Combined CYP1A1/GSTM1 at-risk genotypes are overrepresented in squamous cell lung carcinoma patients but underrepresented in elderly tumor-free subjects

Purpose: Polycyclic aromatic hydrocarbons are activated by cytochrome P450 1A1 (CYP1A1) and inactivated by glutathione S-transferase mu (GSTM1). Therefore, it is expected that a combination of proficient CYP1A1 genotype with deficient GSTM1 variant would result in particularly elevated lung cancer (LC) risk, especially for squamous cell carcinoma (SCC). This study was aimed to validate whether the CYP1A1-C ³⁸⁰¹ (CYP1A1*2) allele has an unfavorable significance alone and/or in combination with the GSTM1 deficiency. Methods: We compared the distribution of CYP1A1 and GSTM1 genotypes in LC patients (n=141), healthy donors (HD, n=204), and elderly tumor-free smokers and non-smokers (ED, n=246). Results: CYP1A1*2 allele carriers demonstrated a clear-cut association with SCC: the adjusted odds ratios (OR) were 2.22 (95% CI=1.06–4.63) and 2.27 (95% CI=1.14–4.52) when HD and ED were used as referents, respectively. CYP1A1*2(+)/GSTM1(-) combined genotypes were overrepresented in the SCC patients (14/70, 20.0%) and underrepresented in the ED (19/246, 7.7%) as compared to the intermediate prevalence in the HD (26/204, 12.7%); the adjusted OR for SCC versus ED reached 3.85 (95% CI=1.43–10.33). Conclusions: In agreement with some literature data, our results support the concerted role of CYP1A1 and GSTM1 at-risk genotypes in SCC predisposition.     

Surface comparison of active and inactive protein kinases identifies a conserved activation mechanism

The surface comparison of different serine-threonine and tyrosine kinases reveals a set of 30 residues whose spatial positions are highly conserved. The comparison between active and inactive conformations identified the residues whose positions are the most sensitive to activation. Based on these results, we propose a model of protein kinase activation. This model explains how the presence of a phosphate group in the activation loop determines the position of the catalytically important aspartate in the Asp-Phe-Gly motif. According to the model, the most important feature of the activation is a "spine" formation that is dynamically assembled in all active kinases. The spine is comprised of four hydrophobic residues that we detected in a set of 23 eukaryotic and prokaryotic kinases. It spans the molecule and plays a coordinating role in activated kinases. The spine is disordered in the inactive kinases and can explain how stabilization of the whole molecule is achieved upon phosphorylation.     

LncRNA PVT1 is increased in renal cell carcinoma and affects viability and migration in vitro

BackgroundRenal cell carcinoma is difficult to diagnose and unpredictable in disease course and severity. There are no specific biomarkers for diagnosis and prognosis estimation feasible in clinical practice. Long non‐coding RNAs (lncRNAs) have emerged as potent regulators of gene expression in recent years. Aside from their cellular role, their expression patterns could be used as a biomarker of ongoing pathology.MethodsIn this work, we used next‐generation sequencing for global lncRNA expression profiling in tumor and non‐tumor tissue of RCC patients. The four candidate lncRNAs have been further validated on an independent cohort. PVT1, as the most promising lncRNA, has also been studied using functional in vitro tests.ResultsNext‐generation sequencing showed significant dysregulation of 1163 lncRNAs; among them top 20 dysregulated lncRNAs were AC061975.7, {"type":"entrez-nucleotide","attrs":{"text":"AC124017.1","term_id":"21327652","term_text":"AC124017.1"}}AC124017.1, {"type":"entrez-nucleotide","attrs":{"text":"AP000696.1","term_id":"6705905","term_text":"AP000696.1"}}AP000696.1, AC148477.4, LINC02437, GATA3‐AS, LINC01762, LINC01230, LINC01271, LINC01187, LINC00472, {"type":"entrez-nucleotide","attrs":{"text":"AC007849.1","term_id":"5091570","term_text":"AC007849.1"}}AC007849.1, LINC00982, LINC01543, {"type":"entrez-nucleotide","attrs":{"text":"AL031710.1","term_id":"3821014","term_text":"AL031710.1"}}AL031710.1, and {"type":"entrez-nucleotide","attrs":{"text":"AC019197.1","term_id":"6648325","term_text":"AC019197.1"}}AC019197.1 as down‐regulated lncRNAs; and SLC16A1‐AS1, PVT1, LINC0887, and LUCAT1 as up‐regulated lncRNAs. We observed statistically significant dysregulation of PVT1, LUCAT1, and LINC00982. Moreover, we studied the effect of artificial PVT1 decrease in renal cell line 786–0 and observed an effect on cell viability and migration.ConclusionOur results show not only the diagnostic but also the therapeutic potential of PVT1 in renal cell carcinoma.     

