Neutral Effect Research Articles

Assessment of the nutritional impact of substituting fishmeal with enzymatically hydrolyzed jojoba meal (Simmondsia chinensis) in the diets of Nile tilapia, Oreochromis niloticus

Enzymatic hydrolysis is a technology that is widely used to enhance the biological and functional characteristics of plant proteins and deactivate anti-nutritious components. In order to tackle this pressing issue, considerable research has been conducted on plant-based proteins as a potential substitute. This study sought to comprehensively examine the impacts of replacing fish meal with enzymatically hydrolyzed jojoba meal (EHJM) in the Nile tilapia, Oreochromis niloticus diets. After the enzymatic hydrolysis process for jojoba meal, hydrolyzed amino acid content increased in enzymatically hydrolyzed jojoba meal (EHJM), while fiber content, phytic acid, saponins, and trypsin inhibitors dramatically decreased. A thorough evaluation of key parameters included growth, gene expression, digestive enzyme activity, digestibility, intestinal histomorphometry, biochemical indicators, and antioxidant enzyme activity. Fish (n = 360, 6.41 ± 0.05 g fish−1) were allocated at random to four groups consisting of four replacement levels of fishmeal protein content with EHJM: 0 % (control diet), 25 % (EHJM25), 50 % (EHJM50) and 75 % (EHJM75) for 74 days. The results showed both the control diet and the EHJM25 groups showed remarkable improvements in performance metrics such as final body weight, specific growth rate, weight gain, feed conversion ratio, and protein efficiency ratio, with no discernible variations between them. The inclusion of EHJM up to 50 % produced a noteworthy (P < 0.05) rise in the intestinal lipase and amylase activities. Furthermore, a high apparent digestibility coefficient for crude lipid and protein values was observed in EHJM25% and EHJM50%. The presence of EHJM up to 25 % in tilapia diets, as a substitute for FM, had a significant impact (P < 0.05) on various parameters in the intestine, including goblet cells, muscularis mucosa, villi height, villi width, absorption area, and height muscularis. Moreover, the inclusion of EHJM with different levels in tilapia diets resulted in a noticeable reduction (P < 0.05) in alkaline phosphatase (ALP) and serum alanine aminotransferase (ALT) levels, with fish given the EHJM75% diet showing the lowest amounts in their plasma. In addition, the amount of plasma total protein, albumin, and globulin were found to rise sharply with the escalating replacement of EHJM in fish diet, reaching the highest values in the EHJM25% and EHJM50% diets. Interestingly, the catalase (CAT) and superoxide dismutase (SOD) levels in fish that were given the control diet and EHJM25% showed a remarkable increase compared to the other treatments (P < 0.05). Fish fed EHJM50% diet had the highest glutathione peroxidase and the lowest malondialdehyde levels. In addition, the fish that were fed the control diet exhibited the highest expression of insulin-like growth factor and growth hormone genes, compared to those fed the EHJM25% diet. Based on the results, the EHJM25% and EHJM50% diets were more effective than the EHJM75% diet in improving the efficiency of Nile tilapia. The inclusion of EHJM in Nile tilapia diets had a neutral effect on growth indices compared to a fishmeal diet (control diet). The optimal replacement level is 55.3 %.

Construction of engineered probiotic that adhere and display nanobody to neutralize porcine reproductive and respiratory syndrome virus.

Pathogenic blue ear disease caused by porcine reproductive and respiratory syndrome virus (PRRSV) bring severe loss to breeding industry due to high infectivity and mortality. L. plantarum serves as the probiotic host strain, known for its beneficial properties in the gut microbiota. E. coli is used as a cloning host for the initial genetic engineering steps, facilitating the construction and amplification of the desired genetic constructs. In this study, using synthetic biology technology, we constructed engineered probiotics which could adhere and display nanobody on the surface to neutralize virus. Firstly, we screen an optimal nanobody to effectively bind with PRRSV by building library, expression and purification. Then, the integration of adhesion protein and nanobody into the genome of probiotics significantly improved its adhesion to IPEC-J2 cells. In addition, this engineered probiotic is almost non-toxic to cells with good safety, which can be used as a daily probiotics to prevent virus fecal transmission. Our study proposed this novel construction strategy of engineering probiotics with both adhesion and neutralization effects, which provided a new therapeutic view for intestinal virus clearance.

Outcomes of guidelines from health technology assessment organizations in community-based primary care: a systematic mixed studies review.

