Deciphering the Molecular Mechanisms of Reactive Metabolite Formation in the Mechanism-Based Inactivation of Cytochrome p450 1B1 by 8-Methoxypsoralen and Assessing the Driving Effect of phe268.

This study provides a comprehensive computational exploration of the inhibitory activity and metabolic pathways of 8-methoxypsoralen (8-MP), a furocoumarin derivative used for treating various skin disorders, on cytochrome P450 (P450). Employing quantum chemical DFT calculations, molecular docking, and molecular dynamics (MD) simulations analyses, the biotransformation mechanisms and the active site binding profile of 8-MP in CYP1B1 were investigated. Three plausible inactivation mechanisms were minutely scrutinized. Further analysis explored the formation of reactive metabolites in subsequent P450 metabolic processes, including covalent adduct formation through nucleophilic addition to the epoxide, 8-MP epoxide hydrolysis, and non-CYP-catalyzed epoxide ring opening. Special attention was paid to the catalytic effect of residue Phe268 on the mechanism-based inactivation (MBI) of P450 by 8-MP. Energetic profiles and facilitating conditions revealed a slight preference for the C4'=C5' epoxidation pathway, while recognizing a potential kinetic competition with the 8-OMe demethylation pathway due to comparable energy demands. The formation of covalent adducts via nucleophilic addition, particularly by phenylalanine, and the generation of potentially harmful reactive metabolites through autocatalyzed ring cleavage are likely to contribute significantly to P450 metabolism of 8-MP. Our findings highlight the key role of Phe268 in retaining 8-MP within the active site of CYP1B1, thereby facilitating initial oxygen addition transition states. This research offers crucial molecular-level insights that may guide the early stages of drug discovery and risk assessment related to the use of 8-MP.

Dietary supplementation with Dunaliella salina microalga promotes quail growth by altering lipid profile and immunity.

The goals of the current research are to ascertain the impacts of Dunaliella salina (DS) on quail growth, carcass criteria, liver and kidney functions, lipid profile, and immune response. Two hundred and forty 7-day-old quail chicks were divided equally into 4 separate groups with 6 replicates with 10 birds each. The groups were as follows: 1) control diet (the basal feed without DS), 2) control diet enriched with 0.25 g DS/kg, 3) control diet enriched with 0.50 g DS/kg, and 4) control diet enriched with 1.00 g DS/kg. Results elucidated that the birds which consumed 0.5 and 1 g DS/kg diet performed better than other birds in terms of live body weight (LBW), body weight gain (BWG), and feed conversion ratio (FCR). There were no significant changes in feed intake (FI) and carcass characteristics due to different dietary DS levels. Compared to the control group, DS-treated groups had better lipid profile (low total cholesterol and LDL values and high HDL values) and immune response (complement 3 values). The quails consumed feeds with different levels of DS had greater (P < 0.038) C3 compared to control. Adding 0.5 and 1 g DS/kg lowered blood concentrations of triglycerides and total protein (TP) values. The high level of DS (1 g/kg) had higher albumin values and lower AST values than other groups (P < 0.05). The creatinine values were at the lowest levels in the group consumed 0.50 g DS/kg feed. No changes (P > 0.05) were demonstrated among experimental groups in the ALT, urea, and lysozyme values. In conclusion, adding D. salina to growing quail diets enhanced growth, immune system, blood lipid profile, and kidney and liver function.

Effects of supplemental vitamin A on reproduction and antioxidative status of aged laying hens, and growth, blood indices and immunity of their offspring.

