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Serum microRNAs (miRNAs) have emerged while potential non-invasive biomarkers to diagnose prostate malignancy (PCa), the most typical noncutaneous malignancy among Western males. -337-5p, -331-3p, and -409-3p had been expressed at different frequencies (75%, 50%, 50%, and 50%, respectively). Importantly, Scano-miR expression evaluation recognized 58 miRNAs, comprising 45 experimentally validated miRNAs and 13 predicted miRNAs (Tables S3 and ?andS4),S4), that have been coexpressed in every 16 samples. Permutation testing had been performed to recognize six differentially expressed miRNAs with significant adjustments within their expression amounts between intense and control samples (miR-605, miR-135a*, miR-495, miR-433, miR-371C3p, and miR-106a, with an adjusted worth of 0.0301, 0.0319, 0.0411, 0.0115, 0.0089, and 0.0017, respectively) (Figs. S1 and ?andS2S2). Desk purchase PRT062607 HCL S3. Expression data for coexpressed miRNAs in intense PCa = 8) and control samples (indolent PCa and healthful people, = 8). Samples from patients with intense PCa generally cluster collectively and also have globally up-regulated serum miRNA expression. Open in another home window Fig. S2. Identification of differentially expressed miRNAs using the Scano-miR assay. Boxplots stand for the background-subtracted, normalized distributions of six differentially expressed miRNAs. The reddish colored bar represents the median, whereas the blue bar represents the interquartile selection of distribution (permutation check; miR-106a, = 0.0017; miR-371C3p, = 0.0089; miR-433, = 0.0115; miR = 605, SPN = 0.0301; miR-135a*, = 0.0319; miR-495, = 0.0411). Regardless of the identification of differentially expressed miRNAs, solitary biomarkers might not be accurate diagnostics for intense PCa. Because of this, we calculated purchase PRT062607 HCL the molecular signature rating for the differentially expressed miRNAs to tell apart intense PCa from control samples utilizing a released mathematical formula referred to by Zeng et al. (31) The molecular signature evaluation exposed that the diagnostic dependability was more than doubled (= 0.0036) upon merging the differentially expressed miRNAs (Fig. 1= 0.0036). (= 16). Correlation between miRNA expression and individual risk was analyzed using the Wilcoxon rank-sum purchase PRT062607 HCL check. ?Correlation value; 0.1, Trend; 0.05, *; simply no correlation, C. Prostate needle biopsies frequently undergrade the real tumor aggressiveness, with up to 20% of individuals having more intense tumors on subsequent biopsy or prostatectomy (32). As a result, we investigated the correlation of our recognized molecular signature to the clinicopathologic top features of PCa following a 2015 National In depth Malignancy Network (NCCN) Recommendations for Prostate Malignancy (Edition 1.2015). To examine such a correlation, prostatic needle biopsy specimens of individuals with GS 8 had been grouped into either VHR or HR PCas. Six individuals out of eight had been informed they have VHR malignancy that progressed into locally advanced or metastatic PCa (GS 9, metastasis, and/or medical stage T3), and the additional two patients got HR PCa (GS 8 and medical stage T3) (Desk S2). Unsupervised hierarchical clustering revealed these six miRNA markers could actually determine a subclass of four individuals out of six, categorized as VHR (Fig. S3). We calculated the purchase PRT062607 HCL statistical correlation of the molecular signature and purchase PRT062607 HCL specific miRNAs to the VHR malignancy using the KaplanCMeier and Wilcoxon rank-sum tests (33, 34). The outcomes show a significant correlation of the identified molecular signature to the clinical pathology of these patients (= 0.041) (Fig. 1 0.05), which supports the notion that these individual biomarkers, considered alone, are not uniquely indicative of disease states. Open in a separate window Fig. S3. Heat map of clustering and clinical association for six differentially expressed miRNAs. Unsupervised hierarchical clustering performed on expression profiles for 16 serum samples reveals that, in general, samples of similar histology are clustered together. Interestingly, a subgroup of four samples identified to be indicative of VHR aggressive PCa cluster together using this molecular signature. Gleason scores are on the range of 9 (black) to 6 (light gray) to healthy (white). Tumor staging is on the range of T3 (black) to.
Introduction Thrombolysis with intravenous tissue plasminogen activator (tPA) may be the only FDA approved treatment for sufferers with acute ischemic stroke, but its make use of is bound by narrow therapeutic screen, selective efficacy, and hemorrhagic complication. tPA provides high affinity and specificity for fibrin. Fibrin binds to tPAs F and K2 domains, AZD4547 ic50 plasminogen binds to tPAs K2 domain, forming a ternary complicated (plasminogen/tPA/fibrin) which catalyzes the transformation of plasminogen to plasmin. Binding of tPA to fibrin may enhance tPA’s catalytic activity by 400-fold8. Intravascular thrombi (bloodstream clots) are comprised of aggregation of activated platelets and fibrin monomers that are cross-connected through lysine aspect chains. Plasmin cleaves fibrin, thus wearing down the meshwork of blood coagulum and leading to recanalization of the blocked vessel. The thrombolytic program could possibly be regulated by 2-antiplasmin and plasminogen activator inhibitor-1 (PAI-1). 2-antiplasmin may be the primary inhibitor of plasmin in the bloodstream, which inhibits plasmin from making fibrin degradation items. PAI-1 may be the primary inhibitor of tPA in the bloodstream, which covalently binds to the C-terminal catalytic domain of tPA and forms an inactive PAI-1/tPA complicated. After that, the inactive PAI-1/tPA complicated could be cleared by liver through low-density lipoprotein receptor-related protein-1 (LRP-1) mediated pathway9. Recombinant human being tPA (alteplase) was authorized by the FDA in 1996 for the treatment of acute ischemic stroke. Standard dose of tPA recommended by FDA is definitely 0.9 mg/kg bodyweight (10% as bolus and remaining as infusion over 60 min; max 90 mg). tPA maintains a rather short therapeutic windowpane of only 3C4.5 hours after symptom onset and may increase risk of symptomatic ICH, therefore only a few individuals could receive (3C8.5%) and benefit (1C2%) from tPA treatment10. Although with limited efficacy and security, tPA remains the only authorized thrombolytic agent for acute ischemic stroke. Open in a separate window Figure 1 tPA-centered thrombolytic pathwayTissue plasminogen activator (tPA), is definitely released by endothelium and circulates in plasma as a inactive complex with plasminogen-activator inhibitor type 1 (PAI-1). Plasminogen and tPA bind to the surface of fibrin clot, forming a ternary complex (plasminogen/tPA/fibrin), which promotes the conversion of plasminogen to plasmin. Plasmin causes lysis of the cross-linked fibrin into fibrin degradation products. PAI-1 could inhibit the activation of plasminogen induced by tPA. 2-antiplasmin inhibits plasmin from creating fibrin degradation products. Number was adapted from earlier publication7 with permission (? 