Enzyme Inhibitors Research Articles

Proteinuria and Progression of Renal Damage: The Main Pathogenetic Mechanisms and Pharmacological Approach

The integrity of the glomerular filtration barrier maintains protein excretion below 150 mg/day. When urinary proteins increase, this indicates damage to the filtration barrier. However, proteinuria is not only a marker of kidney damage but also exacerbates it through various mechanisms involving the glomerular and tubulointerstitial compartments. Therefore, it is essential to intervene with renoprotective action that reduces the proteinuria. In this context, Angiotensin-converting enzyme inhibitors and angiotensin receptor blockers are cornerstone treatments. Recent advancements include sodium–glucose cotransporter 2 inhibitors, initially used for glycemic control, now recognized for their renoprotective properties in both diabetic and non-diabetic populations. Another drug, Finerenone, a selective non-steroidal mineralocorticoid receptor antagonist, has emerged as a promising agent, offering anti-inflammatory and antifibrotic benefits with fewer side effects than traditional steroidal options. Finally, dual inhibition of angiotensin II and endothelin-1 receptors through agents like Sparsentan presents a novel approach with significant antiproteinuric effects in IgA nephropathy and focal segmental glomerulosclerosis. This brief review summarizes the mechanisms by which proteinuria promotes kidney damage and the renoprotective therapeutic approaches available, which can be combined with lifestyle modifications and specific treatments for underlying diseases to mitigate the progression of chronic kidney disease.

Diversity of biological activities of crude venom extracted from five species of South China Sea anemones

Developing novel, efficient, and safe peptide drugs from sea anemones has aroused great interest in countries around the world today. Sea anemones contain complex protein and peptide toxins, which determine the diversity of their biological activities. In this study, a variety of activities were assessed for crude venom extracted from five species of South China Sea anemones, including hemolytic, enzyme inhibition, anticancer, insecticidal, analgesic and lethal activities. The most toxic sea anemone was found to be Heteractis magnifica, which has high lethal activity in mice with an LD50 of 11.0 mg/kg. The crude venom of H. magnifica also exhibited a range of the most potent activities, including hemolytic, trypsin inhibitory, cytotoxic activity against U251 and A549 cells, insecticidal and analgesic activities. In addition, the crude venom of Stichodactyla haddoni was the most effective inhibitor of pepsin, and the crude venom of Heteractis crispa was extremely strong toxicity to HepG2 cells. These findings are of great significance for exploring the potential and application of South China Sea anemone resources, and are expected to provide new directions and possibilities for the development of novel anticancer drugs, analgesics and biopesticides.

Discovery of potential natural therapeutics targeting cell wall biosynthesis in multidrug-resistant Enterococcus faecalis: a computational perspective.

Identifying therapeutic inhibitors of crucial enzymes involved in the peptidoglycan biosynthesis pathway is pivotal for developing new treatments against multidrug-resistant Enterococcus faecalis V583. MurM, an essential enzyme in this pathway, plays a significant role in the bacterium's cell wall synthesis, making it an attractive druggable target for novel antimicrobial strategies. This study explored the potential of natural compounds as inhibitors of MurM, aiming to discover promising drug candidates that could serve as the foundation for future therapeutic development. The three-dimensional structure of MurM was predicted, optimized, and its binding pocket was analyzed by comparing it with related structures. Over 4,70,000 natural compounds from the COCONUT database were subjected to virtual high-throughput screening (vHTS). The top lead candidates were selected based on their Lipinski's profile, ADME profile, toxicity profile, estimated binding free energy (ΔG) and estimated inhibition constant (Ki). Interaction pattern analysis was used to evaluate the non-covalent interactions between the inhibitors and key residues in MurM's binding pocket. Molecular dynamics simulations were performed over 300 ns to assess the structural stability and impact of these inhibitors on MurM's enzyme. Three lead compounds-CNP0056520, CNP0126952, and CNP0248480-were identified and prioritized with estimated ΔG ranging from - 9.35 to -7.9kcal/mol. Molecular dynamics simulations revealed minimal impact on MurM's overall structure and dynamics, with the candidate inhibitors forming stable protein-ligand complexes. These interactions were supported by several non-covalent interactions between the candidate inhibitors and key residues within MurM's binding pocket. These findings suggest that the identified natural product candidates could serve as promising inhibitors of MurM, potentially leading to novel therapeutics targeting cell wall biosynthesis in multidrug-resistant E. faecalis.

