F1 Cells Research Articles

The Effect of Valproate Sodium on the Electrical Activity of Helix aspersa F1 Neurons in a Pentylenetetrazole-Induced Epileptic Model Using an Intracellular Recording System

Background: Epilepsy is a chronic central nervous system disorder with a high prevalence in modern society. Despite using common anticonvulsant drugs, its control is not adequately achieved. Animal models for seizures play the leading role in advancing our understanding of the cellular mechanisms of epilepsy. The present study was an attempt to elucidate the electrophysiological mechanism of the effect of sodium valproate on the cellular model of epilepsy. Understanding the cellular mechanisms of this drug may help clarify the pharmacological screening of other drugs. Methods: The intracellular recording was made from F1 cells of garden Helix aspersa in the presence of Ringer solution. Following the extracellular application of valproate sodium at a concentration of 10 mM after and before the use of (25mM) epileptogenic agent (pentylenetetrazol (PTZ)), we evaluated its effect on paroxysmal depolarization shift (PDS) and electrophysiological characteristics. Results: These results showed that valproate sodium could reduce neuronal excitability. It could significantly hyperpolarize rest action potential by decreasing the frequency of firing rate and increasing the amplitude of afterhyperpolarization (AHP) and can prevent depolarization of rest action potential by PTZ. Conclusions: The results suggested that valproate sodium could reduce the PTZ-induced hyperexcitation by hyperpolarization of resting membrane potential (RMP), a reduction in AHP amplitude, and firing the frequency.

Bifunctional folic-conjugated aspartic-modified Fe3O4 nanocarriers for efficient targeted anticancer drug delivery.

Functionalization of nanocarriers has been considered the most promising way of ensuring an accurate and targeted drug delivery system. This study reports the synthesis of bifunctional folic-conjugated aspartic-modified Fe3O4 nanocarriers with an excellent ability to deliver doxorubicin (DOX), an anticancer drug, into the intercellular matrix. Here, the presence of amine and carboxylate groups enables aspartic acid (AA) to be used as an efficient anchoring molecule for the conjugation of folic acid (FA) (EDC–NHS coupling) and DOX (electrostatic interaction). Based on the results, surface functionalization showed little effect on the physicochemical properties of the nanoparticles but significantly influenced both the loading and release efficiency of DOX. This is primarily caused by the steric hindrance effect due to large and bulky FA molecules. Furthermore, in vitro MTT assay of B16–F1 cell lines revealed that FA conjugation was responsible for a significant increase in the cytotoxicity of DOX-loaded nanocarriers, which was also found to be proportional to AA concentration. This high cytotoxicity resulted from an efficient cellular uptake induced by the over-expressed folate receptors and fast pH triggered DOX release inside the target cell. Here, the lowest IC50 value of DOX-loaded nanocarriers was achieved at 2.814 ± 0.449 μg mL−1. Besides, further investigation also showed that the drug-loaded nanocarriers exhibited less or no toxicity against normal cells.

Synthesize and multi-spectroscopic studies of zinc-naproxen nanodrug as DNA intercalator agent

The zinc-naproxen complex as a nano-drug (NanoD) was synthesized successfully via effective ultrasound-assisted processes. The chemicophysical properties of the NanoD were determined using FT-IR, XRD, SEM, DLS, and EDX mapping analyses. The results confirmed the formation of the 55 nm NanoD laminates. The interaction of the obtained NanoD with calf thymus deoxyribonucleic acid (CT-DNA) was studied as well. Structural and topography changes of DNA in interaction with the NanoD were investigated by atomic force microscopy (AFM). The results of electronic absorption spectroscopy, the DNA-viscosity studies, and competition fluorescence spectroscopy showed that CT-DNA binds to the NanoD through the intercalative binding mode. The data of AFM analysis indicated swollen CT-DNA upon interaction with the NanoD. The in vitro investigation of cytotoxicity of the NanoD on HT-29, Hep G2, and B16–F10 cancer cells as well as normal HFF-1 cells. The obtained results demonstrated high cytotoxicity activity of the NanoD than that of cisplatin in the HT-29 cell line, especially at lower concentrations. On the B16–F10 cell line at lower concentrations (up to 8 μg mL−1), it is comparable to cisplatin and on the Hep G2 cell line and normal HFF-1 cell line at all concentrations, cytotoxicity of cisplatin is more than NanoD.