2014 Bhattacharjee P, Bhattacharyya D. Published in Fibrinolysis and Thrombolysis under CC BY 3.0 Mouse monoclonal antibody to Integrin beta 3. The ITGB3 protein product is the integrin beta chain beta 3. Integrins are integral cell-surfaceproteins composed of an alpha chain and a beta chain. A given chain may combine with multiplepartners resulting in different integrins. Integrin beta 3 is found along with the alpha IIb chain inplatelets. Integrins are known to participate in cell adhesion as well as cell-surface mediatedsignalling. [provided by RefSeq, Jul 2008] license. Available from: http://dx.doi.org/10.5772/57335). Although best known for its part in fibrinolysis, tPA has also been demonstrated to regulate many nonfibrinolytic functions in the central nervous system (CNS). tPA can be synthesized and released by most of the mind cells. Once released, it can bind to these same cells via different receptors or binding partners. The interaction of tPA with these receptors or binding partners AZD4547 ic50 prospects to different effects that can be beneficial or deleterious. Earlier studies possess reported that tPA may boost BBB permeability and mind edema and induce intracerebral hemorrhage in acute ischemic stroke11C14. However, more recent studies have shown potential benefits of tPA for the treatment of ischemic stroke. As evidence of its beneficial effects, tPA offers been shown to play a critical part in inhibiting neuronal apoptosis and advertising practical recovery in late phase after stroke15C18. tPA offers been shown to exert reverse effects at different time points on the same target19 (e.g., extracellular matrix, NMDA receptors). The beneficial and deleterious effects of tPA in the CNS are time-dependent and involve varied mechanisms. For readers who are interested in more details about the pleiotropic effects of tPA in the CNS, please read other reviews20C22. 3. Nanocarriers for tPA Schematic representations of various nanocarriers discussed in this review are shown in Figure 2. The advantages and disadvantages of various nanocarriers are shown in Table 1, and representative tPA-loaded nanocarriers are summarized in Table 2. Open in a separate window AZD4547 ic50 Figure 2 Schematic representation of nanocarriers for tPA-based nanothrombolysis ? Liposomes (A): tPA can be either incorporated into the inner core or adsorbed onto the outer shell of liposomes; tPA can bind to functionalized liposomes by covalent conjugation or by selective non-covalent metallochelation (His-tagged recombinant tPA);? Polymer-based nanoparticles (B): tPA can be encapsulated into PLGA or gelatin nanoparticles, and both nanoparticles can be furtherly coated with chitosan (red) or decorated with PEG or targeting moieties;? Magnetic nanoparticles (C): Most inner core of magnetic nanoparticles is iron oxide (Fe3O4 or -Fe2O3), and the surface is.
The overwhelming most benign lesions from the adrenal cortex resulting in Cushing syndrome are associated with one or another abnormality from the cAMP signaling pathway. cAMP signaling network UK-427857 biological activity marketing leads to tumors in adrenal cortex and various other tissue. Within this review, we summarize all latest data from ours and various other laboratories, helping the watch that Wnt-signaling serves as a significant mediator of tumorigenicity induced by unusual PRKAR1A function and aberrant cAMP signaling. activating mutations lead to constitutive activation of adenylate cyclase and PKA activation, and a variety of manifestations, including the classic triad of polyostotic fibrous dysplasia, caf au lait skin pigmentation, and autonomous endocrine hyperfunction (Weinstein et al., 1991). The most frequent PHF9 affected endocrine tissues are pituitary, ovarian and thyroid, but bilateral macronodular adrenocortical hyperplasia can also be found in the context of MAS (Lee et al., 1986; Stratakis and Kirschner, 1998). These and other molecular findings discussed below demonstrate the strong biological relevance of PKA activation in adrenal tumorigenesis. The first demonstration of PKA involvement with human disease was the finding that inactivating mutations of the gene coding for the 1- regulatory (RI) subunit of protein kinase A (PKA) are responsible for Carney complex (CNC) in the majority of patients (Kirschner et al., 2000a; Kirschner et al., 2000b). CNC is usually a multiple neoplasia syndrome that is inherited in an autosomal dominant manner and is characterized by several UK-427857 biological activity types of skin tumors and pigmented lesions, myxomas, schwannomas, liver and other cancers, and endocrine neoplasms (Carney et al., 1985; Stratakis et al., 2001). In a recent review of 353 UK-427857 biological activity patients with CNC from 185 families, patients from all ethnicities and with a wide spectrum of clinical manifestations were explained (Bertherat et al., 2009; Horvath et al., 2010; Stratakis et al., 2001). defects were found in UK-427857 biological activity 73% of these patients. Main pigmented nodular adrenocortical disease (PPNAD) was the most common endocrine tumor associated with CNC, occurring in 60% of the CNC patients (Bertherat et al., 2009). Isolated PPNAD was the only manifestation in 12% of patients carrying defects. Among the remaining kindreds with micronodular adrenocortical hyperplasia (MAH) and no evidence of mutation, subgroups of patients were recognized by clinical and histopathological criteria (Table 1) (Stratakis, 2009). Genetic defects in cAMP-binding phosphodiesterases (PDEs) have been explained in isolated MAH. Five different mutations were recognized so far in patients with isolated MAH or PPNAD; three of them resulted in premature stop codon generation and the other two were single base UK-427857 biological activity substitutions in the catalytic domain name of the protein, significantly affecting the ability of PDE11A to degrade cAMP (Horvath et al., 2006a; Horvath et al., 2006b). Two missense substitutions, R804H and R867G, were also more frequent among patients with sporadic adrenocortical tumors (Horvath et al., 2006b). In addition, the chromosomal locus harboring the gene encoding phosphodiesterase 8B (PDE8B) was the second