The imbalance of oxidation and antioxidant systems in the biological system can lead to oxidative stress, which is closely related to the pathogenesis of many diseases. Substances with antioxidant capacity can effectively resist the harmful damage of oxidative stress. How to measure the antioxidant capacity of antioxidants has essential application value in medicine and food. Techniques such as DPPH radical scavenging have been developed to measure antioxidant capacity. However, these traditional analytical techniques take time and require large instruments. It is a more convenient method to evaluate the antioxidant capacity of antioxidants based on their electrochemical oxidation and reduction behaviors. This review summarizes the evaluation of antioxidants using electrochemical sensors by bibliometrics. The development of this topic was described, and the research priorities at different stages were discussed. The topic was investigated in 1999 and became popular after 2010 and has remained popular ever since. A total of 758 papers were published during this period. In the early stages, electrochemical techniques were used only as quantitative techniques and other analytical techniques. Subsequently, cyclic voltammetry was used to directly study the electrochemical behavior of different antioxidants and evaluate antioxidant capacity. With methodological innovations and assistance from materials science, advanced electrochemical sensors have been fabricated to serve this purpose. In this review, we also cluster the keywords to analyze different investigation directions under the topic. Through co-citation of papers, important papers were analyzed as were how they have influenced the topic. In addition, the author’s country distribution and category distribution were also interpreted in detail. In the end, we also proposed perspectives for the future development of this topic. In the metabolism process, organisms produce many molecules with high oxidation activity, such as oxygen active free radicals, nitrogen active free radicals, carbon active free radicals, hydrogen peroxide, and singlet oxygen . ExcessiConventional determination methods of antioxidant capacity include chromatography, spectroscopy, and electrochemical methods . ChromatIn recent years, many research groups have reviewed the evaluation of the anti-oxidation ability of electrochemical sensors. However, these reviews generally cover only electrochemical advances in a particular type of methodology or sensors for specific materials. The achievements and development trends of the whole subject have not been scientifically described. Bibliometrics is a method of statistical analysis of published literature. It can summarize the content of a topic and discuss the trend of various directions under the topic according to its development process, and even predict the future direction. In this work, CiteSpace was used for bibliometric analysis. CiteSpace was developed by Dr. Chaomei Chen, a professor at the Drexel University School of Information Science and Technology ,19,20,21The number of publications on a topic each year is a measure of its popularity. The change in publication numbers can reflect the ability of this topic to attract scholars’ attention in different periods. This paper focuses on bone marrow hematopoietic cells . The proBetween 1999 and 2003, fewer than 10 papers were published on the topic. Starting in 2004, the annual number of papers on the topic exceeded 10 for the first time and stayed above 10 until 2008, when it topped 20. The topic began to attract more attention in 2010, with 33 papers published that year and more than 50 papers 2 years later. Since its inception in 2012, the topic has remained hot. The annual number of publications in 2019 reached a historical peak of 74. The trend in the annual publication numbers shows that the topic has been attracting the attention of scientists since it entered academia. This means that the topic’s concerns continue to influence the academic world, which also means that the topic has not been appropriately addressed. Specifically, no electrochemical sensor can fully meet the evaluation of oxidation resistance.On the other hand, we can find some very influential journals, such as Nature and Advanced Materials, on the periphery of significant networks. These journals have a very high impact factor and are involved in the topic, but they do not show the same impact on the topic. There are two possible reasons for this. The first is that the articles in these journals have only recently been published and have not been widely cited. The second possibility is that the evaluation of antioxidants using electrochemical sensors appeals only to scholars in a specific field, so reports published in non-chemical journals would not attract much attention.We further analyzed the categories to which all published papers belonged, which can be used to understand where the topic has cross-influence. Given that so many countries are involved in this topic, as shown in Keyword analysis can be used to understand the different research priorities of the topic and reflect on what other topics are closely related to the topic. Burst detection is a more advanced method than citation counts or downloads for identifying publications receiving significant attention from the research community at various stages of development. Cluster analysis of keywords can understand the research focus formed by different keywords in this topic. s-haplotypes and Hb F levels of oxidative stress markers in sickle cell anemia in Brazil. Rose et al. [2+ oxidation-induced ASF. When ASF was combined with vitamin E or ascorbic acid, the effect was better. It is represented that ASF can act synergistically with vitamins C and E to protect low-density lipoprotein from oxidation. This positive result was also verified in hamsters.#0 (blood cell) The reports in this cluster mainly describe two types of studies. The first is the harm of oxidative damage to the human body. For example, Silva et al.  studied e et al.  examinede et al. . At the e et al.  measurede et al.  reportede et al.  studied e et al.  investigAscophyllum nodosum [Rhizophora apiculata) [Salvia hispanica L.) seeds [#1 (voltametric data) This cluster mainly focuses on the antioxidant properties of plant extracts. Tea ,49,50 an nodosum , mangroviculata) , chia (S.) seeds , and lav.) seeds . Chromat.) seeds ,51,58. E.) seeds . However#2 (enzyme conjugation) This cluster focuses on detecting polyphenols and evaluating antioxidant properties by different detection techniques. Some of this work has focused on specific molecules, such as gallic acid ,61, chlo#3 (DPPH model) This cluster focuses on the mechanisms of antioxidants and the electrochemical detection of antioxidant activity. Brito et al.  used UV-#4 (adsorptive stripping volatammetry) Part of the works in this cluster is focused on the combination of electrochemistry and other analytical techniques, including high-performance liquid chromatography , flow in(Humulus lupulus L.) products [#5 (antioxidative activity assay) The main content of this cluster focuses on investigating hydroxyl radical scavenging by different antioxidants. For example, Ozyurek et al.  studied products , and redproducts .#6 (quenching studies) This cluster of reports focused on analyzing the antioxidant properties of flavonoids. In contrast to the previous clusters, several of these reports involve using carbon nanotubes ,81,82,83#7 (various pH) The reports in this cluster mainly investigated the electrochemical behavior of antioxidants, such as catechin , bis-coe#8 (phenolic acid) The main content of this cluster is the synthesis, characterization, and performance analysis of some derivatives of antioxidant molecules. Therefore, the analytical methods of this group are not limited to traditional analytical methods but also use computational chemistry. It is worth noting that the silhouette value in this cluster is only 0.869, so the content concentration within this cluster is relatively low.#9 (using xanthine myeloperoxidase) Some of the work in this cluster is the preparing of plant extracts and preventing the oxidation of metallic materials ,95,96,97#10 (antioxidative defense mechanism) This cluster contains only two works. Both are case-specific studies. Nia et al.  investig#11 (superoxide ion) This cluster contains three reports which analyze the properties of different macromolecules. Feroci and Fini  reported#12  The work of this cluster is also mainly to analyze the properties of some antioxidants. For example, Susana et al.  investig#13 (micro coulometric titration) This cluster consists of establishing and updating some primary methodologies. For example, Kanyanee et al.  reported2O2 was a sensor reported by Emir et al. [#14 (bone marrow) There are only three papers in this cluster. The keywords that cluster them together are hydrogen peroxide and in vivo. The paper on Hr et al. , and ther et al. ,112.#15  The two works in this cluster investigated the properties of binuclear transition metal complexes  and a tr#16 (microsensor) Only one paper in this cluster reports a quantitative assay for limonin . Cerium We further use the frequency of occurrence of keywords to make the confusion matrix between keywords . As can (1)Cyclic voltammetry and differential pulse voltammetry are the electrochemical techniques most commonly used by electrochemical sensors to analyze the antioxidant capacity of target samples.(2)The electrochemical behavior of antioxidants can be used to understand the mechanism of these substances during redox.(3)The primary sources of antioxidants are plants.(4)Boron doped diamond electrodes, screen printed electrodes, and glassy carbon electrodes are most commonly used as working electrodes for electrochemical sensors.(5)Carbon nanotubes are the most commonly used nanomaterials for electrode surface modification.Based on the above analysis of keywords, the investigation directions of the evaluation of antioxidants using electrochemical sensors can be summarized as follows:Co-citation analysis is a critical way to understand the development of the whole topic. Co-citation analysis can learn from the citations of selected data which papers push the topic forward in each direction. There are smaller sub-networks at the top and bottom right of the figure, and these networks contain older papers. In the lower right corner of the network, Chevion et al.  reportedPunica granatum, Myrtus communis, and Itriphal formulations. Tashkhourian et al. [2 NPs-modified carbon paste electrode. Similar works were conducted by Luo et al. [2, and a dinuclear copper(II) octaazamacrocyclic complex, respectively. Kahl and Golden [Among all sub-networks, the most influential paper reported that the antioxidant capacity of wine was detected by a voltametric scanning technique with carbon nanotube-modified electrodes . Anothern et al.  reportedo et al. ,198 and o et al. , but thed Golden  depositeThe paper of Ziyatdinova et al.  directed(1)The topic started in 1999 and did not attract much attention until 2010. After 2010, the topic became popular, and the trend continues today. This means that the topic has received much attention so far. At the same time, the problems faced by this theme have not been solved perfectly so far.(2)In the early stages of this topic, electrochemistry was a quantitative analysis technique, often used in conjunction with chromatographic and spectroscopic techniques for the separation and quantitative detection of complex samples. After that, cyclic voltammetry technology began to attract attention. The electrochemical behavior of antioxidants was used to measure their redox mechanism and quantitatively analyze their antioxidant capacity.