In this work, artificial neural network coupled with multi-objective genetic algorithm (ANN-NSGA-II) has been used to develop a model and optimize the conditions for the extracting of the Mentha longifolia (L.) L. plant. Input parameters were extraction temperature (40-70 °C), extraction time (4-10 h), and extract concentration (0.25-2 mg/mL) while total antioxidant status (TAS) and total oxidant status (TOS) values of extracts were output parameters. The mean absolute percentage error (MAPE) of selected ANN model was determined as 1.434% and 0.464% for TAS and TOS, respectively. The results showed that the optimum extraction conditions were as follows: extraction temperature of 54.260 °C, extraction time of 7.854 h, and extract concentration of 0.810 mg/mL. The biological activities and phenolic contents of the extract obtained under determined optimum extract conditions were determined. TAS and TOS values of extract were determined as 6.094 ± 0.033 mmol/L and 14.050 ± 0.063 µmol/L, respectively. Oxidative stress index (OSI) as 0.231 ± 0.002, total phenolic content (TPC) as 123.05 ± 1.70 mg/g and total flavonoid content (TFC) as 181.84 ± 1.97 mg/g. Anti- acetylcholinesterase value and anti-butyrylcholinesterase value of the extract was determined as 42.97 ± 0.87 and 60.52 ± 0.80 µg/mL, respectively. In addition, 11 phenolic compounds, namely acetohydroxamic acid, gallic acid, catechin hydrate, 4-hydroxybenzoic acid, caffeic acid, vanillic acid, syringic acid, 2-hydoxycinamic acid, quercetin, luteolin and kaempferol, were determined. It was observed that the extract of M. longifolia produced under optimum conditions exhibited strong biological activities. These results indicate that ANN coupled NSGA-II was an effective method for the optimization extraction conditions of M. longifolia.

Treatment of () infections is challenged by antibiotic resistance. The urease enzyme contributes to colonization in the gastric acidic environment by producing a neutral microenvironment. We hypothesized that urease inhibition could affect viability. This work aimed to assess the effects of acetohydroxamic acid (AHA), ebselen and baicalin on urease activity, bacterial viability and urease genes expression in isolates.

Catheter associated urinary tract infections (CAUTI) caused by urease-positive organisms can lead to catheter blockage: urease metabolizes urea in urine to ammonia causing an increase in pH and hence precipitation of struvite and apatite salts into the catheter lumen and bladder leading to blockage. Acetohydroxamic acid (AHA) is the only urease inhibitor currently approved for patient use, however, it is rarely used owing to its side effects. Here, we report the identification and development of new urease inhibitors discovered using a rational drug design approach. A series of compounds were designed, the compounds were screened and filtered to identify three compounds which were tested in urease activity assays. ,'-Bis(3-pyridinylmethyl)thiourea (Bis-TU) outperformed AHA in activity assays and was tested in an bladder model, where it significantly extended the lifetime of the catheter compared to AHA. Bis-TU was delivered a diffusible balloon catheter directly to the site of activity, thus demonstrating localized drug delivery. This cost-effective drug design approach allowed the identification of a potent urease inhibitor, which could be improved through iterative repeats of the method, and the process of design could be utilized to target other diseases.

Immune checkpoint inhibitors (ICIs) are widely used in cancer treatment; however, the emergence of ICI-associated myocarditis (ICI-MC) presents a severe and potentially fatal complication with poorly understood pathophysiological mechanisms. This study aimed to identify crucial immune-related genes in ICI-MC and uncover potential therapeutic targets using bioinformatics.