Development, optimization, and in vitro evaluation of atorvastatin calcium and vinpocetine codelivery by solid lipid nanoparticles for cancer therapy

BackgroundThe main objective of the present study was to formulate, optimize and characterize solid lipid nanoparticles (SLNs) loaded with Atorvastatin Calcium (ATS) and Vinpocetine (VIN) as a potential drug delivery system to improve its solubility and assess its anti-tumor activity on cell lines. The SLNs were formulated by emulsification with high speed homogenization followed by probe sonication. Central composite design was selected for optimization. Drug: lipid ratio, surfactant: co-surfactant ratio and homogenization speed were considered critical process parameters (CPP) to study the effects on critical quality attributes (CQA) of SLNs i.e. particle size, percent entrapment efficiency (% EE) and percent drug loading (% DL).ResultsThe optimized (F3) SLNs formulations were characterized by transmission electron microscopy (TEM), X- ray diffraction (X-RD), in vitro drug release by dialysis bag method and stability studies. In vitro cell line studies were performed on HepG2, MCF 7 and melanoma B16 F10 cell line. The optimized F3 formulation showed a particle size of 323 ± 6 nm, poly dispersity index (PDI) 0.333 ± 0.02, Zeta potential (ZP) − 30.4 ± 0.66 emv with % EE 64.69 ± 1.1; 65.98 ± 0.91 of ATS and VIN respectively. In vitro release (F3) of ATS and VIN in PBS pH 7.4 was found to be 89.45% and 91.86%, respectively, up to 24 h.ConclusionsIn vitro cell line study demonstrated that SLNs enhanced the anti-cancer activity of ATS, VIN on all the stated cell lines when compared with free drugs. Combination index (CI) for HEPG2 was 0.8, which signified synergistic effect. The results exhibited that SLNs is effective, stable and had enhanced activity against HepG2, MCF 7 and melanoma B16 F10 cell lines.

W/O/W double emulsion-loaded alginate capsules containing Lactobacillus plantarum and lipophilic sea buckthorn (Hippophae rhamnoides L.) pomace extract in different phases.

In this study, double emulsion containing L. plantarum F1 cells and prebiotic mannitol in the inner water phase, lipophilic sea buckthorn pomace extract as an antioxidant in the oil phase, and alginate in the outer water phase showed high encapsulation yield (82.19%), good cell survival rate (76.99%) and low chemical degradation of the oil (peroxide value - 3.8 meq O2/kg fat) after 42 days of storage. Gelation of the outer water phase enhanced the viability of L. plantarum F1 cells both during storage and under gastrointestinal conditions due to strong physical barrier formation. Encapsulated L. plantarum F1 viability throughout the 30-day storage period decreased to the value meeting the minimum required dose for probiotics. In vitro digestion of the loaded alginate capsules showed high survival rate of encapsulated cells under gastric conditions and significant reduction at the end of the duodenal phase of digestion.

The role of Notch ligand Jagged1 in osteosarcoma proliferation, metastasis, and recurrence

BackgroundOsteosarcoma is the most common primary bone cancer occurring in young adults and the 5-year survival rate of patients with metastatic osteosarcoma is less than 30% due to high metastatic recurrence and drug resistance. Notch is a highly conserved cell to cell signaling pathway in evolution, and Jagged1 is an important ligand of Notch. Although some studies have found that Notch receptors and ligands including Jagged1 were highly expressed in osteosarcoma tissues and osteosarcoma cells, the role of Jagged1 in osteosarcoma progression and metastasis are still not clear.MethodsTumor tissues were collected from 68 patients and immunohistochemical staining was employed to group these patients by expression of Jagged1. Real-time quantitative PCR and Western blotting were used to detect the expression of Jagged1. We used siRNA to knockdown the expression of Jagged1 in F5M2 cells. Colony formation assay and MTT were employed to detect and analyze the proliferation of F5M2 cells with or without knockdown of Jagged1. Transwell assay were used to detect the migration and invasion of F5M2 cells.ResultsIn this study, we found that the high expression of Jagged1 is closely related to the metastasis and recurrence of osteosarcoma in 68 clinical specimens. The expression of Jagged1 in F5M2 cells with high metastasis was significantly higher than that in F4 cells with low metastasis. Knockdown of Jagged1 led to lower ability of proliferation, migration, and invasion in F5M2 cells.ConclusionThe high expression of Jagged1 is closely related to the metastasis and recurrence of osteosarcoma. Knockdown of Jagged1 significantly reduced the proliferation, migration, and invasion of osteosarcoma cells. Our results suggested that knockdown of Jagged1 may be a potentially effective treatment for metastatic osteosarcoma.