Intrathecal enalapril reduces adhesion formation in experimentally induced digital flexor tendon sheath injuries in horses.

The objectives of the study were to describe a standing percutaneous adhesion induction model in the digital flexor tendon sheath (DFTS) of horses and to evaluate the effect of intrathecal administration of the angiotensin-converting enzyme (ACE) inhibitor enalapril on tendon healing and adhesion formation. Randomized, blinded, controlled experimental study. Eight healthy horses. A collagenase-induced adhesion model was implemented in the deep digital flexor tendon (DDFT) of both forelimbs under standing ultrasonographic guidance. Daily intrathecal injections of 5 mg enalapril (the treatment condition) were administered to a randomly assigned forelimb for 5 days, with the contralateral limb receiving an equivalent volume of 0.9% NaCl (the control). Lameness and limb circumference were recorded weekly. Horses were euthanized after 8 weeks and evaluated for gross digital flexor tendon sheath (DFTS) adhesions. Tendons were collected for histopathologic scoring of DDFT healing. Paired data were analyzed using a one-sided alternative sign test and longitudinal regression. Multiple DFTS adhesions were formed in control limbs of all horses. The median number of gross DFTS adhesions in treated limbs was less than in control limbs (p = .0039). The average reduction in limb circumference and lameness scores over time occurred faster in treated versus control limbs (p < .025). There were no differences in DDFT histopathologic scores between groups. The standing percutaneous DFTS adhesion induction model demonstrated that intrathecal enalapril reduced DFTS adhesion formation, lameness scores, and limb circumference over time. Intrathecal enalapril administration may reduce morbidity in horses with naturally occurring tendon injuries.

Enzyme (α-Glucosidase, α-Amylase, PTP1B &amp; VEGFR-2) Inhibition and Cytotoxicity of Fluorinated Benzenesulfonic Ester Derivatives of the 5-Substituted 2-Hydroxy-3-nitroacetophenones

The prevalence of small multi-target drugs containing a fluorinated aromatic moiety among approved drugs in the market is due to the unique properties of this halogen atom. With the aim to develop potent antidiabetic agents, a series of phenylsulfonic esters based on the conjugation of the 5-substituted 2-hydroxy-3-nitroacetophenones 1a–d with phenylsulfonyl chloride derivatives substituted with a fluorine atom or fluorine-containing (-CF3 or -OCF3) group were prepared. Their structures were characterized using a combination of spectroscopic techniques complemented with a single-crystal X-ray diffraction (XRD) analysis on a representative example. The compounds were, in turn, assayed for inhibitory effect against α-glucosidase, α-amylase, protein tyrosine phosphatase 1 B (PTP1B) and the vascular endothelial growth factor receptor-2 (VEGFR-2) all of which are associated with the pathogenesis and progression of type 2 diabetes mellitus (T2DM). The antigrowth effect of selected compounds was evaluated on the human breast (MCF-7) and lung (A549) cancer cell lines. The compounds were also evaluated for cytotoxicity against the African Green Monkey kidney (Vero) cell line. The results of an in vitro enzymatic study were augmented by molecular docking (in silico) analysis. Their ADME (absorption, distribution, metabolism and excretion) properties have been evaluated on the most active compounds against α-glucosidase and/or α-amylase to predict their drug likeness.

Patterns and factors associated with electrolyte abnormalities among patients with heart failure in Uganda.

