Immunohistochemical characteristic of C cells in European bison thyroid gland

Introduction. C cells constitute a small percentage of thyroid gland parenchyma. The number, morphology and distribution of C cells differ among species; however, data regarding their characteristics in European bison are sparse. The aim of this study was to evaluate the morphology, distribution pattern and percentage of C cells in European bison thyroid gland together with morphometric analysis.Material and methods. Thyroid glands from 28 European bisons of different sex and age were collected either in autumn-winter (13/28) or in spring-summer (15/28) periods and analyzed by immunohistochemistry.Results. The mean total C cell number per all endocrine (follicular and C cells) cell number (C cell concentration) was 7.33%. The tendency to increase the C cell number from periphery to the central region of thyroid lobe was observed with the mean C cell concentration of 3.95%, 7.89% and 9.97% in peripheral, intermediate and central areas, respectively. Most frequently, C cells were situated intrafolliculary whereas epifollicular and interfollicular positions were observed less often. C cells were polymorphic with long cytoplasmic processes. The mean C cell area was 61.97 μm2 and the mean C cell perimeter, length and width were: 34.92 μm, 12.85 μm and 4.91 μm, respectively. In the majority of C cells, strong immunohistochemical cytoplasmic reaction was observed with the mean color intensity of 78.32. In autumn-winter period, C cells were significantly larger with lower color intensity than during spring and summer.Conclusions. This study leads to deeper characteristics of thyroid gland C cells in European bison. The histomorphometric data suggest that in European bison production of calcitonin by C cells may differ depending on the time of the year

Circulating serum miR-362-3p and miR-6721-5p as potential biomarkers for classification patients with adult-type diffuse glioma

According to the fifth edition of the WHO Classification of Tumours of the Central Nervous System (CNS) published in 2021, grade 4 gliomas classification includes IDH-mutant astrocytomas and wild-type IDH glioblastomas. Unfortunately, despite precision oncology development, the prognosis for patients with grade 4 glioma remains poor, indicating an urgent need for better diagnostic and therapeutic strategies. Circulating miRNAs besides being important regulators of cancer development could serve as promising diagnostic biomarkers for patients with grade 4 glioma. Here, we propose a two-miRNA miR-362-3p and miR-6721-5p screening signature for serum for non-invasive classification of identified glioma cases into the highest-grade 4 and lower-grade gliomas. A total of 102 samples were included in this study, comprising 78 grade 4 glioma cases and 24 grade 2–3 glioma subjects. Using the NanoString platform, seven miRNAs were identified as differentially expressed (DE), which was subsequently confirmed via RT-qPCR analysis. Next, numerous combinations of DE miRNAs were employed to develop classification models. The dual panel of miR-362-3p and miR-6721-5p displayed the highest diagnostic value to differentiate grade 4 patients and lower grade cases with an AUC of 0.867. Additionally, this signature also had a high AUC = 0.854 in the verification cohorts by RT-qPCR and an AUC = 0.842 using external data from the GEO public database. The functional annotation analyses of predicted DE miRNA target genes showed their primary involvement in the STAT3 and HIF-1 signalling pathways and the signalling pathway of pluripotency of stem cells and glioblastoma-related pathways. For additional exploration of miRNA expression patterns correlated with glioma, we performed the Weighted Gene-Co Expression Network Analysis (WGCNA). We showed that the modules most associated with glioma grade contained as many as six DE miRNAs. In conclusion, this study presents the first evidence of serum miRNA expression profiling in adult-type diffuse glioma using a classification based on the WHO 2021 guidelines. We expect that the discovered dual miR-362-3p and miR-6721-5p signatures have the potential to be utilised for grading gliomas in clinical applications

Inhibition of protein disulfide isomerase induces differentiation of acute myeloid leukemia cells

Acute myeloid leukemia is a malignant disease of immature myeloid cells. Despite significant therapeutic effects of differentiation-inducing agents in some acute myeloid leukemia subtypes, the disease remains incurable in a large fraction of patients. Here we show that SK053, a thioredoxin inhibitor, induces differentiation and cell death of acute myeloid leukemia cells. Considering that thioredoxin knock-down with short hairpin RNA failed to exert antiproliferative effects in one of the acute myeloid leukemia cell lines, we used a biotin affinity probe-labeling approach to identify potential molecular targets for the effects of SK053. Mass spectrometry of proteins precipitated from acute myeloid leukemia cells incubated with biotinylated SK053 used as a bait revealed protein disulfide isomerase as a potential binding partner for the compound. Biochemical, enzymatic and functional assays using fluorescence lifetime imaging confirmed that SK053 binds to and inhibits the activity of protein disulfide isomerase. Protein disulfide isomerase knockdown with short hairpin RNA was associated with inhibition of cell growth, increased CCAAT enhancer-binding protein α levels, and induction of differentiation of HL-60 cells. Molecular dynamics simulation followed by the covalent docking indicated that SK053 binds to the fourth thioredoxin-like domain of protein disulfide isomerase. Differentiation of myeloid precursor cells requires the activity of CCAAT enhancer-binding protein α, the function of which is impaired in acute myeloid leukemia cells through various mechanisms, including translational block by protein disulfide isomerase. SK053 increased the levels of CCAAT enhancer-binding protein α and upregulated mRNA levels for differentiation-associated genes. Finally, SK053 decreased the survival of blasts and increased the percentage of cells expressing the maturation-associated CD11b marker in primary cells isolated from bone marrow or peripheral blood of patients with acute myeloid leukemia. Collectively, these results provide a proof-of-concept that protein disulfide isomerase inhibition has potential as a therapeutic strategy for the treatment of acute myeloid leukemia and for the development of small-molecule inhibitors of protein disulfide isomerase

