Mutagenicity Test Research Articles

Platinum as both a drug and its modulator - Do platinum nanoparticles influence cisplatin activity?

Breast cancer was the most frequent cause of cancer death in females in 2022. Despite the development of personalized therapies, chemotherapy frequently remains the only available treatment method. However, the administration of classic antineoplastic drugs, like cisplatin (CDDP), often causes severe side effects and may lead to drug resistance making the therapy inefficient. Therefore, there is a great need for new, effective treatment regimens development. For this reason, we applied platinum nanoparticles (PtNPs) to verify if they can influence the CDDP activity with particular emphasis on the differences due to nanoparticles' sizes. We employed a broad spectrum of physicochemical methods, including Dynamic Light Scattering, Atomic Force Microscopy, Isothermal Titration Calorimetry, Fourier Transform Infrared Spectroscopy, and Near Infrared Spectroscopy and also Differential Scanning Calorimetry, to characterize the possible interactions between nanoparticles and CDDP. Moreover, the impact of PtNPs on CDDP biological activity was investigated using the Ames mutagenicity test on Salmonella enterica serovar Typhimurium TA102 and MTT assay on two breast cancer cell lines MDA-MB-231 and SKBR3. The obtained results revealed PtNPs direct interactions with CDDP dependent on the nanoparticles' size. Despite the lack of explicit confirmation of PtNPs aggregation by AFM imaging and DLS, further physicochemical methods indicated structural changes between nanoparticles alone and PtNPs-CDDP mixtures. Moreover, the biological assays confirmed that PtNPs decrease CDDP mutagenicity and also slightly increase its cytotoxicity on the chosen cell lines. The latter effects are ambiguous, nevertheless, provide a valuable basis for further research.

Ames mutagenicity of 15 aryl, benzyl, and aliphatic ring N-nitrosamines.

The Ames mutagenicity test is an effective means of screening compounds for their carcinogenic potential. Here, we conducted Ames tests on 15 aryl, benzyl, and aliphatic ring N-nitrosamines. Then, by using two indicators of mutagenicity strength calculated from the Ames test results, namely, maximum specific activity (MSA; number of revertant colonies) and maximum fold increase (MFI; relative ratio of increased colonies), we examined the relationship between Ames mutagenicity strength and Carcinogenic Potency Categorization Approach (CPCA) potency category, which is a structure-activity-relationship-based prediction of the carcinogenic potency of nitrosamines. Eleven of the test compounds were Ames positive and four were negative. Of the 11 positive compounds, three were categorized as strong positive (MSA ≥ 1000), five as medium positive (100 ≤ MSA < 1000), and three as weak positive (MSA < 100). The compounds with an aliphatic ring showed a negative relationship between mutagenicity strength (i.e., MSA or MFI) and carcinogenic potential (i.e., CPCA category), whereas, the alpha-methyl aryl N-nitrosamines did not. Overall, MSA and MFI were found to be detailed indicators of the carcinogenic potency of the N-nitrosamines and can potentially be used to support CPCA categorization.

Comprehensive evaluation of the toxicological effects of commonly encountered synthetic cathinones using in silico methods.

Synthetic cathinones (SCs), a group of new psychoactive substances (NPS), are designer molecules with hallucinogenic and psychostimulatory effects. Although the structural similarities of SCs to amphetamines suggest that they may have similar toxicity profiles to those of amphetamine congeners, little is known about SCs from a toxicological point of view. In the present study, the toxicity profiles of commonly encountered SCs (n = 65), listed in the 2020 Report of the United Nations Office on Drugs and Crime (UNODC), were evaluated using in silico methods. We aimed to gain a deeper understanding of key toxicological endpoints: acute oral toxicity (LD50), mutagenicity, genotoxicity, and carcinogenicity prediction using EPA TEST (v.5.1.2 and 4.2.1), VEGA (v.1.2.3), and ProTox (v.3.0). Physicochemical and pharmacokinetic (ADME) properties were estimated using SwissADME and pkCSM. 2,3-MDMC (41) was predicted to be the most lethal SC by the VEGA KNN and the EPA TEST v.5.1.2 with an oral rat LD50 value of 105.17 and 117.77mg/kg, respectively. 4-BEC (2) was the only molecule with a consensus score of positive prediction greater than 0.90 in both TEST mutagenicity models. 2,3-MDMC (41) and methylone (52) were predicted as carcinogenic by VEGA carcinogenicity CAESAR, ISS, IRFMN-ISSCAN-CGX, oral classification, and ProTox models. These two SCs were predicted to be active by VEGA chromosomal aberration (CORAL) and in vitro micronuclei-inducing activity (IRFMN-VERMEER) models. Our results concluded that given the prolonged exposure duration and age range, the genotoxic and carcinogenic potential of SCs should be considered, among other known toxic effects.