(3)Most antioxidant capacity investigations focus on a specific antioxidant or a group of structurally similar molecules, such as flavonoids and polyphenols. However, due to the availability of plants as an important means for human antioxidant intake, many papers also use electrochemical sensing technology to determine plant samples or plant extracts directly. In addition, antioxidant properties derived from plant extracts can also protect metal materials from corrosion.(4)Since antioxidants tend to have significant electrochemical redox properties, commonly used commercial electrodes are already capable of direct detection. Carbon paste electrodes, screen printing electrodes, glassy carbon electrodes, and boron-doped diamond electrodes are the most commonly used working electrodes for analysis. However, advances in materials science have greatly improved the performance of electrochemical sensors. Nanomaterial modifications on the surface of the working electrode can improve the sensing performance remarkably. Among them, carbon nanotubes are the materials most used for electrode surface modification in this topic.(5)Because the oxidants damage DNA, the immobilized DNA on the electrochemical sensor’s surface can be used to measure the antioxidant capacity of antioxidants. The extent to which DNA has been damaged has been an indicator of such electrochemical DNA sensors.The evaluation of antioxidant capacity has important application value in medicine and food science. The antioxidant capacity of the product is directly related to its commercial value. Therefore, it is necessary to evaluate antioxidation capacity scientifically and quickly. Traditional analytical techniques can achieve this goal, but they have limitations in practical operation and are not suitable for large-scale promotion. The electrochemical oxidation and reduction properties of antioxidants allow them to be quantitatively and qualitatively evaluated by electrochemical techniques. Based on the above bibliometric analysis, the development and content of this topic are summarized, and the following conclusions can be drawn:(1)Direct electrochemical sensors mainly rely on antioxidants’ electrochemical oxidation and reduction behavior. This method helps determine a particular antioxidant, but if the sensor is dealing with a complex sample, the electrochemical behavior is difficult to identify accurately. This is because complex samples contain a series of electrochemically active molecules whose presence can interfere with the measured current value of the target molecule. Therefore, it is a challenge to improve the specificity of direct electrochemical sensors to determine antioxidant capacity.(2)Choosing suitable electrode modification material is also a significant challenge. The current trend is to modify the electrode by using binary, ternary, or even multiple nanocomposites. Although nanomaterials have excellent properties, the synergistic effect between multiple materials cannot be explained theoretically. Therefore, the performance stability of these nanocomposites has been a limitation to their widespread use. At the same time, the raw materials of some nanocomposites’ preparation are costly and do not have the prospect of mass synthesis.(3)Although DNA biosensors are methodologically attractive, specially designed DNA requires higher prices. At the same time, such biosensors will be significantly affected by the external environment, so how to ensure their stability is also a meaningful direction.Meanwhile, based on the review of this topic, we believe that the following issues need to be investigated regarding the evaluation of antioxidants using electrochemical sensors:


Dear Editor,3 or in the presence of some defining condition, such as Kaposi sarcoma (KS), which is the most frequent neoplasm in these patients with diagnosis based on clinical and histopathological findings.3In Acquired Immunodeficiency Syndrome (AIDS) there is a deficiency of T-cell-mediated immunity, making the host vulnerable to opportunistic infections and malignancies.3 at the time of the diagnosis. Histopathology of a violaceous skin lesion showed an atypical vascular proliferation affecting the dermis and on immunohistochemistry positivity for CD31 and herpes virus 8 (HHV-8), confirming the diagnosis of KS (3, demonstrating improvement in relation to the first test. However, after approximately eight months, the patient returned to the Infectious Diseases Unit with respiratory distress and hematemesis. He was transferred to the ICU of the Oncology Unit, with rapid progression to death without due clarification of the cause of the bleeding.The present report describes a 31-year-old male patient who presented with a violaceous lesion on the oral cavity, which progressed in size and extension within two months, associated with intense odynophagia. He reported having sex with men. On physical examination, he had a violaceous, infiltrated tumor with areas of leukoplakia, which occupied about 70% of the hard palate . In addiis of KS . Chest rKS is an endothelial malignancy and the epidemic clinical form is associated with HIV infection. Its causes and risk factors include herpesvirus type 8 infection, immunosuppression, genetic predisposition, and the presence of HLA DR5.5It may be the first manifestation of AIDS and indicates late diagnosis. The clinical manifestations of KS include asymptomatic violaceous, erythematous or brownish macules that progress to papules, plaques or tumors, which may bleed or ulcerate. There may be concomitant involvement of the gastrointestinal tract, lungs, lymph nodes, bones, and liver.On histopathology, the initial lesions show proliferation and dilation of the dermal vessels, with large endothelial cells; perivascular infiltrate of lymphocytes and plasma cells, extravasated erythrocytes and hemosiderin deposits. In the plaque and nodular stage, there is a proliferation of blood vessels and atypical spindle cells.After the implementation of ART in AIDS, there has been a reduction in the frequency of KS.Treatment includes both local and systemic therapies and depends on factors such as the KS subtype, the course of the disease, its extent, and the patient's symptoms. The management of localized lesions includes radiotherapy, surgical excision, cryosurgery and laser. If there are multiple skin lesions and/or visceral involvement, chemotherapy is indicated. As for the HIV-related KS subtype, ART is the first treatment option.Despite the reduction in AIDS detection rate in Brazil in recent years,None declared.Camila Gonçalves Pinheiro: Collection of data, drafting of the manuscript.Francisco de Assis Silva Paiva: Collection of data.Isabelle Sousa Medeiros Torres Ferreira: Collection of data.Gabriela Teixeira Viana Suppa Meira: Collection of data.Antônio Carlos Evangelista de Araújo Bonfim: Collection of data.Luciana Cavalcante Trindade: Critical review to obtain intellectual content, and approval of the final version of the manuscript.None declared.


Epicardial adipose tissue (EAT) is known as an important imaging indicator for cardiovascular risk stratification. The present study aimed to determine whether the EAT volume (EV) and mean EAT attenuation (mEA) measured by non-contrast routine chest CT (RCCT) could be more consistent with those measured by coronary CT angiography (CCTA) by adjusting the threshold of fatty attenuation.In total, 83 subjects who simultaneously underwent CCTA and RCCT were enrolled. EV and mEA were quantified by CCTA using a threshold of (N30)  as a reference and measured by RCCT using thresholds of N30, N40 , and N45 . The correlation and agreement of EAT metrics between the two imaging modalities and differences between patients with coronary plaques (plaque ( +)) and without plaques (plaque ( −)) were analyzed.P < 0.001), whereas mEA showed a moderate correlation , P < 0.001). Threshold adjustment was able to reduce the bias of EV, while increase the bias of mEA. Data obtained by CCTA and RCCT both demonstrated a significantly larger EV in the plaque ( +) group than in the plaque ( −) group (P < 0.05). A significant difference in mEA was shown only by RCCT using a threshold of N30 (plaque ( +) vs ( −): − 80.0 ± 4.4 HU vs − 78.0 ± 4.0 HU, P = 0.030). The mEA measured on RCCT using threshold of N40 and N45 showed no significant statistical difference between the two groups (P = 0.092 and 0.075), which was consistent with the result obtained on CCTA (P = 0.204).EV obtained from RCCT showed very strong correlation with the reference , Applying more negative threshold, the consistency of EV measurements between the two techniques improves and a consistent result can be obtained when comparing EF measurements between groups, although the bias of mEA increases. Threshold adjustment is necessary when measuring EF with non-contrast RCCT. The visceral fat located between the myocardial surface and the visceral layer of the pericardium, known as epicardial adipose tissue (EAT), is well known as an important imaging indicator for cardiovascular risk stratification . EvidencCardiac examinations such as echocardiography, coronary CT angiography (CCTA) and the coronary calcium score (CCS) are currently the most common imaging techniques used to assess the amount of EAT , 13. HowAll subjects who underwent coronary CT angiography (CCTA) and non-contrast RCCT simultaneously for health check-ups in our center from January 2019 to August 2020 were retrospectively investigated. Those who underwent surgery or invasive procedures of the lung, mediastinum and heart were excluded. A total of 83 subjects were ultimately enrolled in this study. Information about sex, age, diabetes, dyslipidemia, and hypertension was collected from their records. This retrospective study was approved by the institutional review board. All participants were fully informed and agreed that their medical records would be anonymized for research purposes.CCTA and RCCT scanning were performed on the same 320-slice CT scanner  sequentially without changing the position. The non-ECG-gated RCCT parameters were as follows: 120 kVp; 300 mAs; detector collimation, 160 × 0.5; pitch, 1.0938; rotation time, 0.5 s; matrix size, 1024 × 1024; field of view, 350 mm; and slice thickness, 1.0 mm, covering the scanning range from the lung apices to the bases. Subsequently, CCTA was performed using a breath-hold prospective axial ECG-triggered acquisition protocol. For patients with heart rate > 65 beats/min, metoprolol was taken orally approximately 1–1.5 h before CCTA examination. Sublingual nitroglycerine (0.5 mg) was administered 5 min before scanning, except in the case of contraindications. Intravenous injection of iodinated contrast medium was injected through the right cubital vein with a double cylinder high pressure syringe  followed by saline (25 ml) injection at the same flow rate. The scan was obtained from the carina to the bottom of the heart, and Bolus Tracking automatic trigger scanning technology was used. The monitoring layer was located at the center of the scanning range, the ROI was placed at the center of the descending thoracic aorta, the triggering threshold was set at 120 HU, and the scanner was delayed 6 s to start scanning automatically after reaching the threshold. The parameters were as follows: 100 kVp; 120 mAs; detector collimation, 320 × 0.5; pitch, 1.0938; rotation time, 0.25 s; matrix size, 512 × 512; field of view, 350 mm; and slice thickness, 0.5 mm.All the images were imported from the Picture Archiving and Communication System to the postprocessing workstation . Both the cross-sections and longitudinal reconstructed images were visually inspected to detect coronary plaques by two radiologists with 10 and 15 years of experience in cardiac imaging analysis. The presence of atherosclerotic plaques and stenosis grading were evaluated based on the 18-segment model recommended by Society of Cardiovascular Computed Tomography . Stenosi3) equals the product of the volume of a single voxel and the number of fat-containing voxels. The mEA (reported in HU) was defined