most likely region to be associated with a predisposition to isolated MAH (Horvath et al., 2006a). Sequencing of the = the gene for PDE11A; = the gene for PDE8B, = protein kinase A regulatory subunit 1. Adrenocorticotropic hormone (ACTH)-impartial macronodular adrenal hyperplasia, also known as massive macronodular adrenocortical disease is usually a rare cause of CS (Lacroix, 2009; Zhang et al., 2009). Cortisol production in ACTH-independent macronodular adrenal hyperplasia can be regulated by the aberrant appearance of G-protein-coupled receptors others than ACTH (Lacroix et al., 2004; Lacroix et al., 2001). Somatic loss from the 17q22C24 area and PKA activity adjustments showed that cyclic (c) AMP/proteins kinase A (PKA) signaling is normally changed in ACTH-independent macronodular adrenal hyperplasia much like adrenal tumors with 17q loss or mutations (Bertherat et al., 2003; Bourdeau et al., 2006). A lot more interesting was the discovering that common adrenal lesions (i.e. adrenal adenomas) that didn’t harbor germline or somatic mutations and had been connected with ACTH-independent CS acquired functional abnormalities from the cAMP signaling pathway, as proven by elevated cAMP levels, reduced total PDE activity and/or elevated PKA activity (Bimpaki et al., 2009b). 3.0 C PKA tumorigenicity: Proof from mouse choices and research 3.1 C Mouse types of Carney complicated To characterize the foundation for PRKAR1A-associated tumorigenesis within an pet super model tiffany livingston, we generated mice carrying a floxed duplicate of exon 2 from the murine gene (Kirschner et al., 2005). haploinsufficiency in mice resulted in the introduction of tumors arising in cAMP-responsive tissue, like the bone tissue, Schwann and thyroid follicular cells. down-regulation and augmented cAMP signaling, produced a far more serious phenotype with a substantial decrease in the entire life-span (Griffin.
Supplementary MaterialsS1 File: Fig A, Sequence analysis of BW25113 TA system reduces growth by cleaving transcripts with in-frame sites, and antitoxin DinJ is usually a global regulator that represses its locus as well as controls levels of the stationary sigma factor RpoS. TA system enables the cell to withstand oxidative  and bile acid stress in the gastrointestinal tract . Usually the genes for TA systems occur in pairs, and many antitoxins regulate the TA locus . In addition, some antitoxins such as MqsA of the MqsR/MqsA TA system are intertwined with the general stress response by regulating other loci including levels of the stationary phase sigma factor RpoS [5, 8, 9], and some toxins exhibit a general regulatory effect via post-transcriptional differential mRNA decay [10, 11]. In addition to the general stress response, TA systems also have functions in biofilm formation [9, 10, 12, 13] and in inhibiting the propagation of phage [14C16]. In sites in conjunction with ribosomes . Specifically, YafQ binds the 70S ribosome at the A site via three surface-exposed patches of basic residues that appear to directly interact with 16SrRNA, and YafQ residues H50, H63, D67, and H87 participate in acid-base catalysis during mRNA hydrolysis . Its antitoxin is usually DinJ , and like the locus , the locus is not subject to conditional cooperativity , a form of regulation in which the binding of the 1st toxin to the antitoxin represses the TA locus Gossypol irreversible inhibition whereas additional toxin molecules induce transcription of the locus . DinJ binding to DNA like a dimer is definitely facilitated by its N-terminal ribbon-helix-helix motif, and its C terminus binds YafQ like a heterotetramer [24, 26]. Also, the SOS regulator LexA binds the promoter suggesting a link of manifestation of this operon after DNA damage  although experimentally this has not been seen [27, Mouse monoclonal to A1BG 28]. The physiological functions of the YafQ/DinJ TA system include that it actively participates in the general stress response through the rules of RpoS by antitoxin DinJ via direct repression of ; cold-shock protein CspE enhances translation of RpoS mRNA. YafQ and DinJ will also be involved in regulating persistence in are induced in persisters . The interspecies [35, 36] and interkingdom signal indole  reduces persistence [38, 39], and toxin YafQ raises persistence by Gossypol irreversible inhibition reducing indole by cleaving tryptophanase mRNA . Notably, indole is definitely most active as a signal in at low temps . Critically, aside from erythromycin stress leading to DinJ degradation , little is known about the circumstances that activate YafQ and make it very important to its function in the strain response and persistence. Since a couple of few reviews of temperature impacting the experience of the TA program [41C44], and since small is normally understood in what activates toxin YafQ, we explored the result of temperature over the YafQ/DinJ TA program of and genes in BW25113 using polymerase string reaction (PCR) items  was utilized to develop the dual deletion stress, BW25113 (Desk 1). The kanamycin level of resistance cassette from was taken out through Gossypol irreversible inhibition the use of plasmid pCP20 . Gene deletions were verified by DNA sequencing using primers bacterial strains and plasmids found in this scholarly research. (( KmRBW25113 KmRthis studyBW25113 KmRBW25113 KmRthis studyBW25113 KmRthis studyBW25113 KmRBW25113 KmRPlasmidspCA24NCmR; stress To recognize the proteins that triggered toxicity at 18C in any risk of strain, an EZ-Tn5? KAN-2 Tnp Transposome? Package (Epicentre) was utilized to help make the mutant collection of BW25113 based on the producers instructions. In short, 1 L of transposome was electroporated into 50L of experienced cells (1 L of TypeOne? Limitation Inhibitor was added in to the mixture to improve transduction performance). Warm SOC moderate (1 mL) was utilized as well as the cells had been incubated for 37C for 60 min. The retrieved cells (100 L) had been diluted 10-fold and spread on LB plates with kanamycin (50 g/mL) to enumerate the amount of transposon insertion clones. The rest of the 900 Gossypol irreversible inhibition L of retrieved cells (around 6,000 mutant cells) was inoculated into 20 ml clean moderate and cultured at 18C for 36 hours. After 36 hr, 100 L lifestyle was used in 5 mL of clean moderate and cultured at 18C for 36 hours (this is repeated four even more situations for six total enrichment civilizations). After six rounds of development to enrich for faster-growing BW25113 transposon mutants, colonies had been produced, and 500 unbiased colonies had been cultured in 0.5 mL of fresh medium in 2 mL microcentrifuge tubes for 24 h at Gossypol irreversible inhibition 37C. One L of lifestyle was.