Water assessment of the Itapemirim River/Espírito Santo (Brazil): abiotic and toxicogenetic aspects.

The Itapemirim River is considered one of the most important water resources in the state of Espírito Santo, Brazil. However, environmental problems due to continuous anthropogenic contamination are threatening its potential use. This study assessed water quality by analyzing abiotic and toxicogenetic aspects of the water from four stations along the river. Samples were collected in both dry and rainy seasons. Most of the abiotic variables were below the threshold established by CONAMA Resolution No. 357/2005, and so were most of the metals. However, Al and Cu contents were above those allowed by legislation, ranging from 0.2 to 0.9mg/L. Regarding toxicogenetic aspects, genotoxic effects were observed in meristematic cells of Allium cepa, in micronucleus test and comet assay of Oreochromis niloticus, and CHO-K1 cells. Mutagenic effects were significant at RI 02 (0.34), RI 03 (0.46), and RI 04 (0.12) stations on the first campaign in A. cepa F1 cells, compared to the negative control (0.0). The second campaign revealed the same results, but with the addition of samples from RI 01 (0.17) and RI 03 (0.18) showing mutagenicity in the micronucleus test with fish erythrocytes when compared to the negative control (0.3). Essentially, all the samples evaluated in both campaigns showed damage in A. cepa, O. niloticus, and CHO-K1 cells, thus demonstrating that the water quality of the Itapemirim River is compromised and requires action plans for its recovery.

Homogeneous catalytic hydrogenation of lipids in aqueous dispersions and bacterial cell membranes with an efficient water-soluble Pd(II)-sulfosalan catalyst, Na2[Pd(HSS)

The recently synthesized water-soluble Na2[Pd(HSS)] (Na2HSS = hydrogenated sulfonated salen) was shown to be a non-toxic, active catalyst for modification of model and biomembranes by homogeneous catalytic hydrogenation. Hydrogenation of the unsaturated fatty acyl residues in soy-bean lecithin liposomes and in biomembranes of intact Pseudomonas putida F1 cells was accompanied by substantial cis-trans isomerization around the CC double bonds. The hydrogenations could be run in aqueous media under mild conditions (25 °C, 1 bar H2). Partial saturation (up to 10% conversion) of the membrane lipids of P. putida F1 did not damage the cells.

The anti-Zika virus and anti-tumoral activity of the citrus flavanone lipophilic naringenin-based compounds

Flavonoids are natural products widely recognized for their plurality of applications such as antiviral, antiproliferative, antitumor activities and, antioxidant properties. The flavanone naringenin is presented in citrus fruits and has been studied to combat recurrent diseases that still lack effective treatment. Research groups have been investing efforts to the development of new, safe and active candidates to combat these agents or conditions and despite good results recently reported against the Zika virus (ZIKV) and tumor cells, the use of citrus naringenin is limited due to its low bioavailability. Structural exchanges through functionalization, for example, attaching lipophilic groups instead of hydroxyl groups, can further enhance biological properties.Here, the synthesis and characterization of regioselective naringenin mono-7-O-ethers and both mono and di-fatty acid esters, structurally lipophilic ones were demonstrated. Finally, in vitro studies of anti-ZIKV action and antiproliferative activities against melanoma (B16–F10) and breast carcinoma (4T1) cells showed the ether derivatives were actives, with IC50 ranging from 6.76, 18.5 and 22.6 μM to 28.53, 45.1 and 32.3 μM referring to ZIKV, B16–F10 and 4T1 cell lines, respectively. The lipophilic ethers present the ability to inhibit selectively ZIKV-replication in human cells and inhibitions.This class of modifications in flavonoid molecules could be further explore in the future development of specific anti-ZIKV compounds.

