A Cross-Talk about Radioresistance in Lung Cancer-How to Improve Radiosensitivity According to Chinese Medicine and Medicaments That Commonly Occur in Pharmacies.

Lung cancer is one of the most common cancers in the population and is characterized by non-specific symptoms that delay the diagnosis and reduce the effectiveness of oncological treatment. Due to the difficult placement of the tumor, one of the main methods of lung cancer treatment is radiotherapy, which damages the DNA of cancer cells, inducing their apoptosis. However, resistance to ionizing radiation may develop during radiotherapy cycles, leading to an increase in the number of DNA points of control that protect cells from apoptosis. Cancer stem cells are essential for radioresistance, and due to their ability to undergo epithelial-mesenchymal transition, they modify the phenotype, bypassing the genotoxic effect of radiotherapy. It is therefore necessary to search for new methods that could improve the cytotoxic effect of cells through new mechanisms of action. Chinese medicine, with several thousand years of tradition, offers a wide range of possibilities in the search for compounds that could be used in conventional medicine. This review introduces the potential candidates that may present a radiosensitizing effect on lung cancer cells, breaking their radioresistance. Additionally, it includes candidates taken from conventional medicine-drugs commonly available in pharmacies, which may also be significant candidates.

Combination of Irinotecan and Melatonin with the Natural Compounds Wogonin and Celastrol for Colon Cancer Treatment.

Colorectal cancers are one of the leading cancers worldwide and are known for their high potential for metastasis and resistance to therapy. The aim of this study was to investigate the effect of various combination therapies of irinotecan with melatonin, wogonin, and celastrol on drug-sensitive colon cancer cells (LOVO cell line) and doxorubicin-resistant colon cancer stem-like cells (LOVO/DX cell subline). Melatonin is a hormone synthesized in the pineal gland and is responsible for circadian rhythm. Wogonin and celastrol are natural compounds previously used in traditional Chinese medicine. Selected substances have immunomodulatory properties and anti-cancer potential. First, MTT and flow cytometric annexin-V apoptosis assays were performed to determine the cytotoxic effect and the induction of apoptosis. Then, the potential to inhibit cell migration was evaluated using a scratch test, and spheroid growth was measured. The results showed important cytotoxic effects of the drug combinations on both LOVO and LOVO/DX cells. All tested substances caused an increase in the percentage of apoptotic cells in the LOVO cell line and necrotic cells in the LOVO/DX cell subline. The strongest effect on the induction of cancer cell death was observed for the combination of irinotecan with celastrol (1.25 µM) or wogonin (50 µM) and for the combination of melatonin (2000 µM) with celastrol (1.25 µM) or wogonin (50 µM). Statistically significant improvements in the effect of combined therapy were found for the irinotecan (20 µM) and celastrol (1.25 µM) combination and irinotecan (20 µM) with wogonin (25 µM) in LOVO/DX cells. Minor additive effects of combined therapy were observed in LOVO cells. Inhibition of cell migration was seen in LOVO cells for all tested compounds, while only irinotecan (20 µM) and celastrol (1.25 µM) were able to inhibit LOVO/DX cell migration. Compared with single-drug therapy, a statistically significant inhibitory effect on cell migration was found for combinations of melatonin (2000 µM) with wogonin (25 µM) in LOVO/DX cells and irinotecan (5 µM) or melatonin (2000 µM) with wogonin (25 µM) in LOVO cells. Our research shows that adding melatonin, wogonin, or celastrol to standard irinotecan therapy may potentiate the anti-cancer effects of irinotecan alone in colon cancer treatment. Celastrol seems to have the greatest supporting therapy effect, especially for the treatment of aggressive types of colon cancer, by targeting cancer stem-like cells.

Metabolic profiles and fingerprints for the investigation of the influence of nitisinone on the metabolism of the yeast Saccharomyces cerevisiae.

