Search Results (28)

Background: Primary hepatocytes are excellent models for studying liver functions and liver diseases. However, obtaining high yields of viable hepatocytes remains technically challenging, limiting their broader applications. Most conventional methods rely on a two-step collagenase perfusion technique. Despite its widespread use, this approach has several limitations that reduce the success rate of hepatocyte isolation and culture. The procedure involves multiple parameters that are continually being optimized in order to obtain hepatocytes in high yield and quality that can be used to provide insights into their physiology and pathophysiology. Aim: We aimed to enhance the success rate and reproducibility of hepatocyte isolation with high yield, enabling analysis of diverse physiological and pathophysiological aspects of lipid metabolism. It also establishes an in vitro steatosis model for evaluating therapeutic drugs and molecular interventions. Methods: Rat liver was perfused in situ with EDTA buffer followed by collagenase IV. Liver was then isolated, and hepatocytes were mechanically liberated, filtered, and purified through density-gradient centrifugation. Viable cells were cultured at 700,000 or 1 million cells/well for 24 h. The monolayer was incubated in lipogenic media for an additional 24 or 48 h. Hepatocytes were fixed, neutral lipids were stained using Oil Red O, and the stained area was quantified using Image J software version 1.54. Results: Yield of hepatocytes was ~75–90 million cells/liver, with viability of 86–93%. Cells seeded at 700,000 and 1 million cells/well reached confluences of 60% and 80%, respectively, after 24 h. Steatosis was then induced with lipid accumulation reaching 21% of image area after 24 h and 25% after 48 h. Conclusions: The current protocol presents an efficient and highly reproducible method for isolating primary rat hepatocytes in high yield with high viability. Additionally, the protocol provides a foundation for studying the pathophysiology of fatty liver disease. Full article

Background: Finding the optimal amount of fluid is a major challenge in burn shock. Although there is evidence that a restrictive fluid regime is beneficial, current practice shows fluid resuscitation still well above recommendations. The extent of trauma, pre-hospital care and the patient’s pre-existing conditions influence requirements. Methods: We analysed outcomes and influencing factors of fluid regimes in a retrospective cohort study including 90 severely burnt patients resuscitated with the same protocol. Results: The mean amount of fluids in the first 24 h was 6.5 mL/kg bodyweight (BW)/% total burn surface area (TBSA). A total of 14% received restrictive (<4), 34% received liberal (4–6) and 51% received excessive (>6) mL/kgBW/%TBSA fluids. There was no difference regarding mortality, age, complications, organ failure, inhalation injury or full-thickness burns in the groups. Patients with excessive fluid therapy had a significantly lower ABSI score (9 vs. 11, p = 0.05) and TBSA (35 vs. 51%, p < 0.001), while patients with a restrictive fluid therapy needed fewer incidences of surgery to cover burn wounds (3.5 vs. 9.0 vs. 7.0, p = 0.008). History of liver disease or alcohol abuse tended to indicate excessive fluid administration. Patients with pre-existing heart failure received restrictive fluid therapy (23 vs. 3 vs. 4%, p = 0.03). Conclusions: Individualised, timely therapy monitoring is as essential as identifying patients with a higher or lower fluid requirement. Excessive fluid resuscitation had fewer deleterious consequences in complications than expected but seems to influence wound healing. Awareness of circumstances that prompt deviations from recommended fluid rates remains elementary. Full article

Pediatric Intensive Care Units (PICUs) provide specialized care for critically ill children. Developing and managing these units in South Asia remains challenging. Resource limitations and infrastructural disparities are leading to challenging conditions. Above all, nurses play a pivotal role in delivering quality critical care. Effective nursing practices can curb hospital-acquired infections (HAIs), ensure medication safety, and enable protocols such as the ICU Liberation Bundle. In South Asia, another challenge is the proper management of the nursing workforce. Nurse-to-patient ratios are highly disproportionate, contributing to nurse burnout. This review highlights the country-specific challenges and circumstances. There is no one-size-fits-all solution; effective strategies vary based on each country’s context. With context-specific solutions, nurses can bridge the gap between healthcare teams and families, ultimately improving patient outcomes. Full article

