Characteristic Markers Research Articles

POFUT1 and PLAGL2 are characteristic markers of mucinous colorectal cancer associated with MUC2 expression.

Colorectal mucinous adenocarcinoma (MAC)is one of the most lethal histological types of colorectal cancer, and its mechanism of development is not well understood. In this study, we aimed to clarify the molecular characteristics of MAC via in silico analysis using The Cancer Genome Atlas database. The expression of genes on chromosome 20q (Chr20q) was negatively associated with the expression of MUC2, which is a key molecule that can be used to distinguish between MAC and nonmucinous adenocarcinoma (NMAC).This was consistent with a significant difference in copy number alteration of Chr20q between the two histological types. We further identified 475 differentially expressed genes (DEGs)between MAC and NMAC, and some of the Chr20q genes among the DEGs are considered to be pivotal genes used to define MAC. Both in vitro and in vivo analysis showed that simultaneous knockdown of POFUT1 and PLAGL2, both of which are located on Chr20q, promoted MUC2 expression. Moreover, these genes were highly expressed in NMAC but not in MAC according to the results of immunohistological studies using human samples. In conclusion, POFUT1 and PLAGL2 are considered to be important for defining MAC, and these genes are associated with MUC2 expression.

Exploration of markers in oxidized rancidity walnut kernels based on lipidomics and volatolomics

Walnut kernels are prone to oxidation and rancidity due to their rich lipid composition, but the existing evaluation indicators are not sensitive enough to promote their industrial development. This study aims to investigate the potential markers in oxidative rancidity walnut kernels using lipidomics and volatolomics. The results showed that the antioxidant capacity of walnut kernels significantly decreased after oxidation, with the decreasing of total phenolic content from 36276.34 mg GAE/kg to 31281.53 mg GAE/kg, the DPPH and ABTS free radical scavenging activity from 89.25% to 73.54%, and 61.69% to 43.73%, respectively. The activities of lipoxygenase (LOX) and lipase (LPS) increased by 6.08-fold and 0.33-fold, respectively. By combining volatolomics and chemometrics methods, it was found that significant differences existed in the content of hexanal, caproic acid, 1-pentanol, (E)-2-octenal, and 2-heptanenal before and after walnut kernel oxidation (VIP > 1). Based on the results of lipidomics, it can be concluded that the above five compounds can serve as characteristic markers for walnut kernel oxidative rancidity, mainly produced through glycerol phospholipid (GPL), glyceride, linoleic acid (LA), and α-linolenic acid (ALA) metabolism pathways. Possible mechanisms of lipid degradation in oxidized walnut kernels were also proposed, providing technical support for the storage, preservation, and high-value utilization of walnut kernels.

Induction of lung progenitor cell-like organoids by porcine pluripotent stem cells.

Organoids are invitro 3D models that are generated using stem cells to study organ development and regeneration. Despite the extensive research on lung organoids, there is limited information on pig lung cell generation or development. Here, we identified five epithelial cell types along with their characteristic markers using scRNA-seq. Additionally, we found that NKX2.1 and FOXA2 acted as the crucial core transcription factors in porcine lung development. The presence of SOX9/SOX2 double-positive cells was identified as a key marker for lung progenitor cells. The Monocle algorithm was used to create a pseudo-temporal differentiation trajectory of epithelial cells, leading to the identification of signaling pathways related to porcine lung development. Moreover, we established the differentiation method from porcine pluripotent stem cells (pPSCs) to SOX17+ FOXA2+ definitive endoderm (DE) and NKX2.1+ FOXA2+ CDX2- anterior foregut endoderm (AFE). The AFE is further differentiated into lung organoids while closely monitoring the differentiation process. We showed that NKX2.1 overexpression facilitated the induction of lung organoids and supported subsequent lung differentiation and maturation. This model offers valuable insights into studying the interaction patterns between cells and the signaling pathways during the development of the porcine lung.

Persistence of Chronic Lymphocytic Leukemia Stem-like Populations under Simultaneous In Vitro Treatment with Curcumin, Fludarabine, and Ibrutinib: Implications for Therapy Resistance.

