RESUMEN

The clinical efficacy of adrenergic ß-receptor (ß-AR) blockers in significantly stabilizing atherosclerotic plaques has been extensively supported by evidence-based medical research; however, the underlying mechanism remains unclear. Recent findings have highlighted the impact of lipid-induced aberrant polarization of macrophages during normal inflammatory-repair and regenerative processes on atherosclerosis formation and progression. In this review, we explore the relationship between macrophage polarization and atherosclerosis, as well as the influence of ß-AR blockers on macrophage polarization. Based on the robust evidence supporting the use of ß-AR blockers for treating atherosclerosis, we propose that their main mechanism involves inhibiting monocyte-derived macrophage differentiation towards an M2-like phenotype.

RESUMEN

BACKGROUND & AIMS: Non-selective ß blockers (NSBBs) may negatively influence renal function through decreasing heart rate and cardiac output. This study aimed to systematically investigate their association. METHODS: PubMed, EMBASE, and Cochrane library databases were searched to identify all relevant studies evaluating the association of NSBBs with renal dysfunction in cirrhotic patients. Unadjusted and adjusted data were separately extracted. Odds ratios (ORs) and hazard ratios (HRs) were pooled. Subgroup meta-analyses were performed according to the proportions of ascites and Child-Pugh class B/C and the mean model for end-stage liver disease (MELD) score. Quality of evidence was assessed using the Grading of Recommendations Assessment, Development, and Evaluation framework. RESULTS: Fourteen studies were finally included. Based on unadjusted data, NSBBs significantly increased the risk of developing renal dysfunction (OR = 1.49; p = 0.03), and this association remained significant in subgroup analyses of studies where the proportions of ascites was >70% and Child-Pugh class B/C was 100%. Based on adjusted data with propensity score matching (adjusted OR = 0.61; p = 0.08) and multivariable regression modelling (adjusted HR = 0.86; p = 0.713), NSBBs did not increase the risk of developing renal dysfunction, and this association remained not significant in subgroup analyses of studies where the proportions of ascites was >70% and <70%, the proportion of Child-Pugh class B/C was <100%, and the mean MELD score was <15. The quality of evidence was very low for all meta-analyses. CONCLUSIONS: NSBBs may not be associated with the development of renal dysfunction in liver cirrhosis. However, more evidence is required to clarify their association in specific populations.

Non-selective ß blockers (NSBBs) may negatively influence renal function through decreasing heart rate and cardiac output in liver cirrhosis.Our meta-analysis failed to support the association of NSBBs with an increased risk of developing renal dysfunction after covariate adjustment.

Asunto(s)

RESUMEN

Enantiopure ß-nitroalcohols, as an important class of nitro-containing compounds, are essential building blocks in pharmaceutical and organic chemistry, particularly for the synthesis of ß-adrenergic blockers. In this study, we present the successful protein engineering of halohydrin dehalogenase HHDHamb for the enantioselective bio-nitration of various phenyl glycidyl ethers to the corresponding chiral ß-nitroalcohols, using the inexpensive, commercially available, and safer nitrite as a nitrating agent. The chiral (R)- and (S)-1-nitro-3-phenoxypropan-2-ols were synthesized by the several enantiocomplementary HHDHamb variants through the whole-cell biotransformation, which showed good catalytic efficiency (up to 43% isolated yields) and high optical purity (up to >99% ee). In addition, we also demonstrated that the bio-nitration method was able to tolerate the substrate at a high concentration of 1000 mM (150 g/L). Furthermore, representative synthesis of two optically active enantiomers of the ß-adrenergic blocker metoprolol was successfully achieved by utilizing the corresponding chiral ß-nitroalcohols as precursors.