Health technology assessment (HTA) organizations generate guidelines to inform healthcare practices toward improved health outcomes. This review sought to identify and classify outcomes of guidelines from HTA organizations within published research. We performed a systematic mixed studies review of empirical studies that (a) referred to a published guideline from an HTA organization and (b) reported an outcome resulting from a guideline. We searched the published literature in English or French within seven databases. Outcome types were classified within five dimensions of an existing framework for online health information (e.g., relevance, cognitive/affective impact, and use). Subdimensions were inductively developed. A two-phase sequential data synthesis was performed. Phase 1: a hybrid deductive-inductive thematic analysis identified the types of outcomes and displayed their relationships on a concept map. Phase 2: descriptive statistics were tabulated by the type of outcome. A total of 6,719 records were retrieved through searches on 6 February 2023. After screening, we included 120 observational studies (twenty-one qualitative, ninety-four quantitative, and five mixed methods). Phase 1 identified twenty-nine types of outcomes. The most frequently reported outcomes were within the organizational dimension (reported in ninety-four studies). The most common subdimensions were "Referrals" (thirty-eight occurrences), the "Quality of Prescriptions" (fifteen occurrences), and the "Quality of Diagnosis" (eight occurrences). For Phase 2, we could only generate descriptive statistics on seventeen outcomes. These were almost equally distributed among positive, neutral, and negative effects. Our results contribute to knowledge about the outcomes of HTA guidelines and options for documenting and measuring them in future evaluations.

Development of a Graphene Oxide-Based Aptamer Nanoarray for Improved Neutralization and Protection Effects Against Ricin.

Background/Objectives: Ricin's high toxicity and potential as a bioweapon underscore the need for effective antidotes. Monoclonal antibodies, though effective, are limited by complex production. This study aimed to develop a graphene oxide-based aptamer nanoarray (ARMAN) for improved neutralization and protection against ricin. Methods: High-affinity aptamers targeting ricin's RTA and RTB subunits were selected using SELEX technology and conjugated to graphene oxide (GO) via click chemistry. ARMAN's characteristics, including morphology, stability, and biosecurity, were assessed. Its performance was evaluated in terms of affinity for ricin, neutralization capacity, and therapeutic effects in cellular assays and a mouse model of ricin poisoning. Results: ARMAN exhibited a uniform morphology with an average particle size of 217 nm and demonstrated significantly enhanced affinity for ricin compared to free aptamers. ARMAN showed rapid and effective neutralization ability, significantly increasing cell viability in BEAS-2B, GES-1, and HL7702 cell lines exposed to ricin. In vivo, ARMAN treatment led to a notable prolongation of survival in ricin-poisoned mice, highlighting its potential for both pre- and post-exposure treatment. These findings indicate that ARMAN not only neutralizes ricin effectively but also provides a therapeutic window for treatment. Conclusions: ARMAN's superior binding affinity, serum stability, biocompatibility, and broad therapeutic efficacy make it a promising new antidote against ricin poisoning. This study's findings represent significant progress in the development of rapid-response antidotes, with ARMAN offering a potential solution for both military and civilian emergency response scenarios.

Oxidation of glyoxal with the Mo-oxime complex in a benzalkonium chloride interface: Raghavan and Srinivasan kinetic model

Gaining an understanding of the glucose oxidation product (glyoxal) interaction with the metal–organic complex in a biological environment is pivotal to its usage. Thus, the glyoxal (Gyx) oxidation with Mo-oxime complex (MC) is studied with the spectrophotometric method, following a pseudo-phase approach. The result highlights the inclusion of hydrolysis, ion catalysis, the neutral primary salt effect, and radical generation as the essential factors in Gyx oxidation, with the exclusion of intermediate species formation. The hydrolysis of Gxy is observed to actively engage MC without charge inhibition as charge-neutral reacting species are involved, thus, implicating neutral primary salt effect where the variation of ionic strength of the system keeps the redox rate unchanged. The involvement of charge-neutral specie at the rate controlling step encouraged electrostatic attraction when Mg2+ ion additive is incorporated into the system, leading to ion catalysis. The generation of free radical from Gxy aids the emergence of formic acid. The zero intercept of Michaelis–Menten type plot and the non-appreciable shift in the maximum absorption wavelength of the reaction system and MC rule-out the presence of intermediate species. The contribution of thermodynamic enthalpy is instrumental in the redox process, leading to the formic acid product. The inclusion of benzalkonium chloride (BZC) hastens the Gxy oxidation, which is supported by Raghavan and Srinivasan’s model.

N-Carbamoyl Glutamate Prevents Ammonia Intoxication in Piglets with Reduced Liver Function

Ammonia as the end product of protein catabolism has a depressing effect on neurons. Ammonia neutralization in mammals occurs in the ornithine cycle, the activity of which is regulated mainly at the level of synthesis of N-Carbamoyl phosphate. The Krebs cycle is coupled with the ornithine cycle through a common substrate—arginine succinate. Therefore, the effects of ammonia neutralization and the processes of amino acid and energy metabolism are largely interrelated. This study is aimed at evaluating the efficacy of an N-carbamoyl glutamate additive in terms of optimizing metabolic processes and improving ammonia neutralization in suckling piglets. The experiment was conducted on two groups of piglets (n=15) formed at the age of 24 hours. Piglets in the experimental group were fed with an aqueous solution of the drug at a dose of 10 mg/kg of body weight once daily. The duration of feeding the supplement was 30 days; the average daily gain in body weight was determined at weaning at the age of 30 days. An analysis of the blood biochemical composition was carried out on the 30th day from the onset of the experiment. At the end of feeding the supplement in the experimental group, a decrease in the content of ammonia (p&lt;0.05) and urea in blood plasma, an increase in the concentration of arginine (p&lt;0.05) and triacylglycerols (p&lt;0.05) in comparison with the control group was revealed. The N-carbamoyl glutamate additive under study stimulates the endogenous production of the arginine essential amino acid, neutralizes ammonia formed in metabolic processes, and optimizes the amount of metabolic energy spent on binding ammonia in the urea cycle.

