PE Cell Research Articles

Chitosan/Gold Hybrid Nanoparticles Enriched Electrospun PVA Nanofibrous Mats for the Topical Delivery of Punica granatum L. Extract: Synthesis, Characterization, Biocompatibility and Antibacterial Properties.

PurposeIntending to obtain Punica granatum L. extract (PE)-loaded drug delivery system of better impact and biomedical applicability, the current study reports the use of crosslinked PVA nanofibers (NFs) as platforms incorporating different amounts of biosynthesized PE-CS-gold nanoparticles (PE-CS-Au NPs).MethodsPE-conjugated CS-Au nanoparticles (PE-CS-Au NPs) were synthesized via green chemistry approach. The formation of PE-CS-Au NPs was confirmed by UV spectroscopy, DLS, SEM and STEM. PE-CS-Au NPs were then dispersed into polyvinyl alcohol (PVA) solution at different ratios, where the optimized ratios were selected for electrospinning and further studies. Crosslinking of PE-CS-Au NPs loaded PVA nanofibers (NFs) was performed via glutaraldehyde vapor. The morphology, chemical compositions, thermal stability and mechanical properties of PE-CS-Au NPs loaded NFs were evaluated by SEM, FTIR and DSC. Swelling capacity, biodegradability, PE release profiles, release kinetics, antibacterial and cell biocompatibility were also demonstrated.ResultsBy incorporating PE-CS-Au NPs at 0.6% and 0.9%, the diameters of the nanofibers decreased from 295.7±83.1 nm in neat PVA to 165.6±43.4 and 147.8±42.7 nm, respectively. It is worth noting that crosslinking and incorporation of PE-CS-Au NPs improved thermal stability and mechanical properties of the obtained NFs. The release of PE from NFs was controlled by a Fickian diffusion mechanism (n value ˂0.5), whereas Higuchi was the mathematical model which could describe this release. The antibacterial activity was found to be directly proportional to the amount of the incorporated PE-CS-Au NPs. The human fibroblasts (HFF-1) showed the highest viability (123%) by seeding over the PVA NFs mats containing 0.9% PE-CS-Au NPs.ConclusionThe obtained results suggest that the electrospun PVA NFs composites containing 0.9% PE-CS-Au NPs can be used as antibacterial agents against antibiotic-resistant bacteria, and as suitable scaffolds for cell adhesion, growth and proliferation of fibroblast populations.

Therapeutic effect of SP-8356 on pulmonary embolism-associated cardiac injury is mediated by its ability to suppress apoptosis and inflammation.

The cyclophilin A–CD147 interaction has been reported to be one of the most potential therapeutic targets for the treatment of acute pulmonary embolism. The signalling of extracellular signal‐regulated kinase 1/2 (ERK1/2) was also reported in the pathogenesis of cardiac injury. Since SP‐8356 is regarded as a novel Inhibitor of CD147‐Cyclophilin, the study aimed to evaluate potential therapeutic effects of SP‐8356 for pulmonary embolism‐associated cardiac injury. Western blot and immunohistochemistry were carried out to analyse the expression of MMP‐9, ERK1/2, phosphorylated ERK1/2 (p‐ERK1/2), P65, p‐P65, and CyA protein in PE cell and rat models under distinct conditions. Flow cytometry and TUNEL were carried out to examine the apoptosis of primary rat myocardiocytes and PE rat models under distinct conditions. CyA treatment on primary rat myocardiocytes remarkably raised the expression of MMP‐9, p‐ERK1/2 and p‐P65 protein expression; SP8536 treatment effectively restored the CyA‐induced up‐regulation of MMP‐9, p‐ERK1/2 and p‐P65 protein expression in primary rat myocardiocytes. Besides, flow cytometry analysis showed that SP8536 remarkably suppressed the CyA‐induced elevation of cell apoptosis rate of primary rat myocardiocytes. Moreover, SP8536 notably diminished the abnormal elevation of right ventricular systolic pressure (RVSP), Troponin I and Myeloperoxidase activity in PE rat models. Furthermore, SP‐8536 significantly restored the up‐regulation of MMP‐9, p‐ERK1/2, p‐P65, CyA protein and the cellular apoptosis in the PE rat model. Our study validated that SP‐8356 could suppress cell apoptosis and inflammatory response via down‐regulating the highly expressed MMP‐9, p‐ERK1/2, and p‐P65 and MMP‐9 in PE‐associated cardiac injury in a dose‐dependent manner.