Toxicity assessment of a phytomedicine based on an almond extract from the fruit of Balanites aegyptiaca (L.) DELILE (Zygophyllaceae) intended for helminthiases care in Burkina Faso

Balanites aegyptiaca (L.) Delile (Zygophyllaceae) is a medicinal plant used in traditional medicine in Burkina Faso to treat helminthiasis. Using an ethnopharmacological approach, the "Institut de Recherche en Sciences de la Santé (IRSS)" has developed a phytomedicine based on an almond extract from the fruit of this plant for the treatment of helminthiasis. This study aimed to assess the acute and subacute oral toxicity and in vivo mutagenicity of phytomedicine. The acute toxicity study was carried out following OECD guideline 423 by administering a single dose of 2000 mg/kg body weight (bw) of phytomedicine orally to female mice. For subacute toxicity, OECD guideline 407 was used. Four batches of 10 rats (5 males and 5 females) were used, including a control batch and three treated batches receiving a daily oral administration of the phytomedicine at doses of 250, 500, or 1000 mg/kg bw for 28 days. The mutagenicity test was performed according to OECD guideline 474; five batches of 10 mice (5 males and 5 females) were given oral doses of 500, 1000 or 2000 mg/kg bw of phytomedicine, colchicine as a positive control at 5mg/kg bw and distilled water as a negative control. In the acute toxicity test, the LD50 was estimated at 5000 mg/kg bw. The subacute toxicity study showed no mortality or signs of toxicity in rats. Biochemical analysis revealed no significant differences between control rats and those treated with the phytomedicine for glucose, creatinine, total cholesterol, triglycerides, AST, chlorine, calcium, and potassium. On the other hand, serum levels of ALT, total protein, PO43-, and Na+ were significantly reduced in some treated rats. The mutagenicity test showed no change in micronucleus frequency between treated and control mice. This study showed that phytomedicine based on an extract of B. aegyptiaca fines would present less danger to users. Keywords: Balanites aegyptiaca, acute toxicity, subacute toxicity, micronucleated erythrocytes, phytomedicine.

The transgenic MutaMouse hepatocyte mutation assay in vitro: mutagenicity and mutation spectra of six substances with different mutagenic mechanisms

Mutagenicity testing is a component of the hazard assessment of industrial chemicals, biocides, and pesticides. Mutations induced by test substances can be detected by in vitro and in vivo methods that have been adopted as OECD Test Guidelines. One of these in vivo methods is the Transgenic Rodent Assay (TGRA), OECD test guideline no. 488. An analogous in vitro TGRA has been described, but experience with this test method is limited. In this study, six in vivo TGRA positive mutagens were tested in the in vitro TGRA based on primary MutaMouse hepatocytes. In addition to the functional read-out of the lacZ reporter gene, induced mutations were analysed by next-generation sequencing (NGS). Five of the six in vivo TGRA positive mutagens (N-ethyl-N-nitrosourea (ENU), ethyl methanesulfonate (EMS), mitomycin C (MMC), benzo[a]pyrene (B[a]P), and azathioprine (AZA), but not cyproterone acetate) mutated the lacZ gene in vitro. NGS identified mutations which matched the mutagenic mechanisms described in the literature. The alkylating agent ENU induced a greater proportion of A:T to T:A transversions than did the other alkylating agent, EMS, whereas EMS increased smaller deletions (1-4 bp). G:C to T:A transversions accounted for the majority of mutations identified after treatments with MMC and B[a]P, both of which form monoadducts at the guanine N2 position. AZA induced mainly G:C to A:T transitions, explained by the structural similarity of one of its metabolites to guanine. An increased proportion of mid-size changes (0.3-2.5 kb) was detected only for the crosslinking mutagen MMC. The in vitro TGRA based on primary MutaMouse hepatocytes is a promising in vitro assay for the assessment of mutation induction, reflecting many aspects of the corresponding in vivo TGRA and allowing for mutation spectra analysis to evaluate the induced mutations.

