Top PDF Vascular Adventitia Calcification and Its Underlying Mechanism.

Vascular Adventitia Calcification and Its Underlying Mechanism.

Vascular Adventitia Calcification and Its Underlying Mechanism.

Fibroblasts are the major cell type comprising the adventitia, and they are activated in the pathological state. Fibroblasts undergo phenotype transformation, proliferation, and migra- tion. They can also secrete various cytokines, participating in the development of several dis- eases, including atherosclerosis. Physical damage, hypoxia, and inflammation can cause adventitia fibroblasts to differentiate into myofibroblasts, and this is mainly due to the actions of TGF-1. Myofibroblasts then express smooth muscle actin and collagen. Myofibroblasts con- tain stress fibers and cytoskeletal proteins with systolic and diastolic function. They can migrate, proliferate, and participate in the formation of the new intima. Adventitia fibroblast migration to the intima is the main cause of new intima thickening and luminal stenosis [30]. In addition, myofibroblasts can secrete a variety of cytokines and growth factors, including TGF-ß1, MCP-1, interleukin (IL), endothelin-1 (ET-1), tumor necrosis factor α (TNF-α), MMPs, and HADPH oxidase, which contribute to adventitia inflammation. For example, TGF-ß1 enhances fibroblast phenotype transformation, and MCP-1 promotes the recruitment and activation of macrophages. ET-1 increases the deposition of extracellular matrix compo- nents [31]. Together, these factors promote inflammation. Finally, adventitia fibroblasts, in both physiological and pathological settings, produce reactive oxygen species(ROS), which is involved in oxidative stress.ROS production by adventitia fibroblasts has been shown to be important for cell proliferation and intima regeneration [32–34].
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Lipoic acid, but not tempol, preserves vascular compliance and decreases medial calcification in a model of elastocalcinosis

Lipoic acid, but not tempol, preserves vascular compliance and decreases medial calcification in a model of elastocalcinosis

was a clear decrease of dihydroethidium fluorescence signals consistent with decreased superoxide output and, paradoxically, an increase in DCF-detectable oxidant, consistent at least in part with hydrogen peroxide. Contrarily, the mechanisms underlying LA effects do not seem to involve direct oxidant scavenging, considering the low rate constants of their direct reactions with this cyclic disulfide or its reduction product dihydrolipoic acid (24). Rather, LA may target redox signaling indirectly via Nrf-2-dependent (7) transcriptional activation of antioxidant genes. Another relevant mechanism underlying LA effects could be its interference with endoplasmic reticulum stress, which is known to be associated with atherosclerosis and inflammation (4). In fact, LA decreased, while tempol increased, the nuclear expression of the proapop- totic transcription factor CHOP/GADD153 in cultured vascular smooth muscle cells exposed to the endoplasmic reticulum stressor tunicamycin (unpublished data from our
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Bone metabolism and vascular calcification

Bone metabolism and vascular calcification

Osteoporosis and atherosclerosis are chronic degenerative diseases which have been considered to be independent and whose common characteristic is increasing incidence with age. At present, growing evidence indicates the existence of a correlation between cardiovascu- lar disease and osteoporosis, irrespective of age. The morbidity and mortality of osteoporosis is mainly related to the occurrence of fractures. Atherosclerosis shows a high rate of morbidity and espe- cially mortality because of its clinical repercussions such as angina pectoris, acute myocardial infarction, stroke, and peripheral vascular insufficiency. Atherosclerotic disease is characterized by the accumu- lation of lipid material in the arterial wall resulting from autoimmune and inflammatory mechanisms. More than 90% of these fatty plaques undergo calcification. The correlation between osteoporosis and ath- erosclerosis is being established by studies of the underlying physio- pathological mechanisms, which seem to coincide in many biochemi- cal pathways, and of the risk factors for vascular disease, which have also been associated with a higher incidence of low-bone mineral density. In addition, there is evidence indicating an action of antiresorptive drugs on the reduction of cardiovascular risks and the effect of statins, antihypertensives and insulin on bone mass increase. The mechanism of arterial calcification resembles the process of osteogenesis, involving various cells, proteins and cytokines that lead to tissue mineralization. The authors review the factors responsible for atherosclerotic disease that correlate with low-bone mineral density. Correspondence
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Cellular mechanism underlying formaldehyde-stimulated Cl- secretion in rat airway epithelium.

