Novel sample double dilution calibration method for determination of lithium in biological samples using automatic flow system with in-syringe reaction.

A novel sample double dilution calibration method (SDDCM) and an automatic flow system with in-syringe reaction and spectrophotometric detection were developed for determining lithium in biological samples. The method is based on the reaction of lithium with Thorin in an alkaline medium and the signal was measured at 480 nm. The reaction was performed simultaneously for both standards and samples in three syringes of the automatic flow system. The method was validated and successfully applied to the determination of lithium in synthetic and pharmaceutical samples, with results consistent with the ICP OES method. The novel calibration method, developed for the determination of lithium in biological samples, uses a sample with two dilution degrees. Using the method, the concentration of the analyte is determined by relating the signal for a less diluted sample to the calibration plot for a more diluted sample and vice versa. The implementation of the calibration method was facilitated by preparing solutions directly in the flow system. The use of two sample dilutions makes it possible to determine the analyte in the sample without preliminary preparation. Moreover, obtaining two results based on signals for a sample diluted to different degrees allows them to be verified for accuracy. The proposed approach was successfully verified by the determination of lithium in certified reference materials of blood serum and urine. Using the developed method lithium was determined within the concentration range of 0.06-1.5 mg L-1, with precision (CV, %) less than 6.7, and accuracy (RE, %) better than 6.9. The detection limit was 0.03 mg L-1.

Association between the antioxidant properties of SESN proteins and anti-cancer therapies.

Since the beginning of SESN protein development, they have attracted highly progressive attention due to their regulatory role in multiple signalling pathways. Through their antioxidant activity and autophagy regulation implication, they can function as powerful antioxidants to reduce oxidative stress in cells. SESN proteins received special attention in the field of regulation of reactive oxygen species level in the cell and its interplay with signalling pathways determining energy and nutrient homeostasis. Since perturbations in these pathways are implicated in cancer onset and development, SESNs might constitute potential novel therapeutic targets of broad interest. In this review, we discuss the impact of SESN proteins on anti-cancer therapy based on naturally occurring compounds and conventionally used drugs that influence oxidative stress and autophagy-induced cellular signalling pathways. The significant changes in reactive oxygen species level and nutrient status in cancer cells generate subsequent biological effect through the regulation of SESN-dependent pathways. Thus, SESN may serve as the key molecule for regulating anti-cancer drugs' induced cellular response.

Multiple Cytokine Analysis of Th1/Th2/Th9/Th17/Th22/Treg Cytokine Pathway for Individual Immune Profile Assessment in Patients with Psoriasis.

BACKGROUND Psoriasis is an autoimmune and autoinflammatory disorder that has a significant impact on patient quality of life. The aim of the study was to assess the immune profiles of patients with psoriasis with multiple cytokine analysis. MATERIAL AND METHODS Fifty-two male psoriatic patients and 24 healthy male volunteers were recruited. Granulocyte-macrophage colony-stimulating factor (GM-CSF), interferon gamma (IFN-gamma), interleukin (IL)-1 beta, IL-2, Il-4, IL-5, IL-6, IL-9, IL-10, IL-12p70, IL-13, IL-17A, IL-18, IL-21, IL-22, IL-23, IL-27, and tumor necrosis factor (TNF)-alpha were measured in patients' serum with a Th1/Th2/Th9/Th17/Th22/Treg Cytokine 18-Plex Human ProcartaPlex Panel, based on Luminex xMAP technology. RESULTS The median fluorescence intensities of serum GM-CSF, IL-2, IL-5, IL-10, IL-13, IL-17A, IL-21, and IL-22 were not intensive enough to calculate the cytokine concentration. We observed elevated levels of IL-6 (P=0.001) and IL-9 (P=0.003) in patients, compared with the control group. The levels of IL-1beta (P=0.008) and IL-27 (P=0.006) were decreased. In patients with psoriatic arthritis, we noticed a decreased level of IL-9 compared with that in patients without arthritis (P=0.034). The levels of IL-12 (P<0.05) and IL-18 (P<0.05) correlated positively with the Psoriasis Area and Severity Index. We found negative correlations of IL-9 (P<0.05), IL-12 (P<0.05), and IL-23 (P<0.05) with the age of psoriatic patients; IL-12 (P<0.05) and IL-23 (P<0.05) with psoriasis duration; and IL-6 (P<0.05) and IL-9 (P<0.05) with the Nail Psoriasis Severity Index. CONCLUSIONS Multiple cytokine analysis seems to be an important form of individual immune profile assessment before treatment selection.