Background/Objectives: Anemia is common in critically ill patients, yet red blood cell (RBC) transfusion without active bleeding does not consistently improve outcomes and carries risks such as pulmonary injury, fluid overload, and increased costs. Optimal transfusion thresholds remain debated, with some guidelines recommending a restrictive target of 7 g/dL instead of a more liberal target of 9 g/dL. Methods: We conducted a systematic review and meta-analysis following PRISMA guidelines, searching PubMed, EMBASE, and LILACS from January 1995 to October 2024. Thirteen randomized controlled trials involving 13,705 critically ill adults were included, with 6855 assigned to liberal and 6850 to restrictive transfusion strategies. The risk of bias was assessed using the Cochrane Risk of Bias Tool 2, and the pooled effect sizes were estimated with a random-effects model. We registered the protocol in PROSPERO International Prospective Register of Systematic Reviews (CDR42024589225). Results: No statistically significant difference was observed in 30-day mortality between restrictive and liberal strategies (odds ratio [OR] 1.02; 95% confidence interval [CI], 0.83–1.25; I² = 49%). Similarly, no significant differences emerged for the 90-day or 180-day mortality, hospital or intensive care unit (ICU) length of stay, dialysis requirement, or incidence of acute respiratory distress syndrome (ARDS). However, patients in the restrictive group received significantly fewer RBC units. The trial sequential analysis (TSA) indicated that the evidence accrued was insufficient to definitively confirm or exclude an effect on the 30-day mortality, as the required sample size was not reached. Conclusions: In conclusion, while our meta-analysis found no statistically significant difference in the short-term mortality between restrictive and liberal transfusion strategies, larger trials are needed to fully determine whether any clinically meaningful difference exists in critically ill populations. Full article

Personalizing the management of neovascular age-related macular degeneration (nAMD) poses significant challenges for practicing retina specialists and their patients. This commentary addresses some of these complexities, particularly those that arise in the context of an expanding array of intravitreal agents targeting vascular endothelial growth factor (VEGF) and related retinal disease targets. Many of these newer agents approved by the Food and Drug Administration (FDA) for the treatment of nAMD have labeling that indicates that they can provide non-inferior visual outcomes when compared head-to-head with previously available treatments and can be used at significantly extended dosing intervals in some patients. It can be difficult to know if patients should be transitioned to these agents, especially those who are doing well on existing therapies. Although offering extended intervals may be appropriate for some patients with excellent disease control, retina specialists know that undertreatment risks the loss of visual acuity (VA). It can also be challenging for clinicians to interpret the results delivered by clinical trial treatment protocols compared with what is likely to occur in real-world office settings. Many retina specialists use less liberal treatment paradigms than employed in clinical study protocols and consequently many patients experience shorter injection intervals. Since VA is most closely linked to quality of life, it should be prioritized compared with other endpoints. The authors advocate for maintaining consistent treatment schedules dictated by disease control instead of switching therapies even in the presence of small amounts of macular fluid that may occur with longer injection intervals. Full article

Efficient development of effective functional foods and nutraceuticals requires adequate estimation methods of the bioaccessibility of their bioactive compounds. Specially grain-based nutraceuticals and functional ingredients are often enriched in bound/low bioavailable bioactive phytochemicals. The objective of this work was to evaluate the differences in applying static or dynamic digestion models for the estimation of bioaccessibility of antioxidants present in cereal grain-based/fiber-rich ingredients produced using enzymatic hydrolysis and sprouting processes. Main liberated phenolic compounds, antioxidant activity (ABTS^•+ and ORAC) and ferric reducing capacity were evaluated in the samples following three digestion protocols with differences based on their dynamism: static, semi-dynamic and dynamic. The samples digested with the dynamic method showed higher antioxidant and reducing capacities than those digested with the static and semi-dynamic protocols. The results obtained from the digests with the dynamic model showed a total phenol content (TPs) ranging from 1068.22 to 1456.65 μmol GAE 100 g⁻¹ and antioxidant capacity values from 7944.62 to 15,641.90 μmol TE 100 g⁻¹ (ORAC) and from 8454.08 to 11,002.64 μmol TE 100 g⁻¹ (ABTS^•+), with a reducing power ranging from 2103.32 to 2679.78 mmol Fe reduced 100 g⁻¹ (FRAP). The dynamic character of the protocols used for developing bioactive cereal-based foods significantly affects the estimation of their bioaccessibility, probably giving a better approach to their potential bioavailability in in vivo systems. Full article