Substance depletion of volatile monoterpenes – A confounding factor for toxicity testing in the Ames fluctuation test

In in vitro toxicology, reported test results are typically based on nominal concentrations, i.e., the calculated amounts of a substance added to a defined volume of the test system. Consequently, if a test system does not respond to a certain endpoint, the assay is interpreted as negative and the test substance is deemed to exert no toxicity at the tested nominal concentration. However, depending on the physicochemical properties of the test substance and assay setup, the actual exposure may differ widely from nominal concentrations due to different depletion processes. (R)-(+)-Limonene (RLIM), β-myrcene (βMYR) and linalool (LIN) are naturally occurring terpenes that are permitted as flavoring agents in the European Union without limitations based on their low toxicity. Nevertheless, their hydrophobicity and high volatility classifies them as difficult to test chemicals, which has not been considered in previous in vitro tests. To exclude possible false negative results, in the present study, we assessed the cytotoxic and mutagenic potential of the latter substances toward Salmonella Typhimurium in the Ames fluctuation test using different incubation setups to minimize possible substance losses due to sorption or volatilization. Actual substance concentrations during incubation were verified analytically at different time points via headspace gas chromatography-mass spectrometry (HS-GC-MS). Possible substance depletion due to sorption to well-plate material or volatilization was minimized using a polystyrene-free and headspace-free incubation setup, respectively. The results showed complete volatilization of the monoterpenes RLIM and βMYR in the conventional Ames fluctuation test, which may confound mutagenicity testing. The headspace-free incubation setup greatly improved substance exposure and showed cytotoxicity in low micromolar concentrations, but no signs of mutagenicity were observed.

Optimizing the detection of N-nitrosamine mutagenicity in the Ames test

Accurately determining the mutagenicity of small-molecule N-nitrosamine drug impurities and nitrosamine drug substance-related impurities (NDSRIs) is critical to identifying mutagenic and cancer hazards. In the current study we have evaluated several approaches for enhancing assay sensitivity for evaluating the mutagenicity of N-nitrosamines in the bacterial reverse mutagenicity (Ames) test. Preincubation assays were conducted using five activation conditions: no exogenous metabolic activation and metabolic activation mixes employing both 10% and 30% liver S9 from hamsters and rats pretreated with inducers of enzymatic activity. In addition, preincubations were conducted for both 60 min and 30 min. These test variables were evaluated by testing 12 small-molecule N-nitrosamines and 17 NDSRIs for mutagenicity in Salmonella typhimurium tester strains TA98, TA100, TA1535, and TA1537, and Escherichia coli strain WP2 uvrA (pKM101). Eighteen of the 29 N-nitrosamine test substances tested positive under one or more of the testing conditions and all 18 positives could be detected by using tester strains TA1535 and WP2 uvrA (pKM101), preincubations of 30 min, and S9 mixes containing 30% hamster liver S9. In general, the conditions under which NDSRIs were mutagenic were similar to those found for small-molecule N-nitrosamines.

