Malaysian Journal of Biochemistry
& Molecular Biology
(E-ISSN: 2600-9005)
The Official Publication of the Malaysian Society for Biochemistry & Molecular Biology (MSBMB)
Indexed by SCOPUS and Malaysian Citation Index (MYCITE)
ANNOUNCEMENT
With effect from 15th January 2023, all new submissions will be subjected to a MYR250 publication fee for every accepted paper.
Malaysian Journal of Biochemistry and Molecular Biology (MJBMB) was founded by the Malaysian Society for Biochemistry and Molecular Biology (MSBMB) in the year 1997. It was published two times a year prior to 2010. From 2016 onwards, the journal will be published ONLINE 3 times per year, in April, August and December. The journal publishes research papers in all areas of biochemistry, biophysics and molecular biology, which include DNA and RNA biology, gene expression, glycobiology, enzymology, protein structure and function, lipid chemistry, membranes, immunology, plant biochemistry and physiology, microbiology, immunology, natural product chemistry, biomedical science, research methods and bioinformatics.
Current Issue
December 2024
Malay. J. Biochem. Mol. Biol. (2024) 27 (3)
TABLE OF CONTENTS
Page 1- 8
Bui Hai Ngoc, Nguyen Thi Bich Loan, Pham Thi Luong Hang and Nguyen Dinh Thang
NANOMELANIN AND ANISOMELES INDICA LEAF EXTRACT AS POTENTIAL SUNSCREEN INGREDIENTS
Abstract
Photoprotective agents extracted from natural sources to replace synthetic chemicals are attracting the notice of the scientific community because of their safety and environment friendly. We examined eleven ethanol extracts from medicinal plants and nanomelanin from squid ink sacs for their UV absorption ability and selected the candidates for sunscreen ingredients. Our results revealed that nanomelanin and Anisomeles indica leaf extract exhibited excellent UV absorption capacity at concentrations of 100 µg/mL and 250 µg/mL, respectively. Thus, Anisomeles indica leaves extract and nanomelanin were used as ingredients for sunscreens to protect dorsal mouse skins from the attack of ultraviolet (UV) light. Significantly, sunscreen preparations containing nanomelanin or Anisomeles indica leaf extract could protect dorsal mouse skin from UV attack by reducing skin burn lesions and decreasing skin hyperkeratosis. Remarkably, the mean epidermal thicknesses of mice topically treated with nanomelanin and Anisomeles indica leaf extract were 23.8 μm and 24.5 μm, corresponding with a decrease of 1.70-fold and 1.66-fold, respectively, compared to that of negative control mice (40.6 μm). All the experiments in this study were three-time repeated. Our results suggested that nanomelanin and Anisomeles indica leaf extract could be used as potential sunscreens ingredients.
Page 9 - 16
Nawoda Hewage, Udaya Wijesekara and Rasika Perera
IMPACT OF EXCESSIVE IODINE CONSUMPTION ON THYROID DISORDERS AMONG REPRODUCTIVE AGE FEMALES
Abstract
Thyroid dysfunction due to excessive iodine intake poses a significant risk for women of childbearing age. It is not well known or researched how high iodine consumption affects the development of thyroid diseases in young, reproductive-age females. The current cross-sectional study examined 282 females aged 18-35, assessing thyroid dysfunction and iodine status in both normal weight (controls) and overweight/obese (cases) young females. The study included measurements of height, weight, BMI, thyroid profile, and urine iodine concentration (UIC). Overall prevalence of subclinical hyperthyroidism (SCH) and euthyroidism (EU) among both normal-weight controls and overweight-obese cases were 40.78% and 59.21% respectively. Prevalence of SCH was significantly higher among cases compared to controls (p=0.002). In control group, mean UIC was more than adequate (288.51±150.24 µg/l), whereas in the cases, it was excessive (340.71±147.00 µg/l). There was a statistically significant correlation (p<0.05) between the TSH level and urine iodine concentration in both controls and cases. With an r value of 0.658, the cases further demonstrated a stronger association than the controls, who had an r value of 0.466. Furthermore, UIC demonstrated a significant and strong relationship with SCH in both study groups (r=0.293, p<0.05 and r=0.585, p<0.05, respectively). Further, 57.80% of subjects with elevated UIC were found to have SCH and obese females with high UIC exhibited higher risk of developing SCH (OR=8 [95% CI:6.38-15.71]) when compared to normal controls (OR=2[95% CI:1.38-5.71]). Excess UIC was prevalent among females with SCH and there was a significant association between elevated TSH levels and UIC. Obesity is another risk factor for developing SCH in females who consume more iodine than required. Therefore, it is essential to monitor and control iodine intake and obesity to protect young females from SCH and its associated non-communicable diseases.
