In situ veritas: combining omics and multiplex imaging can facilitate the detection and characterization of cell-cell interactions in tissues

In 1898, microscopic examinations were used to study changes in lymphoid tissues and cell distribution in pathological settings (1). More than 100 years after these first discoveries, the complexity of lymphoid organ composition and the existence of distinct immune-cell subpopulations with a diverse set of functions has been described. For the adaptive immune system to function efficiently, complex series of spatial and temporal interactions between specialized immune cells must take place. This has been a field of vigorous interest, exemplified by the fact that nearly 100.000 papers have been published dealing with the keywords “interaction” and “immune cells” (PubMed search March 2023). Understanding cell-cell communication—resulting in immunological pathways—is being supported by a range of sophisticated analysis pipelines, ranging from in vitro and ex vivo single-cell sequencing analysis to genic analysis of immunological alterations. In this context, it is commonly accepted that cells can communicate through juxtacrine and paracrine processes (2). Signal transmission and reception between neighboring cells is fundamentally involved in the regulation of immunological processes, ranging from tissue homeostasis to defense mechanisms against tumour cells or pathogens. However, the final decryption of immunological programs responsible for coordinated and dynamic immunological adaption is multi-factorial and remains challenging

Characteristics of “Tip-DCs and MDSCs” and Their Potential Role in Leishmaniasis

Since the first description of dendritic cells (DCs) by Steinman and Cohn (1973), the myeloid lineage of leukocytes was investigated intensively. Nowadays it is obvious that myeloid cells, especially DCs, are crucial for the adaptive and innate immune response against intracellular pathogens such as Leishmania major parasites. Based on the overlapping expression of molecules that were commonly used to classify myeloid cells, it becomes difficult to denominate those cell types precisely. Of note, most of these markers used for myeloid cell identification are expressed on a broad range of myeloid cells, and should therefore be handled with care if used for subtyping of myeloid cells. In this mini-review we aim to discuss the relative impact of DCs that release TNF and nitric oxide (Tip-DCs) and myeloid cells with suppressive capacities (myeloid-derived suppressor cells, MDSCs) in infectious diseases such as experimental leishmaniasis. In our point of view it cannot be excluded that the novel subsets that were denominated as “Tip-DCs” and “MDSCs” might not be classical “subsets” but rather represent myeloid cells in a transient maturation stage expressing different genes, in response to the surrounding environment

An Early Reduction in Treg Cells Correlates with Enhanced Local Inflammation in Cutaneous Leishmaniasis in CCR6-Deficient Mice

Resistance to Leishmania major infection is dependent on the development of a cell-mediated Th1 immune response in resistant C57BL/6 mice whereas Th2-prone BALB/c mice develop non-healing lesions after infection. The chemokine receptor CCR6 is shared by anti-inflammatory regulatory T cells and pro-inflammatory Th17 cells. In a recent study we showed that C57BL/6 mice deficient in CCR6 exhibited enhanced footpad swelling and impaired T helper cell migration indicated by reduced recruitment of total T helper cells into the skin after infection and a reduced delayed type hypersensitivity reaction. Based on these findings we tested whether the lack of CCR6 alters Treg or Th17 cell responses during the course of Leishmania major infection. When we analyzed T cell subsets in the lymph nodes of CCR6-deficient mice, Th17 cell numbers were not different. However, reduced numbers of Treg cells paralleled with a stronger IFNγ response. Furthermore, the early increase in IFNγ-producing cells correlated with increased local tissue inflammation at later time points. Our data indicate an important role of CCR6 for Treg cells and a redundant role for Th17 cells in a Th1 cell-driven anti-parasitic immune response against Leishmania major parasites in resistant C57BL/6 mice