The purpose of this investigation was to evaluate the impacts of vitamin A (VA) supplementation in feed at levels of 0 (control), 2,000, 4,000, 6,000, and 8,000 IU VA/kg diet on the reproductive efficiency and antioxidative properties of aged Sinai laying hens at 52 wk of age (n = 300 females and 30 males) in 6 replicates (10 females + 1 male/replicate). As well as blood biochemical indicators, carcass characteristics, growth performance, immunity, and the antioxidative status of their chicks. Results showed that diets supplemented with 2,000 or 6,000 IU/kg of VA increased fertility rate and decreased early embryonic mortality (P < 0.05). Increasing VA from 4,000 to 6,000 IU/kg significantly boosted hatchability rates. All VA levels significantly enhanced glutathione peroxidase (GPx) and reduced malondialdehyde (MDA) and late embryonic mortality. In the shell gland, dietary supplementation of 6,000 or 8,000 IU/kg of VA enhanced actions of GPx actions, catalase (CAT), and superoxide dismutase (SOD). In hatched chicks, all VA levels boosted (P < 0.05) hemoglobin, red blood cell count, and serum concentration of total proteins and IgA while decreasing eosinophils percentage and aspartate aminotransferase activity (AST) concentration. Dietary VA supplementations from 4,000 to 8,000 IU/kg improved lymphocytes, serum total antioxidant capacity (TAC), SOD, and IgM, while decreasing heterophils, heterophils/lymphocytes ratio, and creatinine in hatched chicks. Serum triglyceride concentration was reduced by adding 6,000 or 8,000 IU/kg of VA, while globulin and high-density lipoprotein concentrations were heightened only by 8,000 IU/kg of VA. It could be concluded that the dietary supplementation of VA (6,000 IU/kg) improved reproductive efficiency and antioxidative status in the liver and the shell gland of aged laying hens and improved hemato-biochemicals parameters, antioxidative status, and immunity of their offspring.

Effects of varying dietary microalgae levels on performance, egg quality, fertility, and blood biochemical parameters of laying Japanese quails (Coturnix coturnix Japonica).

This experiment was carried out to investigate the nutritional value of Spirulina and Dunaliella (SD) combination levels (0, 0.5, 1.0, 1.5, and 2.0 g/kg) that affected the laying Japanese quail's efficiency, egg quality, fertility, and blood biological indicators. A total of 150 adult Japanese quails, aged 8 wk, were divided into 5 treatments at random, each consisting of 30 quails. There were 5 duplicates for every treatment, with 2 male and 4 female quails in each. Comparing the addition of various concentrations of a mixture of SD to the control treatment, the results showed no substantial rise in egg production, egg weight, or egg mass. When compared to the control group, final body weight (FBW) was improved with SD supplementation. The quails in control consumed more feed intake (FI) (p < 0.05), and they were different from the groups who got SD therapy in that they had a regular feed conversion ratio (FCR). The percentages of hatchability and fertility increased when SD was added to quail meals at up to 1.00 g/kg. When compared to the control quail, the quail supplemented with SD levels showed a non-significant rise in albumin%, yolk%, Haugh unit, and unit surface shell weight (USSW), as well as an increase in eggshell percentage and a drop in egg shape index (p < 0.05). Renal and hepatic enzyme functioning improved when SD was added to the diets. Additionally, lipid profile indicators were reduced by SD supplementation (except low-density lipoprotein-LDL). Moreover, compared to the control, incorporating SD led to a nonsignificant rise in immunoglobulin concentrations (IgG and IgM). In conclusion, adding SD to the diet can improve body weight, lipid profile, immunological response, and liver and kidney functions in Japanese quail.

Impact of ß-glucan dietary supplementation on productive, reproductive performance and physiological response of laying hens under heat stress conditions.

The exploration for effective in-feed additives is growing owing to the global climatic change trend to alleviate the negative effects of heat stress in laying hens. This research assessed the potential of using B-glucan (G) as an antiheat stress agent in Matrouh laying hens subjected to early heat shock programs during the growing period. Factorial design (3 × 3) was used, including 3 levels of heat stress (control, heat shock at 3 d and at 3 d and 8 wk of age) and 3 levels of ß-glucan (0, 100, and 200 mg ß-glucan /kg diet). During the first 12 wk of egg production (EP), treatments were exposed to heat challenge. The results revealed that heat shock program applications at 3 d and 8 wk of age significantly decreased body weight at 36 wk of age (P < 0.05) and reduced (P < 0.05) feed intake (FI). While significantly (P < 0.05) improved feed conversion ratio (FCR), hemoglobin, RBCs, WBCs, immunoglobulin M (IgM), immunoglobulin G (IgG), and Heat shock protein (HSP70) of the Liver (P < 0.01) as compared with the control group. At the same time, there was a decrease in lymphocyte%, H/L ratio, cortisol, and T3 compared to the thermo-neutral control. When compared to the control group, hens fed a diet containing 200 mg of ßG significantly (P < 0.05) improved body weight at 16 wk and final weight at 36 wk, feed conversion (FCR) (g. feed/g. egg mass), hen-day egg production, and egg mass, as well as the digestibility coefficients of crude protein (CP), dry matter (DM), metabolizable energy (ME), and cortisol. The interactions between heat chock programs and ßG levels were nonsignificant for the most studied traits except daily feed intake. Therefore, the early heat shock exposure 2 times and supplementation of Β-glucan (ßG) at 200 mg/kg diet during the growth period for laying hens that are exposed to heat stress during the reproductive period could improve productive, reproductive performance, HSP70 level and enhance immunity responses.

