Search Results (273)

The transport mechanisms for substrates and nutrients within the heme pathway of the Toxoplasma gondii apicoplast remain poorly understood. However, studies on heme metabolic enzymes have employed disparate genetic manipulation approaches, limiting direct phenotypic comparisons among different enzymes. This research involved screening potential apicoplast proteins in Toxoplasma gondii by cross-referencing and analyzing protein–protein interaction networks. Within the heme enzyme pathway of the apicoplast, eight enzymes were found to be predominantly conserved in the Sarcocystide family. Utilizing the CRISPR-Cas9 system alongside a U1 snRNP-mediated gene-silencing approach, we developed inducible knockdown strains—iKD-PBGD, iKD-UROS, and iKD-UROD—targeting three key metabolic enzymes crucial for the parasite lytic cycle, as demonstrated through replication experiments. To investigate the transport mechanisms for heme-related nutrients or substrates, we knocked down these three enzymes, using TgGRA12 as an initial marker. Continuous fluorescence signals highlighted the parasitophorous vacuole (PV) membrane surrounding tachyzoites during both early and late replication stages, particularly at 48 h post-rapamycin treatment, indicating a transformation of the cyst-like PV resembling that in Toxoplasma gondii. Phenotypically, knockdown of these heme enzymes led to the formation of slowly replicating, cyst-like parasitophorous vacuoles. However, this morphological change did not significantly affect the acute virulence of the parasites in vivo, as determined by mouse survival assays. This study explored the functional roles of the three intermediate metabolic enzymes, offering a novel viewpoint on the gradual demise of Toxoplasma gondii as a potential target for drug development. Full article

In controlled-environment agriculture systems, such as vertical farms, plants can be cultivated under artificial light-dark (L/D) cycles that deviate substantially from the natural 24 h photoperiod. While these regimes may enhance the growth and quality of certain crops, the physiological response is highly species-specific, and long-term implications for many crop species remain insufficiently characterized. This study aimed to evaluate the effects of a shortened L/D cycle (8/4 h, repeated twice daily) on the morphology, photosynthetic activity, and redox status of eggplant (Solanum melongena L., cv. ‘Almaz’) and tomato (Solanum lycopersicum L., cv. ‘Verlioka Plus’) seedlings, compared to a standard 16/8 h photoperiod. Plants were grown in chambers equipped with light-emitting diode (LED) and fluorescent (FLU) lamps under an equivalent daily light integral (DLI: 11.5 mol m⁻² day⁻¹). The shortened 8/4 h L/D cycle induced significant growth suppression, reduced leaf area, and decreased the maximum quantum yield of photosystem II (Fv/Fm), as well as chlorophyll and carotenoid content in both species. Under this regime, eggplants exhibited leaf chlorosis accompanied by necrotic lesions, whereas tomato plants displayed interveinal chlorosis. Notably, significant accumulation of oxidative stress markers—a 37% increase in malondialdehyde (MDA) and a 30% elevation in H₂O₂ levels—occurred exclusively in eggplant. These findings demonstrate that redistributing the same 16 h daily illumination into two 8/4 h L/D cycles induces stress in both eggplant and tomato plants, with eggplant exhibiting greater injury than tomato. The underlying circadian mechanisms and the apparent differences between LED and FLU lighting require further controlled validation. Full article

This study presents a comprehensive investigation of the structural, morphological, and optical properties of aluminum phthalocyanine (AlPc) and silicon phthalocyanine (SiPc) thin films prepared by the close space sublimation (CSS) method, deposited at different evaporator temperatures of 350 °C, 400 °C, and 450 °C, including heterostructures incorporating an indium tin oxide (ITO) layer. Scanning electron microscopy revealed a clear temperature-dependent evolution of surface morphology, with both materials transitioning from isolated crystallites to dense, highly crystalline films. SiPc exhibited higher nucleation density and earlier film densification, while AlPc showed more pronounced grain growth at elevated temperatures, accompanied by crack formation due to internal stress. Optical absorption spectra indicated a red shift in absorption maxima with increasing deposition temperature, associated with improved crystallinity and reduced defect density. The presence of ITO significantly modified the optical response, introducing additional absorption features in the near-infrared region due to interference effects and free-carrier contributions. Fluorescence measurements revealed enhanced emission intensity with increasing temperature for AlPc, while SiPc showed weaker emission overall. The incorporation of ITO led to substantial fluorescence enhancement and the appearance of additional near-infrared emission bands, highlighting the importance of interface engineering. Transmittance spectra demonstrated that ITO-based heterostructures provide a balance between transparency and absorption, with selective attenuation in the 600–800 nm range, enabling band-stop filter behavior. Raman analysis revealed opposite temperature-dependent trends: increasing structural order in AlPc and gradual disorder in SiPc. X-ray diffraction confirmed the crystalline nature of both materials, showing temperature-induced improvements in crystallinity and crystallite size, as well as distinct differences in molecular packing and preferred orientation. These results demonstrate the potential of AlPc and SiPc thin films as functional optical materials with tunable structural and optical properties. Full article

