Analysis of pharmacokinetic profile and ecotoxicological character of cefepime and its photodegradation products.

An important problem is the impact of photodegradation on product toxicity in biological tests, which may be complex and context-dependent. Previous studies have described the pharmacology of cefepime, but the toxicological effects of its photodegradation products remain largely unknown. Therefore, photodegradation studies were undertaken in conditions similar to those occurring in biological systems insilico, in vitro, in vivo and ecotoxicological experiments. The structures of four cefepime photodegradation products were determined by UPLC-MS/MS method. The calculated in silico ADMET profile indicates that carcinogenic potential is expected for compounds CP-1, cefepime, CP-2 and CP-3. The Cell Line Cytomotovity Predictor 2.0 tool was used to predict the cytotoxic effects of cefepime and related compounds in non-transformed and cancer cell lines. The results indicate that possible actions include: non-small cell lung cancer, breast adenocarcinoma, prostate cancer and papillary renal cell carcinoma. OPERA models were used to predict absorption, distribution, metabolism and excretion (ADME) endpoints, and potential bioactivity of CP-2, cefepime and CP-4. The results obtained in silico show that after 96h of exposure, cefepime, CP-1, CP-2, and CP-3 are moderately toxic in the zebrafish model, while CP-4 is highly toxic. On the contrary, cefepime is more toxic to T. platyurus (highly toxic) compared to the zebrafish model, similar to products CP-4, CP-3 and CP-2. In vitro cytotoxicity studies were performed by MTT assay and in vivo acute embryo toxicity studies using Danio rerio embryos and larvae. In vitro showed an increase in the cytotoxicity of products with the longest exposure period i.e. for 8 h. Additionally, at a concentration of 200 µg/mL, statistically significant changes in metabolic activity were observed depending on the irradiation time. In vivo studies conducted with Zebrafish showed that both cefepime and its photodegradation products have only low toxicity. Assessment of potential ecotoxicity included Microbiotests on invertebrates (Thamnotoxkit F and Daphtoxkit F), and luminescence inhibition tests (LumiMara). The observed toxicity of the tested solutions towards both Thamnocephalus platyurus and Daphnia magna indicates that the parent substance (unexposed) has lower toxicity, which increases during irradiation. The acute toxicity (Lumi Mara) of nonirradiated cefepime solution is low for all tested strains (<10%), but mixtures of cefepime and its photoproducts showed growth inhibition against all tested strains (except #6, Photobacterium phoreum). Generally, it can be concluded that after UV-Vis irradiation, the mixture of cefepime phototransformation products shows a significant increase in toxicity.

A comparative study of the predictive performance of different descriptor calculation tools: Molecular-based elution order modeling and interpretation of retention mechanism for isomeric compounds from METLIN database.

In the pharmaceutical industry, the need for analytical standards is a bottleneck for comprehensive evaluation and quality control of intermediate and end products. These are complex mixtures containing structurally related molecules. In this regard, chromatographic peak annotation, especially for critical pairs of isomers and closest structural analogs, can be supported by using a Quantitative Structure Retention Relationship (QSRR) approach. In our study, we investigated the fundamental basis of the reversed-phase (RP) retention mechanism for 1141 isomeric compounds from the METLIN SMRT dataset. Nine different descriptor calculation tools combined with different feature selection methods (genetic algorithm (GA), stepwise, Boruta) and machine learning (ML) approaches (support vector machine (SVM), multiple linear regression (MLR), random forest (RF), XGBoost) were applied to provide a reliable molecular structure-based interpretation of RP retention behaviour of the isomeric compounds. Strict internal and external validation metrics were used to select models with the best predictive capabilities (rtest > 0.73, order of elution > 60 %). For the developed models, mean absolute errors were in the range of 60 to 110 s. Stepwise and GA showed the most suitable performance as descriptor selection methods, while SVM and XGBoost modeling gave satisfactory predictive characteristics in most cases. Validation performed on the published experimental data for structurally related pharmaceutical compounds confirmed the best accuracy of MLR modeling in combination with GA feature selection of general physico-chemical properties. The resulting models will be useful for the prediction of separation and identification of structurally related compounds in pharmaceutical analysis, providing a simultaneous understanding of the interaction mechanisms leading to their retention under RP conditions.

Atmospheric Plasma Supported by TiO2 Catalyst for Decolourisation of Reactive Orange 16 Dye in Water.

