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Combination therapy at appropriately suitable doses presents a promising alternative to monotherapeutic drugs. In this study, Cinnamomum verum and Syzygium aromaticum essential oils and their major compounds have exhibited substantial leishmaniacidal potential against both promastigote and amastigote forms of Leishmania (L.) major. However, they displayed high cytotoxicity against Raw264.7 macrophage cells. Interestingly, when combined with each other or with amphotericin B, they demonstrated a synergistic effect (FIC<0.5) with low cytotoxicity. These combinations are able to modulate the production of nitric oxide (NO) by macrophages. Notably, the combination of S. aromaticum Essential oil with amphotericin B stimulates macrophage cells by increasing NO production to eliminate leishmanial parasites. Furthermore, investigation of the molecular mechanism of action of these synergistic combinations reveals potent inhibition of the sterol pathway through the inhibition of the CYP51 gene expression. The findings suggest that combination therapy may offer significant therapeutic benefits in both food and pharmaceutical fields. [ABSTRACT FROM AUTHOR]

In cattle, Hyalomma scupense serves as an important vector of several pathogens resulting in diseases, subsequently affecting the agricultural field as well as the economy. Resistance to chemical acaricides has become widespread affirming the need for new drugs to tick control. The goal of this study was to investigate the acaricidal, repellent activities as well as the putative mode of action of two essential oils (EOs) from Melaleuca alternifolia (Tea tree) and Chamaemelum nobile (Roman chamomile) on Hyalomma scupense. The chemical composition of EOs was also evaluated. Different concentrations of EOs were tested in vitro for their acaricidal property on adults and larvae of H. scupense using adult immersion test (AIT) and larval packet test (LPT). Additionally, using Ellman's spectrophotometric method, the anticholinesterase (AChE) inhibition activity of M. alternifolia and C. nobile EOs was assessed in order to understand their putative mode of action. The main compounds of C. nobile were α-Bisabolene (22.20%) and (E)-β-Famesene (20.41%). The major components in the analyzed M. alternifolia were Terpinen-4-ol (36.32%) and γ-Terpinene (13.69%). Adulticidal and larvicidal assays demonstrated a promising efficacy of the essential oils against tick H. scupense. The lethal concentration (LC50) values obtained for M. alternifolia and C. nobile oils were 0.84 and 0.96 mg/mL in the AIT and 0.37 and 0.48 mg/mL in the LPT, respectively. Regarding repellent activity, M. alternifolia achieved 100% repellency at the concentration of 1 mg/mL while C. nobile showed 95.98% repellency activity at concentration of 4 mg/mL. Also, M. alternifolia and C. nobile EOs displayed potent AChE inhibition with IC50 value of 91.27 and 100.12 μg/mL, respectively. In the present study, M. alternifolia and, to a lesser degree, C. nobile EOs were found to be effective in vitro acaricides, repellents and acetylcholinesterase inhibitor against H. scupense ticks. These plants may represent an economical and sustainable alternative to toxic synthetic acaricides in the management of ectoparasites of veterinary importance. [ABSTRACT FROM AUTHOR]

The present study aimed to evaluate the leishmanicidal potential of the essential oil (EO) of Micromeria (M.) nervosa and to investigate its molecular mechanism of action by qPCR. Furthermore, in silicointeraction study of the major M. nervosa EO compounds with the enzyme cytochrome P450 sterol 14α-demethylase (CYP51) was also performed. M. nervosa EO was analyzed by gas chromatography-mass spectrometry (GC-MS). Results showed that α-pinene (26.44%), t-cadinol (26.27%), caryophyllene Oxide (7.73 ± 1.04%), and α-Cadinene (3.79 ± 0.12%) are the major compounds of M. nervosa EO. However, limited antioxidant activity was observed, as this EO was ineffective in neutralizing DPPH free radicals and in inhibiting β-carotene bleaching. Interestingly, it displayed effective leishmanicidal potential against promastigote (IC50 of 6.79 and 5.25 μg/mL) and amastigote (IC50 of 8.04 and 7.32 μg/mL) forms of leishmania (L.) infantum and L. major, respectively. Molecular mechanism investigation showed that M. nervosa EO displayed potent inhibition on the thiol regulatory pathway. Furthermore, a docking study of the main components of the EO with cytochrome P450 sterol 14α-demethylase (CYP51) enzyme revealed that t-cadinol exhibited the best binding energy values (−7.5 kcal/mol), followed by α-cadinene (−7.3 kcal/mol) and caryophyllene oxide (−7 kcal/mol). These values were notably higher than that of the conventional drug fluconazole showing weaker binding energy (−6.9 kcal/mol). These results suggest that M. nervosa EO could serve as a potent and promising candidate for the development of alternative antileishmanial agent in the treatment of leishmaniasis. [ABSTRACT FROM AUTHOR]