Competing mechanisms of charge-induced electric field distortion and charge-involved neutralization on surface discharge

Charge-induced surface discharge poses a critical risk to the operational reliability of high-voltage direct current gas-insulated equipment and pulsed power system. In this study, we investigate the effects of charge-induced electric field distortion and charge involvement during surface discharge by separately depositing charge spots on two dielectric layers. The results show that deposited positive charges inhibit positive streamer development, whereas negative charges facilitate it, primarily due to electric field distortion induced by deposited charges. Nevertheless, the involvement of deposited charges in streamer development predominantly exhibits a neutralizing effect, exerting an opposite influence on the streamers. This highlights a competitive relationship between deposited charge involvement and electric field distortion. Additionally, the neutralization of deposited charges with electron avalanches reduces the impact of charge-induced electric field distortion, thereby mitigating its effects on discharge.

Coagulative removal of polyethylene microplastics using polyaluminum chloride in conjunction with laminarin

As emerging pollutants in water, microplastics pose potential risks to aquatic organisms and human health. Previous studies have shown that traditional metal coagulants are effective in removing microplastics from water, but they come with drawbacks such as high chemical dosage and metal residues. Therefore, it is important to explore more efficient coagulation systems. This study utilized laminarin (LA) as a coagulant aid in combination with polyaluminum chloride (PAC) to investigate its enhanced coagulation performance on polyethylene microplastics in water. The performance of coagulation systems for the removal of microplastics was evaluated for different agent dosages, pH, anion and HA content. Experimental results demonstrated that the compounded system (PAC-LA) significantly improved the removal efficiency of PE microplastics compared to the single PAC system, achieving a removal rate of 91.5 % while reducing the dosage of PAC. The enhanced coagulation mechanism of LA was analyzed using various techniques including scanning electron microscope (SEM), Fourier transform infrared spectroscopy (FTIR), X-ray photoelectron spectroscopy (XPS), and zeta potential analysis. The results indicated that charge neutralization and sweep flocculation were the primary coagulation mechanisms in the presence of PAC alone. The effect of charge neutralization and adsorption was strengthened with the introduction of LA. The coagulation system of PAC-LA showed higher removal rates of PE microplastics under different environmental conditions such as varying pH levels, co-existing anions, and humic acid, suggesting a promising application for microplastic control in water.

Recombinant receptor-binding motif of spike COVID-19 vaccine candidate induces SARS-CoV-2 neutralizing antibody response

Introduction: The SARS-CoV-2 pandemic necessitates effective therapeutic solutions. The receptor-binding motif (RBM) is a subdomain of the spike protein's receptor-binding domain (RBD) and is critical for facilitating the binding of SARS-CoV-2 to the human ACE2 receptor. This study investigates the use of the receptor-binding motif (RBM) domain as an immunogen to produce potent neutralizing antibodies against SARS-CoV-2. Methods: The RBM gene was codon-optimized and cloned into the pET17b vector for expression in E. coli BL21 (DE3) cells, induced with 1 mM IPTG. The recombinant RBM protein was purified using Ni-NTA affinity chromatography. After validating the recombinant RBM by Western blotting with anti-His tag antibodies, BALB/c mice were immunized with 20 µg of the purified RBM protein. Anti-RBM IgG was subsequently purified using protein G resin, and its neutralizing capacity was assessed using the Pishtaz Teb Zaman Neutralization Assay Kit. Results: The recombinant RBM protein, with a molecular weight of 10 kDa, was expressed as inclusion bodies. the typical yield of purification was 27 mg/L of bacterial culture. The neutralization test demonstrated a concentration of 36 µg/mL of neutralizing antibodies in the immunized serum, preventing the spike protein from binding to ACE2. Conclusion: Our study demonstrated that anti-RBM antibodies exhibited neutralization effects on SARS-CoV-2. These findings provide evidence for the development of a vaccine candidate through the induction of antibodies against the RBM, necessitating further studies with adjuvants suitable for human use to evaluate its potential for human vaccination.