During herpes virus type 1 (HSV-1) latent infection in individual peripheral sensory ganglia, the main viral gene transcribed may be the latency-associated transcript (LAT) gene. fibroblasts extracted from LAT-expressing transgenic mice in comparison to that in cells extracted from regular mice. HSV-1 DNA quantities in latently contaminated TG of transgenic mice had been comparable to those in regular mice. Reactivation of latent HSV-1 LAT-negative mutants by explant cocultivation of TG from transgenic mice was better than reactivation from normal-mouse TG. Taking into consideration our prior and present outcomes, we suggest that the bigger steady-state degree of the 1 significantly. 5-kb RNA in the TG may hyperlink this transcript to features which by inhibition of trojan replication latency, the LAT gene may protect ganglion cells and raise the possibility of reactivation thereby. Herpes virus type 1 (HSV-1) establishes lifelong latent an infection in individual peripheral sensory ganglia (for testimonials, see personal references 26 and 40), generally in the lack of viral replication (38). From a quiescent, nonreplicating condition, it could reactivate to create recurrent mucocutaneous disease. HSV-1 latency exists in a lot of the adult people (50) and isn’t accompanied by harm to peripheral sensory ganglion cells. During latency, a restricted area of the viral genome, the LAT (latency-associated Cannabiscetin biological activity transcript) gene, is active transcriptionally, and two colinear LATs, 2.0 and 1.5 kb in proportions, gather in latently infected nervous tissues (34, 36, 41). The two 2.0-kb LAT is normally a well balanced intron produced from a much less abundant 8.3-kb RNA (28, 53) and it is further spliced to create the 1.5-kb LAT. As the 2.0-kb transcript is normally noticed during latent and successful infections, the 1.5-kb RNA is apparently particular for neuronal tissues harboring the latent viral genome (13, 42). Both transcripts include several large open up reading frames, have already been discovered in the nucleus and cytoplasm (1, 12, 13, 29), and bind to polyribosomes (12). Although no proteins products have however been discovered, there can be an sign of natural function for the biggest LAT open up reading body (41). Viruses not capable of expressing the LATs possess a faulty reactivation phenotype (15, 37, 45, 47). COLL6 Whether this gene exerts Cannabiscetin biological activity its function during reactivation or the faulty reactivation phenotype is because of impaired capability of LAT-negative infections to determine latent an infection continues to be unsettled. Nevertheless, improved reactivation performance was correlated with HSV-1 capability to create latent an infection in an increased percentage of Cannabiscetin biological activity neurons (45, 46). Neuronal cell lines that exhibit the LAT gene had been produced previously, and it had been showed that HSV-1 replication in these cells is normally partially blocked which the viral immediate-early (IE) genes are repressed in the current presence of the LATs. It had Cannabiscetin biological activity been therefore suggested which the LAT gene may have a job in facilitating the establishment of HSV-1 latency (23). Many questions connected with HSV-1 latent an infection in vivo await elucidation. Is normally splicing from the 1.5-kb RNA particular to the anxious system? What mobile or tissue features, necessary for the latent condition, are mediated with the LAT gene? Can LAT suppress HSV-1 replication and latency improve establishment of, and with what system? Will this gene encode protein? To handle these relevant queries, we’ve produced a transgenic mouse where the HSV-1 DNA fragment that transcribes the two 2.0- and 1.5-kb LATs is normally controlled with the cytomegalovirus (CMV) IE Cannabiscetin biological activity promoter. A recently available study examined the result from the HSV-2 LAT transgene upon HSV-2 an infection and latency in transgenic mice and didn’t recognize any phenotypic difference in the parental mouse stress (49). Today’s work reviews differential digesting of both 2.0- and 1.5-kb LATs in a variety of transgenic-mouse tissues. This implies that splicing of the two 2.0-kb LAT to create the 1.5-kb transcript occurs in every tissues but its steady-state level is normally significantly higher in neuronal tissue. We also examined the effects from the LAT transgene upon the replication from the HSV-1 outrageous type, aswell as an HSV-1 LAT-negative mutant, in principal fibroblasts extracted from the transgenic mice and on the performance of explant reactivation in the trigeminal ganglia (TG). Replication of both infections was suppressed in the.