This study investigated the variability of antioxidant and antibacterial activities of essential oil (EO) from geranium (Pelargonium graveolens) aerial parts, collected from four Tunisian regions. The highest EO yield was obtained in Korba, followed by Beja, Tozeur and Sfax. In all regions, geranium EO was rich in oxygenated monoterpenes mainly due to the predominance of citronellol and geraniol. Antioxidant potential of geranium EO was evaluated by DPPH, ABTS, iron reducing power and β-carotene bleaching assays. Independently of region, geranium EO showed a weak antioxidant potential as compared to the positive control BHT. However, geranium EO was endowed with important antibacterial properties. In fact, Escherichia coli and Staphylococcus aureus were the most sensitive strains than Serratia marcescens, Pseudomonas aeruginosa, Pseudomonas aeruginosa and Enterococcus hirae. These findings suggested that geranium EO could be used as new and rich potential sources of many future multipurpose products for food preservation, pharmaceutical, and industrial applications. [ABSTRACT FROM AUTHOR]

Chlorpyrifos (CPF) poisoning is a public health problem for which there is not currently any effective prophylaxis. In this study, we investigated the protective effect of grape seed extract (GSE) against CPF-induced hepatotoxicity. Rats were daily treated either with CPF (2 mg/kg) or CPF and GSE (20 mg/kg) for 1 week, sacrificed, and their livers dissected for biochemical, molecular, and histopathological analyses. CPF generated liver dysfunction by altering carbohydrate, lipid, amino acid, ammonia and urea metabolism, and provoked mitochondrial impairment through disturbing tricarboxylic acid (TCA) cycle, oxidative phosphorylation (OXPHOS), and mitochondrial viability. CPF also induced cholinergic excitotoxicity along with oxidative stress and histopathological alterations. Interestingly, treatment with GSE prevented all the detrimental effects of CPF through the regulation of cytochrome P450 (CYP450) gene expression. Molecular docking analysis indicated that GSE-containing polyphenols acted as epigenetic modulators through inhibiting DNA (cytosine-5)-methyltransferase 1 (DNMT1), thus favoring the CYP2C6 detoxification pathway. Thereby, GSE might be a promising strategy in the protection of the liver against CPF toxicity. [ABSTRACT FROM AUTHOR]

The essential oils (EOs) of Coriandrum sativum, Carum carvi and Artemisia herba-alba have been screened for their insecticidal activity against Tribolium castaneum, Sitophilus oryzae and Lasioderma serricorne and for their antimicrobial activity against Gram (+), Gram (-) bacteria and against yeast strain. Artemisia herba-alba EO showed good insecticidal activity with 24-h against L. serricorne (LC50 = 29.7) and against T. castaneum (66.1 µg/mL) and antibacterial potentials against Staphyloccocus aureus (MIC = 0.125 mg/mL). The antimicrobial activities of C. carvi EO, rich in D-carvone (72.4%) and D-limonene (23.8%) were particularly remarkable, with LC50 of 27.9 µg/mL against L. serricorne. Similarly, coriander EO, with linalool (64.6%) as the major compound was selected for its antimicrobial activities (Candida albicans, MIC = 1 mg/mL). These findings demonstrated that the tested EOs could be used as a natural source of insecticidal and antimicrobial activity, which may have a variety of uses in the food and pharmaceutical sectors. [ABSTRACT FROM AUTHOR]