Electrolyte abnormalities (EAs) worsen the clinical course of patients with heart failure (HF). The patterns of EAs vary among patients with HF. This study investigated patterns and factors associated with EAs among patients with HF admitted to Hoima Regional Referral Hospital (HRRH) in western Uganda. This hospital-based cross-sectional study used quantitative data of 384 HF patients admitted at HRRH between 21st February and 15th May 2023. Data on sociodemographic, lifestyle, and medical characteristics were collected and presented as descriptive statistics. EAs were considered electrolyte values below or above the reference normal ranges. Bivariate and multiple logistic regression analyses were conducted to establish associations. An association with a p < 0.05 is considered statistically significant. Of 384 HF patients, 342 (89.1%) had EAs. Hypocalcemia was the most common EA, 165 (43.0%). Among the patients, 69 (18.0%) were on diuretics, 185 (48.2%) were on angiotensin-converting enzyme inhibitors (ACEIs)/angiotensin receptor blockers (ARBs), and 105 (27.3%) were on calcium channel blockers (CCBs). Additionally, 264 (68.8%) had a history of hypertension, and 20 (5.2%) demonstrated good drug adherence. Patients with good drug adherence had lower odds of EAs (Adjusted Prevalence Odds Ratio [Adjusted POR] = 0.2, 95% CI: 0.1-0.7, p = 0.009). Those on diuretics had higher odds of EAs compared to those on ACEIs/ARBs and CCBs, with an Adjusted POR of 5.7 (95% CI: 1.3-15.0, p = 0.019). A history of hypertension also increased the odds of EAs (Adjusted POR = 4.0, 95% CI: 1.9-8.4, p < 0.001). The prevalence of EAs in patients with HF at HRRH was high, with hypocalcemia being the most common. Patients with good drug adherence had lower odds of EAs. On the other hand, diuretic use and a history of hypertension were associated with increased odds of EAs.

Clinical profiles and shortterm outcomes of women with peripartum and dilated cardiomyopathies

Background: We aimed to compare the clinical and sociodemographic characteristics, rate of left ventricular reverse remodeling (LVRR), heart failure hospitalization and allcause mortality of women with peripartum and dilated cardiomyopathies (PPCM and DCM respectively) in Nigeria. Methods: This was a prospective longitudinal study and a total of 130 patients (65 for each group) were recruited consecutively and followed up for 6 months. Results: PPCM patients were younger, but the DCM patients had higher frequency of atrial fibrillation and complete left bundle branch block, higher mean left atrial and LV dimensions, higher LV filling pressures, and worse renal dysfunction, respectively, at baseline. At the end of the 6month followup, 15 female DCM vs 21 PPCM patients (p=0.684) had achieved LVRR, 13 DCM vs 11 PPCM patients (p=0.098) were hospitalized for heart failure, and 10 DCM vs 5 PPCM patients (p=0.098) had died. The odds for achieving LVRR was independently increased by systolic blood pressure (SBP) &gt;100 mmHg and tricuspid annular plane systolic excursion &gt;16 mm at baseline in PPCM patients, and by use of angiotensin converting enzyme inhibitors at baseline in female DCM patients. The odds for mortality were increased by tachycardia and pericardial effusion and reduced by the use of loop diuretics at baseline in DCM patients, and SBP &lt;90 mmHg at baseline increased it by 9fold in PPCM patients. Conclusions: Our results suggest that women with DCM and PPCM differ significantly in their demographic and clinical characteristics, and predictors of clinical outcomes.

Structure-Based Design of Covalent SARS-CoV-2 Papain-like Protease Inhibitors.

The COVID-19 pandemic is caused by SARS-CoV-2, a highly transmissible and pathogenic RNA betacoronavirus. Like other RNA viruses, SARS-CoV-2 continues to evolve with or without drug selection pressure, and many variants have emerged since the beginning of the pandemic. The papain-like protease, PLpro, is a cysteine protease that cleaves viral polyproteins as well as ubiquitin and ISG15 modifications from host proteins. Leveraging our recently discovered Val70Ub binding site in PLpro, we designed covalent PLpro inhibitors by connecting cysteine reactive warheads to the biarylphenyl PLpro inhibitors via flexible linkers. Several leads displayed potent enzymatic inhibition (IC50 = 0.1-0.3 μM) and antiviral activity (EC50 = 0.09-0.96 μM). Fumaramide inhibitors Jun13567 (15), Jun13728 (16), and Jun13714 (18) showed favorable in vivo pharmacokinetic properties with intraperitoneal injection. The X-ray crystal structure of PLpro with Jun13567 (15) validated our design strategy, revealing covalent conjugation between the catalytic Cys111 and the fumaramide warhead. The results suggest these covalent PLpro inhibitors are promising SARS-CoV-2 antiviral drug candidates.

Bio-MOFs Based on Natural Phenolic, Hematoxylin Leverages Biomedical Applications: Enzyme Inhibition, Antioxidant, and Antibacterial Properties.