Keratinocytes are key epidermal cells that are highly susceptible to ultraviolet (UV)-induced damage, which can lead to DNA mutations and the development of malignancies. In parallel, UV radiation induces lipid peroxidation (LPO), potentially facilitating the activation of ferroptosis, a form of programmed cell death. We hypothesized that UV-mediated DNA damage, resulting in the formation of cyclobutane pyrimidine dimers (CPDs), occurs preferentially in cells with elevated LPO levels, and that mild induction of ferroptosis in proliferating keratinocytes selectively eliminates cells with high CPD levels. A human keratinocyte cell line was exposed to UVB radiation and subsequently treated with the ferroptosis inducers RSL3 and erastin. Cell death was assessed using LDH analysis, LPO was measured using the fluorescent probe BODIPY™ 581/591 C11, and CPD formation was quantified by ELISA. Using different doses of UVB, we confirmed UVB irradiation simultaneously increases the cell death rate and LPO and CPDs levels in proliferating keratinocytes. Mild induction of ferroptosis in these cells led to a slight increase in the cell death rate and simultaneously to a drastic reduction in CPD levels, suggesting that there is a specific pool of cells predominantly susceptible to UVB in terms of DNA damage and LPO induction. Our findings support our hypothesis that induction of ferroptosis in proliferating keratinocytes exposed to UVB radiation preferentially eliminates cells with elevated CPD levels and may therefore serve as a protective mechanism against UV-induced carcinogenesis. Full article

Igneous intrusions significantly modify the distribution and mobility of elements in coal. In order to explore the influence of igneous intrusions on the elements in coals, the No. 2-2 coal seam of the Shanxi Formation of Juji Coal Mine was selected as the research focus. The No. 2-2 coal was investigated using X-ray fluorescence spectrometry and a high-resolution inductively coupled plasma mass spectrometer. The results show that the volatile gases accompanying the igneous intrusion, together with CO₂ generated by reaction between the intrusion and the coal seam, led to marginally higher MnO and MgO in the thermally altered coals compared to unaltered coal, and also promoted reprecipitation of carbonate and silicate minerals, which increased loss on ignition. The content of P₂O₅ initially rises with increasing distance from the intrusion, indicating that contact metamorphism can lead to a depletion of major elements in thermally altered coals. Meanwhile, the igneous intrusion caused reprecipitation of silicate minerals, but the high temperatures from the intrusion pyrolyzed some of these minerals, resulting in Li, Sb, and U contents that increase with distance from the intrusion. In contrast, Ge, Mo, Ba, Eu, and Pb, together with hydrothermal fluids, entered the thermally metamorphosed coal via fractures and migrated downward, gradually accumulating within it; their contents therefore decrease with increasing distance. Additionally, the intrusion and associated hydrothermal fluids also induced positive Eu, Gd, and Y anomalies and negative Ce anomalies in the thermally altered coals, whereas intermediate-felsic source rocks and oxidizing conditions produced negative Eu, positive Ce, and weakly negative Y anomalies in coals farther from the intrusion. Contact metamorphism results in a significant element depletion, exerting negative effects. Conversely, hydrothermal fluids and fluids from the host rocks promote the redistribution and enrichment of elements in coal, producing positive effects. Overall, the negative effects outweigh the positive ones. Full article