Asunto(s)

RESUMEN

Enantioseparation of five ß-adrenergic blockers was studied using two mobile phases on a cellulose tris(3-chloro-4-methylphenylcarbamate) (Lux-Cellulose-2) chiral column in normal phase mode. The first mobile phase composed of n-hexane: ethanol: diethylamine 60: 40: 0.1 by volume has successfully resolved the chromatographic peaks of three pairs of ß-adrenergic blockers namely, bisoprolol, carvedilol and atenolol. A mixture of n-hexane: ethanol: diethyl amine 75: 25: 0.1 by volume was used as the second mobile phase to separate the four pairs of enantiomers, metoprolol, carvedilol, nebivolol and atenolol with high resolution values. The mobile phases were pumped at a flow rate 1 mL/min with column temperature 25 °C using a UV detector at 230 nm. Molecular docking simulations of the five pairs of enantiomers was carried out in the cavities of the chiral stationary phase to gain a better understanding of the interaction between analyte enantiomers and chiral stationary phase and to better understand the mechanism of chiral recognition. According to the results, hydrogen bond interactions and π-π- interactions were the main types of interaction involved in the chiral recognition. Molecular dynamics simulation was performed to investigate the solvent effect on the interaction of the five pair of enantiomers in the chiral stationary phase cavity under dynamic conditions.

RESUMEN

BACKGROUND: Helicobacter pylori (H. pylori) has been recognized as the class I carcinogen of gastric cancer and several studies have demonstrated that chronic stress may accelerate gastric cancer progression. However, the evidence is not sufficient. METHODS: Here, we developed a mouse model that combined H. pylori infection with chronic stress. Gastric inflammation promotes gastric tumor development progression. To evaluate the number of pro-inflammatory cells through observing the numbers of activated macrophages and neutrophils in mice gastric tumors compared with untreated mice or only treated with one factor. ADRα1d /SerpinA1 expression and localization were assessed under stress conditions and H. pylori infection, and evaluated by analyzing IL-1α, CD8, platelet, and RBC status using α- or ß- blockers against gastritis to prevent gastric cancer. RESULTS: Further mechanism study showed that stress hormones increase the number of CD8+ lymphocytes by activating ADRß2 receptors, leading to IL-1α secretion and tumorigenicity. Gastric carcinogenesis also involves gastric muscle contraction mediated through ADRα1d/Serpina1 interaction. Specifically, we showed that the ADRα1d/SerpinA1 complex increases glucose uptake and the development of hypoxia conditions. These responses promote platelet aggregation and muscle contraction. In turn, gastric cancer cells increase lactate production and promote gastric cell proliferation through Muc-13 and IL-1α stimulation. CONCLUSION: H. pylori infection in combination with chronic stress can lead to gastric cancer, and the synergistic effects of cytokine production (i.e. IL-1α), T lymphocyte dysfunction contributes to gastric carcinogenesis which will offer treatment opportunities for stress-associated gastric cancer and provide new strategies for the prevention and treatment of gastric cancer in clinics.

Asunto(s)

RESUMEN

Bacterial resistance to antibiotics is an increasing public health threat as it has the potential to affect people at any stage of life, as well as veterinary. Various approaches have been proposed to counteract the bacterial resistance development. Tackling bacterial virulence is one of the most promising approaches that confer several merits. The bacterial virulence is mainly regulated by a communication system known as quorum sensing (QS) system. Meanwhile, bacteria can sense the adrenergic hormones and eavesdrops on the host cells to establish their infection, adrenergic hormones were shown to enhance the bacterial virulence. In this study, ß-adrenoreceptor blockers were proposed not only to stop bacterial espionage on our cells but also as inhibitors to the bacterial QS systems. In this context, a detailed in silico study has been conducted to evaluate the affinities of twenty-two ß-blockers to compete on different structural QS receptors. Among the best docked and thermodynamically stable ß-blockers; atenolol, esmolol, and metoprolol were subjected to further in vitro and in vivo investigation to evaluate their anti-QS activities against Chromobacterium violaceum, Pseudomonas aeruginosa and Salmonella typhimurium. The three tested ß-blockers decreased the production of QS-controlled C. violaceum, and the formation of biofilm by P. aeruginosa and S. typhimurium. Additionally, the tested ß-blockers down-regulated the P. aeruginosa QS-encoding genes and S. typhimurium sensor kinase encoding genes. Furthermore, metoprolol protected mice against P. aeruginosa and S. typhimurium. Conclusively, these investigated ß-blockers are promising anti-virulence agents antagonizing adrenergic hormones induced virulence, preventing bacterial espionage, and blocking bacterial QS systems.