Effects of β-blockers on the Outcomes in Patients with Pulmonary Arterial Hypertension Stratified by the Presence of Comorbid Conditions: a Multicenter Prospective Cohort Study (BNP-PL)

Current guidelines do not recommend β-blockers in pulmonary arterial hypertension (PAH) unless indicated by comorbidities. However, the evidence regarding the role of β-blockers in PAH is contradictory. What are the effects of β-blockers on clinical outcomes in patients newly diagnosed with PAH, and how do these outcomes differ based on the presence of cardiovascular comorbidities that are standard indications for β-blocker use? We analyzed data from 806 patients newly diagnosed with PAH enrolled prospectively in the Database of Pulmonary Hypertension in the Polish Population (BNP-PL). The end points were all-cause mortality and a composite of hospitalization due to right heart failure, syncope, or death. Indications for β-blocker use included hypertension, significant arrhythmia, and coronary artery disease. Propensity score matching was used to form a control group based on age, PAH mortality risk variables, and initially introduced PAH-specific therapy. Of the 806 patients, 469 (58.2%) received β-blockers at the time of PAH diagnosis. In propensity score matching, β-blocker treatment showed a higher incidence of the composite end point (hazard ratio, 1.44; 95%CI, 1.04-1.99; P= .03) and had a neutral impact on mortality (hazard ratio, 1.22; 95%CI, 0.87-1.72; P= .25). When stratified according to the presence of comorbidities, β-blockers showed adverse effects on the composite end point in patients without comorbidities and a neutral effect in patients with at least one comorbidity. β-blockers pose significant risks in patients with PAH, especially in patients without coexisting systemic hypertension, coronary artery disease, or arrhythmia. ClinicalTrials.gov; No.: NCT03959748; URL: www. gov.

Impact of Ozone Exposure on the Biochemical Composition of Wheat, Broccoli, Alfalfa, and Radish Seeds During Germination

In recent years, there has been an increasing interest in the use of gaseous ozone (O3) to promote the germination of edible seeds. While its ability to improve seedling vigor and stimulate germination is acknowledged, there has been limited research on the impact of gaseous O3 on the biochemical profile, including phenolic compounds (TPC) and antioxidant activity, of cereals, seeds, and their sprouts. The lack of information has led to the initiation of this study, which aims to assess the impact of ozone treatment duration at a concentration of 50 ppm 1 L min−1, ranging from 1 to 5 h, on the biochemical attributes of broccoli (Brassica oleracea), radish (Raphanus sativus), alfalfa (Medicago sativa) seeds, and sprouts, as well as wheat (Triticum aestivum) grains and sprouts. By optimizing O3 exposure parameters, including duration, this approach has the potential to serve as a valuable tool for enhancing the microbiological and nutritional quality of seeds and cereals. The findings revealed that O3 treatment generally had an adverse impact on TPC in seeds, cereals, and sprouts, resulting in a significant reduction in TPC post O3 treatment. Wheat grains, in particular, displayed the lowest TPC following ozone exposure, with an average decrease of 39.4% compared to the untreated sample. However, it is noteworthy that alfalfa seeds exhibited a positive response to 4 and 5 h O3 treatment, manifesting an average increase in TPC of 13.0% and 27.7%, respectively. In turn, broccoli, radish, and wheat sprouts displayed the lowest TPC, with values of 47.7%, 20.2%, and 18.0% lower than the control samples, respectively. This study revealed that plant responses to O3 exposure varied, and the effects of O3 treatment on TPC levels depended on O3 exposure time. Furthermore, the effect of O3 on the sugar content of the seeds, cereals, and sprouts varied among different plant types, with some showing an increase in content and others showing no substantial changes. This suggests that, depending on the type of seed, O3 may have both positive and neutral effects.

The impact of nickel on plant growth and oxidative balance.

This review summarizes the impact of nickel (Ni) in hydroponics on the growth, basic biochemical parameters and oxidative balance in angiosperms using data from 66 papers (and 181 treatments). Generally, changes in biomass, pigments (chlorophylls and carotenoids) and proteins were negative when comparing concentration (≤100 and >100 μM) and time (≤14 and >14 days). However, we could deduce a higher tolerance to Ni excess in dicots than in monocots. Growth and basic metabolites were often significantly positively correlated. In contrast to proteins, amino acids were positively affected by Ni, indicating proline accumulation and/or protein catabolism. The increase in hydrogen peroxide (H2O2) content was stimulated by time and Ni concentration, and it is higher in dicots and usually negatively correlated with basic metabolites. An increase in Ni concentration stimulates the increase of thiols, but a longer exposure has a neutral or negative effect. On the contrary, the amount of vitamin C (ascorbic acid) is positively influenced by the dose of Ni in roots and the duration of excess Ni in shoots, which points to dynamic changes of this antioxidant in individual organs. Soluble phenols were not as affected, but their importance appears especially in shoots during long-term exposure to Ni with a simultaneous increase in H2O2 content, confirming their antioxidative role. We emphasize that due to the significant quantitative variability in the published studies, we analyze the presented parameters as a percentage change.

Low-Dose Morphine Does Not Cause Sleepiness in Chronic Obstructive Pulmonary Disease: A Secondary Analysis of a Randomized Clinical Trial.