Long non-coding RNA HOTAIR modulates the progression of preeclampsia through inhibiting miR-106 in an EZH2-dependent manner

AimsPreeclampsia (PE) accounts for the foremost cause of maternal and fetal mortality worldwide, whereas, there are no effective treatments for the disease yet. Long non-coding RNAs (lncRNAs) play critical roles in various human disorders, including PE. Here, we identified an up-regulated lncRNA HOTAIR, and explored its underlying mechanisms in PE. Main methodsqRT-PCR analysis was used to examine HOTAIR expression in PE tissues and cell lines. Trophoblast proliferation was examined by colony formation and 5-Ethynyl-2′-deoxyuridine (EdU) incorporation assays. Trophoblast migration and invasion was determined by transwell and wound healing assays. Bioinformatics analysis was performed to verify the regulation HOTAIR on miRNAs. The interaction between HOTAIR and EZH2 was detected using RNA immunoprecipitation assay (RIP). Chromatin immunoprecipitation (CHIP) assay was also performed to verify that the negative regulation of HOTAIR on miR-106a was dependent on the epigenetic repressor EZH2. Key findingsHOTAIR was up-regulated in PE placenta tissues, which repressed the proliferation, migration and invasion of trophoblast cells. HOTAIR significantly repressed miR-106a expression and the reduced miR-106a level was also observed in placentas from PE patients. Additionally, miR-106a mimic enhanced the migration and invasion of trophoblast cells. Further mechanistic analyses implied that the action of HOTAIR is moderately attributable to its repression of miR-106a via association with EZH2. SignificanceHigh level of HOTAIR repressed the proliferation, migration and invasion of trophoblast cells through targeting miR-106 in an EZH2-dependent manner, which may provide new insights into the roles of HOTAIR and miR-106a as potential regulators in PE.

Gastrodin improves preeclampsia-induced cell apoptosis by regulation of TLR4/NF-κB in rats.

To explain gastrodin improved cell apoptosis induced by preeclampsia in vivo and in vitro study. The PE and normal rats were injected with normal saline (Model), low‐dose gastrodin (Gas‐L), medium‐dose gastrodin (Gas‐M), and high‐dose gastrodin (Gas‐H) groups at 50, 100, or 200 mg/kg per day. The rat blood pressure and 24‐hr urine protein level were measured at pregnant days 10, 16, and 20. Evaluating pathology by H&E staining, the cell apoptosis by TUNEL, and MyD88 and NF‐κB (p65) proteins by IHC assay using H/R to simulate PE cell model. Measuring cell proliferation, apoptosis, and MyD88 and NF‐κB (p65) protein expression by MTT, flow cytometry, and WB assay. The SBP, DBP, and 24‐hr urine protein levels were significantly different in PE rats (p < .05). The SBP, DBP, and 24‐hr urine protein levels were significantly improved (p < .05) in vivo and in vitro. The positive apoptosis cells and apoptosis rate were significantly increased with MyD88 and NF‐κB (p65) proteins upregulation (p < .05). The positive apoptosis cells and apoptosis rate were significantly decreased with MyD88 and NF‐κB (p65) proteins depressing in gastrodin‐treated groups with dose‐dependent (p < .05). Gastrodin improves PE‐induced cell apoptosis and pathology changed via MyD88/NF‐κB pathway in vitro and in vivo study.

In vitro and In vivo evaluation of green-hydrothermal synthesized ZnO nanoparticles

ZnO nanoparticles synthesized using Gallic acid isolated from Phyllanthus emblica aqueous extract that designated as Pe ZnO, was subjected for structural and morphological characterization. For in vitro cyto-toxicity assessment of Pe ZnO, Hemolytic assay and Cell Counting Kit assay (CCK 8) on balb 3T3 fibroblasts were conducted while silkworm Bombyx mori larvae were used for in vivo bio-toxicity assay. In dentistry, clinically advocated ZnO (presented as D ZnO) of ∼2 μm size was used as control. Characterization results revealed that Pe ZnO is highly crystalline with hexagonal wurtzite structure and morphologically spherical with an average size of 38 nm as confirmed by X-ray diffraction, electron microscopy and dynamic light scattering techniques. Haemolysis assay at all the tested concentrations exhibit that, lysis of Red Blood Cells by Pe ZnO remained well within toxicity limit of 3% whereas D ZnO showed ∼5.2% hemolysis. Interestingly, Pe ZnO nanoparticles caused less than 36% growth inhibition of 3T3 cells at the highest concentration (2 μl) against 44% by D ZnO. Notably, in vivo study showed 26.67 and 96.67% death of the larvae at highest concentration 0.032 mg/mL of Pe ZnO and D ZnO respectively indicating Pe ZnO is less toxic than commercially available ZnO. Thus, we proved that green hydrothermally synthesized ZnO (PeZnO) is highly biocompatible and therefore propose to consider as potent compound for further evaluation using mammalian models followed by clinical trials.