Leukemic stem cells (LSCs) possess similar characteristics to normal hematopoietic stem cells, including self-renewal capacity, quiescence, ability to initiate leukemia, and drug resistance. These cells play a significant role in leukemia relapse, persisting even after apparent remission. LSCs were first described in 1994 by Lapidot et al. Although they have been extensively studied in acute leukemia, more LSC research is still needed in chronic lymphocytic leukemia (CLL) to understand if reduced apoptosis in mature cells should still be considered as the major cause of this disease. Here, we provide new evidence suggesting the existence of stem-like cell populations in CLL, which may help to understand the disease as well as to develop effective treatments. In this study, we identified a potential leukemic stem cell subpopulation using the tetraploid CLL cell line I83. This subpopulation is characterized by diploid cells that were capable of generating the I83 tetraploid population. Furthermore, we adapted a novel flow cytometry analysis protocol to detect CLL subpopulations with stem cell properties in peripheral blood samples and primary cultures from CLL patients. These cells were identified by their co-expression of CD19 and CD5, characteristic markers of CLL cells. As previously described, increased alkaline phosphatase (ALP) activity is indicative of stemness and pluripotency. Moreover, we used this method to investigate the potential synergistic effect of curcumin in combination with fludarabine and ibrutinib to deplete this subpopulation. Our results confirmed the effectiveness of this ALP-based analysis protocol in detecting and monitoring leukemic stem-like cells in CLL. This analysis also identified limitations in eradicating these populations using in vitro testing. Furthermore, our findings demonstrated that curcumin significantly enhanced the effects of fludarabine and ibrutinib on the leukemic fraction, exhibiting synergistic effects (combination drug index, CDI 0.97 and 0.37, respectively). Our results lend support to the existence of potential stem-like populations in CLL cell lines, and to the idea that curcumin could serve as an effective adjuvant in therapies aimed at eliminating these populations and improving treatment efficacy.

A combinatory genetic strategy for targeting neurogliaform neurons in the mouse basolateral amygdala.

The mouse basolateral amygdala (BLA) contains various GABAergic interneuron subpopulations, which have distinctive roles in the neuronal microcircuit controlling numerous behavioral functions. In mice, roughly 15% of the BLA GABAergic interneurons express neuropeptide Y (NPY), a reasonably characteristic marker for neurogliaform cells (NGFCs) in cortical-like brain structures. However, genetically labeled putative NPY-expressing interneurons in the BLA yield a mixture of interneuron subtypes besides NGFCs. Thus, selective molecular markers are lacking for genetically accessing NGFCs in the BLA. Here, we validated the NGFC-specific labeling with a molecular marker, neuron-derived neurotrophic factor (NDNF), in the mouse BLA, as such specificity has been demonstrated in the neocortex and hippocampus. We characterized genetically defined NDNF-expressing (NDNF+) GABAergic interneurons in the mouse BLA by combining the Ndnf-IRES2-dgCre-D transgenic mouse line with viral labeling, immunohistochemical staining, and in vitro electrophysiology. We found that BLA NDNF+ GABAergic cells mainly expressed NGFC neurochemical markers NPY and reelin (Reln) and exhibited small round soma and dense axonal arborization. Whole-cell patch clamp recordings indicated that most NDNF+ interneurons showed late spiking and moderate firing adaptation. Moreover, ∼81% of BLA NDNF+ cells generated retroaxonal action potential after current injections or optogenetic stimulations, frequently developing into persistent barrage firing. Optogenetic activation of the BLA NDNF+ cell population yielded both GABAA- and GABAB receptor-mediated currents onto BLA pyramidal neurons (PNs). We demonstrate a combinatory strategy combining the NDNF-cre mouse line with viral transfection to specifically target adult mouse BLA NGFCs and further explore their functional and behavioral roles.

First-trimester urinary extracellular vesicles as predictors of preterm birth: an insight into immune programming.