Rationale: Regular, low-dose, sustained-release morphine is frequently prescribed for persistent breathlessness in chronic obstructive pulmonary disease (COPD). However, effects on daytime sleepiness, perceived sleep quality, and daytime function have not been rigorously investigated. Objectives: We sought to determine the effects of regular, low-dose, sustained-release morphine on sleep parameters in COPD. Methods: We conducted prespecified secondary analyses of validated sleep questionnaire data from a randomized trial of daily, low-dose, sustained-release morphine versus placebo over 4 weeks commencing at 8 or 16 mg/d with blinded up-titration over 2 weeks to a maximum of 32 mg/d. Primary outcomes for these analyses were Week-1 Epworth Sleepiness Scale (ESS) and Karolinska Sleepiness Scale (KSS) scores on morphine versus placebo. Secondary outcomes included Leeds Sleep Evaluation Questionnaire scores (end of Weeks 1 and 4), KSS and ESS scores beyond Week 1, and associations between breathlessness, morphine, and questionnaire scores. Measurements and Main Results: One hundred fifty-six people were randomized. Week-1 sleepiness scores were not different on morphine versus placebo (ΔESS [95% confidence interval] versus placebo: 8-mg group, -0.59 [-1.99, 0.81], P = 0.41; 16-mg group, -0.72 [-2.33, 0.9], P = 0.38; ΔKSS vs. placebo, 8-mg group: 0.11 [-0.7, 0.9], P = 0.78; 16-mg group, -0.41 [-1.31, 0.49], P = 0.37). This neutral effect persisted at later time points. In addition, participants who reported reduced breathlessness with morphine at 4 weeks also showed improvement in LSEQ domain scores including perceived sleep quality and daytime function. Conclusions: Regular, low-dose morphine does not worsen sleepiness when used for breathlessness in COPD. Individual improvements in breathlessness with morphine may be related to improvements in sleep. Clinical trial registered with www.clinicaltrials.gov (NCT02720822).

Safety and effectiveness of glycoprotein IIb/IIIa inhibitors in acute coronary syndromes: insights from the SPUM-ACS study

Abstract Aims Data on Glycoprotein IIb/IIIa inhibitors (GPI) use in real world ACS patients following the introduction of potent P2Y12 inhibitors and newer generation stents are scant. Here, we aimed to assess the utilization, effectiveness, and safety of GPI in a large prospective cohort of contemporary ACS patients. Methods SPUM-ACS is a prospective, multicentre study in which ACS patients between 2009 and 2017 were recruited. The primary endpoint of the present study was major adverse cardiovascular events (MACE), a composite of all-cause death, non-fatal myocardial infarction (MI) and non-fatal stroke at one year. Any bleeding events, BARC 3-5 bleeding, and net adverse cardiovascular events (NACE) comprised the secondary endpoints. Results A total of 4395 ACS patients were included in the analysis. GPI-treated patients had more total coronary artery occlusion (56% vs 35%, p&amp;lt;0.001) and thrombus (60% vs 35%, p&amp;lt;0.001) at angiography. Among the propensity score matched (PSM) population (1992 patients equally split into two groups), GPI-treated patients showed lower MACE risk (PSM adjusted HR 0.70, 95% CI 0.49-0.99) but a higher risk of any (PSM adj HR 1.46, 95% CI 1.06-1.99) and major bleedings (PSM adj HR 1.73, 95% CI 1.09-2.76), resulting in a neutral effect on NACE (PSM adj HR 0.87, 95% CI 0.65-1.17). These results remained consistent across all subgroups. When including only patients treated with potent P2Y12, the incidence of MACE was not different between the two groups, (log rank p=0.50 and PSM adjusted HR 0.94, 95% CI 0.60-1.48). In the PSM population, GPI use (HR 0.76, 95% CI 0.59-0.99, p=0.04), TIMI 3 at the end of the procedure (HR 0.16, 95% CI 0.08-0.30, p&amp;lt;0.001) and potent P2Y12 inhibitor use (HR 0.70, 95% CI 0.49-0.99, p=0.04) were protective independent predictors of MACE at follow-up, while oral anticoagulation at discharge (4.48, 95% CI 2.10-9.52, p&amp;lt;0.001), multivessel disease (HR 1.95, 95% CI 1.09-3.48, p=0.02) and KILLIP class ≥ 2 at admission (HR 1.98, 95% CI 0.99-3.95, p=0.05) were adverse independent predictors. Conclusion In ACS patients undergoing PCI and broadly treated with potent P2Y12 inhibitors, GPI use reduced the risk of MACE at 1 year, while increasing the risk of major bleedings, with a neutral effect on NACE. The routine use of these pharmacological agents should be discouraged, while in selected patients GPI use should be considered following personalized balancing between ischaemic and bleeding risk.Events in PSM populationEvents in PSM population

Effects of glucagon-like peptide-1 receptor agonists, sodium-glucose cotransporter-2 inhibitors and their combination on mitochondrial-derived peptide-c and on markers of neurohumoral activation