Multidimensional assessment of the financial position of polish households and its regional diversity.

The article aims to present a multidimensional assessment of Polish households' financial position and its regional diversity in 2018. In the first stage of the assessment, the financial position was compared between voivodships in a one-dimensional approach, taking into account the level of equivalent disposable income and total expenditure, the savings rate, the shares of food, and housing expenditures. In the second stage, a multidimensional assessment of the financial position was performed, based on the value of the synthetic indicator constructed with the TOPSIS method. Additionally, TOPSIS-based typological classes helped account for determinants of the financial position and subjective evaluation of households in each voivodship. The article drew on unpublished individual data from the Household Budget Survey conducted by the Central Statistical Office of Poland in 2018. Analyses from both stages showed that the households in Mazowieckie and Dolnoslaskie voivodships enjoyed the best financial position, followed by Slaskie and Opolskie, while in Podkarpackie, Podlaskie, Warminsko-Mazurskie, and Swietokrzyskie -- the worst. With regards to Zachodniopomorskie and Wielkopolskie voivodships, the financial position of their households was similar to the national average. The reasons for these disparities are social, economic, and demographic and include professional activity (being unemployed), the level of education of household head, location, and meeting debt obligations.

MSCs as Tumor-Specific Vectors for the Delivery of Anticancer Agents-A Potential Therapeutic Strategy in Cancer Diseases: Perspectives for Quinazoline Derivatives.

Mesenchymal stem cells (MSCs) are considered to be a powerful tool in the treatment of various diseases. Scientists are particularly interested in the possibility of using MSCs in cancer therapy. The research carried out so far has shown that MSCs possess both potential pro-oncogenic and anti-oncogenic properties. It has been confirmed that MSCs can regulate tumor cell growth through a paracrine mechanism, and molecules secreted by MSCs can promote or block a variety of signaling pathways. These findings may be crucial in the development of new MSC-based cell therapeutic strategies. The abilities of MSCs such as tumor tropism, deep migration and immune evasion have evoked considerable interest in their use as tumor-specific vectors for small-molecule anticancer agents. Studies have shown that MSCs can be successfully loaded with chemotherapeutic drugs such as gemcitabine and paclitaxel, and can release them at the site of primary and metastatic neoplasms. The inhibitory effect of MSCs loaded with anti-cancer agents on the proliferation of cancer cells has also been observed. However, not all known chemotherapeutic agents can be used in this approach, mainly due to their cytotoxicity towards MSCs and insufficient loading and release capacity. Quinazoline derivatives appear to be an attractive choice for this therapeutic solution due to their biological and pharmacological properties. There are several quinazolines that have been approved for clinical use as anticancer drugs by the US Food and Drug Administration (FDA). It gives hope that the synthesis of new quinazoline derivatives and the development of methods of their application may contribute to the establishment of highly effective therapies for oncological patients. However, a deeper understanding of interactions between MSCs and tumor cells, and the exploration of the possibilities of using quinazoline derivatives in MSC-based therapy is necessary to achieve this goal. The aim of this review is to discuss the prospects for using MSC-based cell therapy in cancer treatment and the potential use of quinazolines in this procedure.

Novel Integrated Flow-Based Steam Distillation and Titration System for Determination of Volatile Acidity in Wines.