Background: The programming of innate and adaptive immunity plays a pivotal role in determining the course of pregnancy, leading to either normal term birth (TB) or preterm birth (PB) through the modulation of macrophage (M1/M2) differentiation. Extracellular vesicles (EVs) in maternal blood, harboring a repertoire of physiological and pathological messengers, are integral players in pregnancy outcomes. It is unknown whether urinary EVs (UEVs) could serve as a non-invasive mechanistic biomarker for predicting PB. Methods: This study investigated first-trimester UEVs carrying M1 messengers with altered immune programming, aiming to discern their correlation to subsequent PB. A birth cohort comprising 501 pregnant women, with 40 women experiencing PB matched to 40 women experiencing TB on the same day, was examined. First-trimester UEVs were isolated for the quantification of immune mediators. Additionally, we evaluated the UEV modulation of "trained immunity" on macrophage and lymphocyte differentiations, including mRNA expression profiles, and chromatin activation modification at histone 3 lysine 4 trimethylation (H3K4me3). Results: We found a significant elevation (p < 0.05) in the particles of UEVs bearing characteristic exosome markers (CD9/CD63/CD81/syntenin) during the first trimester of pregnancy compared to non-pregnant samples. Furthermore, UEVs from PB demonstrated significantly heightened levels of MCP-1 (p = 0.003), IL-6 (p = 0.041), IL-17A (p = 0.007), IP-10 (p = 0.036), TNFα (p = 0.004), IL-12 (p = 0.045), and IFNγ (p = 0.030) relative to those from TB, indicative of altered M1 and Th17 differentiation. Notably, MCP-1 (>174pg/mL) exhibited a sensitivity of 71.9% and specificity of 64.6%, and MCP-1 (>174pg/mL) and IFNγ (>8.7pg/mL) provided a higher sensitivity (84.6%) of predicting PB and moderate specificity of 66.7%. Subsequent investigations showed that UEVs from TB exerted a significant suppression of M1 differentiation (iNOS expression) and Th17 differentiation (RORrT expression) compared to those of PB. Conversely, UEVs derived from PB induced a significantly higher expression of chromatin modification at H3K4me3 with higher production of IL-8 and TNFα cytokines (p < 0.001). Implications: This pioneering study provides critical evidence for the early detection of altered M1 and Th17 responses within UEVs as a predictor of PB and early modulation of altered M1 and Th17 polarization associated with better T-cell regulatory differentiation as a potential prevention of subsequent PB.

A Noncanonical CD56dimCD16dim/- NK Cell Subset Indicative of Prior Cytotoxic Activity Is Elevated in Patients with Autoantibody-Mediated Neurologic Diseases.

Neuromyelitis optica spectrum disorder (NMOSD), myelin oligodendrocyte glycoprotein Ab disease, and autoimmune myasthenia gravis (MG) are autoantibody-mediated neurologic conditions where autoantibodies can induce Ab-dependent cellular cytotoxicity (ADCC), a NK cell-mediated effector function. However, whether ADCC is a pathogenic mechanism in patients with these conditions has not been confirmed. We sought to characterize circulatory NK cells using functional assays, phenotyping, and transcriptomics to elucidate their role in pathology. NK cells from NMOSD patients and MG patients with elevated disease burden exhibited reduced ADCC and CD56dimCD16hi NK cells, along with an elevated frequency of CD56dimCD16dim/- NK cells. We determined that ADCC induces a similar phenotypic shift invitro. Bulk RNA sequencing distinguished the CD56dimCD16dim/- population from the canonical CD56dimCD16hi cytotoxic and CD56hiCD16- immunomodulatory subsets, as well as CD56hiCD16+ NK cells. Multiparameter immunophenotyping of NK cell markers, functional proteins, and receptors similarly showed that the CD56dimCD16dim/- subset exhibits a unique profile while still maintaining expression of characteristic NK markers CD56, CD94, and NKp44. Notably, expression of perforin and granzyme is reduced in comparison with CD56dimCD16hi NK cells. Moreover, they exhibit elevated trogocytosis capability, HLA-DR expression, and many chemokine receptors, including CCR7. In contrast with NMOSD and MG, myelin oligodendrocyte glycoprotein Ab disease NK cells did not exhibit functional, phenotypic, or transcriptomic perturbations. In summary, CD56dimCD16dim/- NK cells are a distinct peripheral blood immune cell population in humans elevated upon prior cytotoxic activity by the CD56dimCD16hi NK cell subset. The elevation of this subset in NMOSD and MG patients suggests prior ADCC activity.