Abstract Background/Introduction Type-2 diabetes mellitus (T2DM) is associated with increased prevalence of cardiovascular disease. Purpose We investigated the effects of insulin, glucagon like peptide-1 receptor agonists (GLP-1RA), sodium-glucose cotransporter-2 inhibitors (SGLT-2i) and their combination on mitochondrial-derived peptide-c (MOTS-c), N-terminal pro B-type natriuretic peptide (NT-proBNP), growth differentiation factor (GDF)-15, oxidant and antioxidant biomarkers and cardiovascular function of T2DM patients. Methods A total of 160 T2DM patients were randomized to insulin (n = 40), liraglutide (n = 40), empagliflozin (n = 40) or their combination (GLP-1RA+SGLT-2i) (n = 40) as add-on to metformin. We measured at baseline, at 4 and at 12 months of treatment: a) plasma levels of MOTS-c, NT-proBNP and GDF-15, b) thiobarbituric acid reactive substances (TBARS), as an oxidant biomarkers, and 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), as an antioxidant biomarker, and c) global left ventricular longitudinal strain (GLS) and global work index (GWI) using speckle-tracking imaging. Results Twelve months after treatment, treatment with SGLT-2i and GLP-1RA+SGLT-2i improved MOTS-c (14.89 ± 11.27 vs. 7.26 ± 4.21 ng/mL, p=0.009, 6.49 ± 3.91 vs. 5.6 ± 3.38 ng/mL, p=0.029, respectively), while treatment with insulin and GLP-1RA had a neutral effect on this biomarker (p &amp;gt; 0.05; Table). GLP-1RA, SGLT-2i and their combination showed a greater reduction of NT-proBNP (-43.1% vs. -54.2% vs. -56.9% vs. -14.7%; Figure) and GDF-15 (-14.8% vs. -15.2% vs. -16.1% vs. -0.9%) than insulin (p &amp;lt; 0.05 for all comparisons), despite the fact that all patient achieved adequate glycemic control. GLP-1RA or GLP-1RA+SGLT-2i provided a remarkable reduction of TBARS and increase of GLS and GWI compared to insulin or SGLT-2i (p &amp;lt; 0.05). In all patients, at 4 and at 12 months, the percentage reduction of NT-proBNP was associated with the improvement of TBARS, GLS and GWI (p &amp;lt; 0.05). The percentage reduction of GDF-15 was correlated with the increase of ABTS and with the improvement of GLS at 4 and 12 months (p &amp;lt; 0.05). In SGLT-2i group, the percentage increase of MOTS-c was related with the percentage reduction of GDF-15 (r = 0.81, p = 0.005) at 12 months. Conclusions Twelve-month treatment with GLP-1RA, SGLT-2i and their combination resulted in reduction of cardiac biomarkers in parallel with a greater decrease of oxidative stress than insulin in T2DM. SGLT-2i appears more effective in the improvement of MOTS-c and markers of neurohumoral activation.TableFigure

Systematic review and meta-analysis on the effects of physical activity-based interventions on frailty status and clinical outcomes in adults with long-term cardiac conditions

Abstract Introduction Frailty is common in patients with long-term cardiac conditions and is associated with poor clinical outcomes, but limited treatment options are available. Previous studies investigating interventions for frailty have shown positive results with physical activity interventions, however, these studies were largely based on general elderly populations. Aim To assess the impact of physical activity-based interventions on frailty and clinical outcomes in frail patients with long-term cardiac conditions. Methods A systematic review and meta-analysis was conducted based on the PRISMA guideline, using Medline, Embase, CENTRAL, PEDro, CINAHL electronic databases for articles published between 2000 and 2023. Studies which recruited adults &amp;gt;18 years with long-term cardiac conditions (including angina, acute coronary syndrome (ACS), valve disease, heart failure (HF), atrial fibrillation, cardiac surgery and congenital heart disease) who underwent physical activity-based interventions were included. The primary outcome was change in frailty (measured using a validated tool) post intervention. We also assessed the effect of interventions on mortality and hospitalisation. Results We screened 2697 records and reviewed 91 full-text articles; 12 studies (14 articles) were included, involving 1394 patients. Median age of participants ranged from 64 to 82 years. Five studies recruited participants with HF, 3 evaluated participants post transaortic valve implantation, 3 recruited cardiac surgical patients and 1 study evaluated participants post ACS. Physical activity-based interventions were multi-domain, including a mixture of aerobic, strength, balance, endurance and respiratory muscle training; the majority (N=10) had individualised exercise prescription. Meta-analysis of 8 randomised controlled trials (RCTs) showed statistically significant improvement in frailty associated with physical activity-based interventions compared to controls [pooled standardised mean difference (95% CI) = -0.34 (-0.59 to -0.08), p = 0.01, I2 = 48%]. Seven RCTs evaluated change in frailty by short physical performance battery (SPPB). Meta-analysis of these studies showed a statistically significant improvement in SPPB associated with physical activity-based interventions compared to controls [pooled mean difference (95% CI) = 0.83 (0.31-1.34), p = 0.002, I2 = 49%]. (Figure) The magnitude of improvement in SPPB exceeded the minimally clinically significant improvement of 0.5. Three studies evaluated the effect of physical activity-based intervention on clinical outcomes, all of which showed neutral effect on mortality and hospitalisation. Conclusion Physical activity-based interventions in patients with frailty and long-term cardiac conditions were associated with significant improvement in frailty status, with neutral effect on mortality and hospitalisation. Figure: Meta-analysis of 7 RCTs assessing effect of physical activity-based interventions on SPPB