The present study deals with the toxicity of Pinus sylvestris essential oil against red poultry mites Dermanyssus gallinae and Hyalomma scupense cattle ticks in laboratory condition. Individual essential oil components identified using a gas chromatograph/mass spectrometer, namely α-pinene (44.92%), β-pinene (17.51%), δ-3 carene (16.18%) and D-Limonene (9.21%) were tested as well as. For this purpose, different doses (0.31; 0.62; 1.25 and 2.5 mg/mL) of essential oil and four monoterpenes using mites contact and spraying in vitro bioassays were carried out. Amitraz and thymol (positive controls), and ethanol and distilled water (negative controls) were also tested each with three replications. Tick larvicidal and ovicidal assays were done testing 0.5; 1; 2 and 4 mg/mL of the P. sylvestris essential oil and its main components. The experiment results from mites revealed that the spraying application method was more effective than the contact bioassay for acaricidal property when P. sylvestris essential oil was toxic to D. gallinae with LC50 values of 0.68 mg/mL. Larvicidal and ovicidal assays revealed the promising activity of the essential oil against tick larvae and eggs. Among all the tested compounds, D-Limonene appears as the most toxic essential oil constituent for both ticks and mites, showing LC50 values lower than the whole essential oil. δ-3-carene was slightly toxic, with LC50 values higher than the whole essential oil, whereas α-pinene and β-pinene showed weak acaricidal activity even at high concentration. On the other hand, P. sylvestris essential oil and its principal components were tested for acetyl-cholinesterase inhibitory activity using Ellman's colorimetric method and compared to a reference galantamine. Interestingly the monoterpene D-Limonene was found the most active AchE inhibitor (IC50 = 27.22 µg/mL), almost comparable to that of positive control. Overall, our research pointed out the importance of the traditional use of P. sylvestris as an ecofriendly alternative against ectoparasites of veterinary importance; D. gallinae and H. scupense. In vivo trials should also be conducted to assure the safe use of natural plants. [ABSTRACT FROM AUTHOR]

Nanoencapsulation is widely considered as a highly effective strategy to enhance essential oils' (EO) stability by protecting them from oxidative deterioration and evaporation. The present study aims to optimize and characterize an efficient technique for encapsulating Cinnamomum (C.) verum essential oil into chitosan nanoparticles using response surface methodology (RSM). Moreover, the optimized C. verum EO nanoparticle was investigated for its antibacterial (against Gram-positive and Gram-negative bacteria), antifungal (against Candida albicans), and antiparasitic activity (against Leishmania parasites). Five parameters were investigated using a Plackett–Burman and Box–Behnken statistical design: the chitosan molecular weight, TPP concentration, C. verum EO/chitosan ratio, mixing method, and the duration of the reaction. Encapsulation efficiency and anti-candida activity were considered as responses. The antibacterial, anticandidal, and anti-leishmanial activities were also assessed using a standard micro-broth dilution assay and the cytotoxicity assay was assessed against the macrophage cell line RAW 264.7. The optimized nanoparticles were characterized using Fourier transform infrared spectroscopy, Zeta potential, and scanning electron microscopy. The study results indicated that under optimal conditions, the nanoencapsulation of C. verum EO into chitosan nanoparticles resulted in an encapsulation efficiency of 92.58%, with a regular distribution, a nanoparticle size of 480 ± 14.55 nm, and a favorable Zeta potential of 35.64 ± 1.37 mV. The optimized C. verum EO/chitosan nanoparticles showed strong antifungal activity against C. albicans pathogens (CMI = 125 µg mL−1), notable antibacterial activity against both Gram-positive and Gram-negative bacteria (ranging from 125 to 250 µg mL−1), high leishmanicidal potential against the promastigotes form of L. tropica and L. major (IC50 = 10.47 and 15.09 µg mL−1, respectively), and a four-fold cytotoxicity reduction compared to non-encapsulated essential oil. These results suggest that C. verum EO-loaded chitosan nanoparticles could be a promising delivery system for the treatment of cutaneous Candida albicans infections. [ABSTRACT FROM AUTHOR]