Simultaneous rapid detection of glycyrrhizin and sennoside A in Daiokanzoto samples by lateral flow immunoassay.

Glycyrrhizin (GLY) and sennoside A (SA) are characteristic bioactive marker compounds of the Kampo medicine Daiokanzoto. Their accurate detection in blends of Rhei rhizoma and Glycyrrhizae radix of several species (4:1 or 4:2) is essential for quality control and to ensure therapeutic efficacy. A rapid, efficient assay can significantly facilitate their detection. To establish a rapid qualitative assay for GLY and SA detection, a lateral flow immunoassay (LFA) was developed using specific monoclonal antibody (mAb) nanoparticles. This assay harnesses the competitive binding of mAb nanoparticles to the immobilized analytes on test strips and free analytes in the samples. Two conjugates for detecting GLY and SA, GLY-bovine serum albumin and SA-human serum albumin, were separately immobilized on the test zones of LFA strips. The detection mechanism is reliant on the visual detection of color changes in the test zones. When GLY and SA were present in samples, they contended with the immobilized conjugates on the strip to bind with the mAb nanoparticles and produced distinct color patterns in the test zones. The limits of detection of the assay for GLY and SA were both 3.13 μg/mL. The capability of the LFA was substantiated using plant samples and Daiokanzoto, and its alignment with indirect competitive ELISA results was confirmed. The introduced LFA is a groundbreaking procedure that offers a rapid, straightforward, and sensitive method for simultaneously detecting GLY and SA in Daiokanzoto samples. It is instrumental in ensuring product quality.

Mesenchymal Stem Cells Cultured in a 3D Microgel Environment Containing Platelet-Rich Plasma Significantly Modify Their Chondrogenesis-Related miRNA Expression.

The aim of this work is to study the effect of platelet factors on the differentiation of mesenchymal stem cells (MSCs) to hyaline cartilage chondrocytes in a three-dimensional environment. MSCs were cultured in a microgel environment with a chondrogenic medium. The microgel consisted of microspheres that combine gelatin and platelet-rich plasma (PRP). The gelatin/PRP microdroplets were produced by emulsion. The gelatin containing the microdroplets was enzymatically gelled, retaining PRP and, just before seeding the cells, platelets were activated by adding calcium chloride so that platelet growth factors were released into the culture media but not before. Platelet activation was analyzed before activation to rule out the possibility that the gelatin cross-linking process itself activated the platelets. The gene expression of characteristic chondrogenic markers and miRNA expression were analyzed in cells cultured in a differentiation medium and significant differences were found between gelation/PRP microgels and those containing only pure gelatin. In summary, the gelatin microspheres effectively encapsulated platelets that secreted and released factors that significantly contributed to cellular chondrogenic differentiation. At the same time, the microgel constituted a 3D medium that provided the cells with adherent surfaces and the possibility of three-dimensional cell-cell contact.

Characterization of volatile organic compounds in walnut oil with various oxidation levels using olfactory analysis and HS-SPME-GC/MS

Walnut oil oxidizes and becomes rancid during storage, that could be significantly affecting flavor and quality. This study aimed to monitor the volatile compounds present in walnut oil during storage, identify the characteristic markers of walnut oil at different oxidation levels, and establish a correlation network analysis based on the relationship between the olfactory analyzer and the characteristic markers to understand their correlation. The results indicated that the oxidation level of walnut oil had a positive correlation with the response of the olfactory analyzer. 219 volatile compounds were identified in walnut oil, with 89 identified as key volatile compounds (VIP >1). Among these, compounds such as (E, E)-2,4-decadienal (6.10%–23.04%)，(E, E)-2,4-heptadienal (2.23%–13.61%)，(E)-2-octenal (0.95%–11.71%)， hexanoic acid (1.63%–4.30%)，1-octen-3-ol (2.53%–19.01%)，(Z)-2-heptenal (5.95%–25.01%)，2,3-dihydro-furan (1.08%–3.20%)，2-pentyl-furan (0.13%–0.54%), pyrazine (0.33%–1.32%), hexanal (24.52%–1.33%)，3-hethylbutylacetate (12.44%–1.29%)， 2-methyl butyl acetate (7.74%–1.56%) and ethenyl hexanoate (4.39%–0.41%) were found to be characteristic volatile compounds in the oxidation process of walnut oil. Furthermore, the correlation network analysis revealed a strong correlation between the olfactory analyzer sensors and the characteristic volatile compounds. The findings of this study can provide valuable data for the development of rapid determination of the oxidation level of walnut oil.