Wtip is required for proepicardial organ specification and cardiac left/right asymmetry in zebrafish.

Wilm's tumor 1 interacting protein (Wtip) was identified as an interacting partner of Wilm's tumor protein (WT1) in a yeast two-hybrid screen. WT1 is expressed in the proepicardial organ (PE) of the heart, and mouse and zebrafish wt1 knockout models appear to lack the PE. Wtip's role in the heart remains unexplored. In the present study, we demonstrate that wtip expression is identical in wt1a-, tcf21-, and tbx18-positive PE cells, and that Wtip protein localizes to the basal body of PE cells. We present the first genetic evidence that Wtip signaling in conjunction with WT1 is essential for PE specification in the zebrafish heart. By overexpressing wtip mRNA, we observed ectopic expression of PE markers in the cardiac and pharyngeal arch regions. Furthermore, wtip knockdown embryos showed perturbed cardiac looping and lacked the atrioventricular (AV) boundary. However, the chamber-specific markers amhc and vmhc were unaffected. Interestingly, knockdown of wtip disrupts early left-right (LR) asymmetry. Our studies uncover new roles for Wtip regulating PE cell specification and early LR asymmetry, and suggest that the PE may exert non-autonomous effects on heart looping and AV morphogenesis. The presence of cilia in the PE, and localization of Wtip in the basal body of ciliated cells, raises the possibility of cilia-mediated PE signaling in the embryonic heart.

Asymmetric distribution of Spalt in Drosophila wing squamous and columnar epithelia ensures correct cell morphogenesis.

The Drosophila wing imaginal disc is a sac-like structure that is composed of two opposing cell layers: peripodial epithelium (PE, also known as squamous epithelia) and disc proper (DP, also known as pseudostratified columnar epithelia). The molecular mechanism of cell morphogenesis has been well studied in the DP but not in the PE. Although proper Dpp signalling activity is required for proper PE formation, the detailed regulation mechanism is poorly understood. Here, we found that the Dpp target gene sal is only expressed in DP cells, not in PE cells, although pMad is present in the PE. Increasing Dpp signalling activity cannot activate Sal in PE cells. The absence of Sal in the PE is essential for PE formation. The ectopic expression of sal in PE cells is sufficient to increase the PE cell height. Down-regulation of sal in the DP reduced DP cell height. We further demonstrated that the known PE cell height regulator Lines, which can convert PE into a DP cell fate, is mediated by sal mis-activation in PE because sal-RNAi and lines co-expression largely restores PE cell morphology. By revealing the microtubule distribution, we demonstrated that Lines- and Sal-heightened PE cells are morphologically similar to the intermediate cell with cuboidal morphology.

ESX/type VII secretion systems of mycobacteria: Insights into evolution, pathogenicity and protection

Pathogenesis of Mycobacterium tuberculosis depends on the secretion of key virulence factors, such as the 6 kDa early secreted antigenic target ESAT-6 (EsxA) and its protein partner, the 10 kDa culture filtrate protein CFP-10 (EsxB), via the ESX-1 secretion system. ESX-1 represents the prototype system of the recently named type VII secretion systems that exist in a range of actinobacteria. The M. tuberculosis genome harbours a total of five gene clusters potentially coding for type VII secretion systems, designated ESX-1 - ESX-5, with ESX-4 being the most ancient system from which other ESX systems seem to have evolved by gene duplication and gene insertion events. The five ESX systems show similarity in gene content and gene order but differ in function. ESX-1 and ESX-5 are both crucial virulence determinants of M. tuberculosis, but with different mechanisms. While ESX-1 is implicated in the lysis of the host cell phagosomes, ESX-5 is involved in secretion of the mycobacteria specific PE and PPE proteins and cell wall stability. Research on type VII secretion systems has thus become a large and competitive research topic that is tightly linked to studies of host-pathogen interaction of pathogenic mycobacteria. Insights into this matter are of relevance for redrawing the patho-evolution of M. tuberculosis, which might help improving current strategies for prevention, diagnostics and therapy of tuberculosis as well as elucidating the virulence mechanisms employed by this important human pathogen.