This work aims to evaluate cinnamon and curcuma essential oils as natural preservatives in minced beef meat. Essential oil chemical compositions and antibacterial activities were studied, and their encapsulation was optimized into nano-emulsions based on droplet size and distribution assessments. Selected formulas were further explored for their physical stabilities and antibacterial activities. Then, their effects on minced beef meat preservation were evaluated. Results showed significant differences in the chemical compositions and the efficiency of the tested essential oils, with cinnamon having a significant antibacterial efficacy. Formulation results showed that cinnamon nanoemulsion, encapsulated by 7.5% Tween 80, possessed an 89 nm droplet size, while the droplet diameter of curcuma nanoemulsion, encapsulated by 5% Tween 80, was 151 nm. Antimicrobial results depicted a significantly higher activity in nanoemulsions as compared to essential oils. For instance, the inhibition diameter of cinnamon essential oils against S. aureus was equal to 35 mm, while that of its nanoemulsion reached 40 mm. The meat preservation results showed that both bulk and nanoencapsulated essential oils significantly inhibited bacterial growth, as well as the formation of methemoglobin and lipid oxidation in meat. Thus, this work draws attention to the enhanced preservation effects of essential oils on the processing of minced beef meat as well as the great potential of nanoemulsions as carriers for essential oils in food industry applications. [ABSTRACT FROM AUTHOR]

This investigation tested the in vitro acaricidal activity of essential oils and raw extracts of Juniperus communis and Origanum majorana compared with the synthetic acaricide amitraz against various stage of Hyalomma scupense cattle tick. The hydrodistilation technique was used to extract essential oils, while chemical components in the plants were obtained using solvents of increasing polarity (hexane, ethyl acetate and methanol). The evaluation on H. scupense was performed with the adult immersion test at concentrations ranging from 1.25 to 10 mg/mL and the larval packet test from 0.625 to 10 mg/mL. Qualitative analysis of J. communis essential oil showed α-pinene (52.31%), δ 3-carene (12.77%), β-phellandrene (7.60%), 1,8 cineole (6.95%) and α- terpinenyl acetate (6.58%) as the major chemical components. O. majorana oil was mainly composed of 4-terpineol (23.05%), Cis-Thujan-4-ol (18.30%), δ-terpinene (13.61%), α-terpinene (9.16%) and sabinene (7.96%). In adult immersion test, J. communis essential oil, at 10 mg/mL concentration exhibited strong acaricidal efficacy and reproductive inhibitory effects on treated ticks (100%) than O. majorana oil (91.95%). In larval packet test, J. communis oil had 100% mortality at 10 mg/mL, 24 h of exposure, whereas O. majorana oil had 90.64% mortality of H. scupense larvae. J. communis essential oil showed greater efficacy than all tested extracts and amitraz on both female adults and larvae of H. scupense. Quantitative analysis showed that J. communis methanolic extract had the greatest ability to extract such phenolic compounds, with a total phenol content of 188.17 (mg gallic acid equivalent/g dry weight) and also had the highest acaricidal activity against engorged females and larvae of H. scupense with LC50 values of 2.46 and 4.12 mg/mL, respectively). The results showed that both plants, particularly J. communis considered as potential candidates for biocontrol of H. scupense in the field. [ABSTRACT FROM AUTHOR]

Biofilm formation of the opportunistic pathogen Pseudomonas (P). aeruginosa is one of the major global challenges to control nosocomial infections due to their high resistance to antimicrobials and host defense mechanisms. The present study aimed to assess the antibacterial and the antibiofilm activities of Peganum (P). harmala seed extract against multidrug-resistant P. aeruginosa isolates. Chemical identification of the active compound and determination of its molecular mechanism of action were also investigated. Results showed that P. harmalan-butanol “n-BuOH” extract exhibited antibacterial activity against multidrug-resistant P. aeruginosa isolates. This extract was even more active than conventional antibiotics cefazolin and vaamox when tested against three P. aeruginosa multidrug-resistant isolates. In addition, P. harmala n-BuOH extract exhibited potent bactericidal activity against PAO1 strain at MIC value corresponding to 500 µg/mL and attained 100% killing effect at 24 h of incubation. Furthermore, P. harmala n-BuOH extract showed an antibiofilm activity against P. aeruginosa PAO1 and exhibited 80.43% inhibition at sub-inhibitory concentration. The extract also eradicated 83.99% of the biofilm-forming bacteria. The active compound was identified by gas chromatography–mass spectrometry as an indole alkaloid harmaline. Transcriptomic analysis showed complete inhibition of the biofilm-related gene pilA when PAO1 cells were treated with harmaline. Our results revealed that P. harmala seed extract and its active compound harmaline could be considered as a candidate for a new treatment of multidrug-resistant P. aeruginosa pathogens-associated biofilm infections. [ABSTRACT FROM AUTHOR]