RESUMEN

BACKGROUND: Currently, it remains unclear whether ß-blockers or nondihydropyridine calcium channel blockers are preferred for the acute management of atrial fibrillation (AF). OBJECTIVE: The objective of this study was to compare the efficacy and safety of intravenous (IV) metoprolol and diltiazem for rate control. METHODS: This was a single-center, retrospective cohort study of patients who presented to the emergency department between 2015 and 2019 with AF with rapid ventricular rate (RVR) and received IV metoprolol or diltiazem. The primary outcome was the percentage of patients who achieved rate control (defined as heart rate < 100 beats per minute). Secondary outcomes included time to rate control, percentage of patients requiring additional agents for rate control, and incidence of cardioversion, bradycardia, and hypotension. RESULTS: A total of 200 patients were included in this study. Rate control was achieved in 35% and 41% of the metoprolol and diltiazem groups, respectively (P = 0.38). Mean time to rate control was not significantly different between the metoprolol and diltiazem groups (35 vs 21 minutes, P = 0.23). One patient developed hypotension, no patient developed bradycardia, and 4 patients required electric cardioversion. No adverse events were observed in patients with ejection fraction ≤40%. CONCLUSION AND RELEVANCE: There was no difference in the achievement of rate control between IV metoprolol and diltiazem. This is the largest study to date comparing the two classes of agents for acute rate control in AF. No patient-specific factors were identified that would influence the preferential use of one medication over the other.

Asunto(s)

RESUMEN

It has become widely accepted that insulin resistance and glucose hypermetabolism can be linked to acute pathologies, such as burn injury, severe trauma, or sepsis. Severe burns can determine a significant increase in catabolism, having an important effect on glucose metabolism and on muscle protein metabolism. It is imperative to acknowledge that these alterations can lead to increased mortality through organ failure, even when the patients survive the initial trauma caused by the burn. By limiting the peripheral use of glucose with consequent hyperglycemia, insulin resistance determines compensatory increased levels of insulin in plasma. However, the significant alterations in cellular metabolism lead to a lack of response to insulin's anabolic functions, as well as to a decrease in its cytoprotective role. In the end, via pathological insulin signaling associated with increased liver gluconeogenesis, elevated levels of glucose are detected in the blood. Several cellular mechanisms have been incriminated in the development of insulin resistance in burns. In this context, the main aim of this review article is to summarize some of the drugs that might interfere with insulin resistance in burns, taking into consideration that such an approach can significantly improve the prognosis of the burned patient.

Asunto(s)

RESUMEN

Interleukin-1ß is a central mediator of innate immune responses and inflammation. It plays a key role in a wide variety of pathologies, ranging from autoinflammatory diseases to metabolic syndrome and malignant tumors. It is well established that its inhibition results in a rapid and sustained reduction in disease severity, underlining the importance of having a repertoire of drugs of this class. At present, there are only three interleukin-1ß blockers approved in the clinic. All of them are biologics, requiring parenteral administration and resulting in expensive treatments. In an exercise to identify small molecule allosteric inhibitors of MAP kinases, we discovered a series of compounds that block IL-1ß release produced as a consequence of a stimulus involved in triggering an inflammatory response. The present study reports the hit-to-lead optimization process that permitted the identification of the compound 13b (AIK3-305) an orally available, potent and selective inhibitor of IL-1ß. Furthermore, the study also reports the results of an in vivo efficacy study of 13b in a LPS endotoxic shock model in male BALB/c mice, where IL-1ß inhibition is monitored in different tissues.

Asunto(s)