Deep active learning with high structural discriminability for molecular mutagenicity prediction

The assessment of mutagenicity is essential in drug discovery, as it may lead to cancer and germ cells damage. Although in silico methods have been proposed for mutagenicity prediction, their performance is hindered by the scarcity of labeled molecules. However, experimental mutagenicity testing can be time-consuming and costly. One solution to reduce the annotation cost is active learning, where the algorithm actively selects the most valuable molecules from a vast chemical space and presents them to the oracle (e.g., a human expert) for annotation, thereby rapidly improving the model’s predictive performance with a smaller annotation cost. In this paper, we propose muTOX-AL, a deep active learning framework, which can actively explore the chemical space and identify the most valuable molecules, resulting in competitive performance with a small number of labeled samples. The experimental results show that, compared to the random sampling strategy, muTOX-AL can reduce the number of training molecules by about 57%. Additionally, muTOX-AL exhibits outstanding molecular structural discriminability, allowing it to pick molecules with high structural similarity but opposite properties.

Evidence for Safety of The Dietary Ingredient Agmatine Sulfate as Assessed by Mutagenicity and Genotoxicity Studies

Agmatine, 1-Amino-4-guanidinobutane, is a ubiquitous naturally occurring molecule present in low amounts in a wide variety of foodstuff. Clinical trials have demonstrated the safety of oral agmatine sulfate and have led to its development as an effective dietary ingredient for promoting resilient nerve functions. Although clearly required, the mutagenic and genotoxic effects of agmatine have not been previously reported. The present study, therefore, undertook to assess the safety profile of agmatine using currently accepted in vitro and in vivo mutagenicity and genotoxicity tests. The test item was G-Agmatine®, a proprietary brand of agmatine sulfate. Using the bacterial reverse mutation assay (Ames test), the study found that G-Agmatine® has no mutagenic effects. It had no clastogenic effects as observed by the in vitro chromosomal aberration test using Chinese Hamster lung cells. And it lacked genotoxic effects as evidenced by the lack of increased frequency of micronucleated polychromatic immature erythrocytes following oral administration in the mouse micronucleus test. Taken together with previously published data, results of the present study further support the safety of agmatine sulfate as a dietary ingredient.

Toxicological profiling of methanolic seed extract of Abutilon indicum (L.) Sweet: in-vitro and in-vivo analysis

Ethnopharmacological relevanceAbutilon indicum, a shrub of the Malvaceae family, is found abundantly in tropical countries like India. A. indicum is widely used for its high medicinal properties. Traditionally, A. indicum seed powder is consumed to treat piles, constipation, chronic cystitis, gonorrhea, gleet, and pregnancy-related problems. Despite having numerous medicinal properties and widespread traditional use of A. indicum seeds, scientific validation, and toxicity studies have yet to be documented. Aims of the studyThe primary objective of this study is to conduct a comprehensive study on phytochemical profiling, in-vitro cytotoxicity, mutagenicity, and in-vivo acute and sub-acute toxicity, and genotoxicity on animal models of methanolic extract of A. indicum seed (MAS). Materials and methodsThe qualitative analysis of MAS was explored through FTIR and HR LC-MS. For in-vitro cytotoxicity, the HEK-293 cell line was used, and the TA100 (Staphylococcus typhimurium) bacterial strain was used for the Ames mutagenicity test. A single oral dose of 250, 500, 1000, or 2000 mg/kg body weight of MAS was given to each male and female rat for acute toxicity study and observed for 14 days for any toxicity signs. In the sub-acute toxicity study, 250, 500, or 1000 mg/kg body weight of MAS was administered orally to each rat for 28 days. The experimental animals were weighed weekly, and general behavior was monitored regularly. After 28 days of the experiment, the rats were sacrificed, and different serum biochemical, hematological, and histological analyses were performed. The blood samples of different doses of MAS were used for genotoxicity study through comet assay. ResultsFTIR analysis found different functional groups, which indicated the presence of phenolics, flavonoids, and alkaloids. HR LC-MS analysis depicts several components with different biological functions. The cell cytotoxicity and Ames mutagenicity results showed minimal toxicity and mutagenicity up to a certain dose. The acute toxicity study conducted in Wistar albino rats demonstrated zero mortality among the animals, and the LD50 value for seed extract was determined to be 2000 mg/kg body weight. Sub-acute toxicity assessments indicated that the administration of seed extract resulted in no adverse effects at dosages of 250 and 500 mg/kg body weight. However, at higher doses, specifically 1000 mg/kg body weight, the liver of the experimental rats exhibited some toxic effects. In the genotoxicity study, minimal DNA damage was found in 250 and 500 mg/kg doses, respectively, but slightly greater DNA damage was found in 1000 mg/kg doses in both male and female rats. ConclusionsThe consumption of A. indicum seed powder is deemed safe; however, doses exceeding 500 mg/kg body weight may raise concerns regarding use. These findings pave the path for the creation of innovative medicines with improved efficacy and safety profiles.