Cellular mechanism underlying formaldehyde-stimulated Cl- secretion in rat airway epithelium.

For a long time, formaldehyde has been considered as a strongly stimulating environmental pollutant. Prolonged exposure to formaldehyde can cause asthma [1], [2], upper respiratory tract inflammation [3], [4], and pneumonedema, etc. Furthermore, formaldehyde has strong carcinogenic effects on human [5]. In the past, formaldehyde was classified mostly as a compound from extraneous contamination [6]. But recent investigations showed that in vivo methylamine can be catalyzed into formaldehyde by semicarbazide-sensitive amine oxidase [7], [8]. The physiological function of this endogenous formaldehyde is still unclear. Studies have demonstrated that endogenous formaldehyde could be related to inflammation responses [9].
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From laboratory manipulations to earth system models: predicting pelagic calcification and its consequences

From laboratory manipulations to earth system models: predicting pelagic calcification and its consequences

well the observed responses can inform us about the net impact of higher CO 2 (lower pH) on diverse, natural assemblages in situ. Mesocosm experiments and shipboard manipulations carried out in a variety of oceanic regions are then vital if we are to improve our predictive capabilities of phytoplankton calcification and their role in the cycling of carbon in a future high-CO 2 world. However, laboratory studies will continue

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Phylogenetic evidence based on Trypanosoma cruzi nuclear gene sequences and information entropy suggest that inter-strain intragenic recombination is a basic mechanism underlying the allele diversity of hybrid strains

Phylogenetic evidence based on Trypanosoma cruzi nuclear gene sequences and information entropy suggest that inter-strain intragenic recombination is a basic mechanism underlying the allele diversity of hybrid strains

Several lines of evidence indicate that TcI and TcII are ‘‘pure’’ lines with great phylogenetic divergence between them. Briones et al. (1999) proposed that T. cruzi I and II should be considered dif- ferent species because of independent long term evolution, distinct ecological niches and epidemiological attributes and with unex- pectedly high phylogenetic distance (rDNA) equivalent to the dis- tance between genera Crithidia and Endotrypanum. The origins of the hybrids are more controversial and two scenarios have been proposed. In the first scenario the different hybrids are generated by two major independent hybridization events (Westenberger et al., 2005, 2006). The first event is the fusion of parental lineages TcI and TcII resulting in a TcI/TcII hybrid. This event was relatively ancient and TcIII and TcIV, originated by it, are homozygous for all non-satellite loci. A more recent back-cross between TcIII and its parental TcII generated the TcV and TcVI which still maintain alle- lic heterozygosity. The association between TcI and TcVI maxicir- cles coding regions is strong evidence that TcI provided the maxicircle to the progeny of TcIII and TcIV thus corroborating the two-hybridization event model and the hybris nature of TcIII (Ruvalcaba-Trejo and Sturm, 2011). The other scenario was postu- lated by De Freitas et al. (2006) based on microsatellite and mul- tilocus mitochondrial genes. In this model three ancestral genotypes are proposed, TcI, TcII and TcIII. The fusion of parental lineages TcII and TcIII originated TcV and TcVI hybrids. The origin of TcIV was not addressed because only one strain from this DTU was used. Analysis using microsatellite (Venegas et al., 2009; Ienne et al., 2010) and nuclear loci (Subileau et al., 2009; Tomazi et al., 2009) supports the two-hybridization model.
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Underlying mechanism of antimicrobial activity of chitosan microparticles and implications for the treatment of infectious diseases.

Underlying mechanism of antimicrobial activity of chitosan microparticles and implications for the treatment of infectious diseases.