Page 17 - 25
Ismiyarto, Martha Tasia Dona, Dwi Susilo, Elmi Pandelaki, Ngadiwiyana, Purbowatiningrum Ria Sarjono, Liswinda Zafira, Marcelinus Christwardana and Abdurohman Zakki
SYNTHESIS OF SUCCINYL CHITOSAN DERIVATIVES AND ITS EVALUATION AS AN ANTIBACTERIAL AGENT
Abstract
Pathogenic bacteria, such as E. coli, S. aureus, and Salmonella, can be found in water and food can cause a variety of ailments; therefore, it is essential to solve these health issues. Among the attempts performed is the modification of chitosan compounds to enhance their antibacterial activity. The purpose of this research was to synthesize numerous chitosan derivatives, including N-Succinyl-Chitosan, Schiff base Chitosan-Salicylaldehyde, and O-Succinyl Schiff Base Chitosan-Salicylaldehyde, in order to produce chitosan derivatives with enhanced an antibacterial activity. N-succinyl chitosan was synthesized with a yield of 89.67wt.% and a substitution degree (DS) of 43.61%. The synthesis of Schiff base chitosan-salicylaldehyde yield of 43.96 wt.% and a substitution degree of 44.44%. The yield of O-succinyl Schiff base chitosan-salicylaldehyde is 56.17 wt.% and the substitution degree is 55.10%. O-succinyl Schiff base chitosan shown better antibacterial activity and minimum inhibitory concentration than unmodified chitosan, Schiff base chitosan salicylaldehyde and N-succinyl chitosan.
Page 26 - 31
Alaa Amer Mohammad, Ali Omar Yassen, Rundk Ahmad Hwaiz and Sardar Nouri Ahmed
LIVER FIBROSIS IN PATIENTS WITH β-THALASSEMIA MAJOR: CORRELATION WITH LIVER ENZYMES AND FERRITIN LEVEL
Abstract
Background: Thalassemia is a genetic blood disorder that affects hemoglobin production. Individuals with β-thalassemia major require regular red blood cell transfusions to survive. Thalassemia causes anemia and abnormal liver function that appears to be related to iron overload over a period of years, may lead to liver fibrosis. Method: The study is retrospective, case control study, biochemical assessments were estimated and evaluated for 79 β-thalassemia major patients including 32 patients diagnosed with liver fibrosis, and 38 control subjects, biochemical tests for AST, ALT, TBIL, ALP and SF were evaluated in Erbil city. Result: The results of this study illustrated that there was a significant difference between β-thalassemia patients and the control group regarding AST, ALP, Bilirubin, and ferritin levels (p<0.0001). Also, Ferritin level was positively correlated with the levels of AST (r=0.3393, P=0.0022), and ALT (r=0.3612, P=0.0011) among β-thalassemia patients. In addition, severe liver fibrosis was encountered in 32 patients with β-thalassemia, which showed higher ALP levels (P=0.0122) in patients with liver fibrosis and no relationship with respect to ferritin level, AST, ALT and TBIL. Conclusion: Patients with β-thalassemia had abnormal liver function. High levels of Ferritin could be the main leading cause of impaired liver function. In this study, β-thalassemia patients with liver fibrosis did not have significant difference in terms of ferritin levels when compared to non-liver fibrosis. We can attribute this inconsistence to the fact that patients with liver fibrosis were under regular chelation therapy. More research is needed to conclude a correlation between them
Page 32 - 40
Muhammad Dawood Shah, Mohammad Amil Zulhilmi Benjamin and Mohammad Iqbal
Orthosiphon stamineus Benth. Impedes Carbon Tetrachloride-Induced Hepatic Damage by Modulating Oxidative Stress
Abstract
Orthosiphon stamineus Benth. (Lamiaceae) is an herb traditionally used in folk medicine for protecting against various diseases such as oxidative stress. This study aimed to assess the antioxidant and hepatoprotective properties of the ethanol extract of O. stamineus (EEOS) in rats with hepatic injury induced by carbon tetrachloride (CCl4). The antioxidant properties of EEOS were evaluated through total phenolic content (TPC), and 2,2-diphenyl-1-picrylhydrazyl (DPPH) and reducing power assays. Sprague-Dawley rats were orally administered EEOS at doses of 100, 500, and 1000 mg/kg b.wt. for two weeks, followed by exposure to CCl4 (1.2 mL/kg). The rats were then euthanised for biochemical analyses. In vitro studies demonstrated the robust antioxidant potential of EEOS, revealing effective scavenging of DPPH free radicals and reducing power. Additionally, EEOS exhibited a high TPC of 127.28 ± 1.57 mg GAE/g db, contributing significantly to its antioxidant activities. Administration of EEOS significantly mitigated CCl4-induced toxicity, as evidenced by reduced alanine