Nitisinone (2-(2-nitro-4-trifluoromethylbenzoyl)-1,3-cyclohexanedione, NTBC) is considered a potentially effective drug for the treatment of various metabolic diseases associated with disorders of L-tyrosine metabolism however, side-effects impede its widespread use. This work aimed to broaden the knowledge of the influence of NTBC and its metabolites 2-amino-4-(trifluoromethyl)benzoic acid (ATFA), 2-nitro-4-(trifluoromethyl)benzoic acid (NTFA), and cyclohexane-1,3-dione (CHD) on the catabolism of L-tyrosine and other endogenous compounds in Saccharomyces cerevisiae. Based on a targeted analysis performed by LC-ESI-MS/MS, based on multiple reaction monitoring, it was found that the dissipation kinetics of the parent compound and its metabolites are compatible with a first-order reaction mechanism. Moreover, it has been proven that formed NTBC metabolites, such as CHD, cause a decrease in L-tyrosine, L-tryptophan, and L-phenylalanine concentrations by about 34%, 59% and 51%, respectively, compared to the untreated model organism. The overall changes in the metabolism of yeast exposed to NTBC or its derivatives were evaluated by non-targeted analysis via LC-ESI-MS/MS in the ion trap scanning mode. Based on principal components analysis, a statistically significant similarity between metabolic responses of yeast treated with ATFA or NTFA was observed. These findings facilitate further studies investigating the influence of NTBC on the human body and the mechanism of its action.

Psychometric Evaluation of the Polish Version of the Caregiver Priorities and Child Health Index of Life with Disabilities (CPCHILD).

Purpose: The assessment of the quality of life is an important element of the clinical examination of the patient. The aim of this study was translation and cross-cultural adaptation of the "Caregiver Priorities and Child Health Index of Life with Disabilities" (CPCHILD) questionnaire into Polish language, and testing of reliability and validity of the CPCHILD-PL for children with cerebral palsy (CP). Material and Methods: A Polish version of CPCHILD was created according to internationally accepted guidelines. Parents (n=77) of 51 boys/26 girls between 3 and 17 years with CP with Gross Motor Function Classification System I-V (GMFCS I-V) participated. To assess the reliability each domain and the total measure was tested for internal consistency and test-retest reliability. Convergent validity was evaluated by correlating the CPCHILD-PL with the CHAQ (Childhood Health Assessment Questionnaire) questionnaire. Results: Test-retest reliability assessed by Spearman correlation coefficient for the final result of CPCHILD-PL and for most of domains were above 0.90. The values of Cronbach's-α coefficient (measuring internal consistency) were high for all domains (except for domain 5: Health) and the entire CPCHILD-PL, with the range 0.88-0.96. The comparison between CPCHILD-PL and the Disability Index (DI) of the CHAQ showed a negative correlation. The higher the DI, the lower the CPCHILD result. The Spearman's rank coefficient was -0.75. Conclusion: The Polish version for the CPCHILD for children with CP seems to be reliable and valid tool for assessing health-related quality of life from the caregiver perspective. It can be used in research and clinical practice for evaluation and comparison of health-related quality of life in children with CP in different countries.

Unraveling the role of Breg cells in digestive tract cancer and infectious immunity.

Over the past two decades, regulatory B cells (Breg cells or Bregs) have emerged as an immunosuppressive subset of B lymphocytes playing a key role in inflammation, infection, allergy, transplantation, and cancer. However, the involvement of Bregs in various pathological conditions of the gastrointestinal tract is not fully understood and is the subject of much recent research. In this review, we aimed to summarize the current state of knowledge about the origin, phenotype, and suppressive mechanisms of Bregs. The relationship between the host gut microbiota and the function of Bregs in the context of the disturbance of mucosal immune homeostasis is also discussed. Moreover, we focused our attention on the role of Bregs in certain diseases and pathological conditions related to the digestive tract, especially Helicobacter pylori infection, parasitic diseases (leishmaniasis and schistosomiasis), and gastrointestinal neoplasms. Increasing evidence points to a relationship between the presence and number of Bregs and the severity and progression of these pathologies. As the number of cases is increasing year by year, also among young people, it is extremely important to understand the role of these cells in the digestive tract.

Passive coupling of membrane tension and cell volume during active response of cells to osmosis.