Here, using natural hematoxylin (HT) as linker, metal-organic frameworks (MOFs) from Cu(II), Fe(II), and Fe(III) ions was prepared. The SEM images and DLS analyses revealed HT-based MOFs are <micrometer sizes with the highest surface area value of 49.2 m2/g for HT-Fe(III) MOFs. Interestingly, HT-based MOFs exhibit fluorescent properties at λem=330 nm with fluorescence intensities of 11485, 2120, and 6790 (a.u) for HT-Cu(II), Fe(II), and Fe(III) MOFs, respectively. Moreover, HT-based MOFs inhibited α-glucosidase enyzme in a concentration-dependent manner e. g., 33.1 %, 69.8 %, and 59.7 % of Cenzyme=500 mg/mL was inhibited by HT-Cu(II)-MOF, HT-Fe(II)-MOF, and HT-Fe(III)-MOFs, respectively. The minimum bactericidal concentration (MBC) values of HT-Cu(II) MOFs for Escherichia coli (gram -), Staphylococcus aureus (gram +), and Candida albicans are determined as 5, 5, and 10 mg/mL, respectively. Also, the antioxidant activities of 250 ppm HT-based MOF based on total phenol content (TPC) tests revealed 279, 208, 124, and 152 mg.gallic acid equivalent/mL (mg GA equivalency/mL) for HT, HT-Cu(II) MOF, HT-Fe(II) MOF, and HT-Fe(III), respectively affirming that antioxidant properties were retained. Moreover, HT-Fe(II) and HT-Fe(III) MOFs (62.5 μg/mL) against human null-1 lung cell line revealed cell viabilities of 98.7±12.2 % and 88.9±11.7 %, respectively as concentration-dependent biocompatibility of MOFs.

Gas Chromatography-Mass Spectrometry and Fourier Transform Infrared Spectroscopy Analysis of Potential α-Glucosidase Inhibitors from Terminalia catappa Leaf Extracts

Aims: Postprandial hyperglyceamia is often caused by insulin insufficiency or cellular glucose uptake complications, and conventional treatments often yield undesirable side effects. The aim of the current work was to assess the α-glucosidase inhibitory activities of T. catappa leaf extracts and use spectroscopic techniques to partially characterize possible inhibitors. Study Design: Phytochemical screening, enzyme inhibition assay and spectrometric and spectroscopic characterization of bioactive compounds from leaf extract and fractions of T. catappa. Place and Duration of Study: The entire work was carried out at the Department of Applied Chemistry and Biochemistry, University for Development Studies, Ghana within nine months. Methodology: T. catappa leaf extract and fractions were qualitatively screened for phytochemicals and their biological activity assessed in α-glucosidase inhibition assay. The potential enzyme inhibitors were characterized by Gas Chromatography-Mass Spectrometry (GC-MS) and Fourier Transform Infrared (FT-IR) Spectroscopy. Results: Phytochemical screening of the extract and the various solvent fractions revealed the presence of alkaloids, flavonoids, saponins, flavanol glycosides, phenolics, and terpenoids. The α-glucosidase inhibition potential of the crude leaf extract was concentration-dependent with a half-maximal inhibitory concentration (IC50) of 337.5 µg/ml. Solvent fractions from the crude extract demonstrated α-glucosidase inhibitory activity with IC50 values ranging from 170.40 µg/ml for ethyl acetate (EtOAc) fraction to 71.90 µg/ml for n-hexane (n-Hex) fraction. Acarbose, the control drug, demonstrated significant α-glucosidase inhibition activity in a similar pattern with IC50 of 6.16 µg/ml. The enzyme inhibition kinetics parameters, specifically the Michaelis constant (KM) and Maximum reaction velocity (Vmax) values, suggested that the inhibition of α-glucosidase by T. catappa extract followed a competitive mode. GC-MS and FT-IR analysis of the n-Hexane fraction identified the compounds Eugenol, Urs-12-en-24-oic acid, 3-oxo-, methyl ester (+)-, phytol, trans-isoeugenol, α-amyrin, and squalene as the potential antidiabetic agents that might be responsible for reducing postprandial blood glucose levels. Conclusion: These findings show that T. catappa leaf extract contains α-glucosidase inhibitors and therefore serves as a valuable resource in the discovery of natural anti-diabetic agents.