as the mean attenuation of all fat-containing voxels. An attenuation histogram was reviewed to show the distribution of fat-containing voxels. A threshold of − 190 to − 30 Hounsfield units (HU)  was applied to extract the fat-containing voxels for CCTA imaging. For the RCCT, the lower threshold was fixed at − 190 HU, and the upper threshold was adjusted and set at − 30 HU (N30), − 40 HU (N40) and − 45 HU (N45). The measured EV and mEA at the corresponding thresholds were recorded as EVN30 and EAN30, EVN40 and mEAN40, EVN45 and mEAN45, respectively.EAT was defined as the visceral fat between the myocardial surface and the visceral layer of the pericardium. Fig. a, b. TheP value < 0.05 was considered statistically significant.Statistical analyses were performed using SPSS version 19.0 for Windows . The Kolmogorov–Smirnov test was used to check whether the data conformed to normal distribution. Continuous variables with normal distribution are expressed as the mean ± standard deviation (SD), and categorical variables are expressed as N (%). The measurement data from CCTA were compared with those from RCCT using the paired t test. Correlations and agreement of EAT measurements between RCCT and CCTA scans were evaluated using Pearson’s correlation test and Bland–Altman analysis. The difference in EAT according to the presence or absence of coronary atherosclerotic plaques was analyzed using Student’s t-test, Mann–Whitney U test, and the dichotomic variables were analyzed using chi-squared test. A Among all 83 subjects enrolled, 49 subjects (59.0%) had no plaques or luminal stenosis detected by CCTA. Plaques were detected in the other 34 41.0%) subjects, in which 9 (26.5%) had minimal stenosis, 18 (52.9%) had moderate stenosis, 5 (14.7%) had severe stenosis, and 1 (2.9%) was occluded. Among the patients with plaques, 24 subjects (70.6%) had calcified plaques, and 10 subjects (29.4%) had noncalcified plaques. Regarding the number of involved coronary artery segments, 17 subjects (50.0%) had only 1 segment, 7 subjects (20.6%) had 2 segments, 5 subjects (14.7%) had 3 segments, and 5 subjects (14.7%) had 4 segments. The general characteristics are shown in Table .0% subjeN30 was − 15.2 (− 29.7 to − 0.6) cm3, suggesting an overestimate of approximately 15% in RCCT compared with CCTA using the same threshold  HU   cm3 and 1.9 (− 11.1 to 14.8) cm3, respectively  correlated moderately with the EV in CCTA   HU, 5.2 (− 0.6 to 11.1) HU, and 7.4 (1.6 to 13.3) HU, respectively  was significantly larger than that of plaque ( −) group  (Table N30 of the plaque ( +) group was significantly larger than that of the plaque ( −) group , but the mEAN30 of the plaque ( +) group was significantly lower than that of the plaque ( −) group  , but the differences in mEA were not significant   group 115.6 ± 44.1 cm4.1 cm3 wThe present study validated that non-contrast RCCT could be used to quantify EV and mEA, though EV might be overestimated if the same upper threshold of − 30 HU was adopted as most previous studies used. By adjusting the upper threshold, the consistency of EV measured by RCCT with that measured by CCTA could be improved substantially, but this was not the case for mEA. Although the bias of mEA increased, the same results were obtained when comparing mEA between the two groups with and without coronary plaques. Interestingly, the mEA measured on non-contrast RCCT using N30 was sensitive enough to detect the differences in EAT characteristics between the groups with or without coronary plaques. The quantification of mEA might be more sensitive for revealing latent pathophysiological characteristics than the quantification of EV.ECG-gated cardiac CT is considered to be the most accurate method to quantify EV because of the high resolution and true volume coverage, allowing also the assessment of mEA , 19. WheAlthough there are abundant reports, the predefined thresholds for fat tissue are frequently inconsistent. The lower threshold is usually set at − 250 HU or − 190 HU, and the upper threshold is set at − 45 HU, − 30 HU, or − 15 HU , 27, 28.The present study demonstrated that adjusting the threshold could improve the consistency between EV measured by RCCT and that measured by CCTA. Bucher, A. M et al. systematically analyzed the influence of technical parameters on the quantification of EV on cardiac CT and found that threshold adjustments, especially the upper level, could make volumetry from different series comparable . As showEA was reported as a measure of fat composition that might indicate the atherosclerotic process . DecreasThe current study had limitations. First, we did not try to determine the optimal threshold or provide a recommended threshold. CT attenuation varies by equipment manufacturer, performance, and scan parameters. There are currently no endorsed guidelines to quantify EAT, even though we defined EV measured on CCTA as a reference because it was widely used in previous studies. We proposed the approach of threshold adjustment to reduce the differences in EAT measurements between the different examination protocols. Second, the number of patients was small, and the patients with coronary plaques were in early stages and asymptomatic. The predictive efficacy of EAT measurements for CAD was not explored. Third, our results may not be applicable to patients with high heart rate because metoprolol was taken for patients with heart rate > 65 beats/min before CT examination in our study.Our study demonstrates that threshold adjustment is necessary when measuring EF with non-contrast RCCT. Comparing with CCTA, the application of more negative threshold improves the consistency of EV measurements and provides a consistent result when comparing EF measurements between groups, although the bias of mEA increases. More studies are needed to reveal the subtle change of EA after contrasted enhancement and using more accurate method such as histogram analysis.


The spent caustic with strong alkali first replaced the alkali activator to prepare the geopolymer. The influence of spent caustic to the geopolymer was characterized through compressive strength measurement, XRD, MIP analysis and NMR, and the immobilization efficiency of organic in geopolymer was evaluated through the measurement of total organic carbon (TOC). The results show that the spent caustic can partially replace the alkali activator to prepare the geopolymer, and it shows a better performance than that which was activated with pure NaOH solution when the alkalinity is between 4 mol and 14 mol. The organic matter in the spent alkali can be effectively fixed in the geopolymer, which will hinder the geopolymerization in the initial stage of the polymerization reaction but has little effect on the chemical structure and mechanical properties of the final product. With the degree of alkalinity increasing, the immobilization efficiency is improved, and the maximum can reach 84.5%. The organics in the spent caustic will hinder geopolymerization at the initial stage but has little effect on the chemical structure and mechanical property of the final product. This study proposes a new method for the recycling of spent caustic, which also reduces the preparation cost of geopolymers. However, treated spent caustic still needs additional treatment, such as acid neutralition, before being discharged to the sewage treatment plant, due to its high COD level and high alkalinity [Spent caustic is a kind of the liquid wastes generated in various chemical industries, especially in oil and gas refineries . It may kalinity . This tr2 footprint in most life cycle categories and was also associated with high costs [2CO3 in the treated spent caustic can reach about 20% [2 emission of geopolymer preparation.Geopolymers are a kind of environmentally friendly aluminosilicate material with a comparable strength performance to the Ordinary Portland Cement, and are considered to be a good potential substitute for the latter . The lowgh costs ,16. Danigh costs . Fortunabout 20% , which hIn this work, we have studied the preparation of geopolymers using spent caustic to partially replace NaOH solution as an alkali activator, and pure NaOH solution with comparable alkalinity was used as a control group. The effect of spent caustic on the compressive strength and microstructure of geopolymers was studied, and the immobilization efficiency of organics in geopolymers was evaluated. This study proposes a new method for the utilization of spent caustic, which also reduces the preparation cost of geopolymers.The class F fly ash used in this paper was purchased from a plant in Hubei province, China, which was used as a raw material in the synthesis of geopolymers. The activator solution was prepared by mixing NaOH with spent caustic or deionized water in different proportions. According to the Chinese Ministry of Environmental Protection Standard HJ/T 300–2007, the immobilization efficiency of organics in geopolymers was evaluated by the leaching toxicity of organics. In this method, the leaching process was performed with the liquid/solid ratio of 20 mL/g for 18 h, which is the same as it in the USEPA TCLP method. However, the extraction solution was prepared by dissolving 17.25 mL acetic acid into 1 L deionized water, and the amount of acid was almost three times that of the latter, so this standard is more severe. In addition, a zero-headspace extraction vessel was used to prevent the volatilization during the leaching process; the specific operation refers to Reference [29Si nuclear magnetic resonance  spectra were used to analyze the microstructure of geopolymer gel. The spectra were collected at 79.5 kHz on a 7 mm probe with a spinning speed of 5 kHz, a pulse width of 6.5 μs and a relaxation delay of 10 s. The 29Si chemical shift was referenced to an external standard of tetramethylsilane.The mechanical property of geopolymers was tested by a mechanical tester  in accordance with the ASTM Test C-39. In the measurement, at least three specimens were tested, and the average value was used. The mineralogical study of geopolymers was characterized by X-ray diffraction  with Cu Ka radiation. The pore distribution of geopolymers was analyzed by mercury intrusion porosimetry analysis . 2CO3, it can be attributed to the combined effect of Na2CO3 and NaOH as alkaline activator in preparation of geopolymers. Zhang et al. [2CO3-activated system showed higher compressive and flexural strengths than the NaOH-activated system. When the alkalinity is further increased to higher than 12 mol/L, the proportion of alkali in the spent caustic is much less than that of sodium hydroxide, which has less of an effect on the geopolymer, resulting in a similar strength.g et al.  investigIn order to study the activation effect of spent caustic on the structure of fly ash-based geopolymers, XRD analyses carried out on NaOH-activated geopolymers and spent caustic-activated geopolymers is shown in 29Si NMR spectra of geopolymers synthesized with different alkali activator after 28 days. The Gaussian peak deconvolution of this spectrum can be used to separate and quantify Q4(mAl) species of silicon  [4(mAl) center with the replacement of each aluminum by silicon is an approximate −5 ppm difference in δ, with Q4(4Al), Q4(3Al), Q4(2Al), Q4(1Al) and Q4(0Al) resonating at approximately −84, −89, −93, −99 and −108 ppm, respectively [2 structure in C-S-H gel [29Si NMR date for the samples after deconvolution. For the samples No. 3 and No. 10, a broad peak ranged from −70 to −120 ppm was shown, which is similar to the spectrum of fly ash. It indicated that a large number of fly ash was not activated to form geopolymer gel. Compared with sample No. 10, the content of Q4(4Al) and Q4(3Al) in sample No. 3 is higher, while that of Q4(0Al) and Crystalline silica is opposite. Considering that the Al-rich gel is the primary reaction product of the alkali activation of fly ash [ ≤ 4, m) . The resectively . It was -S-H gel . The pea-S-H gel . Table 3 fly ash , it suggThe porosity and pore distribution of geopolymers with different alkali activators is shown in The measurement of total organic carbon (TOC) was used to quantify the leaching of organics from geopolymers . The org(1)The spent caustic can partially replace the NaOH to synthesize geopolymers, and the organics in spent caustic can be immobilized in geopolymers. The method can not only make the spent caustic harmless, but also utilize the strong alkalinity of the spent caustic to reduce the cost of preparing the geopolymer.(2)When the degree of alkalinity is higher than 4 mol/L, the geopolymers prepared by the mixed activator of spent caustic and sodium hydroxide have better 28 days compressive strength than that synthesized with pure NaOH solution, and the highest strength can reach 21.86 Mpa.(3)With the degree of alkalinity increasing, the immobilization efficiency of organics in geopolymers is improved, and the maximum can reach 84.5%. The organics in the spent caustic will hinder geopolymerization at the initial stage but has little effect on the chemical structure and phase of the final product.In this study, fly ash-based geopolymers were prepared by using spent caustic as alkali activator and the influence of spent caustic on the formation of geopolymers was studied. The result is as follows:


The peak and decline of the number of news articles are affected by time and related regulations. Conclusion: the main themes of Chinese news content on e-cigarettes are regulations and control, and minor protection. Newspapers should shoulder the responsibilities and play an important role in health communication with balanced coverage.Background: Electronic cigarettes (e-cigarettes) have been a newsworthy topic in China. E-cigarettes are receiving greater consumer attention due to the rise of the Chinese e-cigarettes industry. In the past decade, e-cigarettes have been widely debated across the media, particularly their identity and their health effects. Objective: this study aims to (1) find the key topics in e-cigarette news and (2) provide suggestions for future media strategies to improve health communication. Method: We collected Chinese e-cigarettes news from 1 November 2015 to 31 October 2020, in the Huike (WiseSearch) database, using “e-cigarettes” (Chinese: “电子烟”) as the keyword. We used the Jieba package in python to perform the data cleaning process and the Dirichlet allocation (LDA) topic modeling method to generate major themes of the health communication through news content. Main finding: through an analysis of 1584 news articles on e-cigarettes, this paper finds 26 topics covered with 4 themes as regulations and control ( E-cigarettes are battery-powered nicotine delivery devices, which is an effective and harmless substitute of a smoking cessation program  and coulAs the primary source of information for the public, the function of media goes beyond disseminating information. News coverage is significant given the news media’s ability to shape public awareness and perceptions of the salience of products and policies ,17, whicThe main objectives of the study are to find the key topics in e-cigarette news and to provide suggestions for future strategies to improve health communication. Therefore, we first examined how e-cigarettes are reported in Chinese news and concluded the main themes by collecting a total of 1584 e-cigarettes news and analyze with the LDA topic modeling method, a generative approach for collections of discrete data . Then, wIn order to gain insights about health communication through Chinese media platforms, we collected Chinese e-cigarettes news from 1 November 2015 to 31 October 2020. We chose the keyword “e-cigarettes” and searched the Huike (WiseSearch) database, one of the most professional leading Chinese media databases founded in 1998. This database collected over 96 million news items daily, over 1600 print media, and 50,000 internet media offer news to this database .The reason why we chose LDA topic modeling is because it is a type of topic modeling method that has been utilized in a number of studies for text mining , or in fd|topic) . The basd|topic) .w in a corpus Choose Choose N words (a)θ);Choose a topic (b)Choose a word For each of the LDA assumes the following generative process for each document  M. Blei .To better understand the news data on e-cigarettes, we use LDA topic modeling method to study the main topics of the news content. Our paper applied LDA to generate major themes of health communication through news content. There is a three-level hierarchical Bayesian model in the LDA method, and words are a random mixture of multiple latent topics. Different topics are related to a distribution over words . By applThe screenshot of the Huike database search page in A total of 1584 e-cigarettes news were collected from Huike. We cleaned and prepared the data for LDA topic modeling as illustrated in Choosing a proper topic number as a parameter for further analysis is significant for the LDA topic modeling approach in order to avoid having too many or too few topic numbers that are either difficult to interpret or unable to distinguish one another. Our study looked for the optimum number of topics for LDA by calculating the coherence score . This meWe visualize the topic allocation as circles in To better understand the relationship between different topics, we visualize the 26 topics in As shown theme 3: more harmful substances than combustible tobacco (5.1%), topic 10: addiction and harms (4.3%), topic 12: toxic vapor (4.2%), and topic 18: higher risk of illness influenced by vaping (3.4%) positioned close to each other, thus form theme 3: health effects. As illustrated in According to the visualization and interpretation, we grouped the topics into 4 major themes displayed in To further discuss each topic, we displayed the top 30 keywords as relevant terms, shown in Media sources facilitate us to build the communication map from the origin, where we list news agencies, released most e-cigarette news in In our study, we we discovered that e-cigarettes are becoming more prominent in the health communication media agenda. The result of the topic modeling assists us in comprehending e-cigarette news coverage in the Chinese press. To explore the overall themes of e-cigarette in health communication of media and proposals for media strategies to promote health communication, our study retrieved relevant news from online and print outlets from 2015 to 2020.n = 81, 5.1%), have constructed a relatively negative image of e-cigarettes: e-cigarettes are not conducive to human bodies, especially youth health. Therefore, the nation has issued regulations and taken actions to regulate the sales of e-cigarettes. Such an agenda may change readers’ awareness, perception, and even behavior toward e-cigarette use that people may refuse to vape [Our results revealed that the four major themes emerged: control and regulation, minor protection, industry (brand) development, health effect. Most domestic news covered the negative aspects of e-cigarettes. Accept for theme 1, which mainly involves the development of the industry, the other three themes, and the following topics, especially in topic 3 , consistent with the intent of the WHO’s advocacy and SMAR and STMA’s e-cigarettes deeming regulations. Considering that May 31 is World No Tobacco Day, and its theme in 2020 is “preventing youth from industry manipulation and preventing them from tobacco and nicotine use , news frRegulations and special actions impact the theme of industry (brand) activities which accounted for 25.5% news content shown in . DespiteIn terms of health effects , news media functioned as public health education and an important pathway for preventing smoking , informiThe prevalence of news stories emphasizing the dangers of e-cigarettes may raise public awareness of the dangers of e-cigarettes. The lack of information about the dangers of vaping and the prevalence of young e-cigarette addiction may also contribute to e-cigarette addiction, particularly among adolescents, so news organizations must have the responsibility of providing balanced coverage of health issues.Irrespective from the findings of the news themes, we also noticed that the agenda remains salient. The official news agencies covered and released those e-cigarettes news , shiftinGovernment policymakers, the media, and the general public formed a circle in this process, and one can influence the other. Media serve as a conduit for government information, guiding public debate to some extent and providing politicians with insight into public problems.The news media, which serves as the government’s megaphone, is responsible for e-cigarette health communication, particularly in terms of clarifying associated policies around the world. Different countries have diverse attitudes toward e-cigarettes, and the news content that is disseminated to the public differs as well. Countries such as the United Kingdom are supportive of e-cigarettes, press reports about them frequently focus on the benefits of e-cigarettes and the nation’s support ,55. MoreIn addition, e-cigarette coverage either transmits voices from the government or delivers the voices of other stakeholders. E-cigarette news often covered the attitudes and behaviors of experts, smokers or vapers, youth or parents, e-cigarette practitioners, and other stakeholders on e-cigarettes. Media coverage enhances the transfer of information and facilitates stakeholders better comprehend each other’s perspectives.To ensure research generalizability, our study uses a large sample size in the recent five years. Nevertheless, several limitations warrant a mention. Owing to the fact that Huike (WiseSearch) database only contains text and LDA fails to analyze too short articles, news contents from mass media through new media platforms employing images, and short text, such as microblogs, may be missed. Furthermore, owing to topic model analysis extracts key topics from news texts presenting as keywords, important statements with few coverages could be lost in the analysis process. Based on this coverage’ analysis, we cannot fully understand the extent to which the news coverage affects audiences in China. In this regard, audience-centric studies are necessary. Moreover, there is a tendency for people acquire e-cigarettes information from social media or web forums, future research should study how frames of news stories converted in multiple media channels.This paper intends to explore news coverage concerns for e-cigarettes in China and generated four main themes by LDA analysis, which are regulations and control, minor protection, industry activities, health effects. Based on a comparison of the number of news and related policy changes in China, and further analysis of related e-cigarette policies in countries around the world, we propose the following media strategies suggestions:(1) Skewed coverage of monopolistic e-cigarette brands or one-sided coverage of the pros and cons of e-cigarettes could create an imbalance in health communication, and how to maintain a balanced coverage requires multiple reporting and a combination of opinions. Overwhelming news articles underscore the potential harm to e-cigarette may raise public awareness of the risk of e-cigarettes. The lack of reports on the dangers of vaping and youth e-cigarette addiction may also lead to e-cigarette addiction, especially for adolescents. Therefore, newspapers should shoulder responsibilities and play important roles in health communication with balanced coverage.(2) Newspapers should shoulder the responsibilities of e-cigarette health communication, especially the explanation of changes in important tobacco rules. Newspapers need to play an important role in educating citizens about health literacy and publicizing industry norms.