Hardening of Bimetallic Wires from Secondary Materials Used in the Construction of Power Lines

Copper-sheathed steel wires combine the conductivity of copper and the traction resistance of steel, which makes a bimetallic wire an ideal material for the construction of power lines. Currently, there is a small number of studies devoted to the change in the microstructure of steel-copper wire during its strain. Since steel and copper have different mechanical properties, these metals at the interface can be deformed in different ways. Therefore, the present research is devoted to the study of ECAP-drawing process impacts on the properties of bimetallic steel-copper wire. During the conducted studies, the possibility and efficiency of using the combined strain technology for the formation of ultrafine grained structure and increased strength properties of steel-copper wire have been proved.     

Experimental Analysis of the Stress State of a Prestressed Cylindrical Shell with Various Structural Parameters

The paper presents the results of experimental studies of the features of the operation of prestressed shells, taking into account the various structural parameters of the prestress. It is established that when the winding angle changes from perpendicular to the shell axis to 75° and 65°, the circumferential stresses decrease 1.4 times and 1.2 times, respectively, and the axial stresses increase five and three times, which are two and four times lower than the circumferential, from which it can be concluded that the reduction in the winding angle to the longitudinal the axis of the shell has a positive effect on the stress state of the structure. The study also found that with an increase in the diameter of the winding wire from 1 to 2 mm and a change in the winding angle, the same nature of the stress distribution is observed, but the values of the stress state parameter change, so the efficiency increases up to 25% due to an increase in the winding thickness, depending on the pitch, angle and thickness of the winding, which favorably affects the strength and the bearing capacity of the structure as a whole by increasing the value of the stress state parameter. Thus, the results of the analysis will allow us to use in more detail the possibility of controlling the stress–strain state of the prestressed shell by changing the design parameters, and the results obtained can be used in design or construction, as well as when increasing the strength characteristics of the structure, which allows us to create a high-tech design optimal for these operating conditions, which can positively complement the studies conducted earlier in this direction.     

Triterpene glycosides from the Far Eastern sea cucumber Cucumaria conicospermium

Four new triterpene glycosides, cucumariosides A(2)-5 (1), A(3)-2 (2), A(3)-3 (3), and isokoreoside A (4), along with the previously isolated koreoside A (5), have been found in the sea cucumber Cucumariaconicospermium. Glycoside 1 was isolated as a native substance, while glycosides 2-5 were identified through their desulfated derivatives. Their structures have been deduced by extensive spectral analysis (NMR and MS) and chemical evidence. All the glycosides contain the same branched pentasaccharide carbohydrate chain but differ in the number and positions of the sulfate groups. Glycoside 1 has one, glycosides 2 and 3 have two, and glycosides 4 and 5 have three sulfate groups. Glycosides 2-5 are non-holostane derivatives; their aglycons lack the 18(20)-lactone and are characterized by shortened side chains, which is a very rare feature among the sea cucumber glycosides.     

Switchable targeting of solid tumors by BsCAR T cells

The development of chimeric antigen receptor (CAR) T cell therapy has become a critical milestone in modern oncotherapy. Despite the remarkable in vitro effectiveness, the problem of safety and efficacy of CAR T cell therapy against solid tumors is challenged by the lack of tumor-specific antigens required to avoid on-target off-tumor effects. Spatially separating the cytotoxic function of CAR T cells from tumor antigen recognition provided by protein mediators allows for the precise control of CAR T cell cytotoxicity. Here, the high affinity and capability of the bacterial toxin-antitoxin barnase-barstar system were adopted to guide CAR T cells to solid tumors. The complementary modules based on (1) ankyrin repeat (DARPin)-barnase proteins and (2) barstar-based CAR (BsCAR) were designed to provide switchable targeting to tumor cells. The alteration of the DARPin-barnase switches enabled the targeting of different tumor antigens with a single BsCAR. A gradual increase in cytokine release and tunable BsCAR T cell cytotoxicity was achieved by varying DARPin-barnase loads. Switchable BsCAR T cell therapy was able to eradicate the HER2⁺ ductal carcinoma in vivo. Guiding BsCAR T cells by DARPin-barnase switches provides a universal approach for a controlled multitargeted adoptive immunotherapy.