During osmotic changes of their environment, cells actively regulate their volume and plasma membrane tension that can passively change through osmosis. How tension and volume are coupled during osmotic adaptation remains unknown, as their quantitative characterization is lacking. Here, we performed dynamic membrane tension and cell volume measurements during osmotic shocks. During the first few seconds following the shock, cell volume varied to equilibrate osmotic pressures inside and outside the cell, and membrane tension dynamically followed these changes. A theoretical model based on the passive, reversible unfolding of the membrane as it detaches from the actin cortex during volume increase quantitatively describes our data. After the initial response, tension and volume recovered from hypoosmotic shocks but not from hyperosmotic shocks. Using a fluorescent membrane tension probe (fluorescent lipid tension reporter [Flipper-TR]), we investigated the coupling between tension and volume during these asymmetric recoveries. Caveolae depletion and pharmacological inhibition of ion transporters and channels, mTORCs, and the cytoskeleton all affected tension and volume responses. Treatments targeting mTORC2 and specific downstream effectors caused identical changes to both tension and volume responses, their coupling remaining the same. This supports that the coupling of tension and volume responses to osmotic shocks is primarily regulated by mTORC2.

Synthesis and Characterization of Zeolites Produced from Low-Quality Coal Fly Ash and Wet Flue Gas Desulphurization Wastewater.

This study investigated a low-energy-consuming procedure for the synthesis of zeolite materials from coal fly ash (CFA). Materials containing zeolite phases, namely Na-X, Na-P1, and zeolite A, were produced from F-class fly ash, using NaOH dissolved in distilled water or in wastewater obtained from the wet flue gas desulphurization process, under atmospheric pressure at a temperature below 70 °C. The influence of temperature, exposure time, and alkaline solution concentration on the synthesized materials was tested. In addition, chemical, mineralogical, and textural properties of the obtained materials were determined by X-ray diffraction (XRD), X-ray fluorescence (XRF), scanning electron microscopy (SEM), and cation exchange capacity (CEC). Cd(II), Ni(II), NH4+ cation, and Se(VI) anion sorption experiments were conducted to compare the sorption properties of the produced synthetic zeolites with those of the commercially available ones. Zeolitization resulted in an increase of CEC (up to 30 meq/100 g) compared to raw CFA and enhanced the ability of the material to adsorb the chosen ions. The obtained synthetic zeolites showed comparable or greater sorption properties than natural clinoptilolite and synthetic Na-P1. They were also capable of simultaneously removing cationic and anionic compounds. The structural, morphological, and textural properties of the final product indicated that it could potentially be used as an adsorbent for various types of environmental pollutants.

Synthetic lethality between VPS4A and VPS4B triggers an inflammatory response in colorectal cancer.

Somatic copy number alterations play a critical role in oncogenesis. Loss of chromosomal regions containing tumor suppressors can lead to collateral deletion of passenger genes. This can be exploited therapeutically if synthetic lethal partners of such passenger genes are known and represent druggable targets. Here, we report that VPS4B gene, encoding an ATPase involved in ESCRT-dependent membrane remodeling, is such a passenger gene frequently deleted in many cancer types, notably in colorectal cancer (CRC). We observed downregulation of VPS4B mRNA and protein levels from CRC patient samples. We identified VPS4A paralog as a synthetic lethal interactor for VPS4B in vitro and in mouse xenografts. Depleting both proteins profoundly altered the cellular transcriptome and induced cell death accompanied by the release of immunomodulatory molecules that mediate inflammatory and anti-tumor responses. Our results identify a pair of novel druggable targets for personalized oncology and provide a rationale to develop VPS4 inhibitors for precision therapy of VPS4B-deficient cancers.

Ecological and Health Effects of Lubricant Oils Emitted into the Environment.

