adding the lipoplex to detergent solutions or nucleic acid amplification reactions dissolves the liposomes, releasing 100% of the nucleic acid into the reaction. After initial protocol optimization, it was applied to isolate DNA from Escherichia coli, Staphylococcus aureus, and adenovirus in buffer followed by qPCR detection. This enabled the detection of these pathogens down to concentrations of 1 CFU or 1 PFU/mL, respectively. Comparing it to a standard commercial DNA extraction kit, it was superior, as evidenced by lower Ct-values in the qPCR for all pathogen concentrations. Scaling up to larger volumes, samples containing bacteria were first concentrated through nitrocellulose filters (pore size = 0.45 μm). Tap water, lake water, and rinse water of fresh produce were investigated, leading to relevant limits of detection of 100 CFU in 100 mL of tap water, 1000 CFU in 100 mL of lake water, and 100 CFU in 10 g of iceberg lettuce, respectively. Since the liposome protocol is a homogeneous, simple incubation step, it is a valuable alternative to standard commercial nucleic acid extraction kits.

The role of homogenization cycles and Poloxamer 188 on the quality of mitochondria isolated for use in mitochondrial transplantation therapy

Mitochondrial transplantation (MTx) offers a promising therapeutic approach to mitigate mitochondrial dysfunction in conditions such as ischemia–reperfusion (IR) injury. The quality and viability of donor mitochondria are critical to MTx success, necessitating the optimization of isolation protocols. This study aimed to assess a rapid mitochondrial isolation method, examine the relationship between mitochondrial size and membrane potential, and evaluate the potential benefits of Poloxamer 188 (P-188) in improving mitochondrial quality during the isolation process. Mitochondria were isolated from pectoral muscle biopsies of adult male Sprague–Dawley rats using an automated homogenizer. MitoTracker Deep Red (MTDR) staining and flow cytometry were used to assess mitochondrial purity, while the JC-1 assay evaluated membrane potential. Mitochondrial size groups were compared for membrane potential differences. Homogenization frequency and P-188 supplementation (1 mM) were assessed for their effects on mitochondrial membrane potential and particle size, and particle counts. The rapid isolation method yielded mitochondria that retained sufficient membrane potential to be effectively inhibited by carbonyl cyanide 3-chlorophenylhydrazone (CCCP), a disruptor of mitochondrial membrane potential. Larger mitochondria exhibited significantly higher JC-1 ratios, indicating greater membrane potential. Excessive homogenization (10 cycles) reduced membrane potential compared to 3 cycles homogenization (P = 0.026). P-188 significantly increased the JC-1 ratio from 10.26 ± 2.57 to 33.78 ± 17.78 (P = 0.023). Particle size and count analysis revealed that 10 cycles homogenization significantly increased particle count compared to 3 cycles homogenization (P = 0.0001), but was associated with smaller particle sizes (P = 0.0031). The rapid mitochondrial isolation method produced viable mitochondria, with larger mitochondria exhibiting superior membrane potential. Reducing homogenization frequency and incorporating P-188 improved mitochondrial quality and preserved particle size. These strategies offer promising strategies for optimizing MTx protocols. Further refinement of these techniques is necessary for their clinical application in MTx therapy.

Nose-to-Brain Targeting of Resveratrol Nanoformulations.

Resveratrol [RES] is a polyphenolic stilbene with therapeutic potential owing to its antioxidant, anti-inflammatory, neuroprotective, and cardioprotective properties. However, the very poor oral bioavailability, fast metabolism, and extremely low stability under physiological conditions pose a severe detriment to the clinical use of RES. This newly developed field of nanotechnology has led to the formulation of RES into nanoformulations with the goal of overcoming metabolicpharmacokinetic limitations and enhancing the targeted transport of RES to the central nervous system [CNS]. Among the various routes of administration, the combination of nose-to-brain [N2B] delivery via the intranasal [IN] route has recently garnered attention as a straightforward, noninvasive route for transport to the blood-brain barrier [BBB] for greater effects and less harmful systemic side effects by transporting nano-encapsulated RES into the neural tissues. This review critically summarizes the mechanisms and benefits of the N2B route for the delivery of RES nanoformulations, collating in vivo data demonstrating increased CNS bioavailability and stability and, consequently, improved therapeutic efficacy in animal models of neurodegenerative diseases. Compared with the more 'traditional' routes of administration, IN administration of RES nanoformulations is less toxic, cost-effective, and efficient in crossing the BBB. Therefore, this route represents a promising approach to the management of CNS disorders. Further optimization of nanoformulation design and clinical protocols is required to translate these promising findings into therapeutic strategies aimed at neuroprotection and disease modification in human CNS pathologies.