The beneficial effect of nanomethionine supplementation on growth performance, gene expression profile, and histopathology of heat-stressed broiler chicken.

This study investigated the effects of nanomethionine (nano-meth) on performance, antioxidants, and gene expression of HSP70, HSP90 and Heat Shock factor-1 (HSF-1) from the liver, and TLR4 from the jejunum, of broiler chickens reared under normal temperatures or under heat stress. Three hundred 1-day-old chicks were randomly assigned to 5 treatment groups. Group 1 served as control. Under normal temperature, birds in group 2 received nano-meth (10 mL/L of drinking water) from d1 until the experiment ended. Group 3 birds were heat-stressed (HS) and did not receive any supplementation. Group 4 received nano-meth in the same dose from d1 old until experiment ended, and the birds were exposed to HS. Group 5 birds were HS and received supplementation of nano-meth during the HS period only. Nano-meth improved (P < 0.0001) final body weight, weight gain, feed conversion ratio, and also decreased (P < 0.0001) the effect of HS on growth performance. Reduction (P < 0.0001) in malondialdehyde and changes in antioxidant enzymes GPX and CAT activity indicated the antioxidant effect of nano-meth. Nano-meth supplementation caused an increase in the expression of HSP70 , HSP90 and HSF1, and a downregulation of TLR4 gene expression. Additionally, nano-meth-supplemented groups showed marked improvement in the histological liver structure, intestinal morphology and villus height compared to control or HS groups.

Dietary supplement of fermented grass forage regulates growth performance, antioxidant capacity, and immune response of broiler chickens.

The current study aimed to determine the effects of different levels of fermented Juncao grass (FG) on growth parameters, blood constituents, immunity, and antioxidative properties of broilers. A total of 240 (21-d-old) broiler chicks were randomly distributed to four dietary treatments of sixty birds, with six replicate pens and ten birds in each. Fermented grass was added to the basal diet at four levels with 0, 5, 10, and 15% FG. The results revealed that broilers fed 5% FG had significantly higher (P < 0.05) final body weight (FBW), average daily gain (WG) and average daily feed intake (ADFI). The best conversion ratio (FCR) was recorded for broilers supplemented with 5% FG compared to the group supplemented with 15% FG (P < 0.05). Increasing FG % decreased (P < 0.05) anti-inflammatory cytokines IL-4, IL-6, and IL-10. However, FG increased (P < 0.05) proinflammatory cytokines IFN-γ, IL-1, IL-2, IL-12, and TNF-α (P < 0.05). Moreover, IgA, IgG, and IgM levels increased (P < 0.05) with increasing FG %. In addition, increasing FG % in broiler rations significantly increased (P < 0.05) serum antioxidant levels of T-AOC, GSH-PX, SOD, CAT, NO and GSH, but decreased (P < 0.05) MDA levels compared to the control group. Conclusively, fermented Juncao grass would be considered a novel herbal feed additive for improving broiler performance, immunity, antioxidant, and health status. Nevertheless, further research at the molecular level is needed to quantify the effects of these herbal components on cellular and humoral immune functions in broiler chickens.

How toxic is the COVID-19 drug azithromycin in the presence of Posidonia oceanica? Toxicokinetics and experimental approach of meiobenthic nematodes from a metallically pristine area.

The current study presents the results of an experiment carried to assess the impact of azithromycin, a COVID-19 drug, probably accumulated in marine sediments for three years, since the start of the pandemic, on benthic marine nematodes. It was explored the extent to which a common macrophyte from the Mediterranean Sea influenced the toxic impact of azithromycin on meiobenthic nematodes. Metals are known to influence toxicity of azithromycin. The nematofauna from a metallically pristine site situated in Bizerte bay, Tunisia, was exposed to two concentrations of azithromycin [i.e. 5 and 10 µg l-1]. In addition, two masses of the common macrophyte Posidonia oceanica [10 and 20% Dry Weight (DW)] were considered and associated with azithromycin into four possible combinations. The abundance and the taxonomic diversity of the nematode communities decreased significantly following the exposure to azithromycin, which was confirmed by the toxicokinetic data and behaving as substrate for P-glycoprotein (P-gp). The toxicity of 5 µg l-1 dosage of azithromycin was partially reduced at 10% DW of Posidonia and completely at 20% DW. The results showed that 5 µg l-1 of azithromycin can be reduced by the macrophyte P. oceanica when present in the environment at low masses as 10% DW.

