HERV-W Envelope Triggers Abnormal Dopaminergic Neuron Process through DRD2/PP2A/AKT1/GSK3 for Schizophrenia Risk

An increasing number of studies have begun considering human endogenous retroviruses (HERVs) as potential pathogenic phenomena. Our previous research suggests that HERV-W Envelope (HERV-W ENV), a HERV-W family envelope protein, is elevated in schizophrenia patients and contributes to the pathophysiology of schizophrenia. The dopamine (DA) hypothesis is the cornerstone in research and clinical practice related to schizophrenia. Here, we found that the concentration of DA and the expression of DA receptor D2 (DRD2) were significantly higher in schizophrenia patients than in healthy individuals. Intriguingly, there was a positive correlation between HERV-W ENV and DA concentration. Depth analyses showed that there was a marked consistency between HERV-W ENV and DRD2 in schizophrenia. Studies in vitro indicated that HERV-W ENV could increase the DA concentration by regulating DA metabolism and induce the expression of DRD2. Co-IP assays and laser confocal scanning microscopy indicated cellular colocalization and a direct interaction between DRD2 and HERV-W ENV. Additionally, HERV-W ENV caused structural and functional abnormalities of DA neurons. Further studies showed that HERV-W ENV could trigger the PP2A/AKT1/GSK3 pathway via DRD2. A whole-cell patch-clamp analysis suggested that HERV-W ENV enhanced sodium influx through DRD2. In conclusion, we uncovered a relationship between HERV-W ENV and the dopaminergic system in the DA neurons. Considering that GNbAC1, a selective monoclonal antibody to the MSRV-specific epitope, has been promised as a therapy for treating type 1 diabetes and multiple sclerosis (MS) in clinical trials, understanding the precise function of HERV-W ENV in the dopaminergic system may provide new insights into the treatment of schizophrenia.

Narciclasine induces autophagy-mediated apoptosis in gastric cancer cells through the Akt/mTOR signaling pathway

Background Gastric cancer is a common gastrointestinal cancer and currently has the third-highest mortality rate. Research shows that the natural compound narciclasine has a variety of biological activities. The present study aimed to investigate the effect of narciclasine on gastric cancer cells and its molecular mechanisms and determine whether this compound could be a novel therapy for gastric cancer. Methods MTT and clone assays were employed to detect the proliferation of gastric cancer cells. The cell apoptosis was detected by flow cytometry. The formation of autophagosomes and autophagosomal lysosomes was observed by transmission electron microscopy and laser confocal scanning microscopy. Western blotting was used to detect the expression of apoptosis, autophagy and Akt/mTOR pathway-related proteins. Results In this study, we found that narciclasine could inhibit the proliferation of gastric cancer cells and promote apoptosis in gastric cancer cells. Further experiments showed that narciclasine promoted the levels of autophagy proteins LC3-II, Atg-5 and Beclin-1, reduced the expression of the autophagy transporter p62, and increased autophagic flux. By using the autophagy inhibitors 3-MA and CQ, it was shown that narciclasine could induce autophagy-mediated apoptosis in gastric cancer cells. Finally, we found that narciclasine had no significant effects on the total content of Akt and mTOR in gastric cancer cells, and it involved autophagy in gastric cancer cells by reducing the phosphorylation level of p-Akt and p-mTOR. Conclusions Narciclasine can induce autophagy-dependent apoptosis in gastric cancer cells by inhibiting the phosphorylation level of Akt/mTOR and thus reduce the proliferation of gastric cancer cells.

New sorbents for electrochromatography based on polymer-inorganic dielectric composites

Oligomeric diisocyanate based coatings with different contents of barium titanate (BaTiO3) submicron sized particles as a ferroelectric filler are synthesized on poly(dimethylsiloxane) (PDMS) supports. The study of thus obtained coatings using confocal scanning electron microscopy allowed the characterization of their morphology and features of BaTiO3 particles distribution in the polymer binder, including the determination of threshold filler contents corresponding to the formation of an infinite cluster, matrix-island and chain-like structures as well as the percolation. Dielectric permittivity and dielectric losses of the composites are measured and studied depending on BaTiO3 filler content and relating structural features.