Novel integrated flow-based steam distillation and titration system with spectrophotometric detection was developed for determination of volatile acidity in wines. Using the system, the distillation procedure was carried out in an automatic manner, starting with introducing into a heated steam distillation module a sample and subjecting it to steam distillation. Under selected conditions, all the analyte was transferred to the distillate; therefore, the system did not require calibration. The collected distillate and titrant were introduced into the next monosegments in varying proportions, in accordance with the developed titration procedure, and directed to the detection system to record the titration curve. The titration was stopped after reaching the end point of titration. Procedures for distillation and titration were developed and verified separately by distillation of acetic acid, acetic acid in the presence of tartaric acid as well as acetic acid, tartaric acid, and titratable acidity, with precision (relative standard deviation) and accuracy (relative error) for both procedures lower than 6.9 and 5.6%, respectively. The developed steam distillation and titration systems were used to determine volatile acidity in samples of white and rosé wines separately and as the integrated steam distillation and titration system, both with precision lower than 9.4% and accuracy better than 6.7%.

A Perspective of the Comprehensive and Objective Assessment of Analytical Methods Including the Greenness and Functionality Criteria: Application to the Determination of Zinc in Aqueous Samples.

The recently proposed concept of White Analytical Chemistry (WAC), referring to the Red-Green-Blue color model, combines ecological aspects (green) with functionality (red and blue criteria), presenting the complete method as "white". However, it is not easy to carry out an overall quantitative evaluation of the analytical method in line with the WAC idea in an objective manner. This paper outlines the perspective of the future development of such a possibility by attempting to answer selected questions about the evaluation process. Based on the study consisting in the evaluation of selected model methods by a group of 12 independent analysts, it was shown how well individual criteria are assessed, whether the variability of assessments by different people is comparable for each criterion, how large it is, and whether averaging the scores from different researchers can help to choose the best method more objectively.

Comprehensive Assessment of Flow and Other Analytical Methods Dedicated to the Determination of Zinc in Water.

An original strategy to evaluate analytical procedures is proposed and applied to verify if the flow-based methods, generally favorable in terms of green chemistry, are competitive when their evaluation also relies on other criteria. To this end, eight methods for the determination of zinc in waters, including four flow-based ones, were compared and the Red-Green-Blue (RGB) model was exploited. This model takes into account several features related to the general quality of an analytical method, namely, its analytical efficiency, compliance with the green analytical chemistry, as well as practical and economic usefulness. Amongst the investigated methods, the best was the flow-based spectrofluorimetric one, and a negative example was that one involving a flow module, ICP ionization and MS detection, which was very good in analytical terms, but worse in relation to other aspects, which significantly limits its overall potential. Good assessments were also noted for non-flow electrochemical methods, which attract attention with a high degree of balance of features and, therefore, high versatility. The original attempt to confront several worldwide accepted analytical strategies, although to some extent subjective and with limitations, provides interesting information and indications, establishing a novel direction towards the development and evaluation of analytical methods.

Novel Approach to Automated Flow Titration for the Determination of Fe(III).

A novel approach to automated flow titration with spectrophotometric detection for the determination of Fe(III) is presented. The approach is based on the possibility of strict and simultaneous control of the flow rates of sample and titrant streams over time. It consists of creating different but precisely defined concentration gradients of titrant and analyte in each successively formed monosegments, and is based on using the calculated titrant dilution factor. The procedure was verified by complexometric titration of Fe(III) in the form of a complex with sulfosalicylic acid, using EDTA as a titrant. Fe(III) and Fe(II) (after oxidation to Fe(III) with the use of H2O2) were determined with good precision (CV lower than 1.7%, n = 6) and accuracy ( | RE | lower than 3.3%). The approach was applied to determine Fe(III) and Fe(II) in artesian water samples. Results of determinations were consistent with values obtained using the ICP-OES reference method. Using the procedure, it was possible to perform titration in 6 min for a wide range of analyte concentrations, using 2.4 mL of both sample and titrant.

Novel Approach to Sample Preconcentration by Solvent Evaporation in Flow Analysis.