Background/Objectives: Nobiletin, one of the main components of citrus peel, exhibits potent antioxidant, anti-inflammatory, and metabolic regulatory properties. However, its effect on obesity-associated vasculopathay remains unknown. We aim to investigate the effect of nobiletin in ameliorating oxidative stress and endothelial dysfunction induced by a high-fat diet (HFD). Methods: Male C57BL/6J mice were fed a HFD (60 kcal% fat) or normal chow for four months and orally administered with vehicle or nobiletin (50 mg/kg/day) for 8 weeks. Vasoreactivity in aortas was measured on a wire myograph. Primary rat aortic endothelial cells (RAECs) were isolated from Sprague-Dawley rats for in vitro study. Protein expressions were detected by Western blot. Superoxide production was determined by fluorescence imaging. Results: Exposure to high glucose increased the phosphorylation of JNK (Tyr185) and decreased the protein expressions of Nrf2 and HO-1, as well as downregulated the phosphorylation of AMPK and eNOS (Ser1177) in RAECs. This led to reduced nitric oxide (NO) generation and elevation of oxidative stress. High glucose induction also impaired the endothelium-dependent relaxations (EDRs) in murine aortas. These high glucose-induced impairments were restored by co-treatment of nobiletin (1 μM or 10 μM) whereas effects of nobiletin were abolished by AMPK inhibitor Compound C. The DIO-induced diabetic animal model showed increased body weight and blood pressure, imbalance of glucolipid metabolism, impaired EDRs, and elevated oxidative stress in aortas. AMPK/eNOS and Nrf2/HO-1 pathways were downregulated in aortas from DIO mice. Oral administration of nobiletin could at least partially reverse the above damage. Conclusions: Nobiletin ameliorates endothelial dysfunction by reducing oxidative stress and enhancing NO bioavailability upon activation of AMPK/eNOS and Nrf2/HO-1 pathways in obese diabetic mice. Full article

Background: Terahertz (THz) waves exhibit both photon-like and electron-like properties, showing emerging potential in biomedical applications. Cutaneous squamous cell carcinoma (CSCC) is one of the most common skin tumors. Studies have reported that THz waves can induce apoptosis in cancer cells or ablate tumor tissues. Our previous studies also confirmed that 0.1 THz radiation could significantly promote apoptosis in cutaneous melanoma cells, while it had no apparent effect on fibroblast viability, proliferation, migration, and apoptosis. However, the effects of 0.1 THz radiation on CSCC cells have not yet been explored. Furthermore, there remains a lack of investigation into the structural and functional effects on fibroblasts. Therefore, it is necessary to conduct a systematic study to evaluate the influence of 0.1 THz radiation on both CSCC cells and fibroblasts in order to better understand its potential therapeutic applications in the treatment of skin cancer. Purpose: This study aims to explore the biological effects of 0.1 THz radiation on SCC-7 cells and to uncover the molecular mechanisms underlying THz-induced apoptosis, as well as its potential effect on L-929 cells. Methods: Cell viability was evaluated through the CCK-8 assay, while cell cycle distribution was analyzed with the DNA content detection kit. Wound healing assays were performed to assess cell migration, and Annexin V-FITC staining was used to detect apoptosis. Caspase-3 activity was measured using the caspase-3 activity assay kit. Cell morphology was observed using the Atomic Force Microscope (AFM) and the Transmission Electron Microscopy (TEM). Alterations in membrane potential were detected with the M09 membrane potential probe kit, and intracellular Ca²⁺ levels were quantified using the Fluo-8 AM fluorescent probe. Mitochondrial permeability transition pore (mPTP) opening was assessed with the MPTP detection kit, mitochondrial membrane potential changes were measured using the JC-1 probe kit, and cellular ATP levels were measured with the enhanced ATP assay kit. Subsequently, proteomic analysis was performed. Intracellular reactive oxygen species (ROS) levels were quantified with the ROS detection kit, and cytochrome c (Cyt c) release was quantified using the mouse Cyt c ELISA kit. Apoptosis-inducing factor (AIF) expression was analyzed at both mRNA and protein levels by quantitative real-time PCR (qPCR) and Western blot. AIF expression in CSCC tissues was further evaluated based on the GSE42677 and GSE45164 databases. Finally, cyclosporin A (CsA) was used to inhibit mPTP, and in combination with the iMAC inhibitor, the Aifm1 expression and Cyt c release were examined. Results: Our results showed that THz waves significantly disrupted the membrane integrity of SCC-7 cells and induced mitochondrial structural and functional damage. This resulted in a significant increase in ROS levels and the activation of mPTP and the mitochondrial apoptosis channel (MAC). THz radiation promoted the release of Cyt c and AIF from mitochondria, triggering a noncanonical caspase-3-dependent apoptosis pathway. Notably, L-929 cells did not show significant phenotypic or apoptotic changes under the same irradiation conditions. Bioinformatics analysis of the Gene Expression Omnibus (GEO) database revealed that AIF expression was significantly altered in CSCC tissues compared to normal skin tissues. Conclusions: These findings indicated that 0.1 THz radiation effectively induced apoptosis in SCC-7 cells by triggering mitochondrial dysfunction and ROS generation, which led to the release of AIF. Furthermore, the dysregulation of AIF in CSCC tissues suggested its potential as a promising biomarker. These results provided important molecular insights into the therapeutic potential of THz radiation, particularly for the treatment of cutaneous squamous cell carcinoma. Full article