Abstract 628: Imipramine induced apoptosis and inhibits invasion via suppression of EGFR/ERK/NF-kappa B activation in non-small cells lung cancer

Abstract Imipramine was known as an anti-depression agent; interestingly, recent studies indicated the potential of various tumors growth inhibition. However, the anticancer mechanism of imipramine on non-small cell lung cancer (NSCLC) is remaining ambiguous. Thus, the aim of the study was to determent the effects and underlying mechanism of imipramine on human NSCLC CL1-5/F4 cells. MTT assays were used for testing the cell viability of imipramine. The activation and expression of apoptosis, anti-apoptosis, DNA damage, cellular calcium, and metastasis-associated proteins were observed by flow cytometry, immunofluorescence stain, comet assay and Western blotting assay. The inhibition of nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) after imipramine treatment was performed by NF-κB reporter gene assay and immunofluorescence nuclear translocation assay on CL1-5/F4/NF-κB/luc2 and CL1-5/F4 wild type cells. Moreover, the ability of migration and invasion after imipramine treatment were also tested by trans-well assay and wound healing assay. In animal experiment, CL1-5/F4/NF-κB/luc2 cells were subcutaneously injection into nude mice right flask and treated with imipramine by oral. In addition, the standard treatment erlotinib was also used as a positive control to compare treatment efficacy of imipramine. In vivo NF-κB activation was investigated by optical image. Mice were sacrificed and tumors were extracted for size and IHC staining evaluation on day 14. Our results demonstrated that imipramine significantly reduced cell viability, cell migration and invasion. Reporter gene assay and Western blotting revealed imipramine may decrease NF-κB activation through inhibition of EGFR/ERK/NF-κB pathway. Imipramine also diminished the phosphorylation of ERK, the expression of anti-apoptotic (c-FLIP and XIAP), angiogenesis (VEGF), and metastasis-associated proteins (MMP-9). Additionally, the activity of PARP-1 that induced by imipramine was associated with DNA damage. Furthermore, imipramine was also triggered extrinsic (Fas/FasL, and caspase-8), intrinsic [mitochondrial membrane potential (ΔΨm)] apoptosis pathways, and the accumulation of cellular calcium. Tumor growth and NF-κB activation were both suppressed by imipramine treatment. Tumors IHC stain showed p-EGFR, p-ERK and p-NF-κB expressions level were reduced in imipramine treatment group as compared to vehicle. Most importantly, although body weight was remained unchanged in three treatment group, the cellular necrosis nucleus was found in erlotinib treated liver pathology H&E stain which was not found in control and imipramine groups. Taken together, these results suggested that imipramine can induce NSCLC apoptosis by activating extrinsic/intrinsic apoptosis mechanisms, DNA damage effect and the accumulation of cellular calcium concentration. In addition, imipramine may also suppress tumor proliferation, invasion and migration ability. Most importantly, imipramine diminished tumor progression via inactivation of EGFR/ERK/NF-κB signaling on NSCLC in vitro and in vivo. Citation Format: Po-Fu Yueh, I-Tsang chiang, Fei-Ting Hsu. Imipramine induced apoptosis and inhibits invasion via suppression of EGFR/ERK/NF-kappa B activation in non-small cells lung cancer [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 628.