compared to the neutral pH, suggesting CM have more positive charges at acidic pH that may bind to negatively charged bacterial surface molecules, such as LPS (Fig. 3D). Therefore, we hypothesize that CM bind to E. coli O157:H7 via two distinct mechanisms at neutral pH. First, a hydrogen bond interaction plays a key role in OmpA-mediated binding. Second, ionic interactions contribute to the binding activity. However, positively charged CM at acidic pH bind to negatively charged surface molecules such as LPS, resulting in magnified bacterial cell death. In order to study if CM retain antimicrobial activity in vivo, cows with uterine diseases were used as an animal model. Uterine disease is one of the biggest challenges facing the dairy cattle industries. Economic and revenue losses are largely impacted by these diseases as a result of the infertility, increased culling, milk production decreases, and treatment cost from uterine diseases [38,39,40]. Because the causes of uterine diseases are often linked to a variety of different bacteria, including IUPEC, Trueperella pyogenes, Fusobacterium necrophorum, and Prevotella melaninogenica, treatment of these diseases are often challenging with 30% failure rate [41] using conventional antibiotic treatment. As shown in figure 5, animals treated with CM showed significantly reduced IUPEC numbers in the uterine samples and was even more effective than ceftiofur treatment, suggesting that CM can be used to treat animals with these diseases. In this initial CM treatment trial, we only focused on IUPEC in the uterus, thus additional studies are needed for other pathogens, including Trueperella pyogenes, Fusobacterium necrophorum, and Prevotella melaninogenica. However, CM have a broad-spectrum of antimicrobial activity including Gram negative and positive bacteria (Fig. 4), thus we speculate that CM may have antimicrobial activity against other etiological agents of uterine disease.
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Filtration of aluminum alloys and its influence on mechanical properties and shape of eutectical silicium

Filtration of aluminum alloys and its influence on mechanical properties and shape of eutectical silicium

Progressive materials with high-speed development in wide range of commercial applications are aluminum alloys Al – Si. Future of their further use is connected with their unique characteristic, resp. advantageous relation between mechanical properties and density. Largest use of these alloys is in the transport industry – automobile and aerial industry. Also often used in engineering industry and special role in army industry. Desirable mechanical properties may be considered one of the main reason for wide spectrum efficiency of aluminum alloys, therefore is very important to understand the factors, which have major impact on this properties. Main goal of this work was to execute group of experiments, to prove impact of filtration elements on morphology of eutectical silicon. Shape, deployment and scale of eutectical silicon are one of the factors, that have
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Crotalus durissus cascavella venom toxicity to mammalian cells

Crotalus durissus cascavella venom toxicity to mammalian cells

To evaluate cell morphology, after the incubation period, cells were harvested, transferred to cytospin slides, fixed with methanol for 1 min and stained with hematoxilin and eosin. The morphological characteristics from untreated and treated cells were observed using a light microscope (Motic BA310, Moticam 2000, Causeway Bay, Hong Kong). In order to investigate nuclear alterations, the cells were incubated with 4’,6-diamidine-2’-phenylindole dihydrochloride (DAPI) Life Technologies). Briefly, after the incubation period, cells were fixed with 3.7 % formaldehyde and incubated with DAPI (5 µg/mL). After extensive washing with PBS, images were captured by confocal microscopy (LSM 710 Zeiss) excitation wave length 350 nm.
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Radiol Bras  vol.50 número2

Radiol Bras vol.50 número2

One limitation of our study is that among the subjects with DM or AH in our sample, there was a large number who were at high cardiovascular risk and a small number who were at low cardiovascular risk. Therefore, the risk of BAC might have been underestimated in the latter group. One strength of our study was that we evaluated CKD pa- tients in the pre-dialysis phase. However, although we found BAC to correlate with CKD and with the GFR, those corre- lations did not retain their significance in the multivariate analysis. Another limitation of our study is that, due to its cross-sectional design, we cannot make causal inferences. Therefore, we can state only that BAC might play a role in the pathophysiology of DM.
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Inhibition of bone morphogenetic protein signal transduction prevents the medial vascular calcification associated with matrix Gla protein deficiency.

Inhibition of bone morphogenetic protein signal transduction prevents the medial vascular calcification associated with matrix Gla protein deficiency.

The process of vascular calcification involves the initial loss of VSMC phenotype followed by the development of osteogenic cells and subsequent calcification [38]. Although we observed that BMP inhibition did not prevent loss of VSMC phenotype in the aortas of MGP -/- mice, BMP signaling was required for the induction of osteogenic cells. We next investigated whether calcification of MGP-deficient VSMCs is dependent on BMP signaling in vitro. Calcification was induced in isolated VSMCs by growing the cells in DMEM supplemented with 10% FCS and 2 mM sodium phosphate [38]. Expression of MGP in MGP -/- VSMCs using an adenovirus vector reduced calcification by 70%, as detected by von Kossa stain, compared with adenovirus control-treated cells (Fig. 9A–B, F). Treatment of wild-type VSMCs with siRNA directed against MGP (siMGP) increased calcification compared to treatment with control scrambled siRNA (siSC) (Fig. 9C–D). These results indicate that, similar to in vivo findings, MGP expres- sion modulates the calcification occurring in cultured VSMCs treated with high phosphate- containing media. Treatment with LDN-193189 inhibited the calcification induced by siMGP in wild-type VSMCs by approximately 50% (P = 0.0003; Fig. 9D–E, 9G). Taken together, these observations suggest that MGP inhibits calcification of isolated VSMCs and that calcification of MGP-deficient VSMCs is, at least in part, dependent on BMP signal transduction.
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Int. braz j urol.  vol.36 número1