Previous evidence shows that antibiotic use and antimicrobial resistance (AMR) spread are not always perfectly correlated within and between countries. We conducted an ecological analysis to evaluate how demographic, economic, governance, health, and freedom characteristics of 30 European countries contribute to antibiotic consumption and AMR.Using three sources of data , we created a dataset of: 22 indicators of demographics, health, economic, governance, and freedom; AMR proportions for 25 combinations of pathogens and antibiotics; consumption of antibiotics in the community. We also computed five indexes of demographic, health, economic, governance, and freedom, and an aggregate AMR measure. Relationships between indexes, antibiotic consumption, and AMR proportions were explored using bivariate, multivariable, multivariate, and mediation analyses.p < 0.001), but not necessarily with lower antibiotic consumption. In multivariable models including all five indexes, an increase in the governance index resulted in significant decreases of overall antibiotic consumption (p < 0.001) and AMR proportion (p = 0.006). Mediation analysis showed that the governance index had an indirect effect on AMR via reducing antibiotic consumption, which accounted only for 31.5% of the total effect.Multivariate analysis identified three clusters of countries that mainly differed for demographic, health, governance, and freedom indexes. AMR proportion was lower in countries with better indexes (These findings could be – at least partially – explained by the contagion theory, for which other factors contribute to high levels of AMR in countries with poor governance. As a result of this evidence, reducing antibiotic use alone is unlikely to solve the AMR problem, and more interventions are needed to increase governance efficiency at global level.The online version contains supplementary material available at 10.1186/s12992-023-00913-0. Antimicrobial resistance (AMR) remains a serious public health concern, which imposes a large clinical and economic burden worldwide –3. AlthoObserved disparities between countries may also be explained by social, governance and economic factors contributing to AMR. It is widely known that health outcomes are affected by some of the most common development indicators  . This miInspired by the research by Collignon and colleagues, we developed the hypothesis that other factors are as important – or even more important than antibiotic abuse – to explain disparities in AMR proportions observed across European countries. Although countries consuming higher levels of antibiotics are also those reporting higher AMR levels, the response to any change in antibiotic use is relatively slow . IndirecWe compiled a dataset for 30 European countries  on data related to 2019 or, if this was not available, to the latest year no earlier than 2016. Data for the subsequent years was not used to prevent the analysis being affected by the COVID-19 pandemic. The dataset consisted of: twenty World Bank indicators of demographics, health, economic, and governance ; politicDue to the multiple variables that contribute to antibiotic consumption and AMR, many of which are interrelated, we synthesized the data into aggregated indexes that capture the main probable influences. The aggregation procedure, borrowed from Collignon and colleagues , first rIn a similar way, we created a measure, termed aggregate AMR, to identify the relationships between AMR and indexes described earlier. The aggregate measure was computed as the average of resistance proportions for all the twenty-five combinations of pathogens and antibiotics. Prior to calculating the aggregate measure, missing values for country-specific AMR proportions were replaced with the average value obtained from countries with available data.Descriptive statistics were used to summarize the main features of European countries. Bivariate analyses were performed by the Spearman’s correlation and reported with the correlation coefficient ρ. Bonferroni correction was used to adjust for multiple comparisons. Linear regression models were further applied to identify aggregate indexes that most affected antibiotic use and resistance across European countries. The first model included the consumption of antibiotics for systemic use in the community as the dependent variable, and aggregate indexes described earlier as independent variables. The second model included the aggregate measure of AMR as the dependent variable, with aggregate indexes and antibiotic consumption as independent variables. According to the Shapiro–Wilk Test, all variables were first log-transformed to ensure their normality before being included in the models. All results were reported as β coefficients and their standard error (SE).path a regressed the mediator  on the independent variable ; the path b regressed the dependent variable (i.e.. the aggregate AMR measure) on the mediator; the path c’ regressed the dependent variable on the independent variable, adjusting for the effect of the mediator. All other aggregate indexes were included in the model as covariates. Bootstrapping with 5,000 resamples was used to calculate the bias-corrected and accelerated confidence intervals (CIs) for the indirect effect (a*b). The percentage mediated was expressed as the percentage of the total effect (path c) accounted for by the indirect effect (a*b).A potential mediating effect has been examined for indexes that exhibited linear relationships with both antibiotic consumption and AMR. In particular, a mediation analysis was carried out to evaluate whether antibiotic consumption mediated the relationships between the governance index and the aggregate measure of AMR. In general, the mediation analysis decomposes the total effect between the exposure and the outcome into a direct and an indirect effect through a mediator variable . In partMultivariate analysis was conducted using the K-means algorithm to reveal different clusters of European countries according to aggregate indexes described earlier. The best cluster solution was determined by the number of clusters that maximized the silhouette score . AntibioAll statistical analyses were performed using the SPSS software , with a significance level α of 0.05, unless otherwise indicated.Data on indicators and antibiotic consumption were available for each of the 30 European countries under consideration Table .Table 1IAcinetobacter spp. resistant to fluoroquinolones and carbapenems. Beyond that, health expenditure per capita was strongly and inversely correlated with the proportions of: Acinetobacter spp. and P. aeruginosa resistant to all classes of antibiotics under consideration; K. pneumoniae resistant to fluoroquinolones, third generation cephalosporins, and aminoglycosides; and E. coli resistant to third generation cephalosporins. Similar correlation coefficients were obtained for the GDP per capita, which was included among the economic indicators. With the exception of political stability, all indicators of governance  were strongly and inversely correlated with the proportions of: Acinetobacter spp. and P. aeruginosa resistant to all classes of antibiotics under consideration; K. pneumoniae resistant to fluoroquinolones, third generation cephalosporins, and aminoglycosides; E. coli resistant to fluoroquinolones and third generation cephalosporins; and S. aureus resistant to methicillin. Government effectiveness and regulatory quality were also inversely correlated with the proportion of S. pneumoniae resistant to macrolides, whereas rule of law inversely also correlated with the proportion of E. coli resistant to aminopenicillins. A similar pattern of correlations was evident for indicators of freedom , which were strongly and inversely correlated with the proportion of: Acinetobacter spp. resistant to all classes of antibiotics under consideration; K. pneumoniae resistant to fluoroquinolones, third generation cephalosporins, and aminoglycosides; E. coli resistant to third generation cephalosporins; and P. aeruginosa resistant to piperacillin and tazobactam and ceftazidime. Political rights score was also inversely correlated with the proportions of E. coli resistant to fluoroquinolones and S. pneumoniae resistant to macrolides, whereas civil liberties score inversely correlated with the proportions of P. aeruginosa resistant to fluoroquinolones and carbapenems. Correlation coefficients for all the analyses described earlier are reported in the Table SAMR proportions for specific combinations between pathogens and antibiotics were available for 26 countries or more Table S. The heap = 0.054).Because several indicators – which were also interrelated to each other – might affect antibiotic consumption and resistance, we also performed correlation and multivariable regression analyses using five aggregate indexes related to the demographic, health, economic, governance, and freedom domains. Levels of each index across European countries are depicted in the Figures SIn further analyses, we evaluated the relationship of aggregate indexes and antibiotic consumption with the aggregate measure of AMR, which showed a median value of 22.5% IQR = 21.3%) and some differences between European countries  and could be decomposed into a direct effect of the governance index on AMR  and an indirect effect through antibiotic consumption . These results suggested that antibiotic consumption partially mediated the relationship between the governance index and AMR, and that the mediator accounted for 31.5% of the total effect  and freedom , in fact, showed similar inverse correlation with AMR levels in the bivariate analysis. Higher levels of governance indicators were also associated with lower antibiotic consumption. These findings were in agreement with those reported by previous studies at national and international levels , 13. ForTotal GDP and health expenditure per capita were also inversely correlated with AMR proportions of different combinations between pathogens and antibiotics. However, the effect of country’s expenditures on AMR levels deserves further examination, because economic indicators may reflect other socioeconomic features. Our results from bivariate analysis, in fact, were partially in contrast with the global analysis carried out by Collignon and colleagues, which showed how high GDP per capita was associated with higher AMR levels . AlthougOverall, our analysis confirms AMR as a multifaced issue that requires concerted and informed approaches. To disentangle the multifactorial contribution to antibiotic use and AMR, we developed five indexes and an aggregate measure of AMR. Multivariable analysis showed that higher indexes of governance and health were significantly associated with lower aggregate levels of AMR. This was true even when considering the detrimental effect of antibiotic consumption on AMR. Among all indexes, the one related to governance was also associated with lower antibiotic consumption in the community. Given the dual action of the governance index, we evaluated the mediating effect of antibiotic consumption on AMR levels. Mediation analysis is increasingly being applied in many fields of research, including epidemiological research , and in With so many factors contributing to AMR, conventional methods of data analysis are often inadequate to address the issue. Hence, we grouped countries by socioeconomics, governance, and health characteristics using a multivariate approach. When we combined countries into different clusters, antibiotic consumption and AMR levels were the highest in countries with the lowest health, governance, and freedom indexes . This evidence increased our confidence in the robustness of the findings.Acinetobacter spp. resistance to fluoroquinolones, aminoglycosides, carbapenems). Missing data were imputed with the average values obtained from countries with available data. Although this was considered the best way to solve the issue, we were unable to identify and manage factors that determined the presence of missing values. The hypothesis that countries reporting data have the best surveillance systems suggests that imputing missing values might have introduced a confounding effect. However, the reasons behind the presence of missing values can be various and not always related to the effectiveness of surveillance systems or to the development of the country. For instance, countries with missing data for Acinetobacter spp. were Estonia, Iceland, Luxembourg, and Malta, which are heterogeneous in terms of socio-demographic and organizational features. In the near future, it would be interesting to replicate our analyses taking into account other factors determining the effectiveness of surveillance systems and/or comparing countries from other regions of the world. Additionally, even when complete, AMR data might be underestimated or overestimated without representative surveillance systems. The same applies, presumably, to data on antibiotic consumption, which might be underestimated due to the volume of antibiotics purchased through other sales channels. For the same reasons, Collignon and colleagues' analysis [Our analysis had several limitations to be discussed. First, it was limited to European countries and data for one year. Although a broader approach could provide meaningful evidence at global level, there was the need for using data that were almost complete and consistent between countries. Just considering one year, in our analysis there were some countries with incomplete data on AMR, especially for some combinations between pathogen and antibiotic (e.g., analysis  covered Second, we summarized the overall set of indicators using aggregate indexes that captured domains that may contribute to antibiotic consumption and AMR. The choice of creating ad-hoc aggregate indexes was motivated by the presence of many possible contributing indicators, which were also interrelated to each other. It was not our intention, in fact, to provide evidence on specific indicators, but rather to evaluate what are the main domains associated with antibiotic consumption and AMR. Moreover, the methodology behind the creation of aggregate indexes from available indicators was borrowed from Collignon and colleagues . There iDespite these limitations, the present study was the first applying bivariate, multivariable, and multivariate analyses to study how socio-economic, governance, and health indicators shape AMR levels across European countries. Searching PubMed for articles published from the inception to November 2022, we identified only the study by Collignon and colleagues that applied a multivariable analysis to the above factors contributing to AMR . HoweverAdditional file 1:Table S1. Descriptive statistics of antibiotic consumption. Table S2. Spearman correlation coefficients between indicators and antibiotic consumptions. Table S3. AMR proportions for combinations between pathogens and antibiotics. Table S4. Spearman correlation coefficients between indicators and proportion of AMR for each combination between pathogen and antibiotic. Table S5. Spearman correlation coefficients between aggregate indexes and antibiotic consumption. Table S6. Differences in indexes between clusters. Table S7. Differences in antibiotic use between clusters. Figure S1. Demographic index across European countries. Figure S2. Health index across European countries. Figure S3. Economic index across European countries. Figure S4. Governance index across European countries. Figure S5. Freedom and rights index across European countries.