Introduction: Within the field of periodontology, there has been a proposal for the utilization of noninvasive laser therapy as a potential treatment for persistent periodontitis. The Laser-Assisted New Attachment Procedure (LANAP) employs an Nd:YAG laser as a specific technique. Through its interaction with endogenous chromophores, the Nd: YAG laser exhibits a selective effect on the evaporation of granulation tissue, therefore establishing a correlation with reduced bleeding. The study also examined Oxygen High-Level Laser Therapy (OHLLT). The OHLLT technique employs a high-power diode laser in combination with hydrogen peroxide solutions to facilitate the liberation of singlet oxygen, which possesses antibacterial attributes, within the periodontal pockets. The existing literature indicates their potential to promote the regeneration of tooth support tissues. Objective: The aim of this study is to assess the subjective pain levels reported by patients who have undergone surgery using the OHLLT protocol versus those who have undergone surgery using the LANAP technique. Methods: A total of 20 individuals with a stage III–IV periodontitis diagnosis were recruited for the study. The participants were randomly divided into two groups, each consisting of 10 individuals: Group 1, comprising patients treated according to the LANAP protocol, and Group 2, comprising patients treated according to the OHLLT protocol. After their initial session of nonsurgical periodontal therapy, individuals provided feedback regarding their level of pain, utilizing a Numerical Rating Scale (NRS) comprising time intervals of 0 h (T0), 6 h (T1), 12 h (T2), 24 h (T3), 48 h (T4), and 7 days (T5). The Wilcoxon–Mann–Whitney statistical test was employed to assess the variations in NRS scores between Group 1 and Group 2 at each recording period. (p ≤ 0.05). In addition, a microbiological assessment of the bacterial load in the periodontal region was conducted on all subjects using real-time PCR testing at two time points: prior to treatment (T0) and seven days post-treatment (T5). Results: The findings of this study indicate that the OHLLT group exhibited significantly lower pain levels compared to the LANAP group at all time intervals, except for the preoperative period, where no significant difference was observed (p < 0.05). Group 2 exhibited a more rapid decrease in pain, as demonstrated by a score test approaching zero within 24 h. The quantity of periodontal bacteria seen seven days post-treatment was similar between the two groups and was found to be decreased compared to the pre-treatment levels. Conclusions: The OHLLT and LANAP regimens have demonstrated efficacy in the nonsurgical management of periodontal disease. Nevertheless, it should be noted that the OHLLT approach does not subject the patient to any thermal hazards, unlike the LANAP method. The postoperative discomfort experienced following the OHLLT procedure is indeed reduced, as this technique is characterized by lower invasiveness and reduced dependence on the operator. Full article

The present review discusses restrictive perioperative fluid protocols within enhanced recovery after surgery (ERAS) pathways. Standardized definitions of a restrictive or liberal fluid regimen are lacking since they depend on conflicting evidence, institutional protocols, and personal preferences. Challenges related to restrictive fluid protocols are related to proper patient selection within standardized ERAS protocols. On the other hand, invasive goal-directed fluid therapy (GDFT) is reserved for more challenging disease presentations and polymorbid and frail patients. While the perfusion rate (mL/kg/h) appears less predictive for postoperative outcomes, the authors identified critical thresholds related to total intravenous fluids and weight gain. These thresholds are discussed within the available evidence. The authors aim to introduce their institutional approach to standardized practice. Full article

This study outlines a recycling initiative conducted at Rekular GmbH, focusing on the recycling of 100 refrigerators. The recycling process employed a combination of manual dismantling, depollution, and mechanical processing techniques. Manual dismantling followed a predefined protocol to extract various materials, while the mechanical and physical processes involved shredding, zigzag, magnetic, and eddy current separation (ECS) to liberate and separate different materials. The resulting ferrous, non-ferrous and polymer product fractions were analyzed and categorized, providing valuable insights into the quality of interim products in the refrigerator recycling process. Simulations were then performed using FactSage^TM version 8.2 and HSC Chemistry 10 version 10.3.7.1 software to simulate the recovery of metals from the ferrous and non-ferrous fractions using pyro metallurgical and hydrometallurgical methods. An electric arc furnace (EAF) was utilized for iron (Fe), while a re-smelter process for aluminium (Al), and the black copper route was simulated for copper (Cu) recovery. The recovery rates including metallurgical, mechanical, and physical processes are as follows: Fe (78%), Al (68.4%), and Cu (52.4%). In contrast, the recovery rates through metallurgical processes are as follows: Al (99%), Fe (79%), and Cu (88%). This discrepancy is attributed to losses of these elements resulting from incomplete liberation in mechanical processing. Additionally, a product/centric approach was applied and the recycling index reached 76% for recovery the Al, Cu, and Fe metals in a refrigerator recycling process. Turning to the environmental impact evaluation within the life cycle assessment (LCA), the process unit with the highest emissions per refrigerator in the recycling process was the use of nitrogen during the shredding process, accounting for 3.7 kg CO₂ eq/refrigerator. Subsequently, the consumption of medium voltage electricity from the German grid during mechanical and physical separations contributed to 0.6 kg CO₂ eq/refrigerator. The EAF, and electrolytic refining stages in the metallurgical recovery process also had a notable impact, generating 10.7 kg CO₂ eq/refrigerator. Full article