In vitro effect-based monitoring of water, sediment and soil from a floodplain restoration site in Central Europe

BackgroundFloodplains are biodiversity hotspots and provide numerous ecosystem services. In recent decades, however, 70–90% of Europe’s floodplains have been structurally degraded. Accordingly, many (inter-)national programs aim to restore and protect floodplain ecosystems. The success of such measures also depends on the chemical contamination, especially of floodplain soils and sediments, which serve as sinks and sources for a variety of pollutants. In this study, we assess the current ecotoxicological status of a floodplain restoration site along the Main River (Frankfurt am Main, Germany) and estimate its development potential with respect to the influence of a local industrial plant and potential legacy contaminations. We therefore use in vitro effect-based methods (EBMs) testing for baseline toxicity, mutagenicity, dioxin-like and estrogenic activities, coupled with chemical analysis.ResultsOf all water bodies analyzed, the overall toxicity was highest in two flood depressions. In the respective water phase, estrogenic activities exceeded the environmental quality standard and sediment samples were positive for all tested endpoints. Chemical analysis of these sediments revealed high concentrations of polycyclic aromatic hydrocarbons. Soil samples from frequently flooded areas showed the highest mutagenic potential for both frameshift and point mutations with and without metabolic activation. The industrial effluent showed baseline toxic, mutagenic, and dioxin-like activities, that were highly diluted in the Main River. In turn, most of the sediment samples downstream of the industrial discharge showed significantly elevated baseline toxic, estrogenic and dioxin-like activities as well as increased chemical contamination.ConclusionBased on the results of this study, we rate the overall ecotoxicological status of a recently established tributary and groundwater-fed ponds as good, and identified two flood depressions near the Main River as hot spots of contamination. We assume that the observed mutagenicity in the floodplain soils is related to legacy contaminations from former aniline and azo dye production. In terms of the development potential of the floodplain restoration site, we emphasize considering the remobilization of pollutants from these soils and suppose that, in the long term, pollution of the Main River and the local industrial plant may negatively impact sediment quality in its tributaries. With this study, we confirmed the utility of in vitro EBMs for identifying chemically and ecotoxicologically relevant sites.

Prediction of Deleterious Single Amino Acid Polymorphisms with a Consensus Holdout Sampler.

Single Amino Acid Polymorphisms (SAPs) or nonsynonymous Single Nucleotide Variants (nsSNVs) are the most common genetic variations. They result from missense mutations where a single base pair substitution changes the genetic code in such a way that the triplet of bases (codon) at a given position is coding a different amino acid. Since genetic mutations sometimes cause genetic diseases, it is important to comprehend and foresee which variations are harmful and which ones are neutral (not causing changes in the phenotype). This can be posed as a classification problem. Computational methods using machine intelligence are gradually replacing repetitive and exceedingly overpriced mutagenic tests. By and large, uneven quality, deficiencies, and irregularities of nsSNVs datasets debase the convenience of artificial intelligence-based methods. Subsequently, strong and more exact approaches are needed to address these problems. In the present work paper, we show a consensus classifier built on the holdout sampler, which appears strong and precise and outflanks all other popular methods. We produced 100 holdouts to test the structures and diverse classification variables of diverse classifiers during the training phase. The finest performing holdouts were chosen to develop a consensus classifier and tested using a k-fold (1 ≤ k ≤5) cross-validation method. We also examined which protein properties have the biggest impact on the precise prediction of the effects of nsSNVs. Our Consensus Holdout Sampler outflanks other popular algorithms, and gives excellent results, highly accurate with low standard deviation. The advantage of our method emerges from using a tree of holdouts, where diverse LM/AI-based programs are sampled in diverse ways.

