Processing and characterization of polystyrene nanocomposites based on CoAl layered double hydroxide

AbstractThe present work deals with the development of polystyrene (PS) nanocomposites through solvent blending technique with diverse contents of modified CoAl layered double hydroxide (LDH). The prepared PS as well as PS/CoAl LDH (1–7 wt.%) nanocomposites were characterized by X-ray diffraction (XRD), transmission electron microscopy (TEM), Fourier transform infrared spectroscopy (FTIR), rheological analysis, thermogravimetric analysis (TGA) and differential scanning calorimetry (DSC). The XRD results suggested the formation of exfoliated structure, while TEM images clearly indicated the intercalated morphology of PS nanocomposites at higher loading. The presence of various functional groups in the CoAl LDH and PS/CoAl LDH nanocomposites was verified by FTIR analysis. TGA data confirmed that the thermal stability of PS composites was enhanced significantly as compared to pristine PS. While considering 15% weight loss as a reference point, it was found that the thermal degradation (Td) temperature increased up to 28.5 °C for PS nanocomposites prepared with 7 wt.% CoAl LDH loading over pristine PS. All the nanocomposite samples displayed superior glass transition temperature (Tg), in which PS nanocomposites containing 7 wt.% LDH showed about 5.5 °C higher Tg over pristine PS. In addition, the kinetics for thermal degradation of the composites was studied using Coats-Redfern method. The Criado method was ultimately used to evaluate the decomposition reaction mechanism of the nanocomposites. The complex viscosity and rheological muduli of nanocomposites were found to be higher than that of pristine PS when the frequency increased from 0.01 to 100 s−1

Mechanical, thermal and morphological characteristics of poly(methyl methacrylate) (PMMA) nanocomposites reinforced with Cu-Cr layered double hydroxide

The current work deals with the influence of Cu-Cr layered double hydroxide (LDH) on the properties of PMMA nanocomposites. The Cu-Cr LDH was organically modified with sodium dodecyl sulfate (SDS) in order to obtain LDH layers more compatible with polymer chains. PMMA with various quantities of Cu-Cr LDH nanofiller (1-5 wt%) and 5 wt% of polystyrene-grafted maleic anhydride were first melted and extruded in a twin screw extruder and, finally, processed by injection moulding machine to prepare testing specimens. The nanocomposites were examined by X-Ray diffraction (XRD), transmission electron microscope (TEM) and submitted to thermogravimetric (TGA), tensile and hardness testing. The XRD analysis showed an exfoliated structure of the nanocomposites whereas TEM exhibited the intercalated morphology at higher LDH contents. The TGA results revealed that the thermal degradation temperature was enhanced by the incorporation of Cu-Cr LDH. When 50% of the initial sample weight is established as point of comparison, the decomposition temperature of the nanocomposites was increased about 26 - 32 oC with respect to neat PMMA. From the mechanical analysis, it was also observed that PMMA nanocomposite with 1 wt% LDH presented higher tensile strength and hardness than neat PMMA.Portugal Programa Operacional Regional do Norte de Portugal -NORTE 2020, projeto ref. NORTE-01-0145-FEDER-00015 “TSSiPRO-Technologies for Sustainable and Smart Innovative Products

Dramatic Co-Activation of WWOX/WOX1 with CREB and NF-κB in Delayed Loss of Small Dorsal Root Ganglion Neurons upon Sciatic Nerve Transection in Rats

BACKGROUND:Tumor suppressor WOX1 (also named WWOX or FOR) is known to participate in neuronal apoptosis in vivo. Here, we investigated the functional role of WOX1 and transcription factors in the delayed loss of axotomized neurons in dorsal root ganglia (DRG) in rats. METHODOLOGY/PRINCIPAL FINDINGS:Sciatic nerve transection in rats rapidly induced JNK1 activation and upregulation of mRNA and protein expression of WOX1 in the injured DRG neurons in 30 min. Accumulation of p-WOX1, p-JNK1, p-CREB, p-c-Jun, NF-kappaB and ATF3 in the nuclei of injured neurons took place within hours or the first week of injury. At the second month, dramatic nuclear accumulation of WOX1 with CREB (>65% neurons) and NF-kappaB (40-65%) occurred essentially in small DRG neurons, followed by apoptosis at later months. WOX1 physically interacted with CREB most strongly in the nuclei as determined by FRET analysis. Immunoelectron microscopy revealed the complex formation of p-WOX1 with p-CREB and p-c-Jun in vivo. WOX1 blocked the prosurvival CREB-, CRE-, and AP-1-mediated promoter activation in vitro. In contrast, WOX1 enhanced promoter activation governed by c-Jun, Elk-1 and NF-kappaB. WOX1 directly activated NF-kappaB-regulated promoter via its WW domains. Smad4 and p53 were not involved in the delayed loss of small DRG neurons. CONCLUSIONS/SIGNIFICANCE:Rapid activation of JNK1 and WOX1 during the acute phase of injury is critical in determining neuronal survival or death, as both proteins functionally antagonize. In the chronic phase, concurrent activation of WOX1, CREB, and NF-kappaB occurs in small neurons just prior to apoptosis. Likely in vivo interactions are: 1) WOX1 inhibits the neuroprotective CREB, which leads to eventual neuronal death, and 2) WOX1 enhances NF-kappaB promoter activation (which turns to be proapoptotic). Evidently, WOX1 is the potential target for drug intervention in mitigating symptoms associated with neuronal injury