Infections with viruses, such as hepatitis C (HCV), dengue (DENV), and yellow fever (YFV) viruses, are major public health problems worldwide. In the case of HCV, treatment is associated with drug resistance and high costs, while there is no clinically approved therapy for DENV and YFV. Consequently, there is still a need for new chemotherapies with alternative modes of action. We have previously identified novel 2-hydroxypyrazino[1,2-]indole-1,3(2,4)-diones as metal-chelating inhibitors targeting HCV RNA replication. Here, by utilizing a structure-based approach, we rationally designed a second series of compounds by introducing various substituents at the indole core structure and at the imidic nitrogen, to improve specificity against the RNA-dependent RNA polymerase (RdRp). The resulting derivatives were evaluated for their potency against HCV genotype 1b, DENV2, and YFV-17D using stable replicon cell lines. The most favorable substitution was nitro at position 6 of the indole ring (compound ), conferring EC 1.6 μM against HCV 1b and 2.57 μΜ against HCV 1a, with a high selectivity index. Compound , carrying the acetohydroxamic acid functionality (-CHCONHOH) on the imidic nitrogen, and compound , the methyl-substituted molecule at the position 4 indolediketopiperazine counterpart, were the most effective against DENV and YFV, respectively. Interestingly, compound had a high genetic barrier to resistance and only one resistance mutation was detected, T181I in NS5B, suggesting that the compound target HCV RdRp is in accordance with our predicted model.

The study investigated OH-derived radicals from certain organic acids employed in nuclear fuel processing and separation using EPR spectroscopy and quantum chemistry methods. Hydroxyl radicals were generated through a Fenton-like reaction within the EPR resonator under both acidic and basic conditions, allowing for the detection of neutral and radical anions, respectively. The spectral assignment and analysis were conducted using a combination of literature data and quantum chemical calculations employing DFT theory with B3LYP or LPBE functionals and the L2a_3 basis set. The reaction of the OH radical with lactic and glycolic acids yielded primary C-centered radicals through hydrogen abstraction from these acids. In contrast, the OH radical exclusively generated secondary radicals from oxalic acid, whereas for citric acid, it resulted in both primary and secondary species induced by decarboxylation. The EPR spectrum of acetohydroxamic acid, upon reaction with the OH radical, displayed a complex pattern featuring primary N-type and N-O-type radicals. The decay pathways of the generated radicals were primarily attributed to radical-radical reactions, with the extracted reaction rate constants generally falling within the typical range observed for such reactions. The EPR parameters calculated for potential radicals using B3LYP and LPBE functionals with L2a_3 basis set demonstrated good accuracy for neutral radicals, albeit requiring minor adjustments for radical anions.

A series of novel hydroxamic acid derivatives was designed and synthesized, and their growth inhibitory activity against bloodstream form was evaluated. These compounds are based on conformationally constrained, lipophilic, spiro carbocyclic 2,6-diketopiperazine (2,6-DKP) scaffolds and bear a side pharmacophoric functionality that contains an acetohydroxamic acid moiety (CHCONHOH) linked with the imidic nitrogen atom of the 2,6-DKP ring via an acetamido portion [CHCON(R), R = H, CH]. Most of these analogues were active in the midnanomolar to low micromolar range against . . ()-Isobutyl- or ()-benzyl-substitution on the methylene carbon located between the amine nitrogen atom and carbonyl of the 2,6-DKP ring was studied. The effect of the methyl-substitution on the nitrogen atom of the acetamido portion in the side pharmacophoric functionality was also examined. Compounds and , bearing an isobutyl- or benzyl-substituent, respectively, and concurrently a methyl-substituent, were found to be the most potent hydroxamates of this series (IC = 34 and 53 nM, respectively). Both had promising selectivity over the parasite compared to mammalian cells (SI = 940 and 470, respectively). Moreover, an conformational behavior study on hydroxamic acid and its methyl-substituted counterpart was undertaken using NMR spectroscopy and theoretical calculations.