Eleclazine Suppresses Ventricular Fibrillation in Failing Rabbit Hearts with Ischemia- Reperfusion Injury Undergoing Therapeutic Hypothermia.

Eleclazine is a highly selective late sodium current inhibitor, possibly effective in reducing ventricular fibrillation (VF) in heart failure (HF) with ischemia-reperfusion (IR) injury. The electrophysiological effects of eleclazine at therapeutic hypothermia (TH) are unknown. We investigated the effects of eleclazine in suppressing VF in failing rabbit hearts with IR injury undergoing TH. HF was induced by right ventricular pacing. An IR model was created using coronary artery ligation for 60 min, followed by reperfusion for 30 min. Hearts were excised and Langendorff-perfused for optical mapping and electrophysiological studies. Electrophysiological studies were repeated after TH (33 oC) for 30 min or eleclazine (1 μM) infusion for 20 min. In failing IR-injured hearts, eleclazine reduced action potential duration (APD) dispersion and accelerated intracellular Ca2+ uptake to suppress arrhythmogenic alternans, but also exacerbated rate-dependent conduction slowing, resulting in neutral effects on VF inducibility at normothermia. TH increased VF severity. Eleclazine after TH ameliorated TH-induced APD dispersion and further depressed conduction to reduce VF inducibility and severity. TH after eleclazine also slowed conduction to a greater extent to reduce VF inducibility and severity by extrastimulus pacing. In control IR-injured hearts, eleclazine increased VF severity by dynamic pacing at normothermia, which was counteracted by TH. Eleclazine does not prevent VF at normothermia, but reduces VF inducibility and severity by extrastimulus pacing at TH in isolated failing hearts with regional IR injury.

Effects of beta blockers on outcome in patients with PAH with and without comorbid conditions: a Multicenter Prospective Cohort Study (BNP-PL)

Abstract Background The current guidelines do not recommend the use of beta-blockers (BB) in patients with pulmonary arterial hypertension (PAH) unless necessitated by comorbidities. However, the evidence regarding the role of BB in PAH is contradictory. This study investigates the effects of BB on clinical outcomes in patients newly diagnosed with PAH, stratified by the presence of cardiovascular comorbidities that justify their use. Methods We analyzed data from 806 patients newly diagnosed with PAH, who were prospectively enrolled from March 1, 2018, to August 31, 2023, in the multicentre registry, Database of Pulmonary Hypertension in the Polish population (BNP-PL). The primary endpoints were all-cause mortality and a composite of hospitalization due to right heart exacerbation, syncope, or death. The follow-up period for these patients extended until August 31, 2023. Comorbidities in the study group that indicated the need for BB therapy included hypertension, significant arrhythmia, and coronary artery disease. In the Cox regression analysis, the association between BB use and the predefined endpoints was adjusted for age, baseline European Society of Cardiology risk score, and upfront therapy. Results Among the 806 patients, BBs were administered to 469 (58.2%) at the time of PAH diagnosis. In the entire cohort, BB therapy was linked to increased all-cause mortality (29.6% vs 21.4%; p=0.005, Fig 1.A) and a higher incidence of the composite endpoint (34.1% vs 22.6%; p&amp;lt;0.0001, Fig 1.B). The Number Needed to Harm (NNH) for BB treatment, regarding mortality and the composite endpoint, were 12.1 and 8.7, respectively. In patients without comorbidities, BB use significantly raised the risk of both all-cause mortality (Hazard Ratio [HR] 1.81, 95% Confidence Interval [CI]: 1.01-3.15) and the composite endpoint (HR 1.85, 95% CI 1.09-3.14), after adjustments for age, baseline three-strata PAH mortality risk, and upfront therapy. However, in patients with at least one comorbidity, BB therapy showed a neutral effect on both outcomes. Across the whole group, the use of nonselective BB was associated with a higher risk of the composite endpoint (HR 1.78, 95% CI 1.13-2.82, p=0.01) compared to selective β1 agents, although this did not extend to all-cause mortality (HR 1.34, 95% CI 0.79-2.25, p=0.28). Conclusion BBs, particularly nonselective, pose significant risks in patients with PAH who lack comorbidities that justify their use.