Polygoni cuspidati extract limits the application of their pro-health properties. While nanofibers have attracted increasing attention in nutrition delivery due to their special properties, including an increase in the dissolution and permeability, which affects the bioavailability. Therefore, it is justified to obtain nanofibers from Polygoni cuspidati extract, which showed antioxidant and anti-inflammatory properties as a result of a presence of stilbene analogs in the Polygoni cuspidati extract . In the first stage of the work, using the Design of Experiment (DoE) approach, the Polygoni cuspidati extract  was obtained, which showed the best antioxidant and anti-inflammatory properties. Using the Polygoni cuspidati extract as a substrate, nanofibers were obtained by electrospinning. The identification of nanofibers was confirmed on the basis of the analysis of changes in XRPD diffractograms, SEM picture and FTIR-ATR spectra. Obtaining nanofibers from the Polygoni cuspidati extract significantly improved the solubility of resveratrol and polydatin (approx. 6-fold comparing to pure substance). As a consequence, the penetration coefficients of both tested resveratrol and polydatin also increased. The proposed strategy for the preparation of nanofibers from the Polygoni cuspidati extract is an innovative approach to better use the synergy of biological action of active compounds present in extracts. It is especially during the development of nutraceuticals based on the use of selected stilbenes.The low bioavailability of resveratrol and polydatin obtained from  Epidemiological data indicate that civilization diseases, such as cardiovascular disease, diabetes, overweight, obesity and cancer, are significantly increasing . Key facDiseases develop if there is no homeostasis in the human body, which is determined by the oxidation–antioxidant balance, and free radicals are not neutralized by antioxidants. Therefore, it is extremely important to take antioxidants, whose task is to maintain the above-mentioned oxidative–antioxidant balance and to prevent the development of civilization diseases. Among the valuable natural antioxidants it is worth pointing out polyphenols, which are secondary specialized plant metabolites . An intePolygoni cuspidati rhizoma et radix. It is worth noting that other secondary metabolites also present in the metabolite pathway of resveratrol formation have valuable pro-health properties. Additionally, Polygonum cuspidatum, also known as Hu Zhang in China, has the rich composition of active compounds—epicatechin, resveratrol and polydatin—as well as hydrophobic components, including emodin, physcion, torachrysone and their glycosides . Mo. Mo13]. θ angle and above a structural displacement (network expansion), it remains present by the embed excipient and extract. As we may observe for nanofiber HPβCD-PVP vs. nanofiber no. 5 sample comparison, some of polymer matrix micro-strain could be raveled by adding the extract; however, others will grow by the basic structural orientation of the system components. Further analysis for changeable composition and shares of the system may allow for using structural methods for simple predictions of the reaction  that take place at the nearly amorphous state of the sample.Additionally, it was noted that nanofiber samples spectra show both a polymer matrix and excipient extract presence through the main peak position movement, according to At last, the intramolecular interactions between the extract, PVP and HPβCD were analyzed by FTIR-ATR . To deteAfter confirming that all the analyzed systems have a nanofiber structure, the analysis of the influence of the input parameters  on the pThe first of the analyzed output parameters were the content of active components, which were marked in the prepared nanofibers and are collected in Another critical research was dissolution studies of polydatin and resveratrol from nanofiber , Table 7The total amount of released polydatin and resveratrol from nanofibers at 15 min was assessed to determine the significance of the input factors. Again, while none of the effects was statistically significant , it can An important factor determining the application properties of a buccal product is the ability to bind the polymer to the mucin located in the mucosa. For this purpose, rheological studies of nanofiber mixtures were used, and the results are presented in the N-containing groups on the mucosal protein backbone [When analyzing the significance of the input factors , it can backbone .As previously mentioned, the oral cavity becomes an exciting site for administering polyphenols, especially those undergoing rapid first-pass metabolism, which results in low bioavailability, as in the case of resveratrol . For thiWhile none of the effects was statistically significant , it can Dissolution of a drug in an aqueous environment is almost always a prerequisite for oral absorption, and thus, insufficient water solubility often results in limited oral bioavailability. Intestinal permeability is, in addition to water solubility, a key parameter that governs oral absorption . Thus, s50 = 0.13 ± 0.01 mg/mL. The IC50 for nanofibers in each case remained within the error limits obtained for the W10 extract. Therefore, it confirmed that the electrospinning process does not affect the biological properties of the processed extract.Finally, to confirm the maintenance of the antioxidant properties of the extract, such studies have also been carried out for nanofibers by using the DPPH method. The activity of W10 was assessed and amounted to ICw/w/w, i.e., the design of nanofiber number 5.To summarize the production of electrospun nanofibers step, a model of utility contours of profiles was made for all the tested effects . Based oP. cuspidati rhizome and root.The use of the Design of Experiments (DoE) in the extraction of plant material will be essential in advancing and modernizing the creation of standardized phytotherapy. In our study, the influence of the parameters  on biological properties of the obtained extracts was assessed. It was shown that 70% of methanol in the extraction mixture, temperature 70 °C and 4 cycles are the optimal parameters for extraction of active compounds, such as resveratrol and polydatin from the w/w/w.The DoE approach can also be successfully used to select the optimal electrospinning parameters. In the present study, PVP/HPβCD-loaded blend and core shell nanofibers with smooth and bead-less morphology were successfully fabricated for application as new and controlled drug delivery extract systems. The release rate of active compounds in electrospun nanofibers increased with the increase of HPβCD ratio due to enhanced hydrophilicity of the nanofibers. The best composition of electrospun nanofibers was selected, and it is the ratio of extract/PVP/HPβCD 1:1:1 P. cuspidati extract, suggest the particular benefits of the buccal delivery system as a promising strategy to improve the bioavailability of bioactives.Thus, the PVP/HPβCD-based electrospun nanofibers might be strong enough to be easily inserted within the oral cavity and immediate release the incorporated bioactives, while ensuring patient compliance with the smooth structure of nanofibers and low stiffness during treatment. These characteristics, along with the proven antioxidant and anti-inflammatory properties of


TBT1‐TBT6 which entails 1,4‐dithienylbenzene as the backbone and —SMe groups as the anchoring units, with the scanning tunneling microscope break junction (STM‐BJ) technique, are reported. The molecular conductance of TBT1 with intramolecular O•••S noncovalent interactions is enhanced by about one order of magnitude in comparison to their analogue TBT2 . By replacing the methoxy groups in TBT1 with extending alkoxy chains in TBT3, TBT4, and TBT5, the molecular backbones become twisted and as a consequence the single‐molecule conductance decreases gradually, showing that the intramolecular O•••S noncovalent interaction is influenced by the structural features of alkoxy chains. More importantly, the single‐molecule conductance of TBT3, TBT4, and TBT5 can be boosted by increasing the electric field applied to the molecular junctions. Remarkably, the conductance of TBT3, TBT4, and TBT5 can be reversibly modulated due to the conformational changes between twisted and planar ones by varying the electric field. These results demonstrate that molecules with intramolecular O•••S noncovalent interactions have the potential for in situ control of the electrical properties of molecular‐scale devices.Herein, single‐molecule conductance studies of  Here, single‐molecule conductance studies for molecules with 1,4‐dithienylbenzene as the backbone and —SMe groups as the anchoring units by using the STM‐BJ technique are reported. The results show that the single‐molecule conductance can be tuned by varying side chains and electric field through influencing the intramolecular O•••S interactions and molecular conformation within the metal–molecule–metal junctions. Interestingly, conductance of molecular junctions linked by intermolecular hydrogen bonding, π‐π stacking, coordination interaction and host‐guest interaction was successfully measured and investigated. However, investigations of intramolecular noncovalent interactions through single molecular junction were less conducted.Single‐molecule break junction techniques,1 such Although intramolecular O•••S noncovalent interaction was proved by analysis including single crystal X‐ray diffraction, ultraviolet–visible absorption spectroscopy, nuclear magnetic resonance spectra and density functional theory (DFT) computation, there is still a lack of method by means of the single‐molecule junction. It is significant to explore the effects of structural parameters and external stimuli such as electric field on intramolecular O•••S noncovalent interaction. In addition, molecules with noncovalent conformational interactions have the advantage for constructing molecular devices with responsiveness and reversibility. More importantly, the strong electric fields within the two nanoelectrodes of the break junction technique offer the unique tool to realize high‐performance responsive molecular devices. Thus, it is highly interesting to fabricate molecular level devices by varying the electric fields applied across the junction of molecules with intramolecular noncovalent interactions.Recently, intramolecular O•••S noncovalent interaction have been proposed for designing planar conjugated molecules for high performance organic opto‐electric materials.6, 7TBT1‐TBT6 . This is attributed to the more planar backbone of TBT1 induced by the intramolecular O•••S noncovalent interaction. Interestingly, by replacing the methoxy groups in TBT1 with extending alkoxy chains as in TBT3, TBT4, and TBT5, the single‐molecule conductance decreases gradually under 0.1 V bias voltage. This manifests that the intramolecular O•••S noncovalent interaction are influenced by the structural features of alkoxy groups in TBT3, TBT4, and TBT5. Remarkably, the single‐molecule conductance of TBT3, TBT4, and TBT5 can be boosted by increasing the electric field applied to the molecular junctions. It is worth noting that the conductance of TBT3, TBT4, and TBT5 can be reversibly modulated due to the conformational changes between twisted and planar ones by varying the electric field. We further rationalize these results through characterizations including single crystal X‐ray diffraction, ultraviolet–visible absorption spectroscopy, nuclear magnetic resonance spectra and DFT computations. Our work demonstrates for the first time that molecular conformation can be regulated through intramolecular O•••S noncovalent interactions in single molecular electronics via an external field, and molecules with intramolecular O•••S noncovalent interaction