RESUMEN

Pregnant women with inherited long QT syndrome (iLQTS) are at an increased risk for preterm delivery and intrauterine growth retardation (IUGR) due to their underlying disease. Additionally, they are at a risk of arrhythmogenic events, particularly during the postpartum period because of physiological changes and increased emotional/physical stress. ß-receptor blockers can effectively prevent life-threatening Torsades de Pointes ventricular tachycardia and they are the treatment of choice in iLQTS. Use of ß-receptor blockers in pregnancy is recommended, although IUGR is commonly reported for prenatally exposed infants. IUGR, particularly in preterm infants, can result in adverse neonatal outcomes. This review was performed to support clinicians in their selection of ß-receptor blocker treatment for their pregnant iLQTS women by (i) summarizing the available literature addressing the impact of different ß-receptor blockers on IUGR and (ii) reporting additional aspects which might influence the ß-receptor blocker selection. In general, experts recommend to use nonselective ß-receptor blockers, such as nadolol and propranolol, for iLQTS management as these drugs seem to be superior in effectiveness. However, ß-1-selective receptor blockers, such as bisoprolol or metoprolol, seem to affect less likely uterine contraction, peripheral vasodilation, and are associated with lower IUGR rates and fetal hypoglycemia. They are therefore recommended, except atenolol, as first-line therapy for pregnant women. Additionally, maternal factors such as iLQTS genotype, other underlying comorbidities (e.g., diabetes mellitus type 1, asthma bronchiale), and uteroplacental dysfunction or fetal factors have to be taken into account. Therefore, each woman with iLQTS who wants to become pregnant should be well-advised for a personalized ß-receptor blocker therapy according to the individual risk-benefit evaluation by a multidisciplinary team of cardiologists, gynecologists, pediatric cardiologists, neonatologists, and clinical pharmacologists. During pregnancy, a close monitoring of IUGR and, after birth, monitoring of bradycardia, hypoglycemia, and respiratory depression in the neonate is mandatory. This review summarizes available data on ß-receptor blocker-related risk for IUGR in prenatally exposed infants and illustrates which factors might influence ß-receptor blocker selection with the aim to support clinicians in their pharmacological management of their pregnant iLQTS patients.

Asunto(s)

RESUMEN

INTRODUCTION: Glaucoma, a neuropathy characterized by increased intraocular pressure (IOP), is the major cause of blindness worldwide and its treatment aims at reducing IOP. AREAS COVERED: The authors review the design of the main classes of anti-glaucoma agents. Drugs which interfere with the aqueous humor secretion (adrenergic agonists/antagonists, carbonic anhydrase inhibitors) and with its outflow, by means of both conventional and non-conventional pathways (prostaglandin (PG) analogs, rho kinase inhibitors, nitric oxide (NO) donors) as well as new agents (adenosine receptors modulators, melatonin - fatty acid amide hydrolase hybrids, tyrosine kinase activators, natriuretic peptide analogs) are considered. EXPERT OPINION: The anti-glaucoma drug field has undergone several developments in recent years with the approval of at least three new drugs belonging to novel pharmacological classes, the rho kinase inhibitors ripasudil and netarsudil, and the PG-NO donor hybrid latanoprostene bunod. Eye drops with combinations of two different drugs are also available, allowing for effective IOP control, with once daily administration for some of them, which assures a better patient compliance and ease of administration. Overall, after more than a decade without new anti-glaucoma drugs, the last year afforded interesting new pharmacological opportunities for the management of this disease.

Asunto(s)

RESUMEN

OBJECTIVES: Systematically review the medical literature for the impact of beta-blockers on mortality and functional capacity in patients who suffered severe traumatic brain injury. DATA SOURCES: The search included MEDLINE, EMBASE, and Ovid Evidence-Based Medicine, clinical trial registries, and bibliographies. STUDY SELECTION: All articles that reported outcome in TBI patients treated with beta-blockers. DATA EXTRACTION: Publication year, number of patients, outcome and follow-up. We performed a meta-analysis for each variable for which there were sufficient data to estimate mean differences. DATA SYNTHESIS: 12 studies were included, which involved retrospectively and prospectively collected data on 14,057 patients. The treatment with beta-blockers was associated with a reduction in mortality in patients who were treated with beta-blockers compared to the control group (OR 0.40, 95% CI 0.30-0.54p = <0.00001), with acceptable heterogeneity between studies (I2 = 65% p = 0.00008). Beta-blocker therapy decreases the risk of negative neurological and functional outcomes (OR 0.59, 95% CI 0.38-0.92 p = <0.00001), a very high statistical heterogeneity between the included studies (I2 = 80% p = 0.00004), being able to influence the results. An increase in favorable neurological and functional outcomes is shown (OR 1.19, 95% CI 1.07-1.31 p = 0.001) with acceptable heterogeneity (I2 = 52% p = 0.08). CONCLUSIONS: The beta-blockers therapy is associated with significantly improves outcome in patients with TBI. Treatment with beta-blockers in patients with TBI is a promising frontier in neurotrauma. ABBREVIATIONS: CI: confidence interval; BB: Beta-Blockers; OR = odds ratio; TBI: Traumatic Brain Injury SD: Standard deviation; SNS: Sympathetic nervous system.