Investigation of Algerian Crataegus monogyna Jacq Phenolic Compounds (Using LC-ESI-MS/MS Analysis, Antioxidant Activity, and Enzyme Inhibition) and Their Potential Implications for Food and Nutraceutical Applications

Investigations into the phenolic constituents of the butanolic fraction of Crataegus monogyna were optimized using LC-ESI-MS/MS analysis, identifying and quantifying at least 23 fingerprint phytochemical compounds. The major phenolic compounds were epicatechin (99.916 ± 2.208 mg/g), isoquercetrin (53.31 ± 1.172 mg/g), chlorogenic acid (47.457 ± 1.010 mg/g), quinic acid (37.819 ± 1.406 mg/g), rutin (29.98 ± 0.740 mg/g), hesperidin (5.296 ± 0.177 mg/g, detected for the first time in the C. monogyna species), astragalin (1.774 ± 0.020 mg/g), and nicotiflorin (1.482 ± 0.016 mg/g). The antioxidant properties of the lyophilized butanolic fraction were evaluated using DPPH, GOR, ABTS, CUPRAC, and reducing power assays, all of which demonstrated that there was strong activity. Additionally, the neuroprotective effect was evaluated in vitro, showing a potent inhibitory effect on acetylcholinesterase (AChE) with an IC50 of 43.65 ± 2.10 µg/mL. The antidiabetic effect was investigated through α-amylase inhibition (IC50 = 91.19 ± 0.10 µg/mL), showing high inhibitory activity. In addition, the butanolic extract exhibited significant urease inhibition with an IC50 of 26.36 ± 0.05 µg/mL. These results suggest that Algerian C. monogyna has potential as a therapeutic agent for managing diabetes complications and as a natural source of AChE inhibitors, making it a promising subject for the treatment of urease-related conditions. Its high concentrations of natural antioxidants, such as epicatechin, isoquercetrin, chlorogenic acid, quinic acid, rutin, hesperidin, and astragalin, make it suitable for integration into medicine, pharmaceuticals, cosmetics, and the food sector.

Antidiabetic, anti-inflammatory, antioxidant, and cytotoxicity potentials of green-synthesized zinc oxide nanoparticles using the aqueous extract of Helichrysum cymosum

The current research involved the synthesis of zinc oxide nanoparticles (ZnO-NPs) using an aqueous extract of Helichrysum cymosum shoots, and subsequent characterization via different analytical methods, such as UV–Vis spectroscopy, Scanning electron microscope (SEM), Energy-dispersive X-ray spectroscopy (EDX), X-ray diffraction (XRD), Transmission electron microscope (TEM), and zeta potential. The biological effects of the ZnO-NPs were then tested against C3A hepatocyte cells and L6 myocyte cell lines via series of analysis, including cytotoxicity, antioxidant, anti-inflammatory, and antidiabetic effect via enzymatic inhibition. The UV–Vis analysis showed a maximum absorption spectrum at 360, and the TEM analysis reveals a spherical and hexagonal structures, with an average dimension of 28.05–58.3 nm, and the XRD reveals a crystalline hexagonal structure. The zeta potential evaluation indicated that the ZnO-NPs are relatively stable at − 20 mV, and the FTIR analysis identified some important functional group associated with phenolics, carboxylic acid, and amides that are responsible for reducing and stabilizing the ZnO-NPs. The synthesized ZnO-NPs demonstrated cytotoxic effects on the cell lines at higher concentrations (125 µg/mL and 250 µg/mL), complicating the interpretation of the results of the inflammatory and antioxidant assays. However, there was a significant (p < 0.05) increase in the inhibitions of pancreatic lipase, alpha-glucosidase, and alpha-amylase, indicating beneficial antidiabetic effects.

Angiotensin-Converting Enzyme Inhibitor Washout Period Prior to Angiotensin Receptor/Neprilysin Inhibitor Initiation in the Inpatient Setting.