Lubricating oils used in machines with an open cutting system, such as a saw or harvester, are applied in forest areas, gardening, in the household, and in urban greenery. During the operation of the device with an open cutting system, the lubricating oil is emitted into the environment. Therefore, the use of an oil base and refining additives of petroleum origin in the content of lubricants is associated with a negative impact on health and the environment. The current legal regulations concerning lubricants applicable in the European Union (EU) assess the degree of biodegradability. Legislation permits the use of biodegradable oils at 60% for a period of 28 days. This means that, in practice, lubricating oil considered to be biodegradable can contain up to 50% of the so-called petroleum oil base. The paper aims to draw public attention to the need to reduce the toxicity and harmful effects, due to their composition, of lubricating oils emitted into the environment on health. The authors discuss the impact of petroleum oil lubricants on soils, groundwater, vegetation, and animals, and the impact of petroleum-origin oil mist on health. An overview of test methods for the biodegradability of lubricating oils is presented, including the Organization for Economic Cooperation and Development (OECD) 301 A-F, 310, and 302 A-D tests, as well as their standard equivalents. The current legal regulations regarding the use and control of lubricating oils emitted into the environment are discussed. Legal provisions are divided according to their area of application. Key issues regarding the biodegradability and toxicity of petroleum fractions in lubricating oils are also addressed. It is concluded that lubricating oils, emitted or potentially emitted into the environment, should contain only biodegradable ingredients in order to eliminate the negative impact on both the environment and health. Total biodegradability should be confirmed by widely applied tests. Therefore, a need to develop and implement low-cost and simple control procedures for each type of lubricating oil, ensuring the possibility of an indisputable conclusion about the presence and total absence of petroleum-derived components in oil, as well as the content of natural ingredients, occurs.

The association between night shift work and nutrition patterns among nurses: a literature review.

The shift work system may affect the temporal distribution of eating and diet quality. The paper aimed at reviewing a body of research examining the associations between night shift work and dietary habits among nurses. Data from the PubMed and Google Schoolar databases, as well as references lists in selected papers were searched. The authors used the following keywords: nurses, shift work, diet, nutrition. Papers published in English or Polish were selected for the review, and as many as 19 papers published in 2000-2017 were eventually identified. The studies varied greatly with respect to the study size, subjects' age and the duration of night shift work. The major problem was the heterogeneity of the tools used for dietary assessment. Self-administered questionnaires were used and analyses were rarely adjusted for confounders. Alcohol consumption was the most frequently analyzed aspect (N = 8 studies), followed by the total energy (N = 7), protein, fat (N = 6), and carbohydrate intake, coffee and fruit consumption (N = 5). The results showed quite a consistent association of night work with higher coffee (caffeine) consumption, as well as lower alcohol, and fruit and vegetables consumption. Few studies also reported more frequent snacks consumption, later time of the last meal, eating at night, meals irregularity, and a poorer diet quality among night shift nurses when compared to the reference. The review showed some poor nutritional habits among nurses working night shifts. However, the topic warrants further attention, owing to the relatively small number of the studies performed so far, and their numerous methodological limitations. Med Pr. 2019;70(3):363-76.

Novel coronavirus-like particles targeting cells lining the respiratory tract.

Virus like particles (VLPs) produced by the expression of viral structural proteins can serve as versatile nanovectors or potential vaccine candidates. In this study we describe for the first time the generation of HCoV-NL63 VLPs using baculovirus system. Major structural proteins of HCoV-NL63 have been expressed in tagged or native form, and their assembly to form VLPs was evaluated. Additionally, a novel procedure for chromatography purification of HCoV-NL63 VLPs was developed. Interestingly, we show that these nanoparticles may deliver cargo and selectively transduce cells expressing the ACE2 protein such as ciliated cells of the respiratory tract. Production of a specific delivery vector is a major challenge for research concerning targeting molecules. The obtained results show that HCoV-NL63 VLPs may be efficiently produced, purified, modified and serve as a delivery platform. This study constitutes an important basis for further development of a promising viral vector displaying narrow tissue tropism.

Entry of Human Coronavirus NL63 into the Cell.