have the potential to be used for single‐molecule electrical switches.In this paper, we report the single‐molecule conductance studies of 2TBT1‐TBT3 and investigated the single‐molecule charge transport properties by conductance measurement using STM‐BJ methods. Their molecular structures are shown in Figure TBT1‐TBT3 in Figure TBT1 with strong O•••S noncovalent interactions shows the highest Gaussian fitted conductance peak around ≈10−3.30 G0 at 0.1 V, while the conductance of TBT2 (≈10−4.19 G0) without O•••S noncovalent interactions is around one order of magnitude lower than TBT1. TBT3 is the reference compound with a large alkoxy groups (2‐ethyl‐hexyl group) on the oxygen atom. The conductance value of TBT3 (≈10−4.10 G0) was almost equal with that of TBT2, indicating the interaction of O•••S intensity was weakened by the introduced bulky group. The 2D conductance histograms were constructed by at least 2000 individual traces, as shown in Figure TBT1‐TBT3 were all close to 1.05 nm as shown in the insets of Figure TBT1‐TBT3 should be around 1.55 nm. This value is consistent with the distance between the theoretical distance of the two —SMe anchoring groups (1.4 nm) in molecular backbones, suggesting that these junctions are indeed formed across the molecular backbone. We hence hypothesize that the high conductance observed in TBT1 arises from the intramolecular O•••S noncovalent interactions effect.To verify the intramolecular O•••S noncovalent interactions, we synthesized three molecules TBT1‐TBT3 were obtained by single crystal X‐ray diffraction  Å) are remarkably shorter than the sum of the van der Waals radii of the two atoms (3.25 Å) and such strong intramolecular O•••S noncovalent interaction induces the π‐conjugated block to be a highly planar conformation with a small torsion angle of 1.7°. In contrast, the crystal structure of TBT2 displays a twisted configuration with 52.1° dihedral angle between adjacent thiophene and benzene rings. For molecule TBT3, structural analysis reveals that the distance between oxygen and sulfur is 2.742 (2) Å, which is slightly longer than that in TBT1. However, the existence of alkoxy chains leads to a 20.1° dihedral angle with the molecular backbones. Although the dihedral angle is smaller than TBT2, it is not surprising to predict that the flexible alkoxy chain will result in an increasing torsion angle in the solution. Figure TBT1 < TBT3 < TBT2 and the trend of the molecular conductance variation is opposite. It also suggested that the contribution of conductance enhancement is mainly dominated by the O•••S noncovalent interaction induced coplanarity rather than the charge transport through the O•••S interaction itself. Intramolecular O•••S noncovalent interactions in TBT1 and TBT3 were also identified in the 1H NMR measured under the same condition. As shown in Figure α‐H marked in red, the other hydrogens on thiophenes were defined as β‐H marked in green. TBT1 shows two signals for hydrogens on thiophenes, a broad singlet at δ 7.3636 for α‐H, and a doublet at δ 7.0632 for β‐H consistent with the structure.TBT3 (δ 7.3583 and 7.0515) show two doublets of hydrogens on thiophenes. In contrast, TBT2 shows two high‐field signals in the 1H NMR spectrum at δ 7.0612 and 6.9039  are redshifted compared with TBT2 (319 nm), as shown in Figure π‐electron systems with high charge transport capability.To further support the hypothesis that the intramolecular O•••S noncovalent interactions induced conductance enhancement, the precise configurations of molecules n Figure 3a and 1Htructure.12TBTFigure TBT1 with methoxyl groups, the conductance of TBT3 increased remarkably from ≈10−4.10 G0 to ≈10−3.17 G0 when changing the applied bias voltage from 0.1 to 0.5 V. By contrast, there is no apparent change in the conductance of the molecule with ethyl chains at different bias voltages. Furthermore, 2D conductance histograms were constructed to acquire more information from the stretching process . As shown in the insets of Figure To clarify intramolecular O•••S noncovalent interactions at high bias, we carried out the conductance measurements at 0.5 V. As shown in TBT4‐TBT6  and 0.5 V (≈10−4.38 G0). The conductance modulation trend for these molecules further implies that the electric field affects the intramolecular O•••S noncovalent interactions and thus the molecular conductance. We therefore hypothesize that the twisted molecules containing large alkoxy groups become more planar under a high electric field, thus yielding a high conductance comparable with that of TBT1 with more planar conformation.To further explore the impact of an electric field on the conductance of molecules with intramolecular O•••S noncovalent interactions, we synthesized the other three molecules TBT3 under different bias voltages, we examined the conductance by applying 0.1 V and 0.5 V alternatively using the STM‐BJ technique  than TBT2 . For TBT3, the dihedral angles (‐6.6°and 4.5°) are also smaller than TBT2 but larger than TBT1 due to the steric effect caused by the bulky alkoxy groups. Additionally, we calculated the transmission functions using the nonequilibrium Green's function (NEGF) formalism to further understand the charge transport. Figure TBT1 > TBT3 > TBT2, which is consistent with the experimental results.To further explore the impact of intramolecular O•••S noncovalent interactions on the charge transport in single molecular junctions, we performed density functional theory (DFT) calculations on the junctions formed through Au‐S contacts. As shown in −1 , while the conformations of TBT1 and TBT2 do not show obvious changes. This conformational planarization under a high bias is attributed to the existence of a relatively larger dipole in TBT3, and is responsible for the observed conductance increases under a high applied bias voltage (as reflected by the increase of transmissions). The calculation qualitatively explains the conductance enhancement for TBT3 with intramolecular O•••S noncovalent interaction induced under high electric field.To further explore the effect of the electric field, we performed similar calculations under an applied electric field of 1 Vnm1 Figure . Notably3TBT3, TBT4, and TBT5 and the electric field applied to the molecular junctions. Interestingly, it was found that molecules with intramolecular O•••S noncovalent interactions enable conductance modulation by varying the applied electric field between two electrodes, especially those with large alkoxy groups. In particular, the single‐molecule conductance of TBT3 can be modulated reversibly by one order of magnitude. It is noted that the regulation of molecular conformation through intramolecular O•••S noncovalent interactions in single molecular electronics via external field was never reported before. Consequently, molecules with intramolecular O•••S noncovalent interactions offer an efficient and in situ approach for establishing single‐molecule electrical switches.In conclusion, we show the enhancement of single‐molecule conductance for molecules with intramolecular O•••S noncovalent interactions. Furthermore, the results reveal that the intramolecular O•••S interactions are influenced by the structural features of alkoxy groups in 4TBT1, CCDC No. 2070565 for compound TBT2, CCDC No. 2103035 for compound TBT3).The reagents and starting materials were commercially available and used directly without further purification unless otherwise specified. Gold wires  were purchased from Beijing Jiaming Platinum Nonferrous Metal Co. Ltd. Crystallographic data (excluding structure factors) reported in this paper were deposited in the Cambridge Crystallographic Data Centre (CCDC No. 2070566 for compound 2(dba)3] (0.1 eq.), K3PO4 (3.0 eq.), SPhos (0.2 eq.) and toluene. The mixture was stirred at 110 °C for 10 h under N2 atmosphere. Then the reaction mixture was concentrated under vacuum and the residue was purified by silica gel column chromatography with hexane and CH2Cl2 as eluent.To a Schlenk tube equipped with a magnetic stir bar was charged with 1, 4‐dibromo‐2,5‐dialkoxybenzene (1.0 eq.), 5‐(methylthio)thiophene‐2‐boronic acid pinacol ester (3.0 eq.),  (0.1 eq.), K3PO4 (3.0 eq.), SPhos (0.2 eq.) and toluene. The mixture was stirred at 110°C for 10 h under N2 atmosphere. Then the reaction mixture was concentrated under vacuum and the residue was purified by silica gel column chromatography with hexane and CH2Cl2 as eluent.To a Schlenk tube equipped with a magnetic stir bar was charged with 1, 4‐dibromo‐2,5‐dialkylbenzene (1.0 eq.), 5‐(methylthio)thiophene‐2‐boronic acid pinacol ester (3.0 eq.), [PdTBT2 (113 mg) was obtained as an yellow solid in 28% yield. For compound TBT2, m.p. 71.2–72.1 °C. 1H NMR : δ (ppm) 7.27 , 7.08 , 6.92 , 2.80 , 2.55 , 1.20 .13C NMR : δ(ppm) 144.99, 139.68, 136.87, 133.31, 131.31, 131.12, 126.71, 26.05, 22.17, 15.65. HR‐MS(MALDI‐TOF): calcd for C20H22S4 (M+) 390.0599; found: 390.0599. Calcd for C20H22S4: C, 61.49; H, 5.68; S, 32.83; found: C, 61.19; H, 5.69; S, 33.11.TBT6 (179 mg) was obtained as a yellow liquid in 29% yield. 1H NMR : δ (ppm) 7.17 , 7.06 , 6.86 , 2.66 , 2.53 , 1.47–1.37 , 1.25–1.13 , 0.81 , 0.74 .13C NMR: δ (ppm) 145.52, 137.56, 136.71, 133.61, 133.15, 131.21, 126.83, 40.11, 37.25, 32.45, 28.67, 25.67, 22.94, 22.32, 14.05, 10.73. HR‐MS (MALDI‐TOF): calcd for C32H46S4 (M+) 558.2477; found: 558.2477. Calcd For C32H46S4: C, 68.76; H, 8.30; S, 22.94; found: C, 68.81; H, 8.26; S, 23.03. In brief, a stepper motor and a piezo stack were used to control the distance between the gold tip and substrate. By repeating forming and breaking Au point contacts, molecular junctions were formed. All measurements were carried out in 0.1 x 10−3m solutions of the molecules in 1,2,4‐trichlorobenzene (TCB) at room temperature.Single‐molecule conductance was measured with the homemade STM‐BJ setup.2a In  The geometries of conjugated molecules were optimized first to obtain optimal molecular configurations. The single Au atoms were attached to the S atoms at SMe anchors of the molecules to optimize the geometries further. The Au pyramids with 60 atoms were attached to S atoms, replacing the Au atoms for calculating the transmission across the junctions using the nonequilibrium Green's function (NEGF) formalism. To model the molecular configurations under an electric field, an electric field of 1 V nm−1 was applied first to optimize the geometries. After attaching single Au atoms to the two sides of the optimized molecules, the molecular geometries were relaxed under the electric field. After geometry optimization, the Au pyramids with 60 atoms were attached to S atoms replacing the Au atoms for calculating the transmission across the junctions.DFT calculations were carried out using the Perdew–Burke–Ernzerhof (PBE) exchange‐correlation functional implemented by the Fritz Haber Institute ab initio molecular simulation (FHI‐aims) packages.13 The For statistical analysis, thousands of conductance traces were collected to construct logarithmically binned conductance histograms. These conductance histograms show peaks at the quantum of conductance G0 (2e2/h), corresponding to gold−gold atomic contact. After the rupture of Au point‐contacts, molecular conductance below 1 G0 showed the formation of molecular junctions. The junction length was calibrated by the direct tunneling distance distribution in pure TCB and the final displacements were obtained by calibrating the snap‐back distance (0.5±0.1 nm). In Figures y = y0 + A/(w*sqrt(π/(4*ln(2)))) * exp(−4*ln(2)*(x − xc)^2/w^2)), which were analyzed with the Goodness of fit (R2) and F‐test, significance was defined as p < 0.05. The statistical analyses were performed using Origin 9.0.The data analysis was carried out based on the previously reported methods.14 ForThe authors declare no conflict of interest.Supporting InformationClick here for additional data file.