transaminase (1–91-fold recovery) and aspartate transaminase (1–28-fold recovery) levels of hepatic damage in rats. Moreover, EEOS alleviated the heightened malondialdehyde levels (40–80% recovery) and elevated reduced glutathione levels (30–80% recovery) in CCl4-induced rats, while also restoring levels of various antioxidant enzymes: catalase (12–20% recovery), glutathione peroxidase (23–41% recovery), glutathione reductase (6–30% recovery), glutathione S-transferase (11–25% recovery), and quinone reductase (8–35% recovery). The study conclusively demonstrates the strong antioxidant potential of EEOS, with 1000 mg/kg b. wt. exhibiting efficacy in restoring antioxidant enzymes. These compelling findings highlight the potential of EEOS as a promising candidate for preventing liver damage associated with reactive oxygen species.
Page 41 - 49
Lee Sin Jye, Nur Shaheen Azlan, Yong Yoke Keong, Rhun Yian Koh and Ooi Yin Yin
MESENCHYMAL STEM CELLS-DERIVED EXOSOMAL MIRNAS TARGETING MICROGLIA IN TREATING NEUROINFLAMMATION AND NEUROLOGICAL DISEASES
Abstract
Neurodegenerative diseases (NDD) are projected to exceed cardiovascular disease as the top 1 leading cause of death in the next 20 years. However, existing treatments merely manage symptoms or slow down disease progression without offering a definite cure. Recognising the advantages of MSC-exosomal miRNAs, which can transverse the blood-brain barrier (BBB) as a promising treatment for various incurable disorders, including neuroinflammation, neurological diseases including NDD and central nervous system (CNS) injuries. The study highlights the widespread use of bone marrow-derived MSC and identifies miR-21a-5p, miR-26b-5p, and miR-146a-5p as the most exploited miRNAs by researchers. Nevertheless, the supremacy of this therapeutic approach over existing treatments remains vague. Overall, the study establishes that the targeted modulation of microglia cells with MSC-derived exosomal miRNAs has a positive impact on neurological diseases by reducing neuronal death and mitigating neuroinflammation. Consequently, MSC-derived exosomal miRNAs emerge as a promising avenue for NDD therapy. In this present article, it provides a succinct review of the therapeutic potential of exosomal microRNAs (miRNAs) derived from mesenchymal stem cells (MSC) that target microglia in treating NDD, CNS injuries and neuroinflammation.
August 2024
Malay. J. Biochem. Mol. Biol. (2024) 27 (2)
TABLE OF CONTENTS
Page 1- 15
Mohd Iftar Johwan Johny @ Hasbullah, Bellericter Binjamin, Kimberly Ador, Januarius Gobilik, Clament Chin Fui Seung, Lum Mok Sam, Nurul’Azah Mohd Yaakub, Jamilah Lee Nyuk Choon, and Suzan Benedick
VOLATILE ORGANIC COMPOUNDS (VOCS), DIHYDROXYACETONE (DHA) AND METHYLGLYOXAL (MGO) IN TROPICAL MULTIFLORAL HONEY (APIDAE; APIS CERANA) FROM SITES OF DIFFERENT GEOGRAPHICAL ORIGIN IN SABAH, BORNEO
Abstract
The content of volatile organic compounds (VOCs), dihydroxyacetone (DHA) and methylglyoxal (MGO) in honey depends on the geographical origin but has little been studied in Sabah. The aim of this study was to determine the content of VOCs, DHA and MGO in raw honey of Apis cerana at six study sites that differ in their geographical origin. Each of the study sites contains 3 replicates of honey samples: Ulu Kiulu, Tuaran (adjacent to lowland rainforest park), Nadau, Tamparuli (adjacent to highland forest park), Membatu Laut, Kudat (coconut farm), BHBC, Kudat (Acacia and secondary forest), Kg. Gana, Kota Marudu (rubber and orchard) and FSA, Sandakan (oil palm and orchard). Samples were subjected to liquid–liquid extraction process, followed by gas chromatography-mass spectrometry (GC–MS) for identification and characterisation of VOCs. The DHA and MGO in Apis cerana honey were analysed by high performance liquid chromatography (HPLC). The results indicate that a higher number of VOCs and unique VOCs were identified in honey from forest and mixed forest sites than in honey from other study sites. The study found that the content of DHA and MGO varied according to geographical origin (p < 0.001). The honey sample from Gana, Kota Marudu had the highest DHA concentration (mg/kg), followed by FSA, Sandakan and Membatu Laut, Kudat. The honey sample from BHBC had the highest MGO concentration (mg/kg), followed by Membatu Laut, Kudat and Ulu Kiulu, Tuaran. This study concluded that geographical origin with different botanical sources plays a crucial role in the bioavailability of bioactive nutraceutical/functional therapeutic compounds in multifloral honey, A. cerana in Sabah..