Effects of mineralocorticoid receptor antagonists on new-onset or recurrent atrial fibrillation: a Bayesian and frequentist network meta-analysis of randomized trials

Abstract Aims Clinical and translational research suggests that mineralocorticoid receptor antagonists (MRAs) may prevent atrial fibrosis and electrical remodeling associated with atrial fibrillation (AF). This study aimed to consolidate existing evidence from randomized controlled trials (RCTs) evaluating the effect of MRAs on incident or recurrent AF. Methods Medline, Cochrane Library and Scopus were searched until February 12, 2024. Triple-independent study selection, data extraction and quality assessment were performed. Evidence was pooled using both pairwise and Bayesian and frequentist network meta-analyses. Results Twenty-three RCTs (13,358 participants) were identified. Based on the pairwise random effects meta-analysis, MRAs were associated with a significant reduction in AF events compared to placebo or usual care (risk ratio {RR}= 0.75; 95% confidence interval {CI}= [0.66, 0.87]; P&amp;lt; 0.001; I2= 3%). This protective effect was robust both for new-onset and recurrent AF episodes (subgroup p-value= 0.69), while the baseline HF status was not a significant effect modifier (subgroup p-value= 0.58). MRAs demonstrated a significantly higher reduction in AF events for patients with chronic renal disease compared to placebo (RR= 0.78; 95% CI= [0.62, 0.98]; P= 0.03; I2= 0%). The network meta-analyses revealed that only spironolactone was associated with a significant reduction in AF events (Bayesian RR= 0.76; 95% CI= [0.65, 0.89]; P&amp;lt; 0.001; level of evidence moderate; SUCRA 0.731), while eplerenone and finerenone showed a neutral effect. Conclusion MRAs confer a significant benefit in terms of reducing incident or recurrent AF episodes, irrespective of HF status. In this context, spironolactone may be preferable compared to eplerenone or finerenone.

MiRNA-519e promotes cardiomyocyte cell division accompanied by pro-angiogenic and anti-apoptotic effects

Abstract Background MicroRNAs have the therapeutic potential for heart repair after myocardial infarction(MI). Using a functional high-throughput screening approach, we identified microRNA-519e(miR-519e) as an inducer of cell-cycling in human-derived iPSC-cardiomyocytes(hiPSC-­CM). Purpose This study examines miR-519e for cell division in hiPSC-­CM, additional beneficial functions in heart-related non-CMs, and direct target identification and validation in vitro and in vivo. Methods MiR-519e-mimics or miR-scrambled(miR-scr) were transfected into hiPSC-­CM, human aortic endothelial cells(HAEC) and human cardiac fibroblasts(cFB). Proliferative markers (EdU(5-Ethynyl-2'- deoxyuridine), H3P(Phospho-Histone H3) and AURKB(Aurora B-kinase)) and apoptotic response using MTT assay after exposure to doxorubicin were assessed in hiPSC­-CMs. Angiogenic capacity of HAEC was evaluated by tube formation assay and fibrotic response of cFBs using EdU and α-SMA(smooth muscle actin) staining. In vivo, a mouse MI model with ligation of the left anterior descending followed by intramyocardial injection of miR-519e or miR-scr was used. Direct target identification of miR-519e was assessed using in silico analysis combined with RNA-Seq of miR-519e-mimic treated hiPSC-CMs and validated using Ago2-RNA immunoprecipitation(RIP) followed by qPCR. Results After transfection of miR-519e-mimics in hiPSC-­CM, a significant proliferative response indicative of cell division was observed in immunofluorescence staining and on protein level as compared with miR-scr treated hiPSC-CMs (EdU, H3P and AURKB, p&amp;lt;0.01). Apoptosis was blunted in miR-519e-treated hiPSC-CMs (p&amp;lt;0.05). MiR-519e-treatment of HAEC resulted in more meshes (p&amp;lt;0.01) and longer tube length (p&amp;lt;0.05), indicating enhanced pro-angiogenic capacity. Importantly, EdU-uptake and α-SMA were similar in miR-519e-treated cFB as compared to miR-scr, suggesting neutral effects of miR-519e on myofibroblast transition. In mice undergoing MI, injection of miR-519e-mimics significantly downregulated CDKN1A/P21(Cyclin dependent kinase inhibitor 1A) (p&amp;lt;0.05 vs. miR-scr), a direct target of miR-519e and negative master regulator of cell-cycling. For further novel target identification of miR-519e, we performed in silico analysis combined with RNA-Seq data from miR-519e-treated hiPSC-CM and validation using Ago2-RIP followed by qPCR. Herein, we identified PTEN(Phosphatase and tensin homolog) and TGFBR2(Transforming growth factor β receptor 2) that are involved in cardiomyocyte proliferation and hypoxic response, as direct targets of miR-519e. Conclusion Collectively, miR-519e is a potent inducer of cell division in human cardiomyocytes, with anti-apoptotic and pro-angiogenic capacity in the absence of pro-fibrotic effects. We show that miR-519e treatment downregulates key negative cell-cycle regulators in vivo and we identified novel direct targets of miR-519e that are involved in regulatory pathways for heart regeneration.