PURPOSE: Every advanced oxidation process (AOP) has its limitations in water purification. Novel designs with simultaneous application of different AOPs can offer better solutions for cleaner water. METHODS: We have comparatively studied two advanced oxidation processes (AOPs) on decolourisation of Reactive Orange 16 (RO 16) azo dye pollutant from water: gas plasma treatment by low power atmospheric pressure plasma using novel plasma needle configuration, and semiconductor heterogeneous photocatalysis using titanium dioxide (TiO2) nanopowders. Additionally, simultaneous application of two advanced oxidation processes on azo dye decolourisation was studied. RESULTS: It was found that plasma treatment is very efficient system for the dye removal even for low flow rates (1 slm) of the Ar as feed gas. The presence of 10% of O2 in Ar flow intensified dye oxidation process and shortened required time for total decolourisation. When plasma and catalyst were simultaneously applied, TiO2 was activated with a few Watts plasma source as well as 300 W UV lamp source. The synergic effect of two AOPs was more pronounced for higher feed gas flow rates, resulting in improved decolourisation efficiency. CONCLUSION: Plasma needle can efficiently remove Reactive Orange 16 azo dye from water with a power consumption of only few Watts. With the addition of TiO2 the removal efficiency is significantly improved.

Non-thermal plasma needle as an effective tool in dimethoate removal from water.

Intensive use of pesticides requires innovative approaches for their removal from the environment. Here we report the method for degradation of dimethoate in water using non-thermal plasma needle and analyze kinetics of dimethoate removal and possible degradation pathways. The effects of dimethoate initial concentration, plasma treatment time, Argon flow rate and the presence of radical promoters on the effectiveness of proposed method are evaluated. With argon flow rate of 0.5 slm (standard litres per minute) 1 × 10-4 M dimethoate can be removed within 30 min of treatment. Using UPLC analysis it was confirmed that one of the decomposition products is dimethoate oxo-analogue omethoate, which is in fact more toxic than dimethoate. However, the overall toxicity of contaminated water was reduced upon the treatment. The addition of H2O2 as a free radical promoter enhances dimethoate removal, while K2S2O8 results with selective conversion to omethoate. Using mass spectrometry in combination with the theoretical calculations, possible degradation pathways were proposed. The feasibility of the proposed method for dimethoate degradation in real water samples is confirmed. The proposed method is demonstrated as a highly effective approach for dimethoate removal without significant accumulation of undesirable toxic products and secondary waste.

Non-thermal plasma and ultrasound-assisted open lactic acid fermentation of distillery stillage.

Stillage is the main by-product of bioethanol production and the cost of its treatment significantly affects the economy of bioethanol production. A process of thermal sterilization before lactic acid fermentation (LAF) is energy demanding and is causing deterioration of valuable compounds in stillage. In this study, ultrasound (UT) and plasma (PT) treatments were used for microbial inactivation, and a significant reduction in the number of viable microorganisms in the stillage after PT and UT was observed. After application of treatment, LAF by Lactobacillus rhamnosus ATCC 7469 was initiated. The concentration of LA is used to quantify the efficiency of the stillage revalorization. The highest LA productivity of 1.21 g/Lh and yield of 0.82 g/g were obtained after PT, while UT of 10 min provided productivity of 1.02 g/Lh and LA yield of 0.69 g/g. The results were benchmarked against closed LAF. Around 20% better revalorization of stillage by PT was achieved when compared with conventional sterilization. In addition, an excellent L (+) LA stereoselectivity of 95.5% was attained after PT. From the aspect of energy efficiency, that of PT was three times lower than UT and almost ten times lower than thermal sterilization, but it is the most expensive due to the high consumption of gas which could reduce application of closed Ar atmosphere on larger scales. This way, a simpler and energy efficient process for LA production on stillage was accomplished by "open" fermentation.

Virtual water quality monitoring at inactive monitoring sites using Monte Carlo optimized artificial neural networks: A case study of Danube River (Serbia).

Rationalization of water quality monitoring stations nowadays is applied in many countries. In some cases, missing data from abandoned/inactive stations, spatial and temporal, could be very important, hence the use of artificial neural networks (ANNs) for virtual water quality monitoring at inactive monitoring sites was investigated. The aim was to develop single-output and simultaneous ANNs for the spatial interpolation of 18 water quality parameters at single- and multi-inactive monitoring sites on Danube River course through Serbia. Those different modeling approaches were considered in order to determine the most suitable combination of models. The variable selection and sensitivity analysis in the case of simultaneous models were performed using a modified procedure based on Monte Carlo Simulations (MCS). In general, the multi-target models tend to be more accurate than single target ones, while single output models outperform the simultaneous ones. Hence, for particular monitoring network and set of water quality parameters the optimal combination of models must be defined based on model's accuracy and computational effort needed. The MCS selection procedure has proved to be efficient only in the case of simultaneous multi-target model. MCS based analysis of input-output interactions has shown all significant interactions in the case of simultaneous single-target are grouped as a complex cluster of interactions, where majority of inputs influence on several outputs. In the case multi-target model those interactions were portioned in five separate clusters, there majority of them mimic the input-output interactions that are present in single output models. The modeling strategy for study area was proposed on the basis of the performance of created models (mean average percentage error < 10%): simultaneous multi-target model for pH, alkalinity, conductivity, hardness, dissolved oxygen, HCO3-, SO42- and Ca, single-output multi-target models for temperature and Cl-, simultaneous single-target models for Mg and CO2, single output single target models for NO3-.