Cytotoxic, acute oral toxicity, genotoxic and mutagenic assessment of the essential oil from fresh leaves of Croton argyrophyllus (Kunth.)

Ethnopharmacological relevanceCroton argyrophyllus Kunth., commonly known as “marmeleiro” or “cassetinga,” is widely distributed in the Brazilian Northeast region. Its leaves and flowers are used in traditional medicine as tranquilizers to treat flu and headaches. Aim of the studyThis study was conducted to determine the chemical composition and toxicological safety of essential oil from C. argyrophyllus leaves using in vitro and in vivo models. Materials and methodsThe chemical composition of the essential oil was determined using a gas chromatograph coupled to a mass spectrometer. Cytotoxicity was tested in the HeLa, HT-29, and MCF-7 cell lines derived from human cells (Homo sapiens) and Vero cell lines derived from monkeys (Cercopithecus aethiops) using the MTT method. Acute toxicity, genotoxicity. Mutagenicity tests were performed in Swiss mice (Mus musculus), which were administered essential oil orally in a single dose of 2000 mg/kg by gavage. ResultsThe main components of the essential oil were p-mentha-2-en-1-ol, α-terpineol, β-caryophyllene, and β-elemene. The essential oil exhibited more than 90% cytotoxicity in all cell lines tested. No deaths or behavioral, hematological, or biochemical changes were observed in mice, revealing no acute toxicity. In genotoxic and mutagenic analyses, there was no increase in micronuclei in polychromatic erythrocytes or in the damage and index in the comet assay. ConclusionsThe essential oil was cytotoxic towards the tested cell lines but did not exert toxic effects or promote DNA damage when administered orally at a single dose of 2000 mg/kg in mice.

Spatial and Temporal Evolution Patterns of Droughts in China over the Past 61 Years Based on the Standardized Precipitation Evapotranspiration Index

Based on the data of 2254 daily meteorological stations in China from 1961 to 2021, this study calculated the standardized precipitation evapotranspiration index (SPEI) of the national multi-time scale by using the FAO Penman–Monteith model to quantify the changes in dry and wet conditions. The Mann–Kendall mutation test, wavelet analysis, and other methods were used to study the spatial pattern and temporal evolution of drought. The results showed: (1) In the past 61 years, there were obvious spatial and temporal differences in drought in China, and the interannual variation in drought severity in SPEI-1, SPEI-3, and SPEI-12 gradually decreased at a rate of 0.005/10a, 0.021/10a, and 0.092/10a, respectively. (2) The time point of dry and wet mutation was 1989 according to the MK mutagenicity test. (3) Wavelet analysis showed that the drought cycle on the annual scale and the seasonal scale was consistent, and the main period was about 30 years. (4) In the past 61 years, the drought intensity of different degrees in China has shown a weakening trend, and the drought intensity reached the highest value in 61 years in 1978, at 1836.42. In 2020, the drought intensity was the lowest, at 261.55. (5) The proportion of drought stations has shown a decreasing trend. The proportion of drought-free stations has fluctuated greatly, ranging from 42.12% to 89.25%, with 2020 being the highest. This study provides a scientific basis for further research on the causes and coping strategies of drought and is of great significance for strengthening China’s drought monitoring, early warning ,and adaptation capabilities.

Advanced oxidation of polycyclic aromatic hydrocarbons in tropical soil: Self-catalytic utilization of natural iron contents in an oxygenation reactor supported with persulfate