The first steps of human coronavirus NL63 (HCoV-NL63) infection were previously described. The virus binds to target cells by use of heparan sulfate proteoglycans and interacts with the ACE2 protein. Subsequent events, including virus internalization and trafficking, remain to be elucidated. In this study, we mapped the process of HCoV-NL63 entry into the LLC-Mk2 cell line and ex vivo three-dimensional (3D) tracheobronchial tissue. Using a variety of techniques, we have shown that HCoV-NL63 virions require endocytosis for successful entry into the LLC-MK2 cells, and interaction between the virus and the ACE2 molecule triggers recruitment of clathrin. Subsequent vesicle scission by dynamin results in virus internalization, and the newly formed vesicle passes the actin cortex, which requires active cytoskeleton rearrangement. Finally, acidification of the endosomal microenvironment is required for successful fusion and release of the viral genome into the cytoplasm. For 3D tracheobronchial tissue cultures, we also observed that the virus enters the cell by clathrin-mediated endocytosis, but we obtained results suggesting that this pathway may be bypassed.IMPORTANCE Available data on coronavirus entry frequently originate from studies employing immortalized cell lines or undifferentiated cells. Here, using the most advanced 3D tissue culture system mimicking the epithelium of conductive airways, we systematically mapped HCoV-NL63 entry into susceptible cells. The data obtained allow for a better understanding of the infection process and may support development of novel treatment strategies.

The Thin-Layer Microchromatography (µTLC) and TLC-FID Technique as a New Methodology in the Study of Lubricating Oils.

This paper concerns the possibility of using TLC coupled with a flame ionization detector (FID) and micro-TLC (µTLC) as precursors for microfluidized devices of analytical techniques to identify and determine the presence and content of the petroleum/vegetable oil base in the lubricating oils applied in cutting devices (chainsaws). This research is related to the problem of ensuring, in compliance with the requirements of environmental protection, a sufficient level of biodegradability of lubricating oils emitted to the environment during operation of equipment lubricated with these oils. Such oils include those mainly used in cutting devices and emitted in the form of a mist into the environment during the operation of those devices. When oil components are eco-toxic, contamination of the environment occurs. New methodologies for the identification and determination of the petroleum oil base, which is very difficult to biodegrade, as well as the easily biodegradable ingredients of vegetable origin in the lubricating oils, are presented. The described procedures indicate in an indisputable way whether the oil contains the oil base originating from crude oil and whether it contains adequate enriching additives. The procedures also allow the assessment of the content of particular groups of constituents (µTLC) or the determination of the group composition (TLC-FID).

The nucleocapsid protein of human coronavirus NL63.

Human coronavirus (HCoV) NL63 was first described in 2004 and is associated with respiratory tract disease of varying severity. At the genetic and structural level, HCoV-NL63 is similar to other members of the Coronavirinae subfamily, especially human coronavirus 229E (HCoV-229E). Detailed analysis, however, reveals several unique features of the pathogen. The coronaviral nucleocapsid protein is abundantly present in infected cells. It is a multi-domain, multi-functional protein important for viral replication and a number of cellular processes. The aim of the present study was to characterize the HCoV-NL63 nucleocapsid protein. Biochemical analyses revealed that the protein shares characteristics with homologous proteins encoded in other coronaviral genomes, with the N-terminal domain responsible for nucleic acid binding and the C-terminal domain involved in protein oligomerization. Surprisingly, analysis of the subcellular localization of the N protein of HCoV-NL63 revealed that, differently than homologous proteins from other coronaviral species except for SARS-CoV, it is not present in the nucleus of infected or transfected cells. Furthermore, no significant alteration in cell cycle progression in cells expressing the protein was observed. This is in stark contrast with results obtained for other coronaviruses, except for the SARS-CoV.

Human coronavirus NL63 utilizes heparan sulfate proteoglycans for attachment to target cells.

UNLABELLED: Human coronavirus NL63 (HCoV-NL63) is an alphacoronavirus that was first identified in 2004 in the nasopharyngeal aspirate from a 7-month-old patient with a respiratory tract infection. Previous studies showed that HCoV-NL63 and the genetically distant severe acute respiratory syndrome (SARS)-CoV employ the same receptor for host cell entry, angiotensin-converting enzyme 2 (ACE2), but it is largely unclear whether ACE2 interactions are sufficient to allow HCoV-NL63 binding to cells. The present study showed that directed expression of angiotensin-converting enzyme 2 (ACE2) on cells previously resistant to HCoV-NL63 renders them susceptible, showing that ACE2 protein acts as a functional receptor and that its expression is required for infection. However, comparative analysis showed that directed expression or selective scission of the ACE2 protein had no measurable effect on virus adhesion. In contrast, binding of HCoV-NL63 to heparan sulfates was required for viral attachment and infection of target cells, showing that these molecules serve as attachment receptors for HCoV-NL63. IMPORTANCE: ACE2 protein was proposed as a receptor for HCoV-NL63 already in 2005, but an in-depth analysis of early events during virus infection had not been performed thus far. Here, we show that the ACE2 protein is required for viral entry but that it is not the primary binding site on the cell surface. Conducted research showed that heparan sulfate proteoglycans function as adhesion molecules, increasing the virus density on cell surface and possibly facilitating the interaction between HCoV-NL63 and its receptor. Obtained results show that the initial events during HCoV-NL63 infection are more complex than anticipated and that a newly described interaction may be essential for understanding the infection process and, possibly, also assist in drug design.