Int. braz j urol. vol.36 número1

Most previous open surgical techniques for the treatment of nephroptosis and most of the minimally invasive techniques have been performed by a retroperitoneal approach (7-13). The presently proposed pathogenic mechanism is not revealed and cannot be corrected with a retroperitoneal approach and could thus be an explanation for the poor results in the past. None of the authors who have used a transperitoneal technique have mentioned the find- ing of an incomplete rotation of the colon (2-6). It should, however, be noted that Hübner and Plas (4,5) offer the following description: “Owing to the absence of the fatty capsular tissue, the kidney was easily identified”, a condition, which is very similar to what we describe in Figure-1. In describing the operative procedure, they do not mention mobilizing the colon but rather add that, directly after creating the pneumoperitoneum: “The kidney [is] easily identi- fied. The peritoneum and Gerota’s fascia are opened in a T-shaped incision.” Fornara et al. (6) give the following description: “The line of Toldt was incised and the ureter was identified. The perirenal fat was then dissected and the kidney was completely mobi- lized.” The authors do not describe the necessity of Table 2 – Postoperative course.
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Role of chemokines in promoting instability of coronary atherosclerotic plaques and the underlying molecular mechanism

Role of chemokines in promoting instability of coronary atherosclerotic plaques and the underlying molecular mechanism

The stability of AS plaques has been investigated by researchers around the world using IVUS. Sillesen et al. (14) reported that plaques with thin fibrous caps and large lipid cores were easy to rupture when using IVUS. Purushothaman et al. (15) compared ruptured and non- ruptured plaques in the same artery in ACS patients with IVUS and found that the ruptured plaques showed obvious irregularity and the artery appeared to have positive remodeling. Ueda (16) found that the plaques in unstable angina were mainly irregularly distributed lipid plaques with positive vascular remodeling. By comparing IVUS, serol- ogy, and molecular biology of SAP patients with those of UAP patients, we evaluated the general mechanisms underlying vulnerable plaques: 1) the levels of hs-CRP, MCP-1, RANTES, and fractalkine in the serum are sensitive markers for observing the conditions in UAP patients; 2) unstable plaques show irregularly distributed lipid plaques with low echo and have a larger PA and obvious positive remodeling; and 3) mRNA expression of MCP-l, RANTES, and fractalkine in UAP patients increases significantly, which promotes inflammation and further leads to the formation of vulnerable plaques (17,18).
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THE LEGAL RESERVE AREAS COMPENSATION MECHANISM AND ITS ECONOMIC AND ENVIRONMENTAL IMPLICATIONS

THE LEGAL RESERVE AREAS COMPENSATION MECHANISM AND ITS ECONOMIC AND ENVIRONMENTAL IMPLICATIONS

Finally, it is worth emphasizing that compensation, as it is established by Legislation n. 12.651/2012, is not only criticized by academics, but also from a legal point of view. The Prosecutor General Office (PGO) considers the re-composition of vegetation in areas of the same biome when there is no ecological identity between areas to be insuf- ficient as a compensation mechanism. This is one of the key points of direct actions of unconstitutionality put forward by the Prosecutor’s Office to the Supreme Federal Court (SFC), against the provisions established by the current legislation (BRASIL, 2013; PGO, 2013). The PGO argues that by determining compensation, this legislation goes against the fundamental duty of preserving and restoring essential ecological processes. It also results in the adulteration of legal reserves (BRASIL, 2013). Forms of compensation in- volving the leasing or the donation of an area within a governmental conservation unit are also considered to be unconstitutional. The former because it does not fully comply with the idea of “compensation”, given that there is no legal security with regard to the perpetuity of this protection, and the latter because it is an option created to circumvent the government’s administrative capacity to regularize the legal situation of conservation units, undermining the ecological functions of legal reserves (BRASIL, 2013).
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Low-molecular weight heparin increases circulating sFlt-1 levels and enhances urinary elimination.