The 2022 AHA-ACC HFSA Guideline for Management of Heart Failure recommend initiating an angiotensin receptor/neprilysin inhibitor (ARNI) in patients with heart failure with reduced ejection fraction (HFrEF) who can tolerate an angiotensin-converting enzyme inhibitor (ACEi). The manufacturer recommends initiating a 36-hour washout period when switching from ACEi to ARNI due to an increased risk of adverse effects, including angioedema. This study investigated the adherence to the washout period when transitioning from ACEi to ARNI at a community hospital. The primary objective was to assess the rate of adherence to the 36-hour washout when transitioning patients from ACEi to ARNI. Secondary outcomes included heart failure exacerbation readmission rates within 90 days and the rate of adverse effects (angioedema, hypotension, acute kidney injury, and hyperkalemia). This was a retrospective cohort study including patients with HFrEF who were transitioned from ACEi to ARNI during their hospital stay between March 1, 2016 and December 31, 2022. Patients were excluded if they did not receive an ACEi or ARNI during their admission or if they had an ejection fraction >40%. Pearson chi-square was used to analyze categorical data. Of 33 patients included in this study, 67% received the full 36-hour washout period when transitioning from ACEi to ARNI. There were no significant differences between the rates of hospital readmissions or adverse effects between the groups. No patients experienced hyperkalemia or angioedema. This is the first study to our knowledge to describe real-world prescribing practices when transitioning patients from ACEi to ARNI for the treatment of HFrEF. Larger, multicenter studies are needed to provide more data on prescribing practices outside this single center. Future research should also include pharmacist's role in adhering to the recommended washout.

Phytochemical composition, antioxidant activities and anti‐obesity potential of selected vegetables as affected by drying methods

AbstractObesity has reached epidemic statistics worldwide and there is an urgent need to explore more available options in addressing the condition. The potential of vegetables as natural sources of enzyme inhibitors that can regulate the progression of obesity is of great interest particularly due to possible synergistic antioxidant activities. In this study, we evaluated the phytochemical composition, antioxidant activities, and in vitro enzyme inhibition properties of selected vegetables as influenced by sun and cabinet drying methods. While a significant increase was observed in the phytochemical composition of most of the vegetables, the two drying methods had varying effects on all parameters. Cabinet drying yielded higher saponin (2.86–4.67 mg/100 g), phenolics (19.30–57.02 mg/100 g), and alkaloids (31.58–37.61 mg/100 g) contents while sun drying gave better results for flavonoids (88.13–234.15 mg/100 g). The DPPH (11.45%–29.61%) and ABTS (22.70%–35.48%) antioxidant activities varied significantly. Cabinet‐dried cabbage and sun dried carrot flours displayed the highest inhibition against alpha amylase and pancreatic lipase, respectively. A strong positive correlation was observed between phenolics and amylase inhibition and also between saponin and pancreatic lipase inhibition. Consequently, vegetable flours could serve as versatile products for the therapeutic management of obesity and related diseases.

Phytochemical Composition and Functional Properties of Brassicaceae Microgreens: Impact of In Vitro Digestion

The aim of this study was to compare the concentration of phenolic compounds, glucosinolates, proteins, sugars and vitamin C between kohlrabi (Brassica oleracea var. acephala gongylodes), Savoy cabbage (B. oleracea sabauda), Brussels sprouts (B. oleracea gemmifera), cauliflower (B. oleracea botrytis), radish (Raphanus sativus) and garden cress (Lepidium sativum) microgreens for their antioxidant and hypoglycemic potential. In addition, we applied an in vitro-simulated system of human digestion in order to track the bioaccessibility of the selected phenolic representatives, and the stability of the microgreens’ antioxidant and hypoglycemic potential in terms of α-amylase and α-glucosidase inhibition after each digestion phase. Using spectrophotometric and RP-HPLC methods with statistical analyses, we found that garden cress had the lowest soluble sugar content, while Savoy cabbage and Brussels sprouts had the highest glucosinolate levels (76.21 ± 4.17 mg SinE/g dm and 77.73 ± 3.33 mg SinE/g dm, respectively). Brussels sprouts were the most effective at inhibiting protein glycation (37.98 ± 2.30% inhibition). A very high positive correlation (r = 0.830) between antiglycation potential and conjugated sinapic acid was recorded. For the first time, the antidiabetic potential of microgreens after in vitro digestion was studied. Kohlrabi microgreens best inhibited α-amylase in both initial and intestinal digestion (60.51 ± 3.65% inhibition and 62.96 ± 3.39% inhibition, respectively), and also showed the strongest inhibition of α-glucosidase post-digestion (19.22 ± 0.08% inhibition). Brussels sprouts, cauliflower, and radish had less stable α-glucosidase than α-amylase inhibitors during digestion. Kohlrabi, Savoy cabbage, and garden cress retained inhibition of both enzymes after digestion. Kohlrabi antioxidant capacity remained unchanged after digestion. The greatest variability was seen in the original samples, while the intestinal phase resulted in the most convergence, indicating that digestion reduced differences between the samples. In conclusion, this study highlights the potential of various microgreens as sources of bioactive compounds with antidiabetic and antiglycation properties. Notably, kohlrabi microgreens demonstrated significant enzyme inhibition after digestion, suggesting their promise in managing carbohydrate metabolism and supporting metabolic health.