A preconcentration module operated in flow mode and integrated with a sequential injection system with spectrophotometric detection was developed. Using the system, preconcentration was performed in continuous mode and was based on a membraneless evaporation process under diminished pressure. The parameters of the proposed system were optimized and the system was tested on the example of the spectrophotometric determination of Cr(III). The preconcentration effectiveness was determined using the signal enhancement factor. In the optimized conditions for Cr(III), it was possible to obtain the signal enhancement factors of around 10 (SD: 0.9, n = 4) and determine Cr(III) with precision and intermediate precision of 8.4 and 5.1% (CV), respectively. Depending on the initial sample volume, signal enhancement factor values of about 20 were achieved. Applicability of the developed preconcentration system was verified in combination with the capillary electrophoresis method with spectrophotometric detection on the example of determination of Zn in certified reference materials of drinking water and wastewater. Taking into account the enhancement factor of 10, a detection limit of 0.025 mg L-1 was obtained for Zn determination. Zn was determined with precision less than 6% (CV) and the results were consistent with the certified values.

The function of miR-200 family in oxidative stress response evoked in cancer chemotherapy and radiotherapy.

Since the beginning of the discovery of microRNAs (miRs), these molecules have attracted highly progressive attention due to their powerful regulatory roles in a broad spectrum of biological processes, including proliferation, differentiation, apoptosis and carcinogenesis. With regard to carcinogenesis, the miRs regulatory potency has been associated with cancer onset, progression, metastasis, diagnosis and therapeutic response. In this review we discuss the impact of miR-200 family on drug resistance development during anti-cancer therapy. Developing resistance to chemotherapeutic drugs as well as radiotherapy are major clinical obstacles in the successful therapeutic strategies to cancer treatment. Acquired cancer chemoresistance is a multifactorial phenomenon involving such factors as tumor type, tumor stage, cellular reactive oxygen species (ROS) level or ROS-responsive miRs profile. ROS level could influence the miRs expression level, which changes the cellular profile of the content of miRs. Such significant changes in the cellular miRs profile generate subsequent biological effects through the regulation of their target genes. This review outlines the interactions between ROS and miR-200 family in different kinds of cancers in response to chemotherapy.

Inhibition or Reversal of the Epithelial-Mesenchymal Transition in Gastric Cancer: Pharmacological Approaches.

Epithelial-mesenchymal transition (EMT) constitutes one of the hallmarks of carcinogenesis consisting in the re-differentiation of the epithelial cells into mesenchymal ones changing the cellular phenotype into a malignant one. EMT has been shown to play a role in the malignant transformation and while occurring in the tumor microenvironment, it significantly affects the aggressiveness of gastric cancer, among others. Importantly, after EMT occurs, gastric cancer patients are more susceptible to the induction of resistance to various therapeutic agents, worsening the clinical outcome of patients. Therefore, there is an urgent need to search for the newest pharmacological agents targeting EMT to prevent further progression of gastric carcinogenesis and potential metastases. Therapies targeted at EMT might be combined with other currently available treatment modalities, which seems to be an effective strategy to treat gastric cancer patients. In this review, we have summarized recent advances in gastric cancer treatment in terms of targeting EMT specifically, such as the administration of polyphenols, resveratrol, tangeretin, luteolin, genistein, proton pump inhibitors, terpenes, other plant extracts, or inorganic compounds.

Interactions between microRNA-200 family and Sestrin proteins in endometrial cancer cell lines and their significance to anoikis.

In the present study, we intend to determine whether Sestrin proteins 1, 2, and 3 (SESN1-3) are targets of microRNA-200 family (miR-200) in endometrial cancer (EC) Ishikawa, AN3CA, KLE, and RL 95-2 cell lines and to investigate how these potential interactions influence anoikis resistance of EC cell lines. The luciferase reporter assay, qRT-PCR, and western blotting assays were used to verify whether SESN1-3 are direct targets of miR-200. Moreover, the anoikis assay and transient transfections of miR-200 mimics or inhibitors into EC cell lines were performed to evaluate the modulatory role of miR-200 and SESN proteins on anoikis resistance. We demonstrated that SESN2 protein is a direct target of mir-141 in KLE and RL-95-2 EC cell lines and the functional interaction of miR-141 and SESN2 protein has a downstream effect on anoikis resistance and SESN2 expression level in Ishikawa and AN3CA cell lines. Moreover, we have shown that SESN3 protein is a direct target of miR-200b, miR-200c, and miR-429 in Ishikawa, AN3CA, and KLE cell lines. Our results show that manipulation of miR-200b, miR-200c, and miR-429 expression patterns also has an influence on anoikis resistance in EC cell lines. In conclusion, we identified new interactions between miR-200 and the oxidative stress response SESN proteins that affect anoikis resistance in human EC cells.