Perekhodya granitsy. Rossiyskiy stendap: lokalnye i globalnye smysly

Stand-up comedy is one of the brightest youth scenes of modern Russia, rapidly gaining popularity over the last decade. This genre of modern laugh culture rarely gets the attention of either foreign or domestic scholars and researchers. This article focuses on the career strategies of stand-up comedians, the peculiarities of their communication with audiences in the context of the formation of the stand-up scene in contemporary Russia in comparison with the formats of humor and laugh culture characteristic of the Soviet and post-Soviet times. The uniqueness of the Russian stand-up experience will be revealed through the analysis of characteristic style markers associated with the genre: the liminality / transboundary nature of the emerging culture, the high mobility of stand-up artists, creative collaborations, and touring exchanges within and between cities in their native regions. As a theoretical framework, the construct of youth cultural scene is used as the most adequate to the case under the study. The empirical base of the article is 15 semi-structured interviews with participants in the stand-up scene, non-participant observation of stand-up performances, and the textual analysis of blogs of stand-up comedians. The case study material was collected as part of a large-scale research project aimed at tracing the migration strategies and life plans of talented young people in the Northwestern Federal District of Russia.

Comparative Analysis of Vascular Cell Differentiation From Peripheral Blood Mononuclear Cell‐ and Urine‐Derived Induced Pluripotent Stem Cells

Background and Objectives: Peripheral blood mononuclear cells (PBMCs) and urine‐derived epithelial cells have both emerged as valuable sources for induced pluripotent stem cell (iPSC) generation, each presenting unique advantages in terms of accessibility and reprograming efficiency. This study aimed to assess and compare the potential of PBMC‐derived iPSCs (PiPSCs) and urine‐derived iPSCs (UiPSCs) in generating functional endothelial cells (ECs) and vascular smooth muscle cells (VSMCs), which are critical for vascular tissue engineering and disease modeling. Phenotypic characteristics, differentiation efficacy, and functional properties of iPSC‐derived ECs and VSMCs from these distinct sources were investigated to reveal variations attributed to cellular origin.Methods and Results: PiPSCs and UiPSCs both successfully differentiated into functional ECs and VSMCs. EC differentiation efficiency was similar, yielding about 45% mature ECs with characteristic morphology, marker expression, and tube formation abilities, showing no significant differences between cell types. Transcriptomic analysis revealed upregulation of key endothelial markers (platelet endothelial cell adhesion molecule 1 [PECAM1], cadherin 5 [CDH5], and melanoma cell adhesion molecule [MCAM]) and downregulation of lymphatic markers (Fms‐related tyrosine kinase 4 [FLT4], prospero homeobox protein 1 [PROX1], and podoplanin [PDPN]), confirming blood EC identity. The upregulation of collagen type IV alpha 1 chain (COL4A1) and collagen type I alpha 1 chain (COL1A1) indicated a mature endothelial state with enhanced extracellular matrix (ECM) production. VSMC differentiation resulted in high percentages of α‐smooth muscle actin (α‐SMA) positive cells for both PiPSCs (96%) and UiPSCs (94%). These VSMCs exhibited typical spindle‐shaped morphology, expressed VSMC markers, and responded to carbachol. Transcriptomic analysis showed significant upregulation of VSMC markers (actin alpha 2 [ACTA2], caldesmon 1 [CALD1], calponin 1 [CNN1], transgelin [TAGLN], tropomyosin 2 [TPM2]), with concurrent downregulation of ACTA1 and upregulation of ACTA2, confirming their vascular smooth muscle phenotype. The upregulation of COL6A1 in VSMCs indicated a mature phenotype with enhanced ECM production, crucial for vascular tissue integrity and function. Gene set enrichment analysis highlighted the upregulation of multiple hallmark pathways, delineating a distinctive transcriptional profile.Conclusions: This study presents a comprehensive comparative analysis of functionally differentiated ECs and VSMCs derived from PiPSCs and UiPSCs, providing critical insights into the expression patterns and phenotypic transitions during differentiation. Our findings enhance the understanding of distinct molecular signatures in iPSCs from different sources and their progeny.