Page 16 - 25
Adam Raihan Priambada, Putra Santoso, and Resti Rahayu
IN SILICO STUDY ON THE POTENTIAL OF BIOACTIVE COMPOUNDS FROM COLOCASIA ESCULENTA VAR. MENTAWAI IN INTERVENING TOLL-LIKE RECEPTOR 4 SIGNALING PATHWAY TO MANAGE INFLAMMATORY BOWEL DISEASE
Abstract
Inflammatory bowel disease (IBD) is one of the diseases that is closely associated with the emergence of colorectal cancer. While Non-Steroidal Anti-Inflammatory Drugs (NSAIDs) are a preventive measure against IBD, they come with serious side effects. We aim to assess the potential of bioactive compounds from Mentawai taro (Colocasia esculenta Var. Mentawai) in intervening the cellular signaling system of Toll-like receptor 4 (TLR4) which is closely associated with the progression of IBD. In silico study was conducted using PASS online and Lipinski's five rules followed by molecular docking. The ligands and receptor were obtained from the Protein Data Bank (PDB) and PubChem. Validation and visualization were performed using BIOVIA and PyMol. Based on the PASS online test, all bioactive compounds from C. esculenta have biological activity as anti-inflammatory and anti-ulcerative agents. Furtermore, the results of molecular docking revealed that the compounds from C. esculenta namely astaxanthin, lauric acid, and octadecanoic acid have potential to interact with TLR4 signaling system. However, astaxanthin appears as the most potent compound with the highest binding affinity to TLR4 (-9.0 kcal/mol) and the predominant conventional hydrogen bond. In addition, astaxanthin outperforms the binding affinity of standard drug namely dexamethasone (-8.4 kcal/mol) towards TLR4. Therefore, astaxanthin from Mentawai taro could be considered a promising candidate for drug development in managing IBD.
Page 26 - 30
Thyviaah Ananthan, Nurzafirah binti Mazlana, Norazalina binti Saad, BA Venmathi Maran
CYTOTOXICITY OF COLLAGEN EXTRACTED FROM TOMATO JELLYFISH (CRAMBIONE MASTIGOPHORA)
Abstract
A comprehensive study was conducted to assess the cytotoxicity of collagen extracted from the tomato jellyfish (Crambione mastigophora) across various of the jellyfish regions: bell, whole body, and oral arm. The extracts were tested using 3T3 mouse embryonic fibroblast cells. The results showed that collagen extracted from the bell, whole body and oral arm exhibit no IC50 value up to 500µg/ml. Similarly, standard collagen (bovine tendon type II collagen) also showed no cytotoxic effect on the 3T3 cells. These findings provide insights into the safety and potential applications of tomato jellyfish collagen in various industries.