Low-molecular weight heparin increases circulating sFlt-1 levels and enhances urinary elimination.

Knowing that renal elimination of sFlt-1 can be increased after heparin therapy in patients with intact glomerular barrier, we set out to further characterize the molecular basis of this effect in biochemical studies. We tested the hypothesis that due to their opposite charges, heparin liberates sFlt-1 from larger multi- molecular complexes by charge dependent interaction. Mono- meric sFlt-1 could then potentially be filtered via glomerular barrier or be secreted into the proximal tubule. In order to experimentally substantiate changes in complex size we employed velocity gradient ultra-centrifugation on serum samples to separate native complexes of serum proteins according to their density before and after administration of heparin (Fig. 5A). After separation of up to 22 fractions on SDS-PAGE and immunoblot analysis using anti-Flt-1 specific antibody, sFlt-1 was detected in the fractions 4–17 in samples of both before and after treatment. This led us to infer that sFlt-1 complex size is not affected by heparin administration.
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Uncoupling of glomerular IgA deposition and disease progression in alymphoplasia mice with IgA nephropathy.

Uncoupling of glomerular IgA deposition and disease progression in alymphoplasia mice with IgA nephropathy.

To confirm this idea, we further did BMT with different clones of grouped ddY mice line B, which we recently established by cross-breeding [15]. This prone mouse shows a more severe prognosis of murine IgAN with an increase of serum levels of IgA- IgG IC [13,17] that may be partly due to the pattern of glycosylation in IgA. In this additional model, we indeed found a greater increase of IgA-IgG IC and clear IgG deposition. This finding suggests that secondary lymphoid tissues may be not required for at least the circulating IC formation. However, even though we selected aly/aly recipients showing similar amounts of glomerular IgG co-deposition, full progression of murine IgAN in aly/aly recipients was absent, suggesting other possible underlying mechanisms. Interestingly, this uncoupling in aly/aly mice is linked to a lack of CD4 +
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Enhanced glutathione content allows the in vivo synthesis of fluorescent CdTe nanoparticles by Escherichia coli.

Enhanced glutathione content allows the in vivo synthesis of fluorescent CdTe nanoparticles by Escherichia coli.

i) Absorption and fluorescence spectroscopy: to get absorbance and fluorescence spectra, purified NPs were diluted 1/10 with ultrapure water. Absorbance spectra were recorded with a Per- kin-Elmer Lambda 11 UV-vis spectrophotometer using MiliQ water as blank. Fluorescence spectra were obtained with an ISS- PC photocounting spectrofluorimeter. An excitation spectra was constructed recording fluorescence emission at 515 nm while exciting the sample at different wavelengths (300–500 nm); ii) Dynamic light scattering (DLS): dynamic light scattering of purified fluorescent fractions was used to evaluate NP size as described before [19]. Data were acquired using a Zetasizer nano S90 (Malvern Instruments Limited, UK) instrument with a refraction index of 2.6 and 4-optical sides disposable cuvettes; iii) Atomic force microscopy (AFM): AFM measurements were conducted in a NT- MDT NTEGRA prima equipment using silica-etched tips. Typical samples were investigated at scan rates of 1–3 Hz with 2566256 pixel resolution during image capturing. Samples were dropped onto a mica sheet. Size determination was performed individually recording the z-axis height of each particle and subtracting the surrounding z-axis mica sheet height; iv)Elemental analysis by inductive coupled plasma atomic emission spectrometry: de- termining and quantifying Cd and Te metal species in both cell pellets and purified NPs was carried out using Spectro CIROS Vision ICP-AES. Strains pCA24NgshA and pCA24NgshB were grown as described and treated with CdCl 2 , K 2 TeO 3 or both for
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Topical  effect of  arborea baker on acute corneal pain in mice