The gut microbial composition in polycystic ovary syndrome with hyperandrogenemia and its association with steroid hormones.

Background: Polycystic ovary syndrome (PCOS) is characterized by excess androgens, ovulatory dysfunction, and polycystic ovaries. The mechanisms underlying ovulatory and metabolic disorders in PCOS remain elusive, hampering therapeutic development. Enhanced metabolic health correlates with increased microbiota gene content and microbial diversity. We aimed to explore the impact of gut microbiota and serum steroids on PCOS regulation associated with androgen excess. Methods: The fecal samples of patients with hyperandrogenic PCOS (n = 14) and control group with PCOS (n = 14) were analyzed by 16S rRNA gene sequencing. The peripheral venous blood of all subjects was collected to detect serum hormones. The association between gut microbiota and serum hormones was analyzed with the R language. Results: Our findings reveal that the hyperandrogenic PCOS group exhibits lower richness and diversity of gut microbiota compared to the control group. Characteristic genera in PCOS patients with hyperandrogenism include Bifidobacterium, Enterobacteriaceae_unclassified, Streptococcus, Saccharimonadaceae, Enterococcus, and Eubacterium_nodatum_group. Five hormones, including 5β-androsterone, deoxycorticosterone, corticosterone, 11-dehydrocorticosterone, and cortexolone, emerge as potential serum biomarkers for identifying patients with hyperandrogenic-PCOS (HA-PCOS). Furthermore, a lower vitamin D3 level may act as a susceptibility factor, suggesting that vitamin D3 supplementation could serve as a potential intervention for PCOS with hyperandrogenism. Conclusion: Specific fecal microbiota and serum steroids may be used as characteristic markers for clinical diagnosis of hyperandrogenic-PCOS. This research enhances our understanding of the intricate interplay among hormones, gut microbiota, and hyperandrogenemia in patients with PCOS.

ИЗУЧЕНИЕ ГЕПАТОТОКСИЧЕСКОГО ВОЗДЕЙСТВИЯ ГИДРОКСИДА АЛЮМИНИЯ, ВВОДИМОГО В ЖЕЛУДОЧНО-КИШЕЧНЫЙ ТРАКТ КРЫС, В ПОДОСТРОМ ЭКСПЕРИМЕНТЕ

Aluminum (Al) has various toxic effects on biosystems. Essential properties of aluminum have not been established, the mechanisms of impairment are insufficiently studied. The negative impact of aluminum on the human body leads to liver damage, increased oxidative stress and development of hepatotoxicity. Blood enzymes are characteristic markers of liver function and are used to assess the toxicity of environmental pollutants. The aim of the study was to investigate the activity of enzyme markers of hepatotoxicity, aspartate aminotransferase (AST), alanine aminotransferase (ALT), lactate dehydrogenase (LDH) and alkaline phosphatase (ALP) in the blood of laboratory animals after subacute intoxication with aluminum hydroxide (Al(OH)3). The experiment was conducted on 80 white rats divided into 4 groups. Three groups of animals orally received Al(OH)3 solution of different concentrations for a month, the control group was administered distilled water. At the end of the experiment, enzyme activity in the blood serum was measured. The increase in the AST level in the first group is associated with changes in hepatocytes. The decrease in ALT activity in the experimental groups is associated with the peculiarities of aluminum excretion. By analogy with AST, an increase in the concentration of LDH was registered in the first group, while a decrease in the level of the indicator was shown in the second and third groups. A decrease in the activity of alkaline phosphatase indicates a violation of metabolic processes in cells. The effect of Al(OH)3 for a month caused certain changes in liver markers in the blood serum of experimental animals. For a more complete understanding of the hepatotoxic effects of aluminum, additional biochemical studies are needed to assess the activity of antioxidant defense enzymes.