Ecotoxicity of chrysene and phenanthrene on meiobenthic nematodes with a case study of Terschellingia longicaudata: Taxonomics, toxicokinetics, and molecular interactions modelling.

Polycyclic aromatic hydrocarbons (PAHs) are toxic for humans and marine fauna alike. The current study assessed the impact of PAHs on the migratory behaviour of meiobenthic nematodes collected from the Bizerte lagoon, Tunisia. The experiment lasted for 15 days and was carried in open microcosms, which comprised a lower, contaminated and an upper, uncontaminated compartment. Three treatments were used, for each of them an untreated control was set up: sediment contaminated with chrysene (116 ng g-1 dry weight (DW), with phenanthrene (116 ng g-1 DW) and a mixture of both. The results showed a significant decrease in diversty and abundance in the lower, contaminated compartments compared to the upper zones. The results also highlighted that under an increased stress some species progressively increased in number, these were considered PAH-tolerant species such as Odontophora villoti, some others had an occasionally increased in number were considered as opportunistic species, such as Paracomesoma dubium and the species that showed a progressive decreased in number, such as Metoncholaimus pristiurus and Steineria sp., Terschellingia. longicaudata, and Oncholaimellus sp. were classified as PAH-sensitive. Moreover, an increase in the activity of biochemical biomarkers was observed following the exposure of males and gravid females of T. longicaudata to 29, 58 and 87 ng g-1 DW of chrysene and phenanthrene paralleled by a higher vulnerability of the latter demographic category. Besides, a significant decrease in fertility of females and an increase in pharyngeal sucking power were observed for both types of PAHs considered. The sex ratio was also significantly imbalanced in the favor of males, which suggest that chrysene and phenanthrene affect also the hormone system of T. longicaudata. The high affinities of these PAHs and their molecular interactions with both germ line development protein 3 (GLD-3) and sex-determining protein (SDP) may justify these results and explain the toxicokinetic attributes.

Chemical profile, antiproliferative and pro-apoptotic activities of essential oils of Pulicaria arabica against A549 lung cancer cell line.

Pulicaria arabica has been traditionally utilized in folk medicine for various purposes such as ulcer treatments as well as antidiarrheal agent. Herein, the chemical profiles of Pulicaria arabica essential oils (PAEOs) and the in vitro antiproliferative effect of PAEOs were investigated. Hydrodistillation was employed to prepare PAEOs which were then characterized by GC/MS, while the antiproliferative effects were investigated by MTT assay as well as flow cytometric and RT-PCR analysis. Sixty-four (99.99 %) constituents were recognized from PAEOs. Carvotanacetone (36.97 %), (-)-carvomenthone (27.20 %) and benzene, 2-(1,1-dimethylethyl)-1,4-dimethoxy- (6.92 %) were the main components. PAEOs displayed IC50 values ranging from 30 to 50 µg/mL. DNA content analysis revealed that A549 cells exposed to PAEOs exhibited an increase in G1 cells population. The flow cytometry analysis results also showed that the PAEOs antiproliferative effect was mediated via apoptosis induction. Furthermore, a modulation in the pro-apoptotic markers (caspase-3 and Bax) and anti-apoptotic (Bcl-2) was also observed. In conclusion, PAEOs exhibited a moderate anti-proliferative effect on A549 cells through modulating the cell cycle progression and apoptosis initiation. These findings could offer a potential therapeutic use of PAEOs in lung cancer treatment.

Mosquito larvicidal activity of pyrrolidine-2,4-dione derivatives: An investigation against Culex quinquefasciatus and molecular docking studies.

The pyrrolidine-2,4-dione derivatives were used to conduct a larvicidal test on Culex quinquefasciatus larvae of the second instar. Mannich base condensation method was used to synthesis the pyrrolidine-2,4-dione derivatives by grindstone method. The reaction conditions were mild, resulting in high yields. An analysis of the synthesized compounds was carried out using FTIR, 1H NMR, 13C NMR, mass spectrometry, and elemental analysis. Synthesized compounds (1a-h) were evaluated for larvicidal activities. Compound 1e (LD50: 26.06 µg/mL), and 1f (LD50: 26.89 µg/mL), and were notably more active against Culex quinquefasciatus than permethrin (LD50: 26.14 µg/mL). The docking studies also demonstrated that 1e, and 1f are potent larvicides with higher binding energy (-12.6 kcal/mol) than the control in the mosquito odorant binding protein (PDB ID: 3OGN). The larvicidal properties of lead molecules have made them important for use as insecticides.