Background Irreversible electroporation (IRE) is a tissue ablation method, which relies on the phenomenon of electroporation. can influence the electric field distribution. We present a method for treatment planning of IRE which takes the influence of blood vessels on the electric field into account; this is illustrated on a treatment of 48-year-old patient with a metastasis near the remaining hepatic vein after a right side hemi-hepatectomy. Results Output of the numerical treatment planning method shows that a 19.9 cm3 irreversible electroporation lesion was generated and the whole tumor was protected with at least 900 V/cm. This compares well with the quantity from the hypodense lesion observed in comparison enhanced CT pictures taken following the IRE treatment. A substantial temperatures raise occurs close to the electrodes. Nevertheless, the hepatic vein continues to be open following the treatment without proof tumor recurrence after six months. Conclusions Treatment preparing using accurate pc models was named very important to electrochemotherapy and irreversible electroporation. A significant locating of the scholarly research was, that the top of electrodes significantly warm up. Therefore the medical consumer should generally 3-Methyladenine biological activity prevent putting the electrodes significantly less than 4 mm from risk constructions when following suggestions of the maker. there is certainly significant heating within the vicinity of electrodes, the full total treatment volume can be considerably higher than the amount predicated on thermal results would be anticipated. A limitation from the model can be, that people assumed that pulses had been shipped at 1 Hz continuously, while the truth is, the pulses had been shipped in synchronization using the individuals ECG, which may be up to 100 beats each and every minute realistically, and would create a higher temperatures rise consequently. In Shape 4, some areas in instant vicinity from the electrodes are warmed to temps greater than 100C, because there was no term for boiling included in the numerical model. In fact, these high temperatures could indicate that some 3-Methyladenine biological activity tissue boiling actually occurs near the electrode tips. This could explain the gas bubbles visible in the post-treatment CT images, and which are consistent with reports in the literature.32 3-Methyladenine biological activity Another reason for these findings could be a gas formation due to electrolysis.33 However, the temperature drops to below 70C approximately 4 mm away from the electrodes in this specific case. The coverage of the target tumor with electric fields was very high. The IRE threshold electric field, which depends on the type of number, duration of pulses, and tissue types of liver tumors has not been firmly established yet. 6 In this work, we assumed a value of 800 V/cm for tumor tissue. Nevertheless it can be seen in Figure 2, that almost the whole tumor is covered by electric field of this strength already in the first two pulse trains between electrode pairs 3C4 and 1C2. That, and the very high temperatures achieved in the model seem to indicate, that the used voltages and pulse numbers34 were considerably higher than necessary to achieve complete treatment of the tumor. When liver tumors are surgically treated, at least a protection margin of 0.5 to at least one 1 cm of liver cells across the tumor is resected to make sure removal of any micrometastases encircling the tumor and thereby to avoid local tumor recurrence. For the same cause there’s a need to attain a similar protection margin across the tumor in IRE remedies as well, as well as the IRE lesion in the shown case can be bigger than this objective. Presuming an elliptical approximation from the tumor, the volume of an ellipsoid with the semi-axes increased by 1 cm relative to the tumor, the required lesion volume would be 18.05 cm3. Rabbit polyclonal to ACTL8 This corresponds also to the presented case and should be accounted for every treatment planning. Conclusions Treatment planning using accurate computer models was recognized as important for electrochemotherapy and irreversible electroporation.2,9,23,35,36 On the one hand appropriate numerical treatment planning assures sufficient coverage of the clinical target volume with electric field sufficiently high for efficient tumor treatment also in the vicinity of blood vessles15 and thereby to prevent local recurrences. On the other hand it enables the prediction and control of temperature thus avoiding thermal tissue damage in critical structures, such as nerves or bile ducts. Regarding the strength of the electric field in the presented case, significant overtreatment can be assumed, since electric fields in the tumor were higher than 900 V/cm. In the future monitoring of electric field in real time37.
Supplementary MaterialsPicciani-Suppl. further supported by TNT and promoter mutation TNT assays. CONCLUSIONS These results support the finding that the observed increased cochlin expression in glaucomatous TM is due to relative elevated abundance of TFs. The results also demonstrate the utility of combinatorial bioinformatic and biochemical analyses for genes with uncharacterized promoter regions. Glaucoma is a group of irreversible blinding eye diseases associated with optic neuropathy. Primary open-angle glaucoma (POAG) is often associated with elevated intraocular pressure (IOP), which is due to an imbalance between aqueous humor production and outflow in the anterior chamber of the eye.1 Aqueous humor is a clear liquid produced by the ciliary epithelium that exits through the trabecular meshwork (TM) after bathing the anterior segment structures, such as the cornea and lens, with nutrients. Aqueous outflow is believed to encounter increased resistance at the level of the TM in glaucoma. The mechanisms that impede aqueous outflow elevate IOP are poorly understood. Cochlin, a secretory extracellular matrix (ECM) protein of unknown function, was identified by proteomic analyses to be differentially expressed in glaucomatous compared with normal TM.2 Cochlin is the product of the gene3 located on human chromosome 14, region q12C13.3,4 The cochlin protein sequence is highly conserved, with 94% and 79% amino acid identity with human to mouse and chicken sequences, respectively. 5 Cochlin contains a short signal peptide, an N-terminal factor C homology, and two von Willebrand factor A-like domains. 2,4 In situ hybridization has shown that cochlin mRNA is expressed in the TM, suggesting that the protein is likely expressed and deposited locally.2 Elevated IOP is a significant risk factor for optic nerve damage. Changes in fluid dynamics and incremental fluctuations in IOP results in stress and stretch on TM cells and are thought to trigger early biochemical responses.6 Stress- and stretch-induced modulation of protein expression are mediated by transcription factors (TFs).6,7 Increased cochlin expression has been reported in the TM8 and in the inner ear4; however, the promoter region of cochlin and the details of cochlin gene expression remain to be characterized. Deciphering mechanisms that lead to transcriptional regulation of cochlin expression in the TM is critical for understanding the role cochlin may play in glaucomas pathogenesis. We used a combinatorial approach of bioinformatics and molecular and biochemical analyses to determine whether an increased abundance of transcription factors with the potential to bind and enhance transcription in the promoter region of cochlin was present in nuclear extracts of glaucomatous TM compared with the control. MATERIAL AND METHODS Tissue Procurement and Preparation of Nuclear Extracts Glaucomatous and normal control eyes were obtained from the National Disease Research Institute (Philadelphia, PA) and the Lions Eye Bank (Miami, FL), respectively. The eyes had been enucleated within 10 hours of death and placed in a moisture chamber at 4C and transported. They were dissected within 48 hours, and the TM was carefully excised for study. The available details of the donor were recorded. Relating to available info, all glaucomatous donor eyes experienced POAG (observe Supplementary Table S1; all Supplementary Furniture are online at LBH589 small molecule kinase inhibitor http://www.iovs.org/cgi/content/full/50/7/3106/DC1) Bioinformatic Analyses The human being cochlin upstream promoter Anxa5 gene region was analyzed up to 5000 bp upstream of the translational start site (ATG; accession quantity “type”:”entrez-nucleotide”,”attrs”:”text”:”BC007230″,”term_id”:”13938218″,”term_text”:”BC007230″BC007230; National Center for Biotechnology Info [NCBI], Bethesda, MD). The cochlin DNA sequence was from the UCSC genome internet browser (http://genome.ucsc.edu/ provided in the public website by the University or college of California at Santa Cruz). Putative TF binding sites were identified with commercial software (MatInspector; Matrix Family Library Version 6.3; Genomatix, Munich, Germany). The analysis parameters used have been offered in respective tables. TFs related to the eye (observe Supplementary Table S2) were further investigated for his or her correlation with glaucoma, the TM, the anterior chamber, or the eye, as reported in the literature and for LBH589 small molecule kinase inhibitor its manifestation in the eye per the UniGene database (http://www.ncbi.nlm.nih.gov/UniGene; offered in the public website by NCBI; observe Supplementary Table S3). Tissue-specific associations of TFs in putative cochlin promoter areas for ear, mind, central nervous system (CNS), embryonic cells and liver were also analyzed (MatInspector; Genomatix), and LBH589 small molecule kinase inhibitor a list of tissue-specific TFs was generated (not shown). Only those TFs present in at LBH589 small molecule kinase inhibitor least two search terms were investigated further. For the selected TFs, 5 biotin end-labeled oligonucleotide sequences were generated for the relevant consensus binding sites as well as for their respective complementary sequences. These results were confirmed and/or.