Page 39 - 50
Redzuan Nul Hakim Abdul Razak, Mohd. Azri Bin Abd. Jalil, Nor Farihatasha Isa, Amynah Noor Zahurein Binti Hazidan, Mohamad Farqhan Bin Kelana, Nik Abd Rahman Fahmi Losman, Muhammad Alif Mazlan, Muhammad Lokman Bin Md Isa and Moustafa Ibrahim
SYSTEMATIC REVIEW ON THE APPLICATION OF CELLULAR CULTURE TECHNOLOGY APPROACH IN CONSERVATION OF ENDANGERED SPECIES
Abstract
The extinction of wildlife species is concerning. This issue has posed a threat to the whole biodiversity system significantly. Nevertheless, the extinction of wildlife species can be controlled and stabilized through a conservation program of in situ and ex situ approach respectively but in need of additional programs to save the species. This situation emphasizes the importance of ex situ conservation strategies, particularly cellular technology, in supplementing in situ efforts, which are becoming increasingly challenging. Cellular technology has an important role in conserving genetic diversity through optimized protocols such as assisted reproductive technology (ART). This paper conducts a systematic review and critical analysis of the current understanding of cellular technology in conservation. The study conducted an electronic search in three databases such as PubMed, ScienceDirect, and Scopus for articles published between 2010 and 2021. The search used keywords such as “cellular technology,” “assisted reproductive technology,” “cryopreservation,” “conservation,” “rescue,” “save,” and “endangered species.” The findings suggest that assisted reproductive technologies are identified as cellular technologies for ex situ conservation efforts. However, the effectiveness of these technologies depends on various parameters specific to each endangered species. These parameters include genetic diversity, estrous cycle length, timing and method of semen collection, and the sperm quality and quantity. This review explores the application of cellular technology for conservation of endangered species and their future impact in conservation programs. More exploration is needed to fully harness the potential of cellular technologies in saving endangered species.
Page 38 - 59
Muhammad Amal Zulkipli, Aisyah Jaafar, Yuslina Zakaria, Elhassane Anouar, Mizaton Hazizul Hasan
ANTI-INFLAMMATORY AND ANTIOXIDANT PROPERTIES OF NATURALLY DERIVED IRIDOIDS: A SCOPING REVIEW OF IN SILICO STUDIES
Abstract
This scoping review explores the current state of molecular docking studies on naturally derived iridoid compounds, focusing specifically on their engagement with anti-inflammatory and antioxidant targets. The review searches the PubMed and Scopus databases to locate relevant published studies. Following a systematic search strategy, identified citations were compiled, screened by two independent reviewers, and evaluated against the inclusion criteria. Data extraction was conducted using a reviewer-developed instrument, and appropriate details were captured. The extracted data was stored and analysed. Results were discussed, supplemented by data tables and reported adhering to the PRISMA-ScR checklist. The studies included were characterised as in silico, with a total of 364 studies initially retrieved. After removing duplicates, 143 studies remained, out of which 40 were subjected to a full read, and ultimately, 29 studies were deemed eligible for qualitative synthesis. Iridoids, particularly swertiamarin, amarogentin, geniposide, gentiopicroside, and loganic acid, surfaced as the predominant subjects of examination, spotlighting their pharmacological anti-inflammatory and antioxidant effect. Other iridoids such as aucubin, genipin, verminoside and sweroside were also examined. This review highlights the most potent naturally produced iridoids, each exhibiting the highest affinity to different target proteins, providing a basis for choosing substances and directing more experimental and clinical research. The data from these studies is essential for figuring out how these chemicals could be used in biotechnology and medicine, especially to fight diseases related to inflammation and oxidative stress.
Page 60 - 69
Norliza binti Julmohammad, Kathi Ann Iran Money, Salma Samidin and Nurul Huda
EFFECT OF pH ON THE PHYSICOCHEMICAL, MICROSTRUCTURE, AND FUNCTIONAL PROPERTIES OF DRY HEATED WHEY PROTEIN ISOLATE-LACTOSE AND WHEY PROTEIN ISOLATE
Abstract
This study aimed to investigate the effects of pH on the physicochemical and functional properties of whey protein isolate (WPI) conjugated with lactose through dry heating. Whey proteins are prone to heat and oxidation, leading to changes in physicochemical properties and limiting their application. Conjugation can improve their stability and functionality. WPI was conjugated with lactose through the Maillard reaction at pH 5.0, 6.0, 7.0, and 8.0 using dry heating (60°C, 8 h). Physicochemical and functional properties were analyzed using SDS-PAGE, FTIR, zeta potential, SEM, and functional tests. Conjugation of WPI with lactose significantly improved emulsifying capacity, solubility, and water-holding capacity, with the most pronounced effects observed at pH 7.0, where the degree of glycation was highest. The physicochemical properties, including SDS-PAGE and OPA assay results, showed successful conjugation, with the highest glycation occurring at pH 7.0. The functional properties, particularly emulsifying activity, water-holding capacity, and solubility, were significantly enhanced by conjugation, especially at pH 7.0, while oil-holding capacity showed minimal improvement. Conjugation at pH 7.0 provided the optimal balance between improved protein functionality and stability, suggesting its potential application in food products requiring enhanced emulsifying and solubility properties. This study shows that conjugating WPI with lactose via the Maillard reaction significantly enhances its functional properties, with pH 7.0 being the most favorable condition for improving protein functionality.