Measurements of basal d-glucose transport through GLUT1 across the intact plasma membrane of isolated segments from single fast- and slow-twitch skeletal muscle fibres of rat.

AIM: To develop a method for direct measurement of the fluorescent d-glucose analogue 2-NBDG transport across the plasma membrane of single skeletal muscle fibres and derive the theoretical framework for determining the kinetic parameters for d-glucose transport under basal conditions. METHODS: A novel method is described for measuring free 2-NBDG transport across plasma membrane of single rat muscle fibres at rest. The 2-NBDG uptake was >90% suppressed by 100 µM cytochalasin B in both fast-twitch and slow-twitch fibres, indicating that the 2-NBDG transport is GLUT-mediated. Fibres were identified as fast-twitch or slow-twitch based on the differential sensitivity of their contractile apparatus to Sr2+ . RESULTS: The time course of 2-NBDG uptake in the presence of 50 µM 2-NBDG follows a one-phase exponential plateau curve and is faster in fast-twitch (rate constant 0.053 ± 0.0024 s-1 ) than in slow-twitch fibres (rate constant 0.031 ± 0.0021 s-1 ). The rate constants were markedly reduced in the presence of 20 mM d-glucose to 0.0082 ± 0.0004 s-1 and 0.0056 ± 0.0002 s-1 in fast-twitch and slow-twitch fibres respectively. 2-NBDG transport was asymmetric, consistent with GLUT1 being the major functional GLUT isoform transporting 2-NBDG in muscle fibres at rest. The parameters describing the transport kinetics for both 2-NBDG and d-glucose (dissociation constants, Michaelis-Menten constants, maximal rates of uptake and outflow) were calculated from the measurements made with 2-NBDG. CONCLUSION: Free 2-NBDG and d-glucose transport across the plasma membrane of single rat muscle fibres at rest is fast, conclusively showing that the rate-limiting step in d-glucose uptake in skeletal muscle is not necessarily the GLUT-mediated transport of d-glucose.

Metabolism of the spade-headed Amphisbaenian worm lizard, Diplometopon zarudnyi (Nikolsky, 1907), in Saudi Arabia (Reptilia: Trogonophidae).

The oxygen consumption rate [Formula: see text] and lactate production of the Amphisbaenian worm lizard Diplometopon zarudnyi were measured at temperatures ranging from 15 °C to 35 °C at 5 °C intervals. The [Formula: see text] was significantly different between resting and active states at any specified temperature, while the average value at the resting state generally rose with increased temperature from 15 °C (0.05 ml O2/g/h) to 25 °C (0.111 ml O2/g/h). The aerobic respiration scopes at resting and active states were also significantly different. The highest Q10 values (3.24 and 1.69) were obtained at 15 °C-20 °C and 30 °C-35 °C during resting and active states, respectively, with these values being significantly different. Lactate concentrations were significantly higher during active states than when resting, and the anaerobic scope was found to increase with increased temperature. There was a proportional increase in ATP molecules (µmoles/g/2 min) during aerobic or anaerobic respiration, as well as in total metabolic scope, with increasing temperature, and the anaerobic scope showed significantly higher values than the aerobic scope, confirming the importance of anaerobic behavior for this species.

Morphological characteristics of worm lizard, Diplometopon zarudnyi (Squamata: Trogonophidae), in the central region of Saudi Arabia.

We report morphometric data for 133 specimens of Diplometopon zarudnyi (Squamata: Trogonophidae) collected across its range within Riyadh province of Saudi Arabia. One-way analyses of variance revealed that differences exist in most characters. Snout-vent length and vent-tail length showed slight and inconsistent differences among samples. D. zarudnyi is characterized by 164-175 body annuli; 165-178 dorsal annuli; 13-17 caudal annuli with absence of caudal autotomy. In addition, 2-4 lateral annuli; 45-54 mid-body segments; 39-50 posterior segments; 4-5 head's plates and 4-6 pre-cloacal pores were recorded in both males and females without gender difference. The present study revealed the widespread distribution of this species in the studied region, since land topography in this area is characterized by the absence of any natural barriers which could restrict the spread of this amphisbaenian lizard D. zarudnyi.