Integrating Sports Massage into Pre-training Routines Can Enhance Athletic Preparedness and Performance Capacity in Kabaddi Players

The purpose of the study. The primary objective of this investigation is to assess the efficacy of pre-training sports massage in enhancing warm-up protocols for kabaddi athletes, focusing on its influence on various physiological parameters, performance metrics, and psychological preparedness. Materials and methods. A quasi-experimental design was employed involving a cohort of 20 elite kabaddi athletes (15 males and 5 females) aged between 18 and 30 years, hailing from professional teams in Medan City. The intervention spanned 8 weeks and consisted of a 15-minute pre-training sports massage regimen administered prior to standard warm-up sessions. Data were collected at baseline, after 4 weeks, and following 8 weeks of intervention, encompassing measurements of muscle temperature, range of motion, reaction time, performance indicators, and perceived readiness, utilizing standardized assessment instruments. Results. The intervention yielded statistically significant enhancements across multiple evaluated parameters. Muscle temperature exhibited an average increase of 2.3°C, while range of motion demonstrated a 12.5% improvement in hip flexibility; additionally, performance metrics revealed a 22% augmentation in defensive maneuver efficiency. Reaction time was reduced by 0.3 seconds, and participants indicated a decrease in perceived exertion alongside an enhancement in psychological readiness. All observed improvements were statistically significant (p < 0.001) and characterized by large effect sizes. Conclusions. The implementation of pre-training sports massage emerges as a potentially beneficial intervention for the optimization of warm-up protocols among kabaddi athletes. The extensive advantages noted across physiological, biomechanical, and psychological realms imply that the incorporation of sports massage into pre-training routines may significantly bolster athletic preparedness and performance capacity, particularly in high-intensity contact sports such as kabaddi.

Fumaria officinalis Dust as a Source of Bioactives for Potential Dermal Application: Optimization of Extraction Procedures, Phytochemical Profiling, and Effects Related to Skin Health Benefits

Fumaria officinalis (fumitory), in the form of dust, was employed as a source of bioactive extracts whose chemical profile and biological potential were investigated. According to the results of the optimization of the extraction protocol, the extract with the highest polyphenol yield was prepared using fumitory dust under the optimal conditions determined using the statistical tool, 23 full factorial design: 50% ethanol and a 30:1 mL/g ratio during 120 s of microwave extraction (22.56 mg gallic acid equivalent/g of plant material). LC-MS and spectrophotometric/gravimetric analyses quantified the polyphenol, flavonoid, tannin, alkaloid, and protein contents. Caffeoylmalic acid, quercetin dihexoside, quercetin pentoside hexoside, rutin, and methylquercetin dihexoside were the most dominant compounds. The highest total flavonoid, condensed tannin, alkaloid, and protein yields were determined in the extract prepared using microwaves. In addition to the proven antioxidant potential, in the present study, the anti-inflammatory activity of fumitory extracts is also proven in the keratinocyte model, as well as a significant reduction of H2O2-induced reactive oxygen species production in cells and the absence of keratinocyte cytotoxicity. Thus, detailed chemical profiles and investigated biological effects related to skin health benefits encourage the potential application of fumitory dust extracts in dermo-cosmetic and pharmaceutical preparations for dermatological circumstances.

Fetal Radiation Dose in Common Diagnostic Radiology Procedures for Pregnant Patients: Findings from In-Phantom Measurements

The diagnosis of emergent conditions during pregnancy can be delayed due to insufficient knowledge of fetal radiation doses in different imaging modalities. The aim of this article is to investigate the ranges of fetal doses in most common diagnostic and interventional radiology procedures. Procedures were carried out on an anthropomorphic phantom, Tena, representing a pregnant woman in the 18th week of pregnancy with the fetus in breech position. Different clinical scenarios using computer tomography (CT), radiography, fluoroscopy and digital subtraction angiography were selected in three teaching hospitals. Measurements were performed using radiophotoluminescent glass dosimeters placed in dedicated holes in the fetal head and fetal body. Measured fetal doses were below 1 mGy when the fetus was not in the primary beam. The highest fetal doses, up to 47 mGy, were measured after a CT scan for polytrauma and up to 24 mGy after a CT scan of the abdomen and pelvis. Significant variability in fetal doses for the same procedure was found between different hospitals but within the same hospital also. All obtained results are below the threshold for deterministic effects given by the International Commission for Radiation Protection but can be reached with two or more imaging procedures employed. The variability in fetal doses for the same procedures highlights the need for the improved optimization of imaging protocols.

Robust peptide/RNA complexes prepared with microfluidic mixing for pulmonary delivery by nebulisation.