China faces a persistent deficiency in feed protein resources. Enhancing the utilization efficiency of indigenous feed protein resources emerges as a viable strategy to alleviate the current deficit in protein feed supply. Corn gluten meal (CGM), characterized by a high proportion of crude protein and glutamine, is predominantly employed in animal feed. Nonetheless, the water-insolubility of CGM protein hampers its protein bioavailability when utilized as feed material. The aim of this study was to augment protein bioavailability, liberate glutamine peptides from CGM, and produce glutamine-enriched CGM fermented feed. We executed a co-fermentation protocol using Bacillus subtilis A5, Lactobacillus 02002, and acid protease to generate the CGM fermented feed. Subsequent in vivo experiments with broilers were conducted to assess the efficacy of the fermented product. The findings revealed that the soluble protein, glutamine, small peptides, and lactic acid contents in the fermented feed increased by 69.1%, 700%, 47.6%, and 125.9%, respectively. Incorporating 15% and 30% CGM fermented feed into the diet markedly enhanced the growth performance and intestinal health of broilers, positively modulated the cecal microbiota structure, and augmented the population of beneficial bacteria, specifically Lactobacillus. These results furnish both experimental and theoretical foundations for deploying CGM fermented feed as an alternative protein feed resource. Full article

In liberalized markets, consumers can choose their electricity suppliers or be part of an energy community. The problem with communities is that they may not have enough weight to trade in markets, which can be overcome by forming coalitions. Electricity is traded in spot markets or through bilateral contracts involving consumers and suppliers. This paper is devoted to bilateral contracting, modeled as a negotiation process involving an iterative exchange of offers and counter-offers. It focuses on coalitions of energy communities. Specifically, it presents team and single-agent negotiation models, where each consumer has strategies, tactics, and decision models. Coalition agents are equipped with intra-team strategies and decision protocols. It also describes a study of bilateral contracts involving a seller agent and a coalition of energy communities. By allying into a coalition, members of energy communities reduced their average costs for electricity by between 2% (large consumers) and 64% (small consumers) according to their consumption. Their levelized cost reduction was 19%. The results demonstrate the power of coalitions when negotiating bilateral contracts and the benefit of a low-consumption members alliance with larger players. Full article

The crested porcupine is a frequent host species in wildlife rescue centers and no guidelines for its management, liberation and post-liberation monitoring are yet available. Here, captive-grown porcupines’ behavior and survival in the wild after liberation were investigated and described for the first time. Management strategies adopted at the centers could affect porcupine adaptation to the natural environment. The detention of porcupettes in single cages and fed only human-supplied food may not ensure ethological welfare, nor a suitable behavioral development compatible with wildlife. The liberation of captive-grown porcupines should also be carefully planned to promote and increase the possibility of post-liberation success. Full article

Dietary and social behaviour are non-medical factors that influence health outcomes. Non-communicable diseases are related to dietary patterns. To date, little is known about how social behaviour is associated with health-related dietary patterns, and, in particular, we lack information about the role of sex within this possible relation. Our cross-sectional study investigated associations between dietary patterns and social behaviour including personality traits (self-control, risk taking), political preferences (conservative, liberal, ecological, social) and altruism (willingness to donate, club membership, time discounting) in men and women. We performed sex-specific correlation analyses to investigate relationships between dietary patterns based on self-reported protocols from the Mediterranean Diet Adherence Screener (MEDAS) and the validated Healthy Eating Index (HEI) from the EPIC Study and a self-reported social behaviour questionnaire. In linear regression models, we analysed associations between dietary and social behaviour patterns. Sex differences were measured by interaction analysis for each social behaviour item. The study sample consisted of N = 102 low-risk individuals. The median age of the study participants was 62.4 (25th/75th percentile 53.6, 69.1) years, and 26.5% were women. Analyses showed that a lower HEI score was correlated with a higher BMI in both women and men. MEDAS and HEI showed a positive correlation with each other in men. In men, a higher MEDAS showed a positive correlation when they estimated their ability as high, with the same for self-control and preference for ecological politics and MEDAS. A weak negative correlation has been shown between men with a preference for conservative politics and MEDAS. HEI showed a positive significant correlation with age in men. Male participants without club membership scored significantly higher in the HEI compared to non-members. A negative correlation was shown for time discounting in men. Linear regression models showed positive associations between preferences for ecological-oriented politics and nutrition for both HEI and MEDAS. No sex interactions were observed. We faced a few limitations, such as a small sample size, particularly for women, and a limited age spectrum in a European cohort. However, assuming that individuals with a preference for ecological-oriented politics act ecologically responsibly, our findings indicate that ecological behaviour in low-risk individuals might determine, at least in part, a healthy diet. Furthermore, we observed dietary patterns such as higher alcohol consumption in men or higher intake of butter, margarine and cream in women that indicate that women and men may have different needs for nutritional improvement. Thus, further investigations are needed to better understand how social behaviour affects nutrition, which could help to improve health. Our findings have the potential to inform researchers and practitioners who investigate the nature of the relationship between social behaviour and dietary patterns to implement strategies to create first-stage changes in health behaviour for individuals with a low cardiovascular risk profile. Full article