Abstract 1459: Tbx5 Regulates Progenitor Cell Migration During Cardiac Vasculogenesis

Mutations in TBX5 cause Holt-Oram syndrome (HOS) characterized by congenital heart malformation and upper limb deformity. Human TBX5 is expressed not only in a chamber specific myocardial distribution but also in coronary blood vessels. We have shown that mouse and chick Tbx5 are expressed in the proepicardial organ (PEO), a transitory structure from which epicardium and coronary vessels arise. We therefore investigated Tbx5 function during coronary vasculogenesis by manipulating Tbx5 gene dosage at critical points during chick and mouse cardioogenesis. Overexpression or inactivation of Tbx5 in chick PEO and chick epicardial cells leads to impaired migration and blocks incorporation of epicardial derived-cells (EPDCs) into coronary vessels. These data suggest Tbx5 acts during migration of PEO cells to the myocardium and migration of EPDCs into the subepicardium and myocardium. Epicardial restricted inactivation of Tbx5 using Wt1-cre and Tbx5 lox/lox mice reveals impaired migration (E10.5–11.5) of cytokeratin-positive PEO cells to the heart tube. However, at E14.5, mutant hearts display evidence of an epicardial layer suggesting delayed PE cell migration. However, unlike the flattened layer of epicardial cells in wildtype hearts, mutant hearts exhibit epicardial cells with irregular polygonal shapes and epicardial cyst-like structures lined by delaminated epicardium and filled with erythrocytes. At E17.5, we observe partial detachment of epicardium and nests of multiple contiguous subepicardial vessels without penetration into the myocardium. These observations suggest impaired migration of EPDCs into myocardium with consequent abnormally exuberant vasculogenesis restricted to the subepicardium. PECAM-1 expression analyses in E17.5 mutant embryos and neonatal mutant mice reveal marked reduction of myocardial coronary endothelial cells as well as defective coronary arteries and veins. These defects are associated with an upregulation of HIF1a in the myocardium, and, using an oxygen sensing dye, we observe relative tissue hypoxia in mutant myocardium. Our findings support a conserved requirement for Tbx5 in progenitor cell migration for the establishment of the coronary vasculature.

Centriole duplication in PE (SPEV) cells starts before the beginning of the DNA replication

Centrosome includes two centrioles and is a structural basis of mitotic spindle pole. Duplication of this organelle and doubling of chromosomes quantity during DNA replication are two principal events of cell cycle in the course of preparation for cell division. In this work, cells of pig kidney embryonic cell line PE (SPEV) were individually monitored after mitosis and procentriole appearance was detected by electron microscopy as soon as 5–6 h after mitosis. This period was 1–2 h shorter than minimal duration of G1-phase in PE cell line. Ultrastructural analysis of centrosomes in the cells with known “cell cycle age” in combination with autoradiography study of the same cells using 3H-thimidine directly confirmed that duplication of centrioles started earlier than cells entered in S-phase of cell cycle, i.e., preceded the DNA replication.

Simulated low-earth-orbit cycle-life testing of commercial laminated lithium-ion cells in a vacuum

We tested the life cycle of commercial laminated lithium-ion cells with both liquid (LE cells) and polymer (PE cells) electrolytes by simulating a satellite's low earth orbit (LEO) operation under various environmental conditions to develop a power storage system for microsatellites. We completed 4000 cycles, corresponding to about 8 months of LEO satellite operation. The LE cell initially exhibited better performance than the PE cell in a normal atmosphere. However, the LE cell began to expand in a vacuum (about 20 Pa), accompanied by capacity loss and voltage decline at the end of the discharge. In contrast, the PE cell exhibited excellent endurance in a vacuum and maintained high capacity and voltage at the end of the discharge. The reliable cycling of the PE cell in a vacuum was attributed to the adhesive properties of the gel electrolyte that held the laminate film package and active electrode materials together. A comparison of cell performance at various ambient temperatures demonstrated that the proper ambient temperature range for a PE cell is from 10 to 30 °C.

A systolic array implementation of the Feng-Rao algorithm

An efficient implementation of a parallel version of the Feng-Rao algorithm on a one-dimensional systolic array is presented in this paper by adopting an extended syndrome matrix. Syndromes of the same order, lying on a slant diagonal in the extended syndrome matrix, are scheduled to be examined by a series of cells simultaneously and, therefore, a high degree of concurrency of the Feng-Rao algorithm can be achieved. The time complexity of the proposed architecture is m+g+1 by using a series of t+[g-1/2]+1, nonhomogeneous but regular, effective processors, called PE cells, and g trivial processors, called D cells, where t is designed as the half of the Feng-Rao bound. Each D cell contains only delay units, while each PE cell contains one finite-field inverter and, except the first one, one or more finite-field multipliers. Cell functions of each PE cell are basically the same and the overall control circuit of the proposed array is quite simple. The proposed architecture requires, in total, t+[g-1]+1 finite-field inverters and (t+[(g'1)/2])(t+[(g-1)/2]+1)/2 finite-field multipliers. For a practical design, this hardware complexity is acceptable.