Background/Objectives: The frequent use of antibiotics has led to increasing drug resistance in Aeromonas hydrophila; therefore, there is an urgent need to develop novel antimicrobial agents to prevent and control bacterial diseases in aquaculture. Allicin E (ALE) is derived from garlic (Allium sativum L.), a plant extensively used in traditional medicine for treating infections. This study aimed to evaluate the potential of ALE against A. hydrophila, a major aquaculture pathogen, by investigating its antibacterial efficacy, mechanisms of action, and in vivo protective effects. Methods: The minimum inhibitory and bactericidal concentrations (MIC/MBC) were determined by broth microdilution. Antibacterial mechanisms were investigated through ROS detection, electron microscopy, fluorescent staining, and content leakage measurement. In vivo efficacy was evaluated in Carassius auratus gibelio by monitoring survival rates and bacterial loads, analyzing immune and antioxidant biomarkers, and histopathological analysis after A. hydrophila challenge. Results: ALE exhibited potent antibacterial activity (MIC = MBC = 8 μg/mL), achieving complete bacterial elimination within 1 h and showing a low resistance propensity. Mechanistically, ALE induced ROS accumulation, causing oxidative damage that disrupted membrane integrity and facilitated the leakage of cellular contents. In vivo, ALE significantly enhanced fish survival, reduced bacterial loads, modulated inflammatory cytokines, boosted antioxidant enzyme activities (SOD and CAT), and alleviated tissue damage. Conclusions: ALE possesses potent in vitro antibacterial activity and exerts an inhibitory effect on bacteria-induced inflammatory responses, effectively combating A. hydrophila through a multi-target mechanism and enhancing host resistance. Full article

Honey authenticity control remains analytically challenging due to the complexity of its matrix and the increasing sophistication of adulteration practices. While chromatographic, spectrometric, and isotopic methods provide high confirmatory accuracy, their routine application is constrained by cost, time, and infrastructure requirements. In this context, fluorescence spectroscopy has emerged as a rapid, non-destructive, and cost-efficient screening approach capable of capturing subtle matrix-level compositional variations. This review critically evaluates the application of steady-state and excitation–emission matrix (EEM) fluorescence in honey quality and authenticity assessment. Fluorescence is positioned within tiered analytical frameworks as a first-line or intermediate screening tool preceding confirmatory chromatographic or NMR-based analyses. Emphasis is placed on intrinsic fluorophore domains, excitation–emission measurement strategies, and chemometric interpretation, including multiway analysis and supervised classification models. Recent developments in portable LED-based systems, laser-induced fluorescence, nanoparticle-based probes, and data-fusion strategies are discussed alongside key limitations related to matrix effects, spectral overlap, reproducibility, and model transferability. The review provides a structured framework for the strategic integration of fluorescence spectroscopy into contemporary honey authentication workflows. Full article

2-Amino-3-methylhexanoic acid (AMHA), as a naturally occurring α-amino acid, exhibits excellent bioactivity in inducing plant resistance to biotic and abiotic stresses, promoting plant growth and alleviating pesticide damage. However, its low yield in nature has hindered its industrialization. This study aims to address the current challenges in AMHA production, including synthetic difficulties, low yield, and poor production safety. We investigated the production capacity of Serratia marcescens NAU002-4 using high-performance liquid chromatography (HPLC), orthogonal experiments, and ion exchange chromatography (IEC), supplemented by precursor feeding assays. Optimization of the fermentation medium revealed that 10 g/L corn steep liquor as the nitrogen source and 10 g/L threonine as the precursor resulted in maximal yield. Further optimization of fermentation parameters—initial pH 7.2, inoculation volume 6%, stirring speed 180 rpm, and aeration rate 350 L/h—led to an AMHA titer of 504 mg/L. In parallel, field trials were conducted to evaluate the effects of pure, biosynthetic AMHA on tea plant growth and photosynthetic performance, using plant physiological data and chlorophyll fluorescence rise kinetics. The results demonstrated that biosynthetic AMHA significantly enhances tea plant growth and photosynthetic efficiency, providing a scientific basis for its development and application as a novel biostimulant. Full article