Tannic Acid Modified Silver Nanoparticles Show Antiviral Activity in Herpes Simplex Virus Type 2 Infection

The interaction between silver nanoparticles and herpesviruses is attracting great interest due to their antiviral activity and possibility to use as microbicides for oral and anogenital herpes. In this work, we demonstrate that tannic acid modified silver nanoparticles sized 13 nm, 33 nm and 46 nm are capable of reducing HSV-2 infectivity both in vitro and in vivo. The antiviral activity of tannic acid modified silver nanoparticles was size-related, required direct interaction and blocked virus attachment, penetration and further spread. All tested tannic acid modified silver nanoparticles reduced both infection and inflammatory reaction in the mouse model of HSV-2 infection when used at infection or for a post-infection treatment. Smaller-sized nanoparticles induced production of cytokines and chemokines important for anti-viral response. The corresponding control buffers with tannic acid showed inferior antiviral effects in vitro and were ineffective in blocking in vivo infection. Our results show that tannic acid modified silver nanoparticles are good candidates for microbicides used in treatment of herpesvirus infections.This work was supported by the Polish National Science Centre grant No. 2011/03/B/NZ6/04878 (for MK) and Centre for Preclinical Research and Technology (CePT) Project No. POIG.02.02.00-14-024/08-0 (for MG and MD). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscrip

Tannic-acid modified AgNPs reduce HSV-2 infection <i>in vivo</i>.

<p>(A) Schematics of <i>in vivo</i> experiments. C57BL/6 mice were infected with HSV-2 pre-incubated or not with 13, 33 and 46 nm AgNPs or corresponding carriers (5 µg/ml). (B) HSV-2 DNA titers (copies/µg DNA) in the whole vaginal tissues determined by real-time PCR at 48 h p.i. (N = 6). (C) Sizes of infected sites determined by immunohistochemistry of HSV-2 antigens. (D) Schematics of post-infection treatment at 3 and 18 h p.i. with 33 AgNPs or carrier buffer (3×100 µl at 5 µg/ml) (N = 5). (E) HSV-2 DNA titers (copies/µg DNA) in the whole vaginal tissues determined by real-time PCR at 48 h p.i. (N = 6). (F) Sizes of infected sites determined by immunohistochemistry of HSV-2 antigens (N = 5). The bars represent means from three independent experiments ± SEM. * represents significant differences with <i>p</i>≤0.05, while ** <i>p</i>≤0.001.</p

Tannic acid-modified AgNPs block HSV-2 attachment and penetration.

<p>(A) Schematics of attachment and penetration experiments. (B) Viral inhibition (%) for virus attachment and penetration experiments in 291.03C cell cultures with the use of 33 nm and 46 nm AgNPs (5 µg/ml) and 13 nm AgNPs (2.5 µg/ml) and corresponding carriers. At 24 h p.i. cells and supernatants were collected and titrated to determine PFU/ml in comparison to HSV-2 infected cultures. (C) SEM images in EDS mode of HSV-2 incubated with 13 nm, 33 nm and 46 nm AgNPs, white arrows indicate nanoparticles on the viron's surface. White bars indicate 100 nm. (D) Kinetics of AgNPs and HSV-2 interaction expressed as % of HSV-2 infected positive controls. HSV-2 aliquots were mixed with 2.5 µg/ml of 13, 33 or 46 nm AgNPs or corresponding carriers, incubated for indicated time points, then used to infect GMK-AH1 cells and determine PFU/ml in comparison to HSV-2 infected cultures. The data are shown as means from three independent experiments ± SEM. * represents significant differences with <i>p</i>≤0.05, while ** <i>p</i>≤0.001.</p

Transmission electron microscopy (TEM) images of silver nanoparticles and DLS histograms.

<p>(A) 13±5 (13) nm, (B) 33±7 (33) nm, (C) 46±9 (46) nm and (D) 10±1–65±10 nm AgNPs.</p

Antiviral effects of tannic-acid modified AgNPs require direct interaction.

<p>(A) Schematics and results for pre-treatment experiments. The results are expressed as % of HSV-2 infected control in 291.03C cells pre-treated with 2.5 µg/ml of tannic acid-modified 13 nm, 33 nm and 46 nm AgNPs or respective carriers for 2 h, then infected with HSV-2. (B) Schematics for post-treatment experiments. The results are expressed as percentage of viral inhibition in HSV-2 infected 291.03C cell cultures, in which complete medium containing 33 nm and 46 nm AgNPs (5 µg/ml) and 13 nm AgNPs (2.5 µg/ml) or respective carriers were added at the indicated time points for up to 24 h. The data are shown as means from three independent experiments ± SEM. * represents significant differences with p≤0.05.</p

Cytotoxicity and anti-HSV-2 activity of tannic acid-modified 13, 33, 46 nm AgNPs, unmodified 10–65 nm AgNPs or corresponding carriers in 291.03C cells.^*

<p>* The values shown are means from three independent experiments with each treatment performed in triplicate.</p>†<p>Cytotoxic effects were evaluated by neutral red assay to determine the concentration of 50% cellular cytotoxicity (CC₅₀) of the tested compounds.</p>‡<p>Antiviral effects were evaluated by plaque assay to determine the effective concentration that achieved 50% inhibition (EC₅₀) against HSV-2 infection.</p>§<p>SI. selectivity index.</p><p>CC₅₀/EC₅₀.</p