The growing interest in using zebrafish for genetic and functional dissection of malignancy and infection was highlighted by the second international workshop on Zebrafish Models of Cancer and the Immune Response in Spoleto, Italy (July 20C22, 2009). such as tuberculosis have been established that are now amenable to high-throughput drug screens, a much-needed development in the fight against drug-resistant microorganisms. The success of this workshop and the rapidly growing field of cancer and the immune response in zebrafish have spawned follow-up meetings in Boston (June 2010) and Edinburgh (2011). Introduction The highly successful zebrafish workshop on infectious disease and cancer in zebrafish in Leiden (The Netherlands) in 20071 strongly motivated the authors of this report to organize a follow-up meeting. Prompted by the prediction that similarities between defense mechanisms against microbes and cancer cells can reveal new insights into specific determinants of innate immune responses, this workshop centered on cancer infection and models studies. Because of the amenability of zebrafish to large-scale ahead and reverse hereditary displays, this model organism is fantastic for discovery of book gene features in disease procedures at a throughput level that can’t be matched up by rodent versions. Further, due to its little size and optical transparency, disease manifestations and ensuing immune system responses could be studied in the whole-organism level. Especially advantageous with this framework are fluorescence multicolor labeling methods that enable tagging from the players in disease procedures (e.g., tumor cells, immune system cells, and microbes) for easy recognition have bolstered the explanation for using zebrafish like a tumor model system. The presentations on cancer studies particularly focused on cancer and hematopoiesis, tumor models obtained using forward and reverse genetic screens, xenografts, and and tumor models. Melanoma models Five of the 12 cancer talks scheduled for the first day of the meeting dealt with fish melanoma models. M. Mione (Milan, Italy) focused on different transgenic lines expressing the oncogene Harvey-RAS (H-RAS)V12, BML-275 kinase inhibitor and presented work implicating cellular senescence and ubiquitination in modulating the oncogenic activity of H-RAS. Oncogene-induced senescence is a powerful tumor-suppressive mechanism, which restrains the proliferation of oncogene-expressing cells. In zebrafish, oncogene-induced senescence is associated with increased ubiquitination of the oncogene. E. Patton (Edinburgh, United Kingdom) gave the European Association for Cancer ResearchCsponsored lecture on chemical and genetic control of melanocyte and melanoma development. She illustrated the power of a multiple organism approach (yeast, fish, and cancer cells) for screening BML-275 kinase inhibitor chemical compound libraries. Here the zebrafish was used to identify compounds that affected pigmentation in development. Subsequently, a library of yeast genetic deletion mutants was screened with the most promising BML-275 kinase inhibitor compounds, to identify their CD4 mechanisms of action, a process called chemical profiling. One of the active compounds appears to selectively kill developing and adult melanocytes and affects nevus growth in the zebrafish. M. Schartl (Wuerzburg, Germany) reminded the audience that the spontaneous melanomas discovered in the late 1920s in certain strains of generated the first animal model of cancer that clearly illustrated the genetic nature of the disease. BML-275 kinase inhibitor In addition, molecular studies in this system have uncovered for the first time molecules that are implicated in the pathogenesis of melanoma and are also of relevance for the human disease, for instance, signal transducers and activator of transcription (STAT)5, osteopontin, and the importance of the Ras/Raf/mitogen-activated protein (MAP) kinase pathway. The nuclear translocation of Stat5 seems to be a key event (mediated by the oncogenic epidermal growth factor (Egf) receptor, Xmrk) in the activation of antiapoptotic and pro-proliferative pathways. To make use of genetic tools, transgenic medaka expressing the same oncogene in pigment cells have been generated. Here, tumors develop also in the presence of functional p53, the scale can be suffering from whose lack, however, not the timing of melanoma advancement. A. Hurlstone (Manchester, UK) shown the melanoma versions created in his laboratory and predicated on H-RAS oncogene manifestation driven from the promoter. Among the.