Page 70 - 73
Mohd Azwan Jenol, Purkan Purkan, Dwi Kusuma Wahyuni, Sofijan Hadi, Listijani Suhargo, Mohamad Faizal Ibrahim, Lai-Yee Phang, Mohd Azuraidi Osman, Safinaz Abd Gani and Suraini Abd-Aziz
ENHANCING SILAGE PRODUCTION FROM PINEAPPLE LEAVES: A COMPARATIVE ANALYSIS OF MICROBIAL AND ENZYME-ASSISTED APPROACHES
Abstract
In Indonesia, the pineapple industry plays a crucial role in the economy. However, the country faces a significant challenge with pineapple waste, as more than 40% of it remains unused, leading to environmental issues such as pollution, soil degradation, and greenhouse gas emissions. Silage technology, involving the fermentation of forage crops in an anaerobic environment, is a common practice in agriculture to create livestock feed, especially during periods of low forage availability or unfavorable weather conditions. This study is aimed to explore the potential of utilizing pineapple leaves as a livestock feed by assessing the quality and aerobic stability of silage produced from these leaves. The research specifically investigated the impact of enzyme-assisted and microbial-assisted silage production methods on the nutritional and quality aspects of pineapple leaves silage. The findings revealed that both enzyme-assisted and microbial-assisted techniques enhance the pH, moisture content, and lactic acid content of the pineapple leaves silage when compared to the standard silage production method. This study underscored the significance of the assisted silage system in the improvement of the overall quality and nutritional value of pineapple leaves silage. By doing so, it not only creates a valuable feed resource for livestock but also contributes to food security, environmental sustainability, and enhanced livestock performance within the agricultural sector
Page 74 - 81
Hassan Rezazadeh, Mohammad Ramazani, Ahad Bavili-Tabrizi, Siavoush Dastmalchi, Alireza Mohajjel Nayebi, Armin Taheri, Rana Eftekharnahli, Senty Vun-Sang and Mohammad Iqbal
FERRIC NITRILOTRIACETETE AUGMENTS 7,12-DIMETHYLBENZ(A) ANTHRACENE-INITIATED AND CROTON OIL-PROMOTED SKIN CARCINOGENESIS
Abstract
Although Ferric nitrilotriacetete (Fe-NTA) is a known renal carcinogen, its carcinogenic potential in skin remains poorly understood. This study aims to investigate the potential augmentation effects of Fe-NTA on the initiation and promotion stages of DMBA-induced and croton oil-promoted skin cancer development, shedding light on the mechanisms underlying the synergistic carcinogenic action of these agents. We for the first time demonstrates that Fe-NTA enhances skin carcinogenesis in mice initiated by 7,12-dimethylbenz(a)anthracene (DMBA) and promoted by croton oil. This study looked at the impact of Fe-NTA on croton oil-promoted tumour activation in 7,12-dimethylbenz(a)anthracene (DMBA)-initiated mouse cutaneous carcinogenesis. Four groups of mice, each consisting of 20 animals, were selected for the carcinogenicity tests. Group I served as the control and received no treatment. Groups II, III, and IV received daily topical applications of Fe-NTA at doses of 6, 9, and 12 mg iron per mouse, respectively, for a period of 2 weeks. Nine hours after the last administration of Fe-NTA, all groups of mice, including the control group, received a single topical application of 60 μg of DMBA in 200 μl of acetone. One week after DMBA treatment, the mice received twice-weekly topical applications of 1 mg croton oil in 200 µl acetone/mouse for a period of 30 weeks. The mice were then monitored for the occurrence of tumors up to 36 weeks. Higher doses of Fe-NTA induced an increase in tumor occurrence over times as compared to the control (DMBA+ Croton oil) treated group. Tumors appeared earlier in the Fe-NTA group, with a higher incidence number of tumors. We propose that Fe-NTA boosts croton oil, tumor-promoting potential, and that Fe-NTA-induced oxidative stress is effective for croton-oil mediated cutaneous carcinogenesis. In addition, croton oil-mediated lipid peroxide induction and [3H]thymidine uptake was greater in Fe-NTA treated group. We propose that Fe-NTA increases tumor promotion capability of croton oil and oxidative stress induced by Fe-NTA is effective for croton oil mediated cutaneous tumorigenesis.