Isolation and identification of Bengle (Zingiber cassumunar roxb) as a stimulant in phagocytic activity of macrophages

Immunomodulators are pharmacological agents that modify or regulate the immune system through stimulating the functioning of the immune system and, at the same time, inhibiting excessive immune responses. This study was conducted to determine the active compound in Z. cassumunar that is responsible for increasing the immune system based on the parameters of phagocytic activity. The isolation method began with fractionation, which involved extraction with ethanol and successive fractionation with hexane and chloroform. Z. cassumunar extract, hexane fraction, and chloroform fraction were tested on mice macrophage cells for their phagocytic functions. The phagocytic activity of macrophages was measured by active phagocytic cells (averagely 39.194 ±1.597, 27.923 ±2.941, and 62.090 ±6.947) and phagocytic index (in a row, averagely 47.513 ±2.844, 41.129 ±7.195, and 101.527 ±10.555). The results showed that the Z. cassumunar extract, hexane fraction, and chloroform fraction exhibited more significant phagocytic activities of macrophages (p <0.05) compared with the normal group. Since the chloroform fraction showed the best result, this fraction was further separated by column chromatography. This procedure yielded five sub-fractions, namely F1, F2, F2C, F3, and F4. Based on the phagocytic activity testing, the results were as follows: (1) the active phagocytic cells of F1, F2, F2C, F3 and F4 were 18.860 ±3.191, 27.077 ±4.482, 15.749 ±3.026, 64.333 ±1.780, and 44.943 ±2.944, respectively, and (2) the phagocytic indices were 30.0249 ±3.4231, 44.5969 ±8.3646, 24.5597 ±5.4487, 102.7447 ±1.0806, and 76.5007 ±4.7293. Because F3 produced the best result, this subfraction was then identified using 1H-NMR and 13C-NMR. The identification results showed that F3 was (E)-4-(3,4-dimethoxyphenyl)but-3-en-1-ol as an active compound.

Meroterpenoids from the fruits of Psoralea corylifolia

Fourteen new meroterpenoid compounds, corypsoriols A–N (1–14), and eight known ones (15–22), were isolated from the fruits of Psoralea corylifolia. The absolute configurations of these new compounds were confirmed by NOESY spectrum, ECD calculation, and single crystal X-ray diffraction. Corypsoriols A and B were a novel skeleton which possessed a 6/5/6 tricyclic system, and their plausible biosynthetic pathway was proposed. The cytotoxicity of uncyclized meroterpenoids were stronger than those of cyclized ones, and the half maximal inhibitory concentration values of bakuchiol (15) against NCI–N87, HepG2, HCT-116, Hela and B16–F10 cell lines were in the range of 5–20 μM.

Kaji-Ichigoside F1 and Rosamultin Protect Vascular Endothelial Cells against Hypoxia-Induced Apoptosis via the PI3K/AKT or ERK1/2 Signaling Pathway.

As a pair of differential isomers, Kaji-ichigoside F1 and Rosamultin are both pentacyclic triterpenoids isolated from the subterranean root of Potentilla anserina L., a plant used in folk medicine in western China as antihypoxia and anti-inflammatory treatments. We demonstrated that Kaji-ichigoside F1 and Rosamultin effectively prevented hypoxia-induced apoptosis in vascular endothelial cells. We established a hypoxia model, using EA.hy926 cells, to further explore the mechanisms. Hypoxia promoted the phosphorylation of AKT, ERK1/2, and NF-κB. In hypoxic cells treated with Kaji-ichigoside F1, p-ERK1/2 and p-NF-κB levels were increased, while the level of p-AKT was decreased. Treatment with Rosamultin promoted phosphorylation of ERK1/2, NF-κB, and AKT in hypoxic cells. Following the addition of LY294002, the levels of p-AKT, p-ERK1/2, and p-NF-κB decreased significantly. Addition of PD98059 resulted in reduced levels of p-ERK1/2 and p-NF-κB, while p-AKT levels were increased. Pharmacodynamic analysis demonstrated that both LY294002 and PD98059 significantly inhibited the positive effects of Kaji-ichigoside F1 on cell viability during hypoxia, consistent with the results of hematoxylin-eosin (H&E) staining, DAPI staining, and flow cytometry. The antihypoxia effects of Rosamultin were remarkably inhibited by LY294002 but promoted by PD98059. In Kaji-ichigoside F1- and Rosamultin-treated cells, Bcl2 expression was significantly upregulated, while expression of Bax and cytochrome C and levels of cleaved caspase-9 and cleaved caspase-3 were reduced. Corresponding to pharmacodynamic analysis, LY294002 inhibited the regulatory effects of Kaji-ichigoside F1 and Rosamultin on the above molecules, while PD98059 inhibited the regulatory effects of Kaji-ichigoside F1 but enhanced the regulatory effects of Rosamultin. In conclusion, Kaji-ichigoside F1 protected vascular endothelial cells against hypoxia-induced apoptosis by activating the ERK1/2 signaling pathway, which positively regulated the NF-κB signaling pathway and negatively regulated the PI3K/AKT signaling pathway. Rosamultin protected vascular endothelial cells against hypoxia-induced apoptosis by activating the PI3K/AKT signaling pathway and positively regulating ERK1/2 and NF-κB signaling pathways.