Mycobacterium tuberculosis; M. tb) and more recently, the non-tubercular species such as M. abscessus and M. avium. Clinically, tuberculosis stands as the number two infectious disease killer after COVID-19 and M. abscessus is becoming a more prevalent pathogen among the patients with cystic fibrosis. From the basic science perspective, investigation of molecular mechanisms that these mycobacteria use to stay dormant and evade our immune system, as well as identification of novel drug targets/scaffolds, have been and will continue to be crucial in combating the infections. We are delighted to publish this Research Topic on “Antimycobacterial Drug Discovery: Molecular Therapeutics and Target Identification” that highlights some of the more recent contributions in this research field.The genus of mycobacteria includes the causative agent of tuberculosis . The authors reported the isolation of spontaneous M. smegmatis mutants strain mc2155 that were resistant to iridium-L-phenylglycine antibiotic complex. The discovery that these mutant strains were also resistant to clarithromycin guided the isolation and DNA sequencing efforts that ultimately resulted in the identification of the peptidyl transferase domain of the mycobacterial 23S rRNA as one possible target of the iridium-amino acid complex.The Research Topic was launched with an original article from Falkinham III group  describing target identification efforts for a metal-amino acid complex antibiotic against Quang and Jang  review the current drugs and drug candidates in preclinical trials against M. abscessus. The drugs were classified according to their general chemical classes such as macrolides, beta-lactams, tetracyclines, oxazolidinones, and fluoroquinolones. For the more recent drug candidates, potential molecular targets and their shortcomings were also discussed where relevant, e.g., inhibition of MmpL3 by indole-2-carboxamides. This Review will be valuable for the identification of novel drugs/targets for treatment of M. abscessus.Chen et al.  report the synthesis and biological studies of silver nanoparticles as antitubercular agents. The alginate-capped silver nanoparticles were characterised for their size and morphology, prior to testing them in vitro for their bactericidal abilities. The authors found that their silver nanoparticles were effective against both drug-resistant and dormant M. tb as well as in a macrophage infection model. The in vivo mouse TB model results were also indicative of reduced bacterial burden in lungs without significant gross toxicity.Li et al.  describe the ability of nitazoxanide, a broad-spectrum anti-infective agent, to inhibit RANKL-induced osteoclastogenesis. Nitazoxanide was initially known for their antiprotozoal properties and there were instances of their ability to inhibit M. tb proliferation. The authors found that nitazoxanide also inhibits STAT3 pathway that could be exploited in the RANKL-mediated osteoclastogenesis. Using ovariectomised-induced bone loss model, the compound was shown to ameliorate bone loss in vivo accompanied with suppression of transcription factor NFATc1.Ahmed et al.  share their high-content screening of 10,000 small molecules against M. tb within macrophage-like cells. This is of clinical relevance as M. tb stays in an intracellular environment prior to transitioning into an active form. Five chemotypes were identified: benzene-amide ethers, phenylthioureas, phenyl pyrazoles, thiophene carboxamides and thienopyridines. Although the mechanisms of action and drug targets are yet to be identified, these scaffolds serve as useful starting points to yield antitubercular compounds that could be actively killing intracellular M. tb rather than the active form.Bich Hanh et al.  perform synergistic assay in M. abscessus to determine which antibiotic has the optimal effects when combined with the first-line treatment clarithromycin. Amongst the antibiotics tested, omadacycline was found to be the most synergistic using an in vitro checkerboard combination assay. This combination regime was then tested in zebrafish model infected with M. abscessus, where bacterial burden reduction was observed alongside significant lifespan extension of the infected zebrafish. Future testing in rodents model would be beneficial to ascertain the synergism of clarithromycin-omadacycline combination.Lastly, Antimycobacterial Drug Discovery: Molecular Therapeutics and Target Identification” highlights recent advances in this field, ranging from tubercular to non-tubercular mycobacteria. We acknowledge the highly valuable contributions from all authors to make this Research Topic possible. We look forward to the future submissions for the Volume 2 of this Research Topic in the future.Overall, the first volume this Research Topic: “


Ex-vivo molecular profiling has recently emerged as a promising method for intraoperative tissue identification, especially in neurosurgery. The short-term storage of resected samples at room temperature is proposed to have negligible influence on the lipid molecular profiles. However, a detailed investigation of short-term molecular profile stability is required to implement molecular profiling in a clinic. This study evaluates the effect of storage media, temperature, and washing solution to determine conditions that provide stable and reproducible molecular profiles, with the help of ambient ionization mass spectrometry using rat cerebral cortex as model brain tissue samples. Utilizing normal saline for sample storage and washing media shows a positive effect on the reproducibility of the spectra; however, the refrigeration shows a negligible effect on the spectral similarity. Thus, it was demonstrated that up to hour-long storage in normal saline, even at room temperature, ensures the acquisition of representative molecular profiles using ambient ionization mass spectrometry. The metabolism alterations that accompany malignancy allows to implement rapid molecular techniques in clinical practice ,2,3,4. AThe use of ambient mass spectrometry for ex-vivo tissue analysis in the clinical environment requires the implementation of tissue collection and transfer protocols as part of a routine biopsy investigation pipeline . ApplyinThe activity of phospholipases and other enzymes at room temperature may still persist , but in In this study, the cerebral cortexes of rats from the same litter grown in identical conditions since birth were chosen as the model tissue samples. Since the number of samples required to analyze the effects of storage conditions on the brain tissues’ molecular profiles is relatively big, we had to obtain samples from a total of three animals. All of these rats were grown in the vivarium as control animals for various studies. They were fed with a balanced diet, and only donated blood samples throughout their lives. However, even in this case, natural variability may occur in the brain lipid composition of each animal, so it should be specifically noted that the samples from each participating animal were included in consideration to minimize possible individual effects. The samples were stored in different conditions to evaluate the influences of the short-term storage and washing  media composition and temperature on the molecular profiles required for the development of simple sample collection, transportation from an operating room to the laboratory, and treatment before the analysis.A multifactor experimental design  demonstrates substantial sample-to-sample variations. This reflects the effect of cell lysis caused by the osmotic shock during sample storage ,28. As eThe composition of the solution used for sample treatment before the analysis has lower effects compared to that of the storage solution. The SSM regions corresponding to water treatment are more stable, but detailed analyses of the spectra show that the total ion current appears to be lower. The reason for such behavior is also the cells lysis in the deionized water. Moreover, the samples treated with water demonstrate instable spectra for the first 1 to 3 min of measurement, so a delay occurred between cartridge mounting and spectra recording. No such instability was observed for samples treated with normal saline. The excess of normal saline may increase the rate of sodiated lipid adducts forming during ionization compared to water treatment; however, the unification of the sample transfer and treatment procedures will make this effect reproducible and tolerable during further mathematical analysis .The additional set of samples (24 samples) was prepared and analyzed to prove the patterns revealed by the multifactor experiment. The results of the analysis of the enhanced set of data are visualized in Again, the samples treated with water demonstrated higher reproducibility, but this appears to be a result of the lower intensity  of the spectra. The temperature’s effect on the reproducibility is still present ; however, this is comparable with the intraexperimental variability .As expected, no significant alteration in molecular profiles was observed within an hour of brain tissue storage in normal saline at room temperature  the mea. ConsideMethanol (HPLC-grade), acetic acid (HPLC-grade) and sodium chloride (molecular biology grade) were obtained from Merck .3 each) were immediately dissected from each hemisphere  from the same litter were humanely anesthetized and decapitated. The brain was immediately removed, bisected by longitudinal fissure, and the cerebral hemispheres detached from other brain structures. Twelve cerebral cortex samples  extracts the analytes from the samples and delivers them into the ion source, while a glass microfiber filter inside the cartage protects the ESI needle from clogging. Mass spectra were acquired at the FTMS mass resolution set at 30,000 (FWHM at m/z 400 Da) for m/z values ranging from 500 to 1000 in the negative and positive modes, consecutively . After the acquisition the ion source was washed with the solvent until the intensity of the signal decreased by at least 2 orders of magnitude from the level at the end of acquisition to avoid cross-contamination.The freshly thawed samples were treated with deionized water or normal saline directly before the analysis. All samples were analyzed using the ICE technique and an LTQ Orbitrap XL ETD  mass spectrometer as described elsewhere . BrieflySpectra similarity matrices with cosine measure were calculated as described elsewhere . The widThe obtainment of a reliable molecular profile of resected tissue with ambient mass spectrometry requires specific sample handling procedures. First, cell lysis must be avoided, as it drastically decreases the stability, reproducibility, and intensity of mass spectra. The optimal way to reduce cell damage is to maintain the samples in normal saline at all times between dissection and mass spectrometry analysis. Plenty of sterile normal saline is always accessible in an operation room, so no specific efforts are required to keep samples intact. No significant difference in overall molecular profiles was found between samples stored at +4 °C and those stored at +20 °C for up to one hour, so room temperature is suitable for short-term storage during sample transfer between surgery and laboratory.Lipid degradation in brain tissue samples is relatively slow, so we can see that the minor alterations in relative concentrations of various lipids caused by decomposition and oxidation do not affect the molecular profile noticeably, even after one hour of sample storage at room temperature. Moreover, significant changes in molecular profiles were experienced only after three hours of room temperature exposition in normal saline. Thus, the stability of lipid profiles after short-term exposition at ambient conditions facilitates the implementation of molecular profiling techniques in clinical practice. Placing tissue samples in normal saline without refrigeration provides a reliable way to preserve them before their analysis for up to one hour, so only the urgency of the response from the mass spectrometry laboratory limits the sample transfer time.