Background: Ticks and tick-borne diseases are a severe economic and public-health problem for cattle producers. The emergence of acaricide resistance to synthetic chemical treatments has prompted interest in developing alternative tick control methods. Aim: The main objective of the current research was to identify the chemical structure of Carthamus tinctorius and Nasturtium officinale seed oils and to assess their anti-tick properties against Hyalomma scupense ticks both alone and in combination (1:1). Methods: Analytical methods were used to analyze the chemical components. For in vitro assays, adults of H. scupense were immersed in C. tinctorius and N. officinale seed oils at 100, 125, 200, and 300 mg/ml concentrations; for 5 minutes. Larvae of H. scupense were dipped in 25, 50, 100, 125, 200, and 300 mg/ml doses of seed oils; the mortality percentage was determined after 24 hours. Results: The seed oil safflower was mainly composed of linoleic acid (84.48%), followed by palmitic acid (6.54%) and stearic acid (3.77%). Meanwhile, watercress seed oil was mainly composed of linolenic acid (50.78%), gondoic acid (13.57%), linoleic acid (10.58%), palmitic acid (8.02%), and erucidic acid (6.62%). The Adults Immersion Test showed the sensitivity of ticks to C. tinctorius and N. officinale seed oil: C. tinctorius seed oil caused (95%) mortality of H. scupense at 300 mg/ml, while N. officinale seed oil induced (88.68%) mortality at the same concentration. At a 200 mg/ml concentration, C. tinctorius and N. officinale oil combined caused 100% mortality. Tested oils showed larvicidal efficacy. LC50 values for C. tinctorius and N. officinale seed oils were 84.16 and 61.78 mg/ml, respectively, in 24 hours. LC50 value of oils association (50% C. tinctorius: 50% N. officinale) was 47.96 mg/ml. The mixture of seed oils from two plants tested against H. scupense larvae and adult females at a 1:1 ratio showed synergistic interaction. Conclusion: Seed oils tested alone, and the mixture could be used as an alternative solution in the fight against ticks. [ABSTRACT FROM AUTHOR]

Faba bean (Vicia faba L.) is the major food legume crop in Tunisia. However, its growth and yield is strongly affected by water‐limited environments. In this study, osmotic stress exhibited a negative effect on Bachar and Badii cultivar. Nevertheless, the deteriorating effects of osmotic stress were relatively low on studied parameters of Bachar due to its better efficiency to reduce oxidative damage by increasing enzymatic activities such as catalase (CAT), superoxide dismutase (SOD) and ascorbate peroxidase (APX), accumulation of total chlorophyll (Chlt), soluble sugars and leaf relative water content (RWC). GC–MS analysis determined a total of 11 soluble carbohydrates induced by osmotic stress and differentially accumulated in the both cultivars. Bachar showed elevated levels of mannose, glucose, galactose, ribose, rhamnose and myo-inositol which might help to maintain osmotic adjustment, membranes and proteins protection from the damaging effect of reactive oxygen species. Sugar metabolism related genes (VfNINV3, VfPHS2, VfFRK4, VfHXK1, VfGPI1, VfSTP1.1, VfpGlcT1.1, VfSTP5.1, VfpGlcT1.2, VfSWEET2.1, VfVINV2, VfSUS1, VfPGM1, VfSUT1.1, VfGPT1, VfSPS1, VfSPP1, VfPHS1, VfSUT4.1 and VfTMT1.1) were differentially expressed in both cultivars demonstrating their important roles in sugar accumulation. Most of these genes were upregulated in the leaves of Bachar under moderate and severe stress, which could lead to increase glycolysis and tricarboxylic acid cycle in order to accelerate energy production, necessary to increase osmotic regulation and consequently enhancing the osmotic stress tolerance in that cultivar. Overall, sugars accumulation ability can be used as a useful indicator for the osmotic stress tolerant potential in faba bean breeding programs. [ABSTRACT FROM AUTHOR]