Incontinence Associated Dermatitis (IAD) is a type of skin inflammation caused by chronic exposure to urine and/or faeces. Current treatment strategies involve creating a barrier between the skin and urine/faeces rather than targeting specific irritants. Urease expressing pathogens catalyse the conversion of urea, present in urine, into ammonia. The accumulation of ammonia causes an elevation in skin pH which is believed to activate faecal enzymes which damage skin, and opportunistic pathogens, which lead to secondary infections.

and are both known urease producers and have the potential to cause hyperammonemia. We hypothesized that the risk of hyperammonemia is increased by renal failure, burden of cryptococcal infection, and fungal strain characteristics. We performed a retrospective review of plasma ammonia levels in patients with cryptococcal infections. Risk factors for hyperammonemia were statistically compared between patients with and without hyperammonemia (>53 µmol/L). Cryptococcal cells from three patients included in the study were recovered from our biorepository. Strain characteristics including urease activity, ammonia production, growth curves, microscopy, melanin production, and M13 molecular typing were analyzed and compared with a wild-type (WT) strain. We included 29 patients, of whom 37.9% had hyperammonemia, 59% had disseminated cryptococcal infection (DCI), and 41% had isolated central nervous system infection. Thirty-eight percent of patients had renal failure and 28% had liver disease. Renal failure was associated with 4.4 times (95% confidence interval [CI] 1.5, 13.0) higher risk of hyperammonemia. This risk was higher in DCIs (RR 6.2, 95% CI 1.0, 40.2) versus isolated cryptococcal meningitis (RR 2.5, 95% CI, 0.40, 16.0). Liver disease and cryptococcal titers were not associated with hyperammonemia. from one patient with extreme hyperammonemia demonstrated a 4- to 5-fold increase in extracellular urease activity, slow growth, enlarged cell size phenotypes, and diminished virulence factors. Hyperammonemia was strongly associated with renal failure in individuals with DCI, surpassing associations with liver failure or cryptococcal titers. However, profound hyperammonemia in one patient was attributable to high levels of urease secretion unique to that cryptococcal strain. Prospective studies are crucial to exploring the significance of this association.IMPORTANCE produces and secretes the urease enzyme to facilitate its colonization of the host. Urease breaks down urea into ammonia, overwhelming the liver's detoxification process and leading to hyperammonemia in some hosts. This underrecognized complication exacerbates organ dysfunction alongside the infection. Our study investigated this intricate relationship, uncovering a strong association between the development of hyperammonemia and renal failure in patients with cryptococcal infections, particularly those with disseminated infections. We also explore mechanisms underlying increased urease activity, specifically in strains associated with extreme hyperammonemia. Our discoveries provide a foundation for advancing research into cryptococcal metabolism and identifying therapeutic targets to enhance patient outcomes.

The ability of ureolytic bacteria to break down stable urea to alkaline ammonia leads to several environmental and health challenges. Ureolytic bacteria such as and can become pathogenic and cause persistent infections that can be difficult to treat. Inhibiting urease activity can reduce the growth and pathogenicity of ureolytic bacteria. In the present study, we investigated the synergistic effects of tannic acid (TA) and the urease inhibitors fluoride (F) and acetohydroxamic acid (AHA). The concentration of AHA needed for efficient inhibition of the ureolytic activity of can be significantly reduced if AHA is coapplied with tannic acid and sodium fluoride (NaF). Thus, only 1.20 μmol l AHA in combination with 0.30 mmol l tannic acid and 0.60 mmol l NaF delayed the onset of ureolytic pH increase by 95.8 % and increased the growth lag phase by 124.3 % relative to untreated . At these concentrations, without AHA, TA and NaF increased the onset of the ureolytic pH change by only 37.0 % and the growth lag phase by 52.5 %. The strong inhibition obtained with low concentrations of AHA in triple-compound treatments suggests cobinding of F and AHA at the urease active site and could reduce the side effects of AHA when it is employed as a drug against e.g. urinary tract infections (UTIs) and blocked catheters. This study reports the basis for a promising novel therapeutic strategy to combat infections caused by ureolytic bacteria and the formation of urinary tract stones and crystalline biofilms on catheters.