Topical effect of arborea baker on acute corneal pain in mice

his study aimed to assess the possible topical antinociceptive activity of Vanillosmopsis arborea Baker essential oil (EOVA) and to clarify the underlying mechanism, using the acute model of chemical (eye wiping) nociception in mice. EOVA (25 to 200 mg/kg; p.o. and topical) evidenced signiicant antinociception against chemogenic pain in the test model of formalin-induced neuroinlammatory pain. Local application of 5 M NaCl solution on the corneal surface of the eye produced a signiicant nociceptive behavior, characterized by eye wiping. he number of eye wipes was counted during the irst 30 s. EOVA (25, 50, 100, and 200 mg/kg; p.o. and topical) signiicantly decreased the number of eye wipes. Naloxone, yohimbine, L-NAME, theophylline, glibenclamide, and ruthenium red had no efect on the antinociceptive efect of EOVA. However, ondansetron, p-chlorophenylalanine methyl ester (PCPA), capsazepine, prazosin, and atropine prevented the antinociception induced by EOVA. hese results indicate the topical antinociceptive efect of EOVA and showed that 5-HT, �1, TRPV1, and central muscarinic receptors might be involved in the antinociceptive efect of EOVA in the acute corneal model of pain in mice.
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Preparation and characterization of a novel aspirin derivative with anti-thrombotic and gastric mucosal protection properties.

Preparation and characterization of a novel aspirin derivative with anti-thrombotic and gastric mucosal protection properties.

Comparing the gastric damage in rats inflicted by Ca-ASP to that inflicted by ASP, the damage inflicted on the gastric mucosa by Ca-ASP was much less than that inflicted by ASP, and this may be due to a number of possible factors. Firstly, the modification of ASP by Hap converted the free carboxyl (COOH) group of ASP to a salt form, making it less reactive, and the drug therefore caused no local lesion to the gastric mucosa. Secondly, Ca-ASP inhibited the expression of COX-2 but not COX-1, unlike ASP which inhibited the expression of both enzymes. Although COX-1 and COX-2 both synthesize prostaglandins using the substrate arachidonic acid, COX-1 maintains normal gastric mucosa. The conversion of arachidonic acid to PGH2 is catalyzed by both COX-1 and COX-2. PGH2 is a common substrate for further conversion into PGE2, PGI2, PGD2, PGF2 and TXA2 by a series of specific isomerase and synthase enzymes [27–28]. One of these enzymes, the cytoplasmic prostaglandin E synthase (cPGES) is able to convert COX-1-synthesized but not COX-2 synthesized PGH2 into PGE2. PGE2 is a recognized vasodilator factors in vivo, and it can inhibit platelet aggregation and thrombosis, accelerate the flow of the gastric mucosal microcirculation, promote the secretion of bicarbonate, mediate the adaptive immune protective function, increase protein synthesis and cell renewal, and finally enhance the repair ability of the damaged gastric mucosa [29]. Thirdly, Hap increased the synthesis of PGE2, which in turn could counteract its inhibition by ASP. Studies in recent years have shown that impaired microcirculation is one of the pathological Figure 6. Analysis of platelet counts and COX-2 expression. (A) Platelet counts of rats underwent different treatments. (B) Western blot analysis of COX-2 expression in rats underwent different treatments.
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Synthesis of nanoparticeles in ductile iron with small additions of vanadium and niobium and its mechanical properties

Synthesis of nanoparticeles in ductile iron with small additions of vanadium and niobium and its mechanical properties

mechanical grinding and polishing. Back scattered electrons (BSE) were utilized in SEM in order to reveal difference in chemical compositions of microcomponents present in particular samples. The SEM investigations were used to reveal the distribution of graphite and other big particles. Transmission electron microscopy (TEM), on the other hand, was applied for examination of nanosized secondary precipitates, i.e. vanadium or niobium carbides and/or nitrides (or carbonitrides). The thin foil technique was implemented for this purpose. The 3 mm disks were ground down on sand papers and then dimpled to about 0.1 mm thickness. Afterwards the disks were further thinned in an ion mill until a perforation had appeared. The TEM investigation was carried out by means of a JEOL 2010 ARP analytical scanning transmission electron microscope operating at acceleration voltage of 200 kV. Imaging was performed by conventional transmission mode while for chemical analysis (X-ray Energy Dispersive Spectroscopy - EDS) the nanoprobe mode was utilized. The nanoprobe mode enabled to obtain electron probes approaching a few nanometers in diameter (practically about 10 nm because at smaller electron probes the number of X-ray counts is usually too low for analysis). The EDS analysis was performed by Oxford-Link system attached to the microscope. The Oxford- Link system was equipped with Si(Li) detector. This system detects all elements down to boron. In order to examine the crystallography of precipitates the Selected Area Diffraction (SAD) patterns analysis was also performed.
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