Introduction: Synthetic colourants are subject to various allergies. As a result, the use of plant‐based additives has proved promising. Objective: It is in this perspective that our objective was undertaken to investigate natural colourants from edible cactus. Opuntia species populations were single‐sequence repeat (SSR) fingerprinted, the analysis of betalains and phenolic compounds and the evaluation of their biological potentials were also characterised. Results: Photometric quantification of betalains and phenols showed an interspecific variation across Opuntia species. Opuntia ficus indica fruits showed the highest betalain [betaxanthins; 843.67 and betacyanins; 1400 mg/100 g dry weight (DW)] and polyphenol contents. Reversed‐phase high‐performance liquid chromatography (RP‐HPLC) analysis showed that the variation of individual phenols profile was influenced by interspecific and genetic factors. Isorhamnetin‐O‐(di‐deoxyhexosyl‐hexoside) was the major compound and its content varied according to Opuntia species, while catechol was the predominant phenolic compound in O. humifusa with 1.88 μg/g DW. Concerning cactus species, Opuntia colourants exhibited a potent antiradical activity [half maximal inhibitory concentration (IC50) up to 1 μg/mL]. Opuntia species were effective against Gram‐positive and Gram‐negative bacterial strains [inhibition zone (IZ) up to 27 mm]. A high genetic diversity within Opuntia genotypes based on SSR markers was revealed. UPGMA (unweighted pair group method with arithmetic mean) dendrogram and PCoA (principal coordinate analysis) based on natural pigments and antimicrobial profiles indicated significant variation. The correlation approach proved the presence of a probably metabolic relationship between genetic markers, pigments and their biological activities. Conclusion: A possible association between molecular approach and metabolic profile analysis of Opuntia allows tracing the relationship among species for its genetic conservation. Photometric quantification of betalains and phenols showed an interspecific variation across Opuntia species. Opuntia colorants exhibited a potent antiradical activity (IC50 up to 1 μg/mL). A high genetic diversity within Opuntia genotypes based on SSR markers was revealed. UPGMA dendrogram and PCoA analysis based on natural pigments and antimicrobial profiles indicated significant variation. The correlation approach proved the presence of a probably metabolic relationship between genetic markers, pigments and their biological activities. [ABSTRACT FROM AUTHOR]

This study investigated insecticidal activity of Artemisia herba-alba essential oil (EO) against Tribolium castaneum adults and its impacts on wheat flour quality. GC/MS analysis of the EOs of A. herba-alba showed a predominance of β-thujone (27.79%), camphor (22.65%), chrysanthenone (17.97%) and α-thujone (13.56%). Therefore, the median effective concentrations (LC50 value) of Artemisia herba-alba essential oil used for preliminary screening tests were evaluated. Insect mortalities, occupation space conditions and wheat flour quality were investigated after 30 and 60 days of storage period. The results showed that mortality varies depending on occupation space and storage duration, which reached 42.15% and 59.8% after 30 and 60 days of storage, respectively. Furthermore, moisture, ash, gluten, falling number contents were examined according to international standards. Indeed, rheological dough parameters were examined using a Chopin alveograph and according to ISO standards. Results indicated that A. herba-alba essential oil changed wheat flour physicochemical parameters but did not affect dough rheological parameters which correspond to international ISO standards. [ABSTRACT FROM AUTHOR]

The purpose of this study was to compare the essential oil composition of Inula viscosa leaves by hydrodistillation (HDE), ultrasonic (UDE) and solvent (SE) extractions followed by hydrodistillation. The total polyphenol and flavonoid contents and their antioxidant effects were studied by different solvent of extraction: ethanol (ET), ethyl acetate (EA), methanol (ME) and aqueous (AE). The principal compounds for HDE were: 2-hexenal (3.70%), caryophyllene oxide (3.11%), γ-selinene (3.09%), 3-hexen-1-ol (2.00%), eugenol (1.70%) and trans-caryophyllene (1.34%), while for UDE were: γ-selinene (5.68%), caryophyllene oxide (4.87%), trans-caryophyllene (1.99%) and nerolidol (1.74%). The oil obtained by SE was shown to contain tridecane (3.89%), dodecane (3.08%), trans-caryophyllene (2.94%), caryophyllene oxide (2.56%) and nerolidol (2.53%). Significant changes on phenolic contents were found between the different solvent of extraction. ME and AE extracts led to the highest total polyphenol (PHL) and flavonoid (FL) amounts. The anti-radical activity and reducing power were maximal in AE and ME extract. HPLC examination established that the ferulic acid as major phenolic acid in ME and AE fractions, whereas luteolin was the main compound of EA and ET fractions. [ABSTRACT FROM AUTHOR]