Small interfering RNA (siRNA) and messengerRNA (mRNA) have drawn considerable attention in recent years due to their ability to modulate the expression of specific disease-related proteins. However, it is difficult to find safe, robust, and effective RNA delivery systems suitable for pulmonary delivery to treat lung diseases. In this study, two cationic peptides, namely LAH4-L1 and PEG12KL4, were employed as non-viral vectors for siRNA and mRNA delivery. Four formulations (i.e. LAH4-L1/siRNA; PEG12KL4/siRNA; LAH4-L1/mRNA and PEG12KL4/mRNA) were investigated. Microfluidic mixing method was utilised to fabricate RNA complexes in a controllable and reproducible manner. Upon optimisation of the microfluidic mixing protocol, a vibrating mesh nebuliser was employed to aerosolise the RNA complexes, and their transfection efficiency was evaluated on A549 and BEAS-2B cells. Following nebulisation, inhalable mist was generated for all RNA formulations with mass median aerodynamic diameter below 5μm. Although the hydrodynamic particle sizes of the RNA complexes were significantly reduced to around 100nm after nebulisation regardless of the original size of the complexes prior to nebulisation, the RNA binding efficiency and the in vitro RNA transfection ability of all the peptide formulations were successfully preserved with no significant differences compared to the same system before nebulisation. The current result indicates that both LAH4-L1 and PEG12KL4 hold significant potential for future clinical application for pulmonary siRNA and mRNA delivery through nebulisation.

Optimization of the Extraction Protocol for Pacific Ciguatoxins from Marine Products Prior to Analysis Using the Neuroblastoma Cell-Based Assay.

Ciguatera poisoning (CP) is caused by the consumption of marine products contaminated with ciguatoxins (CTXs) produced by dinoflagellates of the genus Gambierdiscus. Analytical methods for CTXs, involving the extraction/purification of trace quantities of CTXs from complex matrices, are numerous in the literature. However, little information on their effectiveness for nonpolar CTXs is available, yet these congeners, contributing to the risk of CP, are required for the establishment of effective food safety monitoring programs. An evaluation of six extraction/purification protocols, performed with CTX3C spiked on fish flesh and a neuroblastoma cell-based assay (CBA-N2a), revealed recoveries from 6 to 45%. This led to the development of an optimized 3-day protocol designed for a large number of samples, with CTX1B and CTX3C eluting in a single fraction and showing recoveries of 73% and 70%, respectively. In addition, a reduction in adverse matrix effects in the CBA-N2a analyses was demonstrated with naturally contaminated specimens, increasing the sensitivity of the method, which now meets the very low guidance level recommended by international agencies. However, efforts are still required to reduce the signal suppression observed in LC-MS/MS analysis. This optimized protocol contributes to the technological advancement of detection methods, promoting food safety and improving CP risk assessment in marine products.

Artificial Liver Support Systems in Acute Liver Failure and Acute-on-Chronic Liver Failure: Systematic Review and Meta-Analysis.

To systematically review the safety and efficacy of nonbiological (NBAL) or biological artificial liver support systems (BAL) and whole-organ extracorporeal liver perfusion (W-ECLP) systems, in adults with acute liver failure (ALF) and acute-on-chronic liver failure (ACLF). Eligible NBAL/BAL studies from PubMed/Embase searches were randomized controlled trials (RCTs) in adult patients with ALF/ACLF, greater than or equal to ten patients per group, reporting outcomes related to survival, adverse events, transplantation rate, and hepatic encephalopathy, and published in English from January 2000 to July 2023. Separately, we searched for studies evaluating W-ECLP in adult patients with ALF or ACLF published between January1990 and July 2023. Two researchers independently screened citations for eligibility and, of eligible studies, retrieved data related to study characteristics, patients and interventions, outcomes definition, and intervention effects. The Cochrane Risk of Bias 2 tool and Joanna Briggs Institute checklists were used to assess individual study risk of bias. Meta-analysis of mortality at 28-30 days post-support system initiation and frequency of at least one serious adverse event (SAE) generated pooled risk ratios (RRs), based on random (mortality) or fixed (SAE) effects models. Of 17 trials evaluating NBAL/BAL systems, 11 reported 28-30 days mortality and five reported frequency of at least one SAE. Overall, NBAL/BAL was not statistically associated with mortality at 28-30 days (RR, 0.85; 95% CI, 0.67-1.07; p = 0.169) or frequency of at least one SAE (RR, 1.15; 95% CI, 0.99-1.33; p = 0.059), compared with standard medical treatment. Subgroup results on ALF patients suggest possible benefit for mortality (RR, 0.67; 95% CI, 0.44-1.03; p = 0.069). From six reports of W-ECLP (12 patients), more than half (58%) of severe patients were bridged to transplantation and survived without transmission of porcine retroviruses. Despite no significant pooled effects of NBAL/BAL devices, the available evidence calls for further research and development of extracorporeal liver support systems, with larger RCTs and optimization of patient selection, perfusion durability, and treatment protocols.