Since the discovery of the inner nuclear transmembrane protein emerin in the early 1990s, nuclear envelope (NE) components and related involvement in nuclei integrity and functionality have been highly investigated. contribution of the LINC complex (for Linker of Nucleoskeleton to Cytoskeleton), hosting KASH and SUN proteins interactions. This close interplay between compartments has been related to diverse functions from nuclear integrity, activity and positioning through mechanotransduction pathways. At the same time, mutations in NE components genes coding for proteins such as lamins or nesprins, had been associated with a wide range of congenital diseases including cardiac and muscular diseases. Although most of these NE associated proteins are ubiquitously expressed, a large number of tissue-specific disorders have been associated with diverse pathogenic mutations. Thus, diagnosis and molecular explanation of this group of diseases, commonly called nuclear envelopathies, is currently challenging. This review aims, first, to give a better understanding of diverse functions of the LINC complex components, from the point of view of lamins and nesprins. Second, to summarize human congenital diseases with a special focus on muscle and heart abnormalities, caused by mutations in genes coding for these two types of NE associated proteins. genes (Behrens et al., 1994; Zhang et al., 2001; Wilhelmsen et al., 2005; Roux et al., 2009). SUN proteins are embedded in the INM and interact using INCB8761 kinase inhibitor their N-terminus domain with nuclear pore complex (NPC) and lamins (Padmakumar et al., 2005; Haque et al., 2006). Conversely, C-terminus domain is located in the perinuclear space and interacts with nesprins, which are embedded in the ONM (Padmakumar et al., 2005; Crisp et al., 2006; Haque et al., 2006; Ketema et al., 2007; Horn et al., 2013). This LINC complex play a role in diverse specialized cellular activities such as nuclear morphology maintenance, nuclear positioning, genes expression and cell signaling (Crisp et al., 2006; Lombardi and Lammerding, 2011; Mellad et al., 2011; Stroud et al., 2014). At the nucleoplasmic side, the LINC complex interacts with the nuclear lamina, a network of intermediate filaments just beneath INM and mainly composed of two Rabbit Polyclonal to p47 phox different groups of lamin: A-type (lamins A and C) and B-type lamins (lamins B1 and B2) (Fisher et al., 1986; McKeon et al., 1986; Peter et al., 1989). In this review, we will summarize functional diversities of proteins associated to the LINC complex with a particular focus on lamin A/C and nesprins. We will recapitulate known muscular and cardiac abnormalities induced by mutations in those genes and will discuss our recent advances in related pathogenesis. Nesprins and Lamins as Components of the LINC Complex In human, full-length nesprin-1 and nesprin-2, which are the ubiquitously expressed giant nesprins isoforms, are respectively, with a molecular weight of 1 1 MDa (146 exons) and 800 kDa (116 exons), the second and the third largest described proteins, after the untouchable titin (4.2 MDa) (Zhang et al., 2001). Nesprins-1 and -2 are ubiquitous proteins, commonly described as ONM components and composed of three major domains: (i) a Calponin Homology (CH) domain, also called Actin-Binding Domain (ABD) located in the N-terminus side that binds to the actin cytoskeleton; (ii) a long central rod domain composed of multiple spectrin repeats (SR) (respectively, 74 and 56 SR in nesprin-1 and -2) and that supports interactions with other proteins such as emerin with the Emerin Binding Domain (EBD) or lamins with the Lamin Binding Domain (LBD) and finally (iii) a C-terminus KASH domain embedded in the ONM (Zhang et al., 2001; Rajgor and Shanahan, 2013; INCB8761 kinase inhibitor Figure ?Figure1A1A). An additional domain has been described and called adaptive domain (AD). This highly conserved domain is located at the C-terminus extremity and is crucial for structural stabilization of SR (Simpson and Roberts, 2008; Zhong et al., 2010). Open in a separate window FIGURE 1 Nesprins and lamins as part of the LINC complex. (A) Nesprins and lamins isoforms structures. AD, adaptive domain; EBD, emerin binding domain; KASH domain, Klarsicht/ANC-1/Syne Homology; LBD, lamin binding domain; NBD, nesprin INCB8761 kinase inhibitor binding domain; SR, spectrin repeats. (B) The healthy LINC complex and its interactors. Nesprins play multiple functions.
Supplementary MaterialsSupplementary Dataset 1 srep19258-s1. design of plant tubulin has also been studied in various species. The gene is only expressed in flowers, and are transcribed in leaves, roots, and flowers10,19. Of the nine of genes, the transcripts of and preferentially accumulate in leaves and petioles12, whereas the transcript is primarily expressed in the roots20, and the other seven are expressed in vegetative tissues. Rice is predominantly expressed in flowers and other seven are differentially expressed during development17. The transcript levels of five cotton are much higher in fibers than that in various other tissues, including pollen13, and 9 of 19 genes are preferentially expressed in cotton fiber cells14. have the highest transcript levels in pollen, whereas the other and are upregulated in the xylem18. Functionally distinct microtubule subtypes are generated in cells through the expression of different tubulin isotypes and through post-translational modifications (PTMs). In animals, tubulins have different homologs that undergo various PTMs such as tyrosination/detyrosination, acetylation, polyglutamylation, and polyglycylation, which in turn lead to the appearance of various tubulin isoforms and classes of MTs21,22,23,24,25,26,27. In vegetation, a lot of tubulin isotypes have already been isolated from different species, whereas investigations on PTMs in plant Cilengitide small molecule kinase inhibitor tubulin are limited. Willow (genes and twenty genes in were used for the identification of – and -tubulin genes, including both of DNA and CDS, via reciprocal BLAST analysis using protein sequence of 20 and 15 tubulin genes29. homologs to the 20 tubulin genes were identified by using BLASTP, with the e-value cut-off set at 1-E03. The same protocol was performed for the detection of willow homologs by using the 15 tubulin genes. DNA cloning and sequencing Total RNA was HYPB extracted and treated with RNase-free Dnase (Promega, Madison, USA) to remove contaminating DNA. Purification of first-strand cDNA was conducted following the protocol of Lu genome. PCR was performed as follows: 94?C for 2?min, followed by 30 cycles of 94?C for 30?s, 56?C for 45?s, and 72?C for 2?min. The PCR products were cloned into the pMD18-T vector (Takara, Japan, Cilengitide small molecule kinase inhibitor http://www.takara.com.cn) and sequenced. The 28 cDNAs (8 TUAs and 20 TUBs) from were designated as SaTUA1CSaTUA8 and SaTUB1CSaTUB20, respectively. Real-time PCR Analysis Stem developing phloem, full expanded leaves, stem developing xylem, shoot tips (1.0?cmC1.5?cm from the top of the plant), and inflorescence were obtained from three 1-year-old identification of and genes The present study identified a total of eight genes, which were designated through genes, namely, to genes ranged from 1,350 bp to 1 1,356 bp, whereas that of the genes ranged from 1,335 to 1 1,356 bp. The eight cDNAs encode eight distinct TUA proteins, whereas the 20 cDNAs encoded 19 TUB proteins, mainly because and encoded the same protein. The length of the TUA proteins ranged from 449 to 451 amino acids, whereas that of the TUB proteins ranged from 444 to 451 amino acids. The shared 73.9% to 94.5% cDNA sequence Cilengitide small molecule kinase inhibitor and 88.6% to 98.4% protein sequence identity?(Supplementary file 2: Figure S1, Table S2), whereas the shared 74.6% to 99.8% cDNA and 86.8% to 99.1% (except for SaTUB7/12) protein sequence identity (Supplementary file 3: Figure S2, Table S3). Three functional domains in TUA and TUB were characterized using electron crystallography: the N-terminal domain, which contained the GTP binding site; the C-terminal domain, which comprised Cilengitide small molecule kinase inhibitor microtubule-associated proteins (MAPs); and the motor protein binding region and an intermediate domain containing the Taxol binding site34. PTMs are essential for the maturation of the tubulin protein, which include modifications such as tyrosination/ detyrosination35, acetylation36, polyglutamylation37, phosphorylation, and polyglycylation38. Except for acetylation, all modifications take place in the hypervariable C-terminal region39. The C-terminal region consisted of about 20 amino acid residues that constitute a.