A novel photoelectrochemical aptasensor for alpha-fetoprotein assay based on a 3D self-supporting TiO2/CdS/BiOI Z-scheme heterojunction

Alpha-fetoprotein (AFP) is a characteristic marker of liver cancer. This study establishes a PEC aptamer sensor using BiOI/CdS/TiO2 heterojunction material and realizes accurate detection of AFP.

Comparative immunological study of children infected with Entamoeba spp. parasites in Thi-Qar province

Background. Amoebiasis is a one of the major public health problems, it is caused by the genus Entamoeba which includes several species, such as E. histolytica, E. dispar and E. moshkovskii. Purposes. The present study aimed to differentiate among Entamoeba spp. by using molecular method (PCR) and to identify characteristic serum markers (IgG and IL-6) according to species. Methods. About 697 stool samples were collected from children under five years old, and blood samples (3 ml) of venous blood were collected from patients with positive microscopic examination to evaluate some immune parameters (IL-6, IgG) for children infected with the parasite using the ELISA test. DNA extraction was carried out from fecal samples using a special stool DNA extraction kit and Multiplex PCR primers were designed to detect E. histolytica, E. dispar and E. moshkovskii. Results. About 100(14.34%) were infected with Entamoeba spp. by microscopic examination and 68 samples DNA-samples were positive. Fifty samples 50% were infected with E. histolytica, and 12% with a mixed infection. A significant increase (p≤0.05) in the concentration of IgG immunoglobulin in the sera of children infected with the E. histolytica parasite. A significant increase (p≤0.05) in the concentration of interleukin-6 in the sera of patients groups infected with Entamoeba spp., the highest significant increase (p≤0.05) was recorded in patients group infected with the E. moshkovskii. Conclusion. Molecular tests have the ability to detect and distinguish between the Entamoeba species infecting humans. Amoebiasis caused an increase in the concentration of IgG and interleukin-6.

Трансплантация суспензии эндотелиальных клеток в эксперименте ex vivo

Relevance. Corneal transplantation with entire cornea or anormal layers replacement remains the only treatment option for patients with corneal endothelium pathology. Selective endothelial replacement was introduced into practice two decades ago, namely posterior lamellar keratoplasty, which has advantages over penetrating keratoplasty. Improvements of this method led to technique creation where isolated Descemet's membrane with endothelium monolayer without stromal layer is transplanted. This technology is associated with risks of intra- and postoperative complications, and is difficult to perform. Nowadays, development is in progress of introducing corneal endothelial cells (CECs) transplantation without stromal layer and even without Descemet's membrane. CECs transplantation in suspension form has potential to change treatment approach of corneal endothelium pathologies giving possibility of rapid eyesight recovery and reducing need for graft material. Purpose. To evaluate effectiveness of corneal CECs suspension transplantation on cadaver eye in ex vivo experiment. Material and methods. CECs suspension was obtained by modified enzymatic method. CECs transplantation experiment consisted of 3 stages. Firstly, CECs loss was calculated depending on cell administration method. Secondly, corneoscleral button was used as recipient for the obtained suspension. The third stage was carried out on cadaver eyeball. Results. The viability of transplanted CECs and their effective adhesion to Descemet's membrane was confirmed in this experiment. The characteristic markers of CECs ZO-1, Na+/K+ -ATPase, Ki67 were determined in corneal samples by immunohistochemistry assay and single cells expressing Vimentin were detected after one-week cultivation. Conclusion. Based on transplantation experiment results, it can be noted this technique is quite promising in surgical rehabilitation of patients with corneal endothelial dysfunction. Thus, it seems relevant to continue research to achieve the main goal – to fully cover the defect of central zone of corneal posterior surface with isolated CECs given that their morphology and functional activity are preserved. Key words: cornea, endothelial corneal dystrophy, endothelial cell suspension, corneal endothelial cell transplantation

Metadiscoursal Realisation of Pragmatic Strategies @ResearchProject Twitter Accounts