Downregulation of uPAR and Cathepsin B Induces Apoptosis via Regulation of Bcl-2 and Bax and Inhibition of the PI3K/Akt Pathway in Gliomas

Glioma is the most commonly diagnosed primary brain tumor and is characterized by invasive and infiltrative behavior. uPAR and cathepsin B are known to be overexpressed in high-grade gliomas and are strongly correlated with invasive cancer phenotypes.In the present study, we observed that simultaneous downregulation of uPAR and cathepsin B induces upregulation of some pro-apoptotic genes and suppression of anti-apoptotic genes in human glioma cells. uPAR and cathepsin B (pCU)-downregulated cells exhibited decreases in the Bcl-2/Bax ratio and initiated the collapse of mitochondrial membrane potential. We also observed that the broad caspase inhibitor, Z-Asp-2, 6-dichlorobenzoylmethylketone rescued pCU-induced apoptosis in U251 cells but not in 5310 cells. Immunoblot analysis of caspase-9 immunoprecipitates for Apaf-1 showed that uPAR and cathepsin B knockdown activated apoptosome complex formation in U251 cells. Downregulation of uPAR and cathepsin B also retarded nuclear translocation and interfered with DNA binding activity of CREB in both U251 and 5310 cells. Further western blotting analysis demonstrated that downregulation of uPAR and cathepsin B significantly decreased expression of the signaling molecules p-PDGFR-β, p-PI3K and p-Akt. An increase in the number of TUNEL-positive cells, increased Bax expression, and decreased Bcl-2 expression in nude mice brain tumor sections and brain tissue lysates confirm our in vitro results.In conclusion, RNAi-mediated downregulation of uPAR and cathepsin B initiates caspase-dependent mitochondrial apoptosis in U251 cells and caspase-independent mitochondrial apoptosis in 5310 cells. Thus, targeting uPAR and cathepsin B-mediated signaling using siRNA may serve as a novel therapeutic strategy for the treatment of gliomas

Activation of adherent vascular neutrophils in the lung during acute endotoxemia

BACKGROUND: Neutrophils constitute the first line of defense against invading microorganisms. Whereas these cells readily undergo apoptosis under homeostatic conditions, their survival is prolonged during inflammatory reactions and they become biochemically and functionally activated. In the present study, we analyzed the effects of acute endotoxemia on the response of a unique subpopulation of neutrophils tightly adhered to the lung vasculature. METHODS: Rats were treated with 5 mg/kg lipopolysaccharide (i.v.) to induce acute endotoxemia. Adherent neutrophils were isolated from the lung vasculature by collagenase digestion and sequential filtering. Agarose gel electrophoresis, RT-PCR, western blotting and electrophoretic mobility shift assays were used to evaluate neutrophil activity. RESULTS: Adherent vascular neutrophils isolated from endotoxemic animals exhibited decreased apoptosis when compared to cells from control animals. This was associated with a marked increase in expression of the anti-apoptotic protein, Mcl-1. Cells isolated 0.5–2 hours after endotoxin administration were more chemotactic than cells from control animals and expressed increased tumor necrosis factor-alpha and cyclooxygenase-2 mRNA and protein, demonstrating that they are functionally activated. Endotoxin treatment of the animals also induced p38 and p44/42 mitogen activated protein kinases in the adherent lung neutrophils, as well as nuclear binding activity of the transcription factors, NF-κB and cAMP response element binding protein. CONCLUSION: These data demonstrate that adherent vascular lung neutrophils are highly responsive to endotoxin and that pathways regulating apoptosis and cellular activation are upregulated in these cells

Guidelines for the use and interpretation of assays for monitoring autophagy (3rd edition)