The impact of concentration and administration time on the radiomodulating properties of undecylprodigiosin in vitro.

Undecylprodigiosin pigment (UPP) is reported to display cytotoxic activity towards various types of tumours. Nevertheless, its efficacy in modifying the cellular response to ionising radiation is still unknown. In this study, the radiomodulating effects of UPP were investigated. The effects of UPP were assessed in vitro by treating cultures of human peripheral blood with UPP and ionising radiation using two treatment regimens, the UPP pre-irradiation treatment and UPP post-irradiation treatment. The activity of UPP was investigated evaluating its effects on the radiation-induced micronuclei formation, cell proliferation, and induction of apoptosis. The redox modulating effects of UPP were examined measuring the catalase activity and the level of malondialdehyde, as a measure of oxidative stress. The results showed that UPP effects on cellular response to ionising radiation depend on its concentration and the timing of its administration. At low concentration, the UPP displayed radioprotective effects in γ-irradiated human lymphocytes while at higher concentrations, it acted as a radiosensitiser enhancing either mitotic catastrophe or apoptosis depending on the treatment regimen. The UPP modified redox processes in cells, particularly when it was employed prior to γ-irradiation. Our data highlight the importance of further research of the potential of UPP to sensitize tumour cells to radiation therapy by inhibiting pathways that lead to treatment resistance.

Biological effects of bacterial pigment undecylprodigiosin on human blood cells treated with atmospheric gas plasma in vitro.

It is known that some bacterial species are more resilient to different kinds of irradiation due to the naturally developed protective mechanisms and compounds such as pigments. On the other hand, reasoned tissue engineering using plasma remains a critical task and requires very precise control of plasma parameters in order to mitigate its potential detrimental effects. Here we isolated a natural protective agent, microbially produced undecylprodigiosin ((5'Z)-4'-methoxy-5'-[(5-undecyl-1H-pyrrol-2-yl)methylene]-1H,5'H-2,2'-bipyrrole), and investigated its effects on human blood cells independently and in combination with plasma. Two approaches were applied; the first, undecylprodigiosin (UP pigment) was added to the blood cultures, which then were exposed to plasma (pre-treatment); and the second- the blood cultures were exposed to plasma and then treated with pigment (post-treatment). The interactions of plasma and UP pigment with blood cells were investigated by conducting a series of biological tests providing the information regarding their genotoxicity, cytotoxicity and redox modulating activities. The exposure of cells to plasma induced oxidative stress as well as certain genotoxic and cytotoxic effects seen as elevated micronuclei incidence, decreased cell proliferation and enhanced apoptosis. In blood cultures treated with UP pigment alone, we found that both cytotoxic and protective effects could be induced depending on the concentration used. The highest UP pigment concentration increased lipid peroxidation and the incidence of micronuclei by more than 70% with maximal suppression of cell proliferation. On the contrary, we found that the lowest UP pigment concentration displayed protective effects. In combined treatments with plasma and UP pigment, we found that UP pigment could provide spatial shielding to plasma exposure. In the pre-treatment approach, the incidence of micronuclei was reduced by 35.52% compared to control while malondialdehyde level decreased by 36% indicating a significant mitigation of membrane damage induced by plasma. These results open perspectives for utilizing UP pigment for protection against overexposures in the field of plasma medicine.

Microplasma Induced Cell Morphological Changes and Apoptosis of Ex Vivo Cultured Human Anterior Lens Epithelial Cells - Relevance to Capsular Opacification.

Inducing selective or targeted cell apoptosis without affecting large number of neighbouring cells remains a challenge. A plausible method for treatment of posterior capsular opacification (PCO) due to remaining lens epithelial cells (LECs) by reactive chemistry induced by localized single electrode microplasma discharge at top of a needle-like glass electrode with spot size ~3 µm is hereby presented. The focused and highly-localized atmospheric pressure microplasma jet with electrode discharge could induce a dose-dependent apoptosis in selected and targeted individual LECs, which could be confirmed by real-time monitoring of the morphological and structural changes at cellular level. Direct cell treatment with microplasma inside the medium appeared more effective in inducing apoptosis (caspase 8 positivity and DNA fragmentation) at a highly targeted cell level compared to treatment on top of the medium (indirect treatment). Our results show that single cell specific micropipette plasma can be used to selectively induce demise in LECs which remain in the capsular bag after cataract surgery and thus prevent their migration (CXCR4 positivity) to the posterior lens capsule and PCO formation.