The catalysts derived from natural iron minerals in the advanced oxidation process offer several advantages. However, their utilization in soil remediation is restricted due to the presence of soil impurities, which can inhibit the catalytic activity of these minerals. The soils in tropical regions exhibit lower organic matter content, limited cation exchange capacity, and are non-saline, this enhances the efficiency of utilizing natural iron minerals from tropical soil as a catalyst. In this regard, the catalytic potential of naturally iron-bearing tropical soil was investigated to eliminate phenanthrene (PHE), pyrene (PYR), and benzo[α]pyrene (B[α]P) using an oxygenated reactor supported with persulfate (PS). The system showed an efficient performance, and the removal efficiencies under the optimum conditions were 81 %, 73 %, and 86 % for PHE, PYR, and B[α]P, respectively. This indicated that the catalytic activity of iron was working efficiently. However, there were changes in the soil characteristics after the remediation process such as a significant reduction in iron and aluminum contents. The scavenging experiments demonstrated that HO• had a minor role in the oxidation process, SO4•− and O2•− emerged as the primary reactive species responsible for the effective degradation of the PAHs. Moreover, the by-products were monitored after soil remediation to evaluate their toxicity and to propose degradation pathways. The Mutagenicity test showed that two by-products from each PHE and B[α]P had positive results, while only one by-product of PYR showed positive. The toxicity tests of oral rat LD50 and developmental toxicity tests revealed that certain PAHs by-products could be more toxic from the parent pollutant itself. This study represents a notable progression in soil remediation by providing a step forward in the application of the advanced oxidation process (AOP) without requiring additional catalysts to activate oxidants and degrade pollutant PAHs from the soil.

A Comprehensive Review of the Nutritional Composition and Toxicological Profile of Date Seed Coffee (Phoenix dactylifera)

Approximately 8 million tons of dates (Phoenix dactylifera) are produced globally each year. The seeds of the fruit, which make up 10–15% of its weight, are typically discarded. Date seed coffee is a sustainable food system innovation rooted in the traditions of high date-producing regions. Dating back to the late 19th century, date seed coffee has evolved from a historical coffee substitute to a modern caffeine-free alternative. Date seed coffee has a long history of consumption in the European Union (EU). This indicates that it may not require novel food authorization. The composition of date seeds is evaluated in this review and a toxicological risk assessment for date seed coffee is conducted. Subchronic studies show that consuming date seed or date seed coffee has no adverse effects. Therefore, currently unavailable chronic toxicity, carcinogenicity, and reproductive toxicity studies may be unnecessary. However, for a comprehensive evaluation, it is recommended to conduct an in vitro mutagenicity test. This review provides information on the safety of date seed coffee and highlights the need for further research.

MOLECULAR DOCKING OF THE KERUING's (DIPTEROCARPUS) GENUS, SECONDARY METABOLITES OF THE DIPTEROCARPACEAE FAMILY'S AS ANTI-INFLAMMATION AGAINST CYCLOOXYGENASE-2 (COX-2)

Objective: Kalimantan, Indonesia, has a tropical forest abundant in forest products. One of these products is the Dipterocarp tree, which includes the Keruing genus (Dipterocarpus). Dipterocarpus contains secondary metabolites that may be potential sources for new drug compounds. One of these metabolites has the potential to act as an anti-inflammatory agent. Based on pharmacophore modelling and molecular docking, this study used molecular docking to investigate the inhibitory mechanism and affinity of Dipterocarpus secondary metabolites on the 3N8Y inflammatory receptor. Methods: The study involved multiple stages, such as preparing and optimizing the structure of the test compounds, constructing a 3D receptor structure of 3N8Y, validating the methodology, and performing energy docking simulations to analyze the interactions. From the study that has been done, the results for the test compounds were evaluated for their MolDockScore, Pharmacokinetic parameters (ADME), and toxicity. Results: The results revealed that the oligomer resveratrol compound exhibited the lowest MolDockScore value of-104.7400, comparable to natural ligands. In addition to that, this method produces reliable outcomes through pharmacokinetic predictions such as HIA (88.4794%), Caco2 (5.1917 nm/sec), and PPB (100%). Furthermore, the toxicity profile exhibited negative results for mutagenic, non-mutagenic, and carcinogenic tests, including genotoxic and nongenotoxic substances. Conclusion: The oligomeric resveratrol (3',5',4-trihidroksi-trans-stilben) compounds have potential as anti-inflammatory agents by acting on the 3N8Y receptor, which further needs to be tested in vivo.