To ensure the global communication and visibility of their investigations, international research projects leverage online settings and endorse specific digital academic practices. Twitter as a Social Medium for Research Purposes has become an effective outlet to widely disseminate their project development, knowledge production and research findings. To meet these aims, research groups display pragmatic strategies responding to three overarching communicative intentions –informative, interactional and promotional– as well as metadiscursive markers to establish links through their texts with the audience. This paper analyses these practices by looking into the metadiscoursal realisations of a taxonomy of twenty-seven data-driven pragmatic strategies in ten Horizon2020 research project Twitter accounts. First, we revisit metadiscursive adjustments for the digital environment of Twitter. Then, we identify salient metadiscourse features instantatiating the pragmatic strategies using NVivo. In general, interactional metadiscursive features predominate over interactive ones, being attitude markers, self-mentions and directives characteristic markers in informative, promotional and interactional strategies, respectively. Moreover, some metadiscourse categories are found to rely on non-verbal markers for their realisation. The analysis expands the understanding of complex digital discursive practices developed by researchers to disseminate their results, account for their funding, make themselves visible and engage multiple audiences.

Deciphering the mysteries of Aconitum pendulum: Unique identification of various processed products and characteristic chemical markers

Aconitum pendulum, a member of the Aconitum genus within the Ranunculaceae family, is known for its remarkable anti-inflammatory, anti-tumor, and analgesic properties. However, the application of Aconitum pendulum in traditional medicine is often restricted by its inherent toxicity, necessitating specific processing methods to ensure safe usage. Tibetan medicine utilizes distinctive processing techniques such as the barley wine, Hezitang, and zanba frying systems to render the herb safe for medicinal use. Despite the known pharmacological importance, scant research exists on how these processing methods affect the phytochemical profile of Aconitum pendulum, indicating a necessity for comparative analysis of the resulting concoctions. In this study, we employed 1H nuclear magnetic resonance (1H NMR) spectroscopy and ultra-performance liquid chromatography coupled with quadrupole time of flight mass spectrometry (UPLC-Q-TOF-MS) to explore the metabolomic variations among different processed products of Aconitum pendulum. Multivariate data analysis techniques, including pattern recognition, allowed us to investigate the intricacies of the compound profiles. To discover signature chemical markers that can differentiate the various processed products, we conducted paired t-tests and generated heat maps highlighting the detected differences. Our integrated analysis of the 1H NMR and UPLC-Q-TOF-MS data revealed distinct chemical signatures unique to each method of Aconitum pendulum processing. By employing chemometric analysis with variable importance in projection (VIP) scores greater than 1.0, and coupled with the statistical rigor of paired t-tests (P < 0.05), we successfully identified significant chemical markers in various processed products, including ferulic acid (FA), caffeic acid (CA), 3-acetylaconitine (AAC) and benzoylaconitine (BAC). Our findings provide a rapid and robust analysis of the chemical constituencies within raw and processed Aconitum pendulum. The discovery of potential chemical markers through an array of advanced pattern recognition techniques offers a substantial contribution to the pharmaceutical analysis. The results of this research pave the way for future comparative studies and facilitate the recognition of distinctively processed Aconitum pendulum products in traditional medicine.

Reassessing endothelial-to-mesenchymal transition in mouse bone marrow: insights from lineage tracing models

Endothelial cells (ECs) and bone marrow stromal cells (BMSCs) play crucial roles in supporting hematopoiesis and hematopoietic regeneration. However, whether ECs are a source of BMSCs remains unclear. Here, we evaluate the contribution of endothelial-to-mesenchymal transition to BMSC generation in postnatal mice. Single-cell RNA sequencing identifies ECs expressing BMSC markers Prrx1 and Lepr; however, this could not be validated using Prrx1-Cre and Lepr-Cre transgenic mice. Additionally, only a minority of BMSCs are marked by EC lineage tracing models using Cdh5-rtTA-tetO-Cre or Tek-CreERT2. Moreover, Cdh5+ BMSCs and Tek+ BMSCs show distinct spatial distributions and characteristic mesenchymal markers, suggestive of their origination from different progenitors rather than CDH5+ TEK+ ECs. Furthermore, myeloablation induced by 5-fluorouracil treatment does not increase Cdh5+ BMSCs. Our findings indicate that ECs hardly convert to BMSCs during homeostasis and myeloablation-induced hematopoietic regeneration, highlighting the importance of using appropriate genetic models and conducting careful data interpretation in studies concerning endothelial-to-mesenchymal transition.