Indirectly Activated Treg Allow Dominant Tolerance to Murine Skin-grafts Across an MHC Class I Mismatch After a Single Donor-specific Transfusion.

Tolerance induced in stringent animal transplant models using donor-specific transfusions (DST) has previously required additional immunological manipulation. Here, we demonstrate a dominant skin-allograft tolerance model induced by a single DST across an major histocompatibility class I mismatch in an unmanipulated B6 host. C57BL/6 (H-2) (B6) mice were injected intravenously with splenocytes from B6.C.H-2 (H-2k) (bm1) or F1 (B6 × bm1) mice before skin transplantation. Mice were transplanted 7 days postinjection with donor (bm1 or F1) and third-party B10.BR (H-2) skin grafts. B6 hosts acutely rejected skin grafts from B6.C.H-2 (bm1) and F1 (B6 × bm1) mice. A single transfusion of F1 splenocytes into B6 mice without any additional immune modulation led to permanent acceptance of F1 skin grafts. This graft acceptance was associated with persistence of donor cells long-term in vivo. The more rapid removal of DST bm1 cells than F1 cells was reduced by natural killer-cell depletion. Tolerant grafts survived an in vivo challenge with naive splenocytes. Both CD4CD25 and CD4CD25 T cells from F1 DST treated B6 mice suppressed alloproliferation in vitro. Tolerance was associated with expansion of peripheral Foxp3CD4CD25 regulatory T cells (Treg) and increased forkhead box P3 (Foxp3) expression in tolerant grafts. In tolerant mice, Foxp3 Treg arises from the proliferation of indirectly activated natural Foxp3 Treg (nTreg) and depletion of Foxp3 Treg abrogates skin-graft tolerance. This study demonstrates that the persistence of transfused semiallogeneic donor cells mismatched at major histocompatibility class I can enhance tolerance to subsequent skin allografts through indirectly expanded nTreg leading to dominant tolerance without additional immunological manipulation.

Proteomic alteration of porcine intestinal epithelial cells after pretreatment with Lactobacillus plantarum followed by infection with enterotoxigenic Escherichia coli F4

Enterotoxigenic Escherichia coli (ETEC) F4 causes diarrhea in infants and weaned piglets. The technique of isobaric tags for relative and absolute quantitation (iTRAQ) was used in this study to determine the differentially expressed proteins in porcine intestinal epithelial cells (IPEC-J2) after pretreatment with Lactobacillus plantarum (LP) followed by challenge with ETEC F4. A total of 4771 proteins were identified in IPEC-J2 cells, with 90, 105, and 134 differentially expressed proteins in cells exposed to ETEC, LP, and LP + ETEC, respectively. The COG analysis divided the identified proteins into 20 categories. The GO and KEGG annotation indicated that most of the differentially expressed proteins were enriched in various biological metabolism including cell cycle control, cell division and differentiation. Additionally, western blotting analyses confirmed the reduced abundance of selected proteins of the mTOR and MAPK signal pathways affected by ETEC F4. Moreover, LP pretreatment increased JNK activation in IPEC-J2 cells infected with ETEC F4. These results may provide further insights into the mechanisms involved in the interaction between ETEC F4 and intestinal epithelial cells, and broaden the understanding of the protective effects of LP in alleviating ETEC-provoked diarrhea of piglets.