Effect of fullerene C60 thermal and tribomechanical loading on Raman signals

Fullerene C60 powder was loaded by 1 N normal force and exposed to sliding under different frequencies for 15 min. It is shown that the velocity of the sliding movement determines the stability of the fullerene C60 powder. At slow velocity of movement with a frequency of 1 Hz under 1 N normal force, the fullerene C60 structure remains undamaged after 15 min sliding. On the contrary, high sliding velocities of 10 Hz and 50 Hz affected fragmentation of the fullerene C60, which resulted in a reduction of the coefficient of friction (COF). During sliding with 1 Hz, the friction reached the highest level with an average COF of 0.59 ± 0.03. The faster relative motion under 1 N normal force gave a lower average COF with 0.39 ± 0.03. The initial fullerene C60 powder formed a thick compressed layer in the tribomechanical loaded zone. As proven by Raman spectroscopy, operating the tribomechanical sliding test at 50 Hz stimulated the re-attraction of fresh C60 fullerene island onto the fragmented layer from outside of the loaded powder regions. The COF was increasing again up to 0.44 ± 0.04 for 1 N normal force and 50 Hz frequency. The fragmentation and decomposition of fullerene C60 with increasing sliding velocity is attributed to thermal heating up during fast relative movement. Raman spectra of the tribomechanical loaded fullerene C60 are compared with Raman spectra from slowly heated up C60 in air and with Raman spectra of laser irradiated fullerene C60

{Bis[2-(diphenyl­phosphan­yl)phen­yl] ether-κ2 P,P′}(1,1′-dibenz­yl-1H,1′H-4,4′bi-1,2,3-triazole-κ2 N3,N 3′)copper(I) hexa­fluorido­phosphate dichloro­methane hemisolvate

In the crystal structure of the title compound, [Cu(C18H16N6)(C36H28OP2)]PF6·0.5CH2Cl2 or [Cu(DPEPhos)(Bn-bta)]PF6·0.5CH2Cl2 {DPEPhos = bis­[(diphenyl­phosphan­yl)phen­yl] ether and Bn-bta = 1,1′-dibenzyl-1H,1′H-4,4′-bi-1,2,3triazol­e}, the Cu atom is coordinated by two N and two P atoms of the ligands in a strongly distorted tetra­hedral environment. There are two crystallographically independent complex cations present, which differ significantly in their geometrical parameters. The solvent molecule is disordered but satisfactory atomic positions could not be determined

Water-soluble pristine C60 fullerene attenuates acetaminophen-induced liver injury

Introduction: Oxidative stress has been suggested as the main trigger and pathological mechanism of toxic liver injury. Effects of powerful free radical scavenger С60 fullerene on rat liver injury and liver cells (HepG2 line) were aimed to be discovered. Methods: Acute liver injury (ALI) was simulated by single acetaminophen (APAP, 1000 mg/kg) administration, on a chronic CLI, by 4 weekly APAP administrations. Pristine C60 fullerene aqueous colloid solution (C60FAS; initial concentration 0.15 mg/mL) was administered per os or intraperitoneally at a dose of 0.5 mg/kg (ALI) or 0.25 mg/kg (CLI) daily for 2 or 28 days, respectively, after first APAP dose. Animals were sacrificed at 24th hour after the last dose. Biochemical markers of blood serum and liver autopsies were analyzed. EGFR expression in HepG2 cells after 48-hour incubation with C60FAS was assessed. Results: Increase of serum conjugated and unconjugated bilirubin (up to 1.4-3.7 times), ALT (by 31-37%), and AST (by 18%) in non-treated ALI and CLI rats were observed, suggesting the hepatitis (confirmed by histological analysis). Liver morphological state (ALI, CLI), ALT (ALI and CLI), bilirubin (CLI), α-amylase, and creatinine (ALI) were normalized with C60FAS administration in both ways, which may indicate its protective impact on liver. However, unconjugated bilirubin sharply increased in ALI animals receiving C60FAS (up to 12 times compared to control), suggesting the augmentation of bilirubin metabolism. Furthermore, C60FAS inhibited EGFR expression in HepG2 cells in a dose-dependent manner. Conclusion: C60FAS could partially correct acute and chronic toxic liver injury, however, it could not normalize bilirubin metabolism after acute exposure

The role of trust in the acceptance of adjuvant endocrine therapy in breast cancer patients