Background: Restriction of oral potassium intake is a necessary dietary intervention for managing chronic hyperkalemia. These dietary changes may have negative impacts on nutrition status, particularly in geriatric cohorts with multiple comorbidities. Sodium zirconium cyclosilicate (SZC) is a newly introduced potassium binder intended for patients with hyperkalemia. We aimed to investigate whether the improvements in hyperkalemia with SZC therapy and the liberation of potassium intake restriction may improve nutrition status in a primarily geriatric patient cohort with chronic hyperkalemia. Methods: Patients who were maintained on SZC therapy for at least 3 months were retrospectively studied. Following the initiation of SZC and improvement in hyperkalemia, instructions on the restriction of potassium intake were loosened according to the institutional protocol. The change in nutrition status during the 3 month therapeutic period using SZC was investigated by referencing the prognostic nutritional index score (PNI), geriatric nutritional risk index score (GNRI), and controlling nutritional status (CONUT) scores. Results: A total of 24 patients (median age 78 years, 58% men, median estimated glomerular filtration rate 29.8 mL/min//1.73 m²) were included. Serum potassium level decreased significantly from 5.4 (5.1, 5.9) to 4.4 (4.2, 4.9) mEq/L without any drug-related adverse events, including hypokalemia. Nutrition-related scores, including the PNI score, the GNRI score, and the CONUT score, improved significantly following 3 months of SZC therapy (p < 0.05 for all). Psoas muscle volume and average days for one movement also improved significantly during the therapeutic period (p < 0.05 for both). Conclusions: Mid-term SZC therapy and liberation of potassium intake restriction might improve nutrition status in geriatric patients with chronic hyperkalemia. Full article

Background and Objectives: The erector spinae plane block (ESPB) is a multimodal opioid-sparing component, providing chest-wall analgesia of variable extent, duration, and intensity. The objective was to examine the ESPB effect on perioperative opioid usage and postoperative rehabilitation when used within a Nociception Level (NOL) index-directed anesthetic protocol. Materials and Methods: This prospective, randomized, controlled, open-label study was performed in adult patients undergoing on-pump cardiac surgery in a single tertiary hospital. Eighty-three adult patients who met eligibility criteria were randomly allocated to group 1 (Control, n = 43) and group 2 (ESPB, n = 40) and received general anesthesia with NOL index-directed fentanyl dosing. Preoperatively, group 2 also received bilateral single-shot ultrasound-guided ESPB (1.5 mg/kg/side 0.5% ropivacaine mixed with dexamethasone 8 mg/20 mL). Postoperatively, both groups received intravenous paracetamol (1 g every 6 h). Morphine (0.03 mg/kg) was administered for numeric rating scale (NRS) scores ≥4. Results: The median (IQR, 25th–75th percentiles) intraoperative fentanyl and 48 h morphine dose in group 2-to-group 1 were 1.2 (1.1–1.5) vs. 4.5 (3.8–5.5) µg·kg⁻¹·h⁻¹ (p < 0.001) and 22.1 (0–40.4) vs. 60.6 (40–95.7) µg/kg (p < 0.001). The median (IQR) time to extubation in group 2-to-group 1 was 90 (60–105) vs. 360 (285–510) min (p < 0.001). Two hours after ICU admission, 87.5% of ESPB patients were extubated compared to 0% of controls (p < 0.001), and 87.5% were weaned off norepinephrine compared to 46.5% of controls (p < 0.001). The median NRS scores at 0, 6, 12, 24, and 48 h after extubation were significantly decreased in group 2. There was no difference in opioid-related adverse events and length of stay. Conclusions: NOL index-directed ESPB reduced intraoperative fentanyl by 73.3% and 48 h morphine by 63.5%. It also hastened the extubation and liberation from vasopressor support and improved postoperative analgesia. Full article