Sequential injection system with in-line solid phase extraction and soil mini-column for determination of zinc and copper in soil leachates.

A sequential injection (SI) system equipped with in-line solid phase extraction column and in-line soil mini-column is proposed for determination of zinc and copper in soil leachates. The spectrophotometric determination (560 nm) is based on the reaction of both analytes with 1-(2-Pyridylazo)-2-naphthol (PAN). Zinc is determined after retaining copper on a cationic resin (Chelex100) whereas copper is determined from the difference of the absorbance measured for both analytes, introduced into the system with the use of a different channel, and zinc absorbance. The influence of several potential interferences was studied. Using the developed method, zinc and copper were determined within the concentration ranges of 0.005-0.300 and 0.011-0.200 mg L-1, and with a relative standard deviation lower than 6.0% and 5.1%, respectively. The detection limits are 1.4 and 3.0 µg/L for determination of zinc and copper, respectively. The developed SI method was verified by the determination of both analytes in synthetic and certified reference materials of water samples, and applied to the determination of the analytes in rain water and soil leachates from laboratory scale soil core column and in-line soil mini-column.

A guide for endometrial cancer cell lines functional assays using the measurements of electronic impedance.

Endometrial cancer cell lines are critical tools to investigate the molecular mechanism of tumorigenesis using the end point cell-based assay such as proliferation, cytotoxicity, apoptosis, anoikis or migration and invasion. The proper assay optimization and performance is essential for physiologically relevant results interpretation. In this study we use label-free real-time cell analysis platform (xCELLigence) to optimize growing conditions for proliferation and migration experiments of two types of endometrial cancer cell lines HEC-1-B, HEC-1-A, KLE, and Ishikawa. Profiling of cell lines by cell index measurement in proliferation and migration experiments was performed. Our experimental approach allowed us to monitor particular stage of the cell growth, to see the relation between seeding density and dynamic cell growth as well as to choose the optimal serum concentration as chemoattractant in migration experiment. The highest rate of proliferation was shown for Ishikawa cells. The rapid pace of cellular migration was observed in case of KLE and HEC-1-B cells as compared to weak migratory activity of Ishikawa cells. The cell index that reflects the cell status characterized real-time cytological profile of each analyzed cell line. These cell profiles were crucial for better planning the classical end-point assays used in further research.

New approach to H-point standard addition method for detection and elimination of unspecific interferences in samples with unknown matrix.

A novel method for correction of unknown unspecific (additive) interferences was developed in sequential injection analysis (SIA) using a Lab on Valve module (LOV) with spectrophotometric detection. The method implements a novel idea to calibrate by the standard addition method in several different chemical conditions created in such a way to measure different signal for an analyte and unchanged signal for interferents causing additive effect. This approach, being an enhancement of the H-point standard addition method (HPSAM), enables to quantify unbiased concentration of an analyte in the presence of unknown interferences. The method was tested on the example of the determination of ascorbic acid in soft drinks and juices basing on reduction of Fe(III) to Fe(II) and reaction of the latter with o-phenanthroline to the ferroin complex absorbing at 512.0nm. The analytical utility of the method has been verified and confirmed by the spectrophotometric determination of total acidity in rose and red wines in the presence of bromothymol blue absorbing at 616.0nm. Calibration solutions were prepared automatically in the designed flow system. Ascorbic acid was determined with LOD of 1.4mgL-1 and LOQ of 4.2mgL-1 within linear working range up to 80mgL-1, while in case of the determination of total acidity the values of 4.2, 11.8 and 100, respectively, were obtained. A sample was consumed in volumes of 400 and 1000µL in both cases. The analyses are simple, "green", and non-expensive. The developed method is readily applicable to analysis of real samples of complex unknown matrices and adaptable to different analytical methods.

In vitro and in vivo activity of miR-92a-Locked Nucleic Acid (LNA)-Inhibitor against endometrial cancer.