The transfusion of genetically engineered autologous T cells has created a revolution in adoptive cell cancer therapy of disseminated hematologic cancers during the last decade (1–3). chimeric antigen receptor (CAR) T cell therapy has proven its efficacy in therapy-resistant chronic lymphocytic leukemia (4, 5). Despite the unequivocal success of the approach, substantial side effects are also well documented (6–10). The administered CAR T cells, constantly stimulated by the target antigen, can rapidly proliferate to large numbers in the recipient and release proinflammatory cytokines, which, in some cases, lead to severe and sometimes fatal side effects such as cytokine release syndrome (CRS) (11), neurotoxicity, and cerebral edema (12, 13). Another concern in CAR-based therapy is the choice of target. CD19 is a suitable target for CAR T cells because it is expressed only by B cell populations. However, in solid tumors, targeted antigens are also expressed at low levels but broadly across various normal tissues. The on-target off-tissue activity of CAR T cells has resulted in severe toxicities in the translation of CAR T therapy to solid tumors in the clinic (14, 15). Therefore, developing approaches to control CAR T cell activity after infusion is a high priority. A promising solution to this problem is the design of switchable CARs. In this case, artificial molecular switchers “bridge” the tumor cell with CAR T cells. By varying the switcher concentration, it became possible to control cytotoxicity and the sensitivity of the CAR T cells to antigen density. The predictable pharmacokinetics and pharmacodynamics of the switcher allow for the control of CAR T cell effector functionality and persistency. Several reports have demonstrated that bifunctional moieties could be used to control CAR T activity in vivo (16–21).Small molecules such as fluorescein isothiocyanate (FITC) (22), folate (23), rimiducid (24), rapamycin (25), proteolysis-targeting chimera compounds (26), and dasatinib (27) are being investigated to design such safety switches. US Food and Drug Administration (FDA)–approved small-molecule HCV-NS3 inhibitors have been successfully adapted to design novel safety switches for CAR T cells (28, 29).The RNA (30–33) together with ribonucleases (RNases) (34, 35) was shown to possess a broad therapeutic potential. RNases are a large family of bacterial or eukaryotic enzymes that has a natural protein partner that can form a tight and specific complex—RNase inhibitor. The RNase superfamily is an excellent source of proteins for generating regulated CAR T cells. The barnase-barstar toxin-antitoxin system is orchestrated by the RNase toxin barnase and inactivated by the cognate barstar antitoxin, representing an outstanding example of molecular switching based on the extraordinary affinity of the barnase-barstar complex (K_D ∼10⁻¹⁴) (36–39). Here, the barnase–barstar interaction was applied to guide barstar-modified CAR (BsCAR) T cells to tumor cells by barnase-based molecular switches. Barnase was fused with designed ankyrin repeat proteins (DARPins) that are specific to tumor antigens HER2 (human epidermal growth factor receptor 2) and EpCAM. The resulting DARPin-barnase (DARPin-Bn) proteins enabled specific targeting of tumor cells by T cells modified with universal BsCAR. The high affinity of barnase-barstar binding provide a unique regulatory potential of CAR T therapeutics in vivo. This approach can reinforce the CAR technology with therapeutic modes, including the redirection of T cell cytotoxicity toward combinations of multiple tumor antigens.     

Endogenous humoral determinants of vascular endothelial dysfunction as triggers of acute poisoning complications

The vascular endothelium is not only the semipermeable membrane that separates tissue from blood but also an organ that regulates inflammation, vascular tone, blood clotting, angiogenesis and synthesis of connective tissue proteins. It is susceptible to the direct cytotoxic action of numerous xenobiotics and to the acute hypoxia that accompanies acute poisoning. This damage is superimposed on the preformed state of the vascular endothelium, which, in turn, depends on many humoral factors. The probability that an exogenous toxicant will cause life-threatening dysfunction of the vascular endothelium, thereby complicating the course of acute poisoning, increases with an increase in the content of endogenous substances in the blood that disrupt endothelial function. These include ammonia, bacterial endotoxin, indoxyl sulfate, para-cresyl sulfate, trimethylamine N-oxide, asymmetric dimethylarginine, glucose, homocysteine, low-density and very-low-density lipoproteins, free fatty acids and products of intravascular haemolysis. Some other endogenous substances (albumin, haptoglobin, haemopexin, biliverdin, bilirubin, tetrahydrobiopterin) or food-derived compounds (ascorbic acid, rutin, omega-3 polyunsaturated fatty acids, etc.) reduce the risk of lethal vascular endothelial dysfunction. The individual variability of the content of these substances in the blood contributes to the stochasticity of the complications of acute poisoning and is a promising target for the risk reduction measures. Another feasible option may be the repositioning of drugs that affect the function of the vascular endothelium while being currently used for other indications.