Objective: Despite adjuvant endocrine therapy (AET) considerably reducing mortality and recurrence in hormonal receptor-positive (HR+) breast cancer (BC) patients, acceptance of AET remains an issue. The reasoning behind the lack of acceptance is complex and multifactorial, and some associated risk factors have been previously analyzed. Our study aims to assess women's beliefs and concerns toward AET and women's trust in the treating physician, focusing on determining the importance of these factors in the acceptance of AET. Methods: Out of n = 539, n = 269 women with HR + BC participated in a cross-sectional online survey. The main study variables were AET necessity beliefs and concerns (Beliefs about Medicines Questionnaire) and the trust in treating physicians. A binary hierarchical logistic regression was applied to predict AET acceptance. Results: We did not observe a meaningful mean difference in the necessity beliefs between women who accepted versus refused AET by the time of study conduct. Women with ongoing AET intake indicated significantly higher trust in their treating physician (d = 0.57) and have lower concerns (d = -1.65) regarding AET than women who had declined or discontinued AET prematurely. Results of the logistic regression demonstrated that after adjusting for clinical factors (e.g., prognosis, age), higher trust and lower concerns significantly increased the likelihood of accepting AET treatment. Conclusions: Our results demonstrate the importance of discussing potential concerns regarding AET and establishing a trustful patient-physician relationship, which outweighs non-modifiable factors such as cancer prognosis

Mode of photoexcited C60 fullerene involvement in potentiating cisplatin toxicity against drug-resistant L1210 cells

Introduction: C60 fullerene has received great attention as a candidate for biomedical applications. Due to unique structure and properties, C60 fullerene nanoparticles are supposed to be useful in drug delivery, photodynamic therapy (PDT) of cancer, and reversion of tumor cells’ multidrug resistance. The aim of this study was to elucidate the possible molecular mechanisms involved in photoexcited C60 fullerene-dependent enhancement of cisplatin toxicity against leukemic cells resistant to cisplatin. Methods: Stable homogeneous pristine C60 fullerene aqueous colloid solution (10-4 М, purity 99.5%) was used in the study. The photoactivation of C60 fullerene accumulated by L1210R cells was done by irradiation in microplates with light-emitting diode lamp (420-700 nm light, 100 mW·cm-2). Cells were further incubated with the addition of Cis-Pt to a final concentration of 1 μg/mL. Activation of p38 MAPK was visualized by Western blot analysis. Flow cytometry was used for the estimation of cells distribution on cell cycle. Mitochondrial membrane potential (Δψm) was estimated with the use of fluorescent potential-sensitive probe TMRE (Tetramethylrhodamine Ethyl Ester). Results: Cis-Pt applied alone at 1 μg/mL concentration failed to affect mitochondrial membrane potential in L1210R cells or cell cycle distribution as compared with untreated cells. Activation of ROS-sensitive proapoptotic p38 kinase and enhanced content of cells in subG1 phase were detected after irradiation of L1210R cells treated with 10-5M C60 fullerene. Combined treatment with photoexcited C60 fullerene and Cis-Pt was followed by the dissipation of Δψm at early-term period, blockage of cell transition into S phase, and considerable accumulation of cells in proapoptotic subG1 phase at prolonged incubation. Conclusion: The effect of the synergic cytotoxic activity of both agents allowed to suppose that photoexcited C60 fullerene promoted Cis-Pt accumulation in leukemic cells resistant to Cis-Pt. The data obtained could be useful for the development of new approaches to overcome drug-resistance of leukemic cells

C60 fullerene against SARS-CoV-2 coronavirus: an in silico insight

Based on WHO reports the new SARS-CoV-2 coronavirus is currently widespread all over the world. So far > 162 million cases have been confirmed, including > 3 million deaths. Because of the pandemic still spreading across the globe the accomplishment of computational methods to find new potential mechanisms of virus inhibitions is necessary. According to the fact that C60 fullerene (a sphere-shaped molecule consisting of carbon) has shown inhibitory activity against various protein targets, here the analysis of the potential binding mechanism between SARS-CoV-2 proteins 3CLpro and RdRp with C60 fullerene was done; it has resulted in one and two possible binding mechanisms, respectively. In the case of 3CLpro, C60 fullerene interacts in the catalytic binding pocket. And for RdRp in the first model C60 fullerene blocks RNA synthesis pore and in the second one it prevents binding with Nsp8 co-factor (without this complex formation, RdRp can’t perform its initial functions). Then the molecular dynamics simulation confirmed the stability of created complexes. The obtained results might be a basis for other computational studies of 3CLPro and RdRp potential inhibition ways as well as the potential usage of C60 fullerene in the fight against COVID-19 disease