Characterization and Novel Activation of 72-kDa Metalloproteinase in Retinal Interphotoreceptor Matrix and Y-79 Cell Culture Medium

Analysis of bovine interphotoreceptor matrix and conditioned medium from human Y-79 retinoblastoma cells by gelatin SDS-PAGE zymography reveals abundant activity of a 72-kDa M r gelatinase. The 72-kDa gelatinase from either source is inhibited by EDTA but not aprotinin or NEM, indicating that it is a metalloproteinase (MMP). The 72-kDa MMP is converted to a 62-kDa species with APMA treatment after gelatin sepharose affinity purification, typical of previously described gelatinase MMP-2. The latent 72-kDa gelatinase from either bovine IPM or Y-79 media autoactivates without APMA in the presence of calcium and zinc after 72 hr at 37°C, producing a fully active mixture of proteinase species, 50 (48 in Y-79 medium), 38 and 35 kDa in size. The presence of inhibitory activity was examined in both whole bovine IPM and IPM fractions separated by SDS-PAGE. Whole IPM inhibited gelatinolytic activity of autoactivated Y-79-derived MMP in a dose-dependent manner. Inhibitory activities are observed in two protein fractions of 27-42 and 20-25 kDa. Western blots using antibodies to human tissue inhibitor of metalloproteinase 1 and 2 (TIMP-1 and -2) reveal the presence of two TIMP-1-like proteins at 21 and 29 kDa in inhibitory fractions of the bovine IPM. TIMP-2 was not detected in the inhibitory IPM fractions, consistent with the observed autoactivation of bovine IPM 72-kDa gelatinase. Potential roles for this IPM MMP-TIMP system include physiologic remodelling of the neural retina-RPE cell interface and digestion of shed rod outer segment as well as pathological processes such as retinal detachment, PE cell migration, neovascularization and tumor progression. Cultured Y-79 cells appear to be a good model for studying the production and regulation of this proteinase system.

Restoration of Leukemia Virus-Suppressed Immunocytes In Vitro by Peritoneal Exudate Cells

Depressed antibody responsiveness to sheep erythrocytes in mice infected with Friend leukemia virus continued in vitro when spleen cell cultures from infected animals were cultured in the presence of antigen. Addition of PE cells from normal donor mice to the immunologically depressed splenocyte cultures resulted in a marked restoration of antibody responsiveness. Restoration of the immune response was PE cell dose-dependent; a ratio of 1 PE cell per 10 splenocytes resulted in the largest numbers of PFC's. These results suggest that impaired antibody responsiveness by spleen cell cultures from FLV-infected mice may be due, in part, to effects on antigen-processing macrophages, since restoration of immune responsiveness occurs by PE cell supplements.

Failure of peritoneal exudate macrophages to reverse immunologic impairment by Friend leukemia virus.

Transfer experiments with peritoneal exudate macrophages from normal donor mice were performed to determine if a defect of normal macrophage function or activity was a major or contributing factor to the immunosuppression characterizing leukemia virus infection of mice. Challenge immunization of Friend leukemia virus-infected mice with sheep erythrocytes resulted in markedly depressed hemolytic antibody responses, as compared to responses of normal noninfected mice. When PE cell suspensions rich in macrophages were transferred from normal donor mice to leukemia virus infected recipients there was no affect on the FLV-induced impairment of the immune response. Similar transfer of PE cells to normal uninfected mice generally resulted in a moderate depression of the expected immune response. In no case did the PE cells enhance the immune responses in normal or virus-infected mice.

Enhanced susceptibility of penicillin-resistant staphylococci to phagocytosis after in vitro incubation with low doses of nafcillin.

SummaryIncubation of penicillin-resistant staphylococci with a subinhibitory concentration of nafcillin increased the susceptibility of the organisms to in vitro phagocytosis by normal mouse peritoneal exudate cells (macrophages). Methicillin and oxa-cillin were effective as inhibitors of bacterial growth only when used in relatively large doses; subinhibitory doses did not significantly affect subsequent phagocytosis by the PE cell. Penicillin G had no detectable effect on the staphylococci used in this system, either as a direct inhibitor of microbial growth or by influencing phagocytosis. The synergistic effect described here between nafcillin and macrophages may be related to the clinical efficacy reported for this antibiotic even when present only in low doses.The authors wish to thank Dr. Kenneth E. Price of Bristol Laboratories for the supplies of sodium methicillin and sodium oxacillin.