Asunto(s)

RESUMEN

OBJECTIVE: To establish an analytical method for determination of 20 kinds of ß-receptor blockers residues in animal foods by ultra high performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS). METHODS: The samples of animal foods were enzymatic hydrolysis by trichloroacetic acid(TCA), purified by MCX column. The separation was performed on a Waters ACQUITYTM BEH C_(18 )column(100 mm×2. 1 mm, 1. 7 µm), then the target compounds were detected by UPLC-MS/MS with ESI positive ion scan in mode of multiple reaction monitoring(MRM) and quantified by matrix matched external standard method. RESULTS: At the spiked level of 1, 2 and 4 µg/kg, the recoveries of each compound were in the range of 61. 9%-119. 1% with the relative standard deviations of 1. 5%-28. 4%(n=6). The qualitative limits of detections were 0. 01-0. 15 µg/kg and the quantitative limits were 0. 03-0. 50 µg/kg for the 20 targets compounds. By using the established method, the target compounds in 30 animal foods were detected, and no excessive veterinary drug residue were detected. CONCLUSION: The established method is simple, rapid, high sensitivity and good stability, with a wide variety and a certain development. It can provide more convenient and fast detection method support for the daily monitoring of veterinary drug residues in animal foods.

Asunto(s)

RESUMEN

Enantiomeric separation of six ß-adrenergic blockers was systematically studied for the first time on a polysaccharide-based chiral stationary phase, i.e. Chiralpak IB, under the normal-phase mode. The effect of alcohol modifiers, alcohol content and basic additive on enantiomeric separation was evaluated and optimized. Under the optimal conditions, the enantiomers of atenolol, bevantolol, cartelol, esmolol, metoprolol and propranolol were all baseline resolved with resolutions of 1.50, 8.56, 2.05, 2.11, 3.56 and 4.02, respectively. Additionally, molecular docking was tested to explain chiral recognition mechanisms of this set of the drug enantiomers on Chiralpak IB. The details of the various interactions affecting enantiomeric separation were confirmed from the molecular level and the modeling data were in agreement with the chromatographic results concerning the enantioselectivity.

Asunto(s)

RESUMEN

A recent study found a decreased risk of Parkinson disease (PD) associated with the ß2 adrenergic agonist (ß2-agonist) salbutamol. However, other mechanisms might explain this apparent association. Using the UK Clinical Practice Research Datalink, we formed a cohort of 2,430,884 patients aged 50 years or older between 1995 and 2016. During follow-up, 8,604 cases of PD were identified and matched to 86,040 controls on sex, age, date of cohort entry, and duration of follow-up, after applying a 1-year latency time window. Incidence rate ratios of PD associated with use of ß2-agonists were estimated using conditional logistic regression. Ever-use of ß2-agonists was associated with a 17% decreased rate of PD (rate ratio = 0.83, 95% confidence interval: 0.75, 0.91) compared with no use. However, this association was limited to early short-term use and was no longer observed after more than 2 years of cumulative duration of use (rate ratio = 0.97, 95% confidence interval: 0.80, 1.17). A similar pattern was observed when stratifying by time since first ß2-agonist prescription and by duration of follow-up. The apparent association of ß2-agonists with a decreased risk of PD is likely the result of reverse causality rather than a biological effect of these drugs on the risk of PD.

Asunto(s)

RESUMEN

BACKGROUND: ß-Adrenergic blockers (BBs) have been associated with increased risk for severe anaphylactic reaction (AR) from contrast media; however, this was shown with intravenous contrast media, before widespread use of low osmolar contrast media, angiotensin-converting enzyme-inhibitors (ACE-Is), and cardioselective BBs. OBJECTIVE: To assess the risk for AR during cardiac catheterization (CC) associated with BB or ACE-I exposure. METHODS: Patients who experienced adverse reactions during CC from January 2004 to December 2013 were identified; 1 to 2 matched controls were assigned for each case. We analyzed AR rates in association with demographic variables, medication exposures (BBs, ACE-Is, angiotensin-receptor blockers, aspirin), and comorbidities: cardiovascular disorders, asthma, and atopy. RESULTS: We analyzed 71,782 CCs. Of these, severe 70 reactors were identified-46 (0.06%) fulfilled AR criteria. There were 35 cases of mild to moderate AR and 11 cases of severe AR (0.015%). There were no significant differences in age (61.3% vs 61.5%), sex (63% vs 64% males), cardiovascular disorder rate (78% vs 93%), and exposure to BBs (46% vs 51%; cardioselective: 77% vs 80%) and ACE-Is (37.0% vs 37.2%) in cases versus controls. Via multivariate logistic regression, BB exposure was not associated with greater AR frequency (P = .35) or severity (P = .40). Neither cardioselective BBs (P = 0.2) nor noncardioselective BBs (P = .5) influenced AR severity. ACE-Is had no effect on AR frequency (P = .35) or severity (P = .14). Lower AR frequency was associated with cardiovascular disorder (P = .01). CONCLUSIONS: In this case-control study, severe AR was rarely observed. Exposure to BBs or ACE-Is did not significantly influence AR frequency or severity; however, most BBs were cardioselective. Our findings imply that cardioselective BB or ACE-I suspension is not warranted in association with CC.