Semi-Automated Live Tracking of Microglial Activation in CX3CR1GFP Mice During Experimental Autoimmune Encephalomyelitis by Confocal Scanning Laser Ophthalmoscopy

Confocal scanning laser ophthalmoscopy (cSLO) is a non-invasive technique for real-time imaging of the retina. We developed a step-by-step protocol for the semi-automatic evaluation of myeloid cells in cSLO images from CX3CR1GFP mice, expressing green fluorescent protein (GFP) under control of the endogenous CX3C chemokine receptor 1 locus. We identified cSLO parameters allowing us to distinguish animals with experimental autoimmune encephalomyelitis (EAE) from sham-treated/naïve animals. Especially cell count (CC) and the total microglial area (SuA) turned out to be reliable parameters. Comparing the cSLO results with clinical parameters, we found significant correlations between the clinical EAE score and the SuA and of the inner retinal layer thickness, measured by optical coherence tomography, with the CC as well as the SuA. As a final step, we performed immunohistochemistry to confirm that the GFP-expressing cells visualized by the cSLO are Iba1 positive and validated the step-by-step protocol against manual counting. We present a semi-automatic step-by-step protocol with a balance between fast data evaluation and adequate accuracy, which is optimized by the option to manually adapt the contrast threshold. This protocol may be useful for numerous research questions on the role of microglial polarization in models of inflammatory and degenerating CNS diseases involving the retina.

Bilateral Corneal Ghost Vessels in an Otherwise Healthy Child

We report a rare case of bilateral corneal ghost vessels in a 6–year-old child with an unremarkable past ocular and past medical history. This study was a single observational case report. A 6-year-old girl was referred to our clinic for further evaluation, due to suboptimal visual acuity in both eyes. Her past medical and ocular history revealed no systemic, inflammatory, infectious, or degenerative disorders. Slit-lamp examination revealed regressed blood vessels (“ghost vessels”) in the anterior and mid-corneal stroma as the only pathologic finding. Confocal scanning microscopy of both corneas demonstrated scattered branching railroad-shaped ghost vessels at the level of the middle and anterior stroma. Complete systemic workup was performed for the patient. No identifiable risk factor for the development of corneal vascularization was found. According to our findings, we assume that in our patient, vasculogenesis occurred due to angioblast invasion to the presumptive cornea due to disequilibrium in mechanisms involved in vascular patterning during embryonic development.

Study of cytotoxic activity of Ti–13Nb–13Zr medical alloy with different surface finishing techniques

The characterization of materials surface is essential, as the initial in vivo response is highly dependent on surface properties. Surface topography is a key aspect that influences the response of cells to products resulting from interaction with the surface of a titanium alloy, including parameters such as adhesion, spread, migration, proliferation, and differentiation of cells. Various surface modifications are used to improve the interface properties between MC3T3 and NHDF cells and the Ti–13Nb–13Zr-based surface. Among the techniques discussed in this paper, scanning electron microscopy, laser confocal scanning microscopy, and computed tomography are adequate to investigate materials topography at different scale levels. Chemical characterization of the outer layers of Ti–13Nb–13Zr samples was performed with X-ray photoelectron spectrometry. Studies have shown that the surfaces resulting from the treatment enabling the formation of titanium oxide and zirconium oxide show the lowest cytotoxicity. Implants made from the new generation of titanium alloy, not containing toxic elements, with the use of surface modification could be an essential innovation in implantology.

Physicochemical properties and antimicrobial activity of calcium hydroxide pastes in association with other compounds

Aim: This study aimed to evaluate the pH, Ca2+ release, solubility, and antimicrobial activity of calcium hydroxide (Ca(OH)2) pastes in association with different substances. Methods: Sixty acrylic teeth (n=10) were filled with pastes that associated Ca(OH)2 with the following substances: benzalkonium chloride 5% (G1) and 50% (G2) both in propylene glycol, arnica glycolic extract (G3), green tea glycolic extract (G4), Calen/PMCC™ (G5), and Calen™ (G6). In the group G1 to G4 were used 1g of Ca(OH)2 powder with 0,8g of vehicle. pH and Ca2+ release was measured after 7, 15, and 30 days. For solubility, micro-CT was used immediately and at the periods of 7, 15, and 30 days. For the antimicrobial analysis, a biofilm of E. faecalis was induced in vitro on bovine dentin discs. Live/dead viability dye and confocal scanning microscopy were used. Results: The highest pH values occurred on the first 7 days, and the G6, G1, G3, and G5 presented the highest pH values at this period (P <0.05). Ca2+ release was higher in all groups at 7 days, with the highest values observed in G1, G5, and G6. The volume of all pastes showed no significant difference in the intragroup analysis at 7 and 15 days (P <0.05). G1 and G2 showed the highest antimicrobial action (P <0.05). For the biovolume, there was difference between the G6 and the other groups (P >0.05) with G1 presenting the lowest values. Conclusion: Benzalkonium chloride 5% increases the antimicrobial action of the Ca(OH2), without impairs physicochemical properties.