This study investigated the impact of the pretreatment of pea plants (Pisum sativum L. Ran 1) for five days by three times higher light intensity (360 μmol m⁻² s⁻¹) than the intensity for their cultivation (120 μmol m⁻² s⁻¹) on the photosynthetic apparatus’s ability to withstand moderately high temperatures. Photosystem II (PSII) performance was assessed by pulse amplitude-modulated (PAM) fluorometry—evaluation of F_v/F_m, Chl fluorescence decrease ratio—R_Fd, excitation pressure on PSII (1 − qP), non-photochemical quenching (NPQ) analysis, and PsbA (D1) abundance. The redox state of P700 was used to examine photosystem I (PSI), and the redox kinetics of P700 was evaluated as an estimate of cyclic electron flow (CEF). The energy distribution and interaction between the two photosystems were assessed by 77 K chlorophyll fluorescence. Diphenylhexatriene (DPH) fluorescence polarization and PsbS accumulation were followed to estimate alterations in thylakoid membrane characteristics. Our data show that pea plants pretreated with a higher level of light intensity showed higher resistance to temperature increase, maintaining R_Fd values similar to control plants, and the effect of high temperature on PSII excitation pressure (1 − qP) was mitigated. A significant difference between the two groups of plants was observed in terms of quantum yields in both types of non-photochemical quenching, with light pretreated plants showing no change in the energy partitioning ratio while the exposure of non-high light pretreated plants to elevated temperatures led to a more significant increase in quantum yield of constitutive non-photochemical quenching. When plants were exposed to higher temperature, the accumulation of PsbS, induced by high light treatment, was accelerated, and stabilization of thylakoid membrane also occurred. A complex mechanism behind the enhanced tolerance to higher temperature includes the reorganization of membrane pigment–protein complexes, which is regulated by the buildup of PsbS and the accompanying redistribution of excitation energy. Full article

Background: In cancer surgery, resection of the primary tumor and regional lymph nodes (LNs) is critical. Adequate LN examination is essential to detect metastasis, which determines the cancer stage. Fluorescence emission allows for visual differentiation and rapid monitoring of LNs. Methods: Cancer tissue is stained with a fluorescent dye (indocyanine green, ICG) to identify LNs. Fluorescence is induced from the stained LNs using LED light, and a photosensor coupled with a speaker detects the fluorescence signal and triggers an audible alarm. Filters are applied to prevent false alarms. Results: Upon LN detection, an alarm is emitted from the speaker, and the results are recorded using the LED and a digital multimeter (DMM). In clinical trials, ICG is injected to induce LN fluorescence staining, followed by LED irradiation to induce the fluorescent wavelength and verify LN imaging. Discussion: In clinical trials, ICG stains both LNs and blood vessels, which may lead to false positives. To address this limitation, artificial intelligence algorithms can be trained to specifically identify LNs. Conclusions: Detection of fluorescence wavelengths via photosensors allows for rapid identification of LNs, confirmed through an audible alarm, thereby reducing surgical time. This method shows potential for broad application in cancer surgery. Full article

Drought is a major environmental threat to agriculture. This study examined the role of anthocyanins in plant drought tolerance by comparing two basil varieties differing in leaf anthocyanin content: green basil (Ocimum basilicum L. Italiano Classico) and purple basil (Ocimum basilicum L. Dark Opal). The impact of the PEG-induced drought stress was assessed by monitoring changes in chlorophyll a fluorescence parameters (JIP and PAM), leaf pigment content, anthocyanin and total phenolic levels, oxidative stress markers (malondialdehyde, hydrogen peroxide and membrane integrity), as well as radical-scavenging capacity (DPPH assay). Drought stress led to a modification on both the donor (Wk) and acceptor (Vj) sides of PSII, which influences Q_A reoxidation and amounts of the closed reaction centers (1-qP). These changes inhibited photosystem II photochemistry, the rate of the electron transport (ETR), and the rate of the photosynthesis (R_Fd) and decreased performance indices (PI_ABS, PItotal), as well as the photosystem I photochemistry. The drought-induced changes were associated with an increase in the dissipated energy per reaction center (DI₀/RC). The results show that photosynthetic functions in purple basil were less affected under drought stress compared to green basil. The reason for better tolerance of purple basil is associated with elevated anthocyanin levels, which correlate with enhanced antioxidant capacity, reduced hydrogen peroxide accumulation, lower membrane lipid peroxidation, improved relative water content and membrane stability. In addition, rapid cyclic electron flow around photosystem I and a higher carotenoid to chlorophyll ratio contribute to drought tolerance in purple basil. After re-watering, purple basil recovers its photosynthetic function almost completely, unlike green basil, which shows further suppression. The increase in the anthocyanin content and radical-scavenging capacity, as well as the smaller oxidative damage under drought stress, are the main reasons for the better recovery in purple basil. Overall, the findings highlight that higher anthocyanin accumulation in purple basil confers greater drought tolerance and recovery capacity by stabilizing photosynthetic processes and reducing oxidative stress. Full article