In 2008 we published the first set of guidelines for standardizing research in autophagy. Since then, research on this topic has continued to accelerate, and many new scientists have entered the field. Our knowledge base and relevant new technologies have also been expanding. Accordingly, it is important to update these guidelines for monitoring autophagy in different organisms. Various reviews have described the range of assays that have been used for this purpose. Nevertheless, there continues to be confusion regarding acceptable methods to measure autophagy, especially in multicellular eukaryotes. For example, a key point that needs to be emphasized is that there is a difference between measurements that monitor the numbers or volume of autophagic elements (e.g., autophagosomes or autolysosomes) at any stage of the autophagic process versus those that measure fl ux through the autophagy pathway (i.e., the complete process including the amount and rate of cargo sequestered and degraded). In particular, a block in macroautophagy that results in autophagosome accumulation must be differentiated from stimuli that increase autophagic activity, defi ned as increased autophagy induction coupled with increased delivery to, and degradation within, lysosomes (inmost higher eukaryotes and some protists such as Dictyostelium ) or the vacuole (in plants and fungi). In other words, it is especially important that investigators new to the fi eld understand that the appearance of more autophagosomes does not necessarily equate with more autophagy. In fact, in many cases, autophagosomes accumulate because of a block in trafficking to lysosomes without a concomitant change in autophagosome biogenesis, whereas an increase in autolysosomes may reflect a reduction in degradative activity. It is worth emphasizing here that lysosomal digestion is a stage of autophagy and evaluating its competence is a crucial part of the evaluation of autophagic flux, or complete autophagy. Here, we present a set of guidelines for the selection and interpretation of methods for use by investigators who aim to examine macroautophagy and related processes, as well as for reviewers who need to provide realistic and reasonable critiques of papers that are focused on these processes. These guidelines are not meant to be a formulaic set of rules, because the appropriate assays depend in part on the question being asked and the system being used. In addition, we emphasize that no individual assay is guaranteed to be the most appropriate one in every situation, and we strongly recommend the use of multiple assays to monitor autophagy. Along these lines, because of the potential for pleiotropic effects due to blocking autophagy through genetic manipulation it is imperative to delete or knock down more than one autophagy-related gene. In addition, some individual Atg proteins, or groups of proteins, are involved in other cellular pathways so not all Atg proteins can be used as a specific marker for an autophagic process. In these guidelines, we consider these various methods of assessing autophagy and what information can, or cannot, be obtained from them. Finally, by discussing the merits and limits of particular autophagy assays, we hope to encourage technical innovation in the field

Cholinergic receptor pathways involved in apoptosis, cell proliferation and neuronal differentiation

Acetylcholine (ACh) has been shown to modulate neuronal differentiation during early development. Both muscarinic and nicotinic acetylcholine receptors (AChRs) regulate a wide variety of physiological responses, including apoptosis, cellular proliferation and neuronal differentiation. However, the intracellular mechanisms underlying these effects of AChR signaling are not fully understood. It is known that activation of AChRs increase cellular proliferation and neurogenesis and that regulation of intracellular calcium through AChRs may underlie the many functions of ACh. Intriguingly, activation of diverse signaling molecules such as Ras-mitogen-activated protein kinase, phosphatidylinositol 3-kinase-Akt, protein kinase C and c-Src is modulated by AChRs. Here we discuss the roles of ACh in neuronal differentiation, cell proliferation and apoptosis. We also discuss the pathways involved in these processes, as well as the effects of novel endogenous AChRs agonists and strategies to enhance neuronal-differentiation of stem and neural progenitor cells. Further understanding of the intracellular mechanisms underlying AChR signaling may provide insights for novel therapeutic strategies, as abnormal AChR activity is present in many diseases

Cross flow microfiltration of oil-water emulsions using clay based ceramic membrane support and TiO2 composite membrane

The main objective of this work is to study the effect of cross flow filtration conditions on the separation of oily wastewater using ceramic support and TiO2 membrane. Firstly, the low cost clay based ceramic membrane support was prepared by uniaxial compaction method using combination of pyrophyllite, quartz, feldspar, kaolin, ball clay and calcium carbonate along with PVA as a binder. Subsequently, TiO2 composite membrane was fabricated via hydrothermal route employing TiO2 sol derived from TiCl4 and NH4OH solution. Cross flow microfiltration investigations were carried out by utilizing oil-water emulsion concentration of 200 mg/L at three distinct applied pressures (69–207 kPa) and three cross flow velocities (0.0885, 0.1327, and 0.1769 m/s). Compared to ceramic support, TiO2 composite membrane demonstrates better performance in terms of flux and removal efficiency of oil and also the rate of flux decline during filtration operation is lower due to highly hydrophilic surface of the TiO2 membrane. TiO2 membrane displays the oil removal efficiency of 99% in the entire range of applied pressures investigation, while ceramic support shows 93–96% of oil removal