Autologous and Heterologous Minor and Major Bone Regeneration with Platelet-Derived Growth Factors.

This review aims to explore the clinical applications, biological mechanisms, and potential benefits of concentrated growth factors (CGFs), autologous materials, and xenografts in bone regeneration, particularly in dental treatments such as alveolar ridge preservation, mandibular osteonecrosis, and peri-implantitis. A systematic literature search was conducted using databases like PubMed, Scopus, and Web of Science, with keywords such as "bone regeneration" and "CGF" from 2014 to 2024. Only English-language clinical studies involving human subjects were included. A total of 10 studies were selected for qualitative analysis. Data were processed through multiple stages, including title and abstract screening and full-text evaluation. The findings of the reviewed studies underscore the potential of the CGF in enhancing bone regeneration through stimulating cell proliferation, angiogenesis, and extracellular matrix mineralization. Autologous materials have also demonstrated promising results due to their biocompatibility and capacity for seamless integration with natural bone tissue. When combined with xenografts, these materials show synergistic effects in improving bone quantity and quality, which are crucial for dental implant success. Future research should focus on direct comparisons of different techniques, the optimization of protocols, and broader applications beyond dental medicine. The integration of CGFs and autologous materials into routine clinical practice represents a significant advancement in regenerative dental medicine, with the potential for improved patient outcomes and satisfaction.

Evaluation of workload distribution and exposure factors in common use in ten general x-ray rooms in Ireland: An initial step towards protocol optimisation.

This study aimed to assess current practice in general x-ray rooms in three Irish hospitals. Evaluating the extent to which x-ray protocols are currently standardised can be considered as a preparatory step towards the larger goal of implementing optimisation in a diagnostic radiology department. Data on various aspects of general x-ray room exposures was manually collected and analysed to highlight variations with exposure factors in use and workload distribution across each of the ten hospital x-ray rooms included in the study. Workload distribution for the x-ray rooms surveyed demonstrated the most commonly performed x-ray, the chest x-ray, to account for 44% of exposures acquired, compared to the most recently published national statistic of 31%1, suggesting a potential change in referral practices over the past decade. A high degree of protocol variation for the same imaging indications exists across the x-ray rooms included in the study, for example there were 14 protocols in routine use for shoulder imaging across the 10 x-ray rooms, spanning a wide range of tube potentials from 55 to 81kVp. The lack of x-ray protocol standardisation (and associated image quality variation) prompts questions around the implementation of optimisation, as required by Irish legislation, for general x-ray protocols. National review of x-ray referral practices or most common general x-ray exposures in use would be welcomed, as deviations from the previously published dataset are locally evident. This body of work considers the variety of x-ray protocols in use for specific anatomical areas and outlines how similar analyses at a local level could provide a foundation for targeted optimisation exercises.

Optimization of protocol for analysis of dicarboxylic acids in ambient aerosol samples using GC-MS: method comparison and application.

Dicarboxylic acids (DCAs), with their deliquescence and hygroscopic nature, can function as cloud condensation nuclei (CCN) and ice nuclei (IN), affecting rainfall patterns. DCA analysis can serve as organic molecular markers for anthropogenic and biogenic sources. Very few studies deal with the optimization of the protocol for qualitative and quantitative analysis of DCAs using gas chromatography-mass spectrometry (GC-MS). In this study, we have optimized the extraction of DCAs from aerosol samples by employing several organic solvents with differing relative polarities. Extraction efficiencies of organic solvents were evaluated at different temperatures and pressures using the advanced energized dispersive extractor. The optimized outcomes demonstrate that extraction using methanol at 105 °C for 5 min resulted in high efficiencies, achieving over 80% recovery for all targeted C3-C10 dicarboxylic acids (DCAs). N,O-bis-(trimethylsilyl) trifluoroacetamide (BSTFA) with 1% trimethylchlorosilane (TMCS) was chosen as the derivatizing reagent, and reaction conditions were optimized to give maximum conversions. The derivatization process, conducted with 30 µL of BSTFA + 1% TMCS in a 200 µL reaction mixture at 70°C for 90min, yielded effective and reliable results for subsequent analysis. Separation of compounds was done on the HP-5 column with Helium as the carrier gas. Protocol was finalized by selecting the operating parameters in SIM mode that reduce the total run time while maintaining a clear resolution of peaks.