Polish population data on 15 autosomal STRs of AmpFlSTR NGM PCR kit.

The objective of the research was to provide a comprehensive database of autosomal microsatellite loci included in AmpFlSTR NGM PCR kit for a population of Poland considering possible genetic differentiation of a forensic interest. Fifteen STR markers were analyzed in 2041 unrelated individuals residing in eight geographically different regions. All the loci were found to be in Hardy-Weinberg equilibrium. The combined probability of match is 3.52 × 10(-19) and the combined Power of Exclusion is 0.9999998. The F(ST) estimate over all 15 STRs is 0.0051 for the Polish population. We established that a combined NGM database may be employed for a Polish population.

Genetic variation of 15 autosomal STR loci in a population sample from Poland.

Fifteen autosomal STR loci included in AmpFlSTR NGM kit were analyzed in 154 unrelated individuals from Poland. This multiplex kit enables simultaneous amplification of 10 standard STR loci included in AmpFlSTR SGM Plus kit (D3S1358, vWA, D16S539, D2S1338, D8S1179, D19S433, TH01, FGA, D21S11 and D18S51) and five new mini- and midi-STR loci (D10S1248, D22S1045, D2S441, D1S1656 and D12S391). Population study was conducted to evaluate usefulness of the loci (especially the five new microsatellite systems) in forensic genetic identification examinations. All 15 markers were found to be in Hardy-Weinberg equilibrium. The combined probability of match for the 15 studied STR loci was 3.998 x 10(-19). The same parameter calculated for five new microsatellite loci equaled 8.83 x 10(-7). Discrimination power was particularly high in case of D1S1656 (0.975) and D12S391 (0.972) STR loci.

A population data for 17 Y-chromosome STR loci in South Poland population sample--some DYS458.2 variants uncovered and sequenced.

Seventeen Y-chromosomal short tandem repeat loci were analyzed in a sample of 435 unrelated healthy male individuals from Southern Poland (including highlanders from Tatra Mountains). One duplication in the locus DYS389II (29,30) and five microvariant alleles in the locus DYS458 were found. The most frequent haplotype, found in three individuals, was as follows (in the order of Yfiler loci): {16, 13, 25, 30, 15, 15, 11/14, 13, 11, 11, 10, 23, 11, 13, 14, 11, 21}. Gene diversity for South Poland population amounts close to 1.000. Performed differentiation test between all pairs of samples, based on haplotype frequencies, represented as non-differentiation exact P values indicates that there is no statistically significant differences in haplotype frequencies between South Poland and Austrian as well as South Poland and Wallachian populations.

Determination of phenotype associated SNPs in the MC1R gene.

Prediction of physical appearance based on genetic analysis is a very attractive prospect for forensic investigations. Recent studies have proved that there is a significant association between some genetic variants of the melanocortin 1 receptor (MC1R) gene and red hair color. The present study focuses on the potential forensic applicability of variation within this pigment-related gene. Sequencing of the complete MC1R gene was performed on a group of red-haired individuals and controls with different pigmentation. A major role in determination of red hair color is played by two MC1R variants--C451T and C478T. The optimized minisequencing assay for genotyping of the above positions and three other important red hair-related MC1R polymorphisms, C252A, G425A, and G880C was successfully applied to analyze typical forensic specimens. Determination of a homozygous or heterozygous combination can be a good predictor of both red hair color and fair skin of a subject.