Asunto(s)

RESUMEN

INTRODUCTION: Metoprolol is the most frequently used ß-receptor blockers; however, the prescribed dose in China is far less than the recommended doses in the guidelines. Based on the Chinese and International guidelines and the Chinese clinical practice, we are conducting this study (NCT03413410) to test the feasibility and tolerability of the metoprolol optimal dosing pathway by observing the percentage of patients achieving target dose in Chinese acute coronary syndrome (ACS) patients during hospitalisation. METHODS AND ANALYSIS: A total of about 1000 patients aged ≥18 years, hospitalised for ACS will be enrolled from ~15 hospital sites in China between February 2018 and April 2019. The percentage of patients achieving the target metoprolol dosage at discharge is the primary endpoint. The secondary endpoints included the following: mean heart rate (HR) and blood pressure (BP) of the patients who have achieved target dose at discharge and during the follow-up period, percentage of patients experiencing bradycardia (HR <50 beats/min), hypotension (BP <90/60 mm Hg) and drug-related temporary heart failure worsening during hospitalisation and 1 month after discharge, respectively. We will also assess the proportion of patients reporting metoprolol-related adverse events and the leading causes for metoprolol discontinuation. ETHICS AND DISSEMINATION: The study protocol has been approved by the Ethics committee of the Chinese PLA General Hospital (number: S2017-112-01). Study findings will be disseminated through presentations at national and international conferences and submitted for publications in peer-reviewed journals. TRIAL REGISTRATION NUMBER: ClinicalTrials.gov registry (NCT03413410).

Asunto(s)

RESUMEN

In this study, ultra performance liquid chromatography-quadrupole electrostatic field orbitrap mass spectrometry was used to establish a high-throughput screening method for 20 ß -blockers and their metabolites in foods of animal origin. The sensitivity and applicability of the established method were improved by optimizing the instrument and pretreatment conditions. The samples were purified by high speed centrifugation at low temperature, and the compounds were separated by a C8 column. Qualitative and quantitative analyses of the compounds were performed in full MS/dd-MS2 (data-dependent MS2) mode. Twenty compounds showed a good linear relationship in the range 0.1-10 µg/L, with correlation coefficients (r2) greater than 0.99. The limits of detection (LODs) ranged from 1 to 5 µg/kg, while the limits of quantification (LOQs) ranged from 2 to 10 µg/kg. The average recoveries were 60.37%-100.84%, with relative standard deviations less than 10%. The method is operationally simple and has good reproducibility and high accuracy, which are essential for ß -blockers and metabolite residue screening in foods of animal-origin.

Asunto(s)

Asunto(s)

RESUMEN

In recent years, ß-adrenergic blockers have become the first choice drugs for glaucoma treatment. Timolol holds the main position among them, being a part of most combined antiglaucoma preparations. The use of timolol maleate in clinical practice may be accompanied by severe side effects affecting different organs and systems. The fact that cells with ß-adrenergic receptors are widely common within the human body explains pharmacodynamic effects of timolol maleate. Because of these undesirable side effects, timolol maleate often evokes negative reaction from doctors and patients, which to certain extent limits its usage in ophthalmological practice. Obviously, efficacy and safety of timolol administration depends on individual characteristics making personalized approach necessary for every patient. Such particular approach, being the foundation of personalized medicine, increases efficacy and safety of timolol while reducing costs by using targeted doses.

Asunto(s)