Bioactivity-Guided Isolation of Cytotoxic Phenanthrenes from Spiranthes sinensis.

Bioactivity-guided separation led to the isolation of six novel phenanthrenes, spiranthesphenanthrenes A-F (1-6), together with 19 known compounds, including seven phenanthrenes (7-13), one bibenzyl compound (14), five flavonoids (15-16 and 20-22), and six simple phenolic compounds (17-19 and 23-25), from the petroleum ether (PE) and ethyl acetate (EtOAc) extracts of Spiranthes sinensis (Pers.) Ames, an edible medicinal plant named "panlongshen" in Chinese that is popularly used in medicinal foods and herbal teas. The structures of the obtained compounds were identified on the basis of extensive NMR spectroscopy and HR-ESI-MS analyses. The cytotoxicities of the phenanthrenes (1-13), the bibenzyl compound (14) , and the flavonoids (15-16 and 20-22) toward SGC-7901, HepG2, and B16-F10 cell lines were examined in vitro. Compounds 1 and 7 exhibited moderate cytotoxic activities toward all of the selected cancer cell lines, and their IC50 values ranged from 19.0 ± 7.3 to 30.2 ± 5.6 μM. Spiranthesphenanthrene A (1) exhibited higher cytotoxic activity than the positive control cisplatin toward the B16-F10 cell line (IC50 = 19.0 ± 7.3 μM). A wound healing assay revealed the inhibition of the migration of B16-F10 cancer cells in a time- and dose-dependent pattern by treatment with 2.5, 5, and 10 μM solutions of compound 1 for 24 and 48 h, respectively. Western blots revealed that compound 1 obviously increased the level of the E-cadherin protein (an epithelial marker) and decreased the levels of the vimentin and N-cadherin proteins (mesenchymal markers). Furthermore, the level of the transcription factor Snail was also obviously decreased by compound 1 in a dose-dependent manner. Taken together, compound 1 inhibits the migration of B16-F10 cancer cells, which may be closely related to the inhibition of the epithelial-mesenchymal transition. Compound 1 represents a promising drug candidate for the prevention of tumor metastasis.

Global Proteomic and Methylome Analysis in Human Induced Pluripotent Stem Cells Reveals Overexpression of a Human TLR3 Affecting Proper Innate Immune Response Signaling.

When considering the clinical applications of autologous cell replacement therapy of human induced pluripotent stem cells (iPSC)‐derived cells, there is a clear need to better understand what the immune response will be before we embark on extensive clinical trials to treat or model human disease. We performed a detailed assessment comparing human fibroblast cell lines (termed F1) reprogrammed into human iPSC and subsequently differentiated back to fibroblast cells (termed F2) or other human iPSC‐derived cells including neural stem cells (NSC) made from either retroviral, episomal, or synthetic mRNA cell reprogramming methods. Global proteomic analysis reveals the main differences in signal transduction and immune cell protein expression between F1 and F2 cells, implicating wild type (WT) toll like receptor protein 3 (TLR3). Furthermore, global methylome analysis identified an isoform of the human TLR3 gene that is not epigenetically reset correctly upon differentiation to F2 cells resulting in a hypomethylated transcription start site in the TLR3 isoform promoter and overexpression in most human iPSC‐derived cells not seen in normal human tissue. The human TLR3 isoform in human iPSC‐NSC functions to suppress NF‐KB p65 signaling pathway in response to virus (Poly IC), suggesting suppressed immunity of iPSC‐derived cells to viral infection. The sustained WT TLR3 and TLR3 isoform overexpression is central to understanding the altered immunogenicity of human iPSC‐derived cells calling for screening of human iPSC‐derived cells for TLR3 expression levels before applications. stem cells 2019;37:476–488

Genome composition and pollen viability of Jatropha (Euphorbiaceae) interspecific hybrids by Genomic In Situ Hybridization (GISH).