The Impact of Heart Failure Chronic Treatment Prior to Cardiac Transplantation on Early Outcomes

Background and Objectives: Cardiac transplantation represents the option for patients with end-stage heart failure (HF), providing the best survival rate. However, the postoperative complications of transplant patients remain a challenge for clinicians. The objective of our study was to evaluate the effect of preoperative chronic HF treatment on the occurrence of in-hospital complications. Materials and Methods: We retrospectively included a total of 50 patients who underwent cardiac transplantation between January 2011 and December 2023 from the Emergency Institute for Cardiovascular Diseases and Transplantation of Targu Mures. We correlated the preoperative chronic HF treatment with the postoperative complications by Spearmen’s correlation coefficient, respectively. With logistic regression, the associations between the treatment and specific complications were determined. Results: Significant negative correlations were found between Carvedilol treatment with 2-month mortality (r = −0.30; 95% CI: −0.53–−0.02; p = 0.03), Ramipril with hospital stay (r = −0.38; 95% CI: −0.60–-0.12; p &lt; 0.01) and intensive care unit (ICU) stay (r = −0.37; 95% CI: −0.59–−0.11; p = 0.01), and Spironolactone usage with hospitalization duration (r = −0.28; 95% CI: −0.52–−0.01; p = 0.04). Furthermore, Carvedilol treatment represented a protective factor against early acute kidney injury (AKI) (OR: 0.22; 95% CI: 0.05–0.91; p = 0.03). Spironolactone treatment was a protective factor against AGR (OR: 0.12; 95% CI: 0.02–0.66; p = 0.01) treatment, in contrast to angiotensin-converting enzyme inhibitor (ACEI) therapy (OR: 5.30; 95% CI: 1.03–27.17; p = 0.04). Conclusions: Pre-transplant Carvedilol treatment was negatively correlated with the 2-month mortality rate. Ramipril and Spironolactone therapy were negatively correlated with hospitalization duration, and Ramipril was additionally correlated with ICU stay. Moreover, Carvedilol therapy represented a protective factor against early AKI. Pre-transplant Spironolactone was associated with lower event rates of AGR, in contrast to ACEI treatment. Prospective studies with larger cohorts are needed in order to draw drastic conclusions.

Design, Synthesis, QSAR Studies, and Molecular Modeling of Some Novel Bis Methyl 2-[3-(benzo[d]thiazol-2-yl)-2-terephthaloyl-bis-4-oxo-thiazolidin- 5-ylidene]acetates and Screening of their Antioxidant and Enzyme Inhibition Properties.

Benzothiazolamine-based bisthiourea precursors were prepared in good yields. These bisthiourea derivatives were cyclized into symmetrical Bis Methyl 2-[3-(benzothiazol- 2-yl)-2-terephthaloyl-bis-4-oxo-thiazolidin-5-ylidene]acetates, by their condensation with (DMAD) dimethyl but-2-meditate in the presence of dry methanol. All these compounds were evaluated for their biological applications. Antioxidant activities were performed by adopting a DPPH radical assay, and an in vitro enzyme inhibition assay was performed to investigate their enzyme inhibitory potential against butyrylcholinesterase (BChE) and acetylcholinesterase (AChE). Molecular modeling and QSAR studies were performed to monitor the binding propensity of imidathiazolidinone derivatives with enzymes and DNA. Also, electronic and steric descriptors were calculated to determine the effect of structure on the activity of imidathiazolidinone derivatives. The characterization of all the synthesized compounds was done by their physical data, FT-IR, NMR and elemental analysis.