Fluorescence emission-guided blood flow and lymph node location detection are important observation methods in cancer removal surgery, where near-infrared LED illumination is used to induce fluorescence emission. However, conventional LED light sources have narrow beam widths, resulting in a limited excitation area and a restricted field of view (FOV). In this study, we propose a balanced optical illumination module that combines a beam-focusing condenser lens and a beam-diffusing lens to expand the beam width while efficiently redistributing optical energy. When only the LED was used, the beam diameter and central irradiance were 4.0 cm and 1.43 mW/cm², respectively. With the condenser lens, the beam diameter remained nearly unchanged (3.98 cm), while the central irradiance decreased to 0.91 mW/cm². When the condenser was combined with the proposed diffuser structure, the beam diameter increased to 14.1 cm, corresponding to an approximately 3.5-fold expansion, while the central irradiance was measured at 0.72 mW/cm², reflecting the redistribution of optical energy from an initially Gaussian-like irradiance distribution into a wider and more uniform illumination area. This irradiance level exceeds the minimum threshold of 0.6 mW/cm² required to induce fluorescence emission, as defined for the experimental working distance of 30 cm and LED power of 200 mW. By integrating the irradiance distributions of both the bare LED and the proposed structure over their respective illuminated surfaces, the measured total power is physically consistent with energy conservation, showing an expected transmission loss of 18.8% due to optical absorption and scattering. These results demonstrate that the proposed beam diffusion-concentration approach provides an effective and practical solution for wide-field fluorescence-guided lesion observation during cancer removal surgery. Full article

Ni–Fe alloys were prepared via electrodeposition from a citrate electrolyte under different magnetic field (MF) strengths (0.1 T, 0.3 T, and 0.5 T) and configurations parallel (B_‖) and perpendicular (B_⊥) to the electrode surface. The magnetohydrodynamics (MHD) induced by the magnetic field influences their structural, compositional, and electrocatalytic properties. The application of an external homogeneous magnetic field significantly altered the alloy morphology, composition, and crystallinity, which we investigated. Scanning electron microscopy (SEM) and X-ray fluorescence (XRF) studies reflected that moderate MF intensity (0.3 T) led to modest variation in Fe incorporation and produced smoother, denser, and more homogeneous Ni–Fe films, particularly under (B_⊥), while high field strength (0.5 T) resulted in surface roughening and compositional nonuniformity. X-ray diffraction (XRD) confirmed that the perpendicular MF enhanced crystallinity and favored (111) and (200) texture growth, while parallel orientation led to weaker and broader diffraction peaks, providing less-ordered growth. Furthermore, slight variations in Fe content were observed with changes in magnetic field strength and direction. Electrochemical measurements demonstrated that the deposited Ni-Fe under various magnetic field conditions modulated both the hydrogen evolution reaction (HER) and the oxygen evolution reaction (OER) in 1 M NaOH. For HER, the lowest overpotential (η₁₀ = −227 mV at 10 mA cm⁻²) and Tafel slope (120 mV·dec⁻¹) were obtained for the deposited Ni-Fe under (B_‖) at 0.1 T, while for OER, the best catalytic performance occurred for fabricated Ni-Fe alloy under (B_⊥) at 0.3 T with an overpotential value of (η₁₀ = 320 mV, Tafel = 202 mV·dec⁻¹). The deposited Ni-Fe alloys under both orientations exhibited reasonable stability and durability during a prolonged operation process. Overall, the findings demonstrate that controlling magnetic field orientation and intensity during electrodeposition provides a versatile route to tailor the microstructure and optimize the bifunctional electrocatalytic performance of Ni–Fe alloys for the water-splitting reaction in an alkaline medium. Full article