Interspecific hybridization is required for the development of Jatropha curcas L. improved cultivars, due to its narrow genetic basis. The present study aimed to analyze the parental genomic composition of F1 and BC1F1 generations derived from interspecific crosses (J. curcas/J. integerrima and J. curcas/J. multifida) by GISH (Genomic In Situ Hybridization), and the meiotic index and pollen viability of F1 hybrids. In F1 cells from both hybrids, 11 chromosomes of each parental was observed, as expected, but chromosome rearrangement events could be detected using rDNA chromosome markers, suggesting unbalanced cells. In the BC1F1, both hybrids had 22 chromosomes, suggesting that only n = 11 gametes were viable in the next generation. However, GISH allowed the identification of three and two alien chromosomes in J. curcas//J. integerrima and J. curcas//J. multifida BC1F1 hybrids, respectively, suggesting a preferential transmission of J. curcas chromosomes for both hybrids. Pollen viability in F1 hybrids derived from J. curcas/J. integerrima crosses were higher (82-83%) than those found for J. curcas/J. multifida (68%), showing post-meiotic problems in these last hybrids, with dyads, triads, polyads, and micronuclei as post-meiosis results. The here presented cytogenetic characterization of interspecific hybrids and their backcross progenies can contribute to the selection of the best genotypes for future assisted breeding of J. curcas.

Characterization of Functional Effects of Two New Active Fractions Isolated From Scorpion Venom on Neuronal Ca2+ Spikes: A Possible Action on Ca2+-Dependent Dependent K+ Channels

Introduction:It is a long time that natural toxin research is conducted to unlock the medical potential of toxins. Although venoms-toxins cause pathophysiological conditions, they may be effective to treat several diseases. Since toxins including scorpion toxins target voltage-gated ion channels, they may have profound effects on excitable cells. Therefore, elucidating the cellular and electrophysiological impacts of toxins, particularly scorpion toxins would be helpful in future drug development opportunities.Methods:Intracellular recording was made from F1 cells of Helix aspersa in the presence of calcium Ringer solution in which Na+ and K+ channels were blocked. Then, the modulation of channel function in the presence of extracellular application of F4 and F6 toxins and kaliotoxin (KTX; 50 nM and 1 μM) was examined by assessing the electrophysiological characteristics of calcium spikes.Results:The two active toxin fractions, similar to KTX, a known Ca2+-activated K+ channel blocker, reduced the amplitude of AHP, enhanced the firing frequency of calcium spikes and broadened the duration of Ca2+ spikes. Therefore, it might be inferred that these two new fractions induce neuronal hyperexcitability possibly, in part, by blocking calcium-activated potassium channel current. However, this supposition requires further investigation using voltage clamping technique.Conclusion:These toxin fractions may act as blocker of calcium-activated potassium channels.

Characterization of Functional Effects of Two New Active Fractions Isolated From Scorpion Venom on Neuronal Ca2+ Spikes: A Possible Action on Ca2+-Dependent Dependent K+ Channels.

Introduction:Based on the literature, unihemispheric concurrent dual-site anodal transcranial Direct Current Stimulation (a-tDCSUHCDS) of primary Motor cortex (M1) and Dorsolateral Prefrontal Cortex (DLPFC) would be more efficient than conventional a-tDCS of M1 to induce larger and longer-lasting M1 corticospinal excitability. The main objective of the present study was to compare the effects of a-tDCSUHCDS and conventional M1 a-tDCS on the extent and durability of the motor sequence acquisition in healthy individuals.Methods:In this randomized sham-controlled study, healthy volunteers were randomly divided into three groups: experimental (a-tDCSUHCDS), control (M1 a-tDCS), and sham stimulation groups. The participants practiced serial response time task over three consecutive days when they simultaneously received a-tDCS. Using the skill measure, we assessed motor learning up to 4 weeks after the completion of experimental conditions.Results:Data analysis revealed that all groups exhibited the improved trend over the training course (P<0.001). There were no significant differences in skill acquisition among groups at post-intervention (P>0.05), while a significant improvement was observed between experimental and sham group at the retention time (P<0.05). Moreover, there were no significant differences between the control and two other groups with regard to the retention time (P>0.05).Conclusion:These results revealed a significant increase in the skill acquisition by a-tDCSUHCDS technique with regard to retention issue, which could be a valuable finding in neuro-rehabilitation field.