Prophages divert Staphylococcus aureus defenses against host lipids

Phages are ubiquitous in bacteria, including clinical Staphylococcus aureus, where Sfi 21/Sa3 phages often integrate into the hlb gene, which encodes Hlb sphingomyelinase. This integration acts as a rapid regulatory switch for Hlb production. Our findings suggest that Sfi 21/Sa3 prophages and Hlb activity influence S. aureus fitness by modulating the incorporation of the toxic linoleic acid (C18:2) from serum into the bacterial membrane. This process relies on C18:2 derived from 1,3-diglyceride, facilitated by the FakB1 kinase subunit. Palmitic acid (C16), primarily released from serum through Hlb activity, competes with C18:2 for FakB1. This mechanism contributes to adaptation to AFN-1252, an antibiotic inhibiting the fatty acid synthesis pathway (anti-FASII). Since S. aureus relies on exogenous fatty acids for growth, AFN-1252 treatment leads to increased proportion of C18:2 in the membrane. Furthermore, Hlb inhibition, whether by prophage insertion, gene inactivation, or enzyme inhibition, delays S. aureus adaptation, resulting in a higher proportion of C18:2 in the membrane. This study sheds light on the role of lipid environments in infections and may contribute to the accurate prediction of infection risks and therapeutic efficacy. Moreover, since both anti-FASII agent and Hlb inhibitor enhance C18:2 incorporation, they represent potential candidates for combined strategies against S. aureus.

Design, synthesis of antipyrine-based Schiff bases and investigation of their cholinesterase and carbonic anhydrase activities by in vitro and in silico approaches

In this study, a series of antipyrine-based Schiff bases (1-10) was designed, synthesized, and evaluated as potential inhibitors of various metabolic enzymes, including acetylcholinesterase (AChE), butyrylcholinesterase (BChE), and human erythrocyte carbonic anhydrase I and II (hCA I and hCA II). The target molecules were characterized by UV-Vis, FT-IR, 1H NMR, 13CNMR, LC-HRMS, and elemental analysis. All tested derivatives demonstrated low nanomolar inhibition with Ki values of in the range of 20.58 ± 0.35 to 53.11 ± 1.02 nM against AChE, 21.84 ± 0.40 to 54.41 ± 1.05 nM against BChE, 27.45 ± 0.41 to 48.22 ± 0.91 nM against cytosolic hCA I isoform associated with epilepsy, and 6.02 ± 0.11 to 29.32 ± 0.54 nM against cytosolic hCA II isoform associated with glaucoma. In general, most these molecules, except for a few, inhibited these enzymes more than acetazolamide (AZA) and neostigmine. Among them, compounds 5, 7, 8, and 9 showed the best inhibitory activities against AChE, BChE, hCA I, and hCA II, respectively. Docking results were calculated for the compounds that showed the best inhibitory activity against these enzymes and for reference compounds. Based on the molecular docking results, they were determined to have high binding energies, including hydrogen bonds, electrostatic interactions, and hydrophobic interactions. Absorption, distribution, metabolism, and excretion (ADME) parameters determined that all the synthesized pyrazolone ring-bearing Schif base derivatives (1-10) have the expected physicochemical properties in terms of drug-likeness and can be evaluated as orally active potential. Properties such as the electrophilicity index and chemical hardness were also investigated by density functional theory (DFT) at the B3LYP/6-311G** level of theory.

Properties and kinetic behavior of free and immobilized laccase from Oudemansiella canarii: Emphasis on the effects of NaCl and Na2SO4 on catalytic activities

Studies have highlighted the great potential of Oudemansiella canarii laccase in degrading synthetic dyes for reducing their toxicity. Immobilization of enzymes improves usability in degradation processes and the present work succeeded in immobilizing this laccase onto MANAE-agarose. Immobilization improved pH, thermal, and storage stabilities. Both, free and immobilized enzymes presented Michaelis-Menten kinetics with the substrate 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) with Km values of 0.056 ± 0.003 and 0.195 ± 0.022 mM, respectively. Immobilization increased Vmax 1.27-fold. NaCl caused incomplete (hyperbolic) inhibition, which was satisfactorily described by the one-substrate one-modifier mechanism. Immobilization reduced the maximal inhibition by NaCl from 80.2 to 55.7 %. The effect of Na2SO4 was predominantly stimulation, but inhibition of the free enzyme occurred at high substrate concentrations. Stimulation of the immobilized enzyme by Na2SO4 was much more pronounced. It strongly depended on the substrate concentration and was much stronger (up to 300 %) at low substrate concentrations. The combined effects of substrate and sulfate on the immobilized laccase could be satisfactorily described by the one-substrate one-modifier mechanism. The modified response of the immobilized O. canarii laccase to NaCl and Na2SO4 considerably favors its use as a tool in bioremediation processes because environmental contamination by salts frequently represents a strong operational challenge.