BACKGROUND: Endometrial cancer is the most common cancer of the female reproductive tract. Based on our previous studies we speculated that miR-92a exhibited pro-oncogenic properties in endometrial cancer, and therefore its inhibition could be used as a therapeutic measure in this disease. Therefore in the present study we aimed to investigate both in vitro and in vivo if inhibition of miR-92a in endometrial cancer would limit cancer cells proliferation. METHODS: miR-92a expression was evaluated in four endometrial cancer cell lines using qPCR. Inhibition of miR-92a activity was obtained in endometrial cancer cell lines by a transient transfection of a custom designed Locked Nucleic Acid (LNA)-Inhibitor, developed to work both in vitro and in vivo. In vitro proliferation studies were performed using xCELLigence RTCA DP system. In vivo experiment was performed in Cby.Cg-Foxn1 < nu>/cmdb mice bearing endometrial cancer xenografts, which were intraperitoneally injected with nine dosages of 25 mg/kg of miR-205-LNA-inhibitor. RESULTS: qPCR revealed increased expression of miR-92a in HEC-1-B, Ishikawa and AN3CA cells. LNA-i-miR-92a inhibited endometrial cancer growth in vitro. It was also demonstrated that systemic administration of LNA-i-miR-92a was feasible and exerted inhibitory effect on endometrial cancer xenograft growth in vivo with only mild toxic effects in treated animals, however the effect was observed until 12th experimental day and the last three dosages did not maintain the attenuating effect with the acceleration of tumor growth observed at the end and after cessation of the intraperitoneal therapy. CONCLUSIONS: Taken together, these results indicate that intraperitoneal delivery of miR-92a-LNA-modified-inhibitor is feasible, devoid of significant toxicity and moderately inhibits endometrial cancer growth in vivo, and therefore warrants further studies investigating other routes of inhibitor delivery possibly in other animal models.

Locked nucleic acid-inhibitor of miR-205 decreases endometrial cancer cells proliferation in vitro and in vivo.

Pathogenesis of endometrial cancer has been connected with alterations of microRNA expression and in particular miR-205 up-regulation was consistently reported in this carcinoma. Presented study aimed to investigate if inhibition of miR-205 expression using LNA-modified-nucleotide would attenuate endometrial cancer cells proliferation in vitro and in vivo.In the course of the study we found that the proliferation of endometrial cancer cells (HEC-1-B, RL-95, KLE, Ishikawa) transfected with LNA-miR-205-inhibitor and evaluated using real time cell monitoring as well as standard cell proliferation assay, was significantly decreased. Next, LNA-miR-205-inhibitor was used to assess the in vivo effects of miR-205 inhibition of endometrial cancer growth. Cby.Cg-Foxn1/cmdb mice bearing endometrial cancer xenografts were intraperitoneally injected with nine dosages of 25mg/kg of miR-205-LNA-inhibitor or scramble control or phosphatase buffered saline and were observed for 32 days. We found that systemic administration of miR-205-LNA-inhibitor was technically possible, and exerted inhibitory effect on endometrial cancer xenograft growth in vivo with only mild toxic effects in treated animals.In conclusion our results suggest that systemic delivery of miR-205-LNA-inhibitor is feasible, devoid of significant toxicity, and could be a promising treatment strategy for endometrial cancer. Therefore it warrants further studies in other animal models.

Solenoid micropump-based flow system for generalized calibration strategy.

Generalized calibration strategy (GCS) is one of the innovative approaches aimed at verification and improvement of accuracy of analytical determinations. It combines in a single procedure the interpolative and the extrapolative calibration approaches along with stepwise dilution of a sample with the use of a dedicated flow system. In the paper a simple solenoid micropump-based flow system designed for implementation of GCS has been described. The manifold consists of several modules fully operated by a computer and connected with each other in a properly designed network. Its performance and usefulness were tested on determination of calcium by FAAS in synthetic and natural samples containing strong interferents. It was shown how GCS can serve for detection, examination and elimination of the interference effects. It was demonstrated that the designed manifold enabled to perform GCS procedure with very good precision, in short time and with very low standard, sample and reagent consumption.