Separation and characterization of hydrolytic degradation product of deucravacitinib by validated high‐performance liquid chromatography method and liquid chromatography‐mass spectrometry

AbstractDeucravacitinib, SOTYKTU was approved recently in the year 2022 to treat psoriasis. The present work attempts to identify and characterize degradation products formed when the drug is exposed to hydrolytic, oxidative, thermal, and photolytic stress conditions as well as to study the kinetics of the drug's degradation under various stress conditions. A high‐performance liquid chromatography method was developed for the separation of deucravacitinib and its degradation product comprising mobile phase of ammonium acetate (pH 4.75) buffer as solvent A and acetonitrile as solvent B and Phenomenex Gemini, C‐18 (250 × 4.6 mm, 5µ) column as stationary phase. Injection volume, flow rate, and detection wavelength for the method were optimized as 10 µL, 1.0 mL/min, and 254 nm, respectively. Accuracy, precision, linearity, robustness, and selectivity were found to be acceptable when validated in the concentration range between 5 and 150 µg/mL of deucravacitinib. The structure of the degradation product was characterized using liquid chromatography‐tandem mass spectrometry which shows a protonated molecular ion peak at m/z 358.1805 in the electrospray ionization positive mode. In silico mutagenicity tests yielded positive results for deucravacitinib and its degradation product, triggering an alarm for mutagenicity for both structures, whereas in silico toxicity studies did not generate any structural alerts.

Biological evaluation of 9-thioansamitocin P3

Highly cytotoxic maytansine derivatives are widely used in targeted tumor delivery. Structure-activity studies published earlier suggested the C9 carbinol to be a key element necessary to retain the potency. However, in 1984 a patent was published by Takeda in which the synthesis of 9-thioansamitocyn (AP3SH) was described and its activity in xenograft models was shown. In this article we summarize the results of an extended study of the anti-tumor properties of AP3SH. Like other maytansinoids, it induces apoptosis and arrests the cell cycle in the G2/M phase. It is metabolized in liver microsomes predominately by C3A4 isoform and doesn't inhibit any CYP isoforms except CYP3A4 (midazolam, IC50 7.84 μM). No hERG inhibition, CYP induction or mutagenicity in Ames tests were observed. AP3SH demonstrates high antiproliferative activity against 25 tumor cell lines and tumor growth inhibition in U937 xenograft model. Application of AP3SH as a cytotoxic payload in drug delivery system was demonstrated by us earlier.

Phenolic compounds occurrence and human health risk assessment in potable and treated waters in Western Cape, South Africa.

Phenolic pollutants from industrial and agricultural activities pose a major threat to the world's potable water supply. The persistent micro-pollutants often find their way into drinking water sources with possible adverse human health implications. In this study, bottled water, tap water, and wastewater treatment plant (WWTP) effluent samples from the Boland region of the Western Cape, South Africa were assessed to determine 4-chlorophenol (4-CP) and 2,4-dichlorophenol (2,4-DCP) levels using HPLC/DAD instrumentation. The selected area is known for its vast agricultural ventures and wineries. Evaluation of the human health risk (cancer risk) for the pollutants was conducted using the hazard quotient (HQ). The Ames mutagenicity test was also conducted using the Salmonella typhimurium T98 and T100 strains and the S9 activation enzyme. Trace levels of the phenolics were detected in the samples with a range of 9.32 × 10-7-1.15 × 10-4mg/L obtained for 4-CP, and 8.80 × 10-7-1.72 × 10-4mg/L recorded for 2,4-DCP. Both compounds had levels below the limit of 0.01mg/L prescribed by South African legislation. The assessed HQ for the phenolic concentrations indicates a low level of potential ecological risk and none of the samples had a cancer risk value that exceeded the regulatory limit. The possibility of the analyzed samples causing cancer is unlikely, but non-carcinogenic adverse effects were found. Strong mutagenicity was observed for the T98 strains with a potential ability to cause mutation toward the insertion or deletion of a nucleotide. The T100 bacterial strain showed very slight mutagenicity potential, however, it is unlikely to cause any mutation. The levels of phenolics in the potable water samples may pose a significant threat to human health. Hence, screening persistent organic chemicals in potable water sources and evaluating their potential human health effects is pertinent to prevent associated health challenges.