domain: health


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The Flutist of Arnhem: A Story of Operation Market Garden. Author: Antonio Gil. Publisher: Dead Reckoning. 152 pages.Â Price $ 24.95 ISBN: 9781682474631 I first encountered the defense of Arnhem bridge in Cornelius Ryanâ€™s book â€œA Bridge Too Farâ€. Later came the movie with a cast of thousands and a number board games covering aspects of the operation. Antonio Gil revisits the events of this battle with his recent graphic novel â€œThe Flutist of Arnhem: A Story of Operation Market Gardenâ€. Gil weaves a good overview of General Montgomeryâ€™s big push to end the war with a fictional tale of an SOE agent trying to deliver important documents to the Allied High Command. The two separate story lines intersect during the events of Operation Market Garden â€“ the combined air assault and ground offensive designed to push the Allies over the Rhine and into Germany. The book retells the story Operation Market Garden with enough detail to understand the events, without getting bogged down within individual combat actions. While the...Read More
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domain: history


Draft Outdoor Dining Policy
Consultation closed 5.00pm, Friday 15 June 2018.
The City of Norwood Payneham & St Peters is seeking feedback on the Council’s draft Outdoor Dining Policy and on the operation of the existing Outdoor Dining Policy.
The Outdoor Dining Policy provides a framework that guides the design, siting and assessment and permit process for outdoor dining areas, and also sets out the obligations of traders when operating an outdoor dining area with a Permit
Download: draft Outdoor Dining Policy 2018-2021
Alternatively, you can view a hard copy of the draft Policy at the Norwood Town Hall or one of the Council’s Libraries.
Have your say
Section 1.6.5 of the existing Policy is being updated to reflect legislative changes introduced by the State Government on 1 July 2016 to ban smoking in all outdoor dining areas in South Australia. No other major changes to the Policy or Permit process are proposed.
The Council is also seeking general feedback on the operation of the existing Outdoor Dining Policy, to assist in finalising the review.
Please provide your written feedback via:
- Email: firstname.lastname@example.org
- Post: Attention: Mark Thomson, Manager, Development Assessment, City of Norwood Payneham & St Peters, PO Box 204, Kent Town, SA, 5071
Any comments that you submit will be taken into consideration by the Council, prior to the adoption and implementation of the Policy.
Council’s Manager, Development Assessment
T: 08 8366 4567


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Tourette Syndrome, also known as Tic Disorders, is a neurological disorder that manifests itself as involuntary movements and vocalizations. This situation, which starts in childhood and usually exacerbates during adolescence, can negatively affect social life and daily activities. In this article we wrote for you, we will try to answer all the questions about the causes, symptoms and types of Tourette Syndrome.
- What Causes Tourette Syndrome (tic Disorders)?
- What Are the Symptoms of Tourette Syndrome (Tic Disorders)?
- What Are The Types of Tourette Syndrome (tic Disorders)?
- How Is Tourette Syndrome (tic Disorders) Diagnosed?
- How Is Tourette Syndrome (tic Disorders) Treated?
- Does Tourette Syndrome (tic Disorders) Go away?
- Tourette Syndrome (tic Disorders) Treatment Prices in Turkey
What Causes Tourette Syndrome (tic Disorders)?
The exact cause of Tourette Syndrome is not yet known. However, it is thought that genetic and environmental factors come together to cause this disorder. Individuals with a family history of Tourette’s Syndrome or tic disorders are more likely to experience this condition. In addition, the imbalance of some chemical substances in the brain and problems with the functionality of brain structures may also play a role in the emergence of tic disorders.
What Are the Symptoms of Tourette Syndrome (Tic Disorders)?
The symptoms of Tourette Syndrome vary from person to person and may worsen and subside over time. In general, there are two main symptom groups: motor tics and vocal tics. Motor tics are involuntary muscle movements and include actions such as blinking, nodding, and shrugging. Vocal tics, on the other hand, include involuntary vocalizations and examples such as coughing, throat clearing, and shouting can be given. These symptoms usually begin in children aged 5-10 years and become more severe during adolescence. In some cases, symptoms may decrease or disappear completely during adulthood.
What Are The Types of Tourette Syndrome (tic Disorders)?
Tourette Syndrome is the most severe and common form of tic disorders. However, tic disorders can be divided into different types;
Transient Tic Disorder; These are tics that usually last 4-6 months and then go away on their own. Transient tic disorder can be seen in approximately 20% of children.
Chronic Motor or Vocal Tic Disorder; It is a type of tic disorder that lasts longer than one year and includes only motor or vocal tics. This condition is less common than transient tic disorder.
Tourette’s Syndrome; It is a type of tic disorder in which both motor and vocal tics are seen together, lasting at least one year and usually starting in childhood.
Tourette Syndrome and other tic disorders are disorders that negatively affect quality of life. Therefore, it is important to start the correct diagnosis and treatment process by consulting a specialist when you notice the symptoms.
How Is Tourette Syndrome (tic Disorders) Diagnosed?
Tourette Syndrome is a neurological disorder in which involuntary movements and sounds, called motor and vocal tics, begin in childhood. The diagnosis of this condition is usually made by child neurology and psychiatry specialists. There are some criteria for diagnosing a tic disorder.
- First, motor and/or vocal tics must occur continuously for at least one year.
- The age of onset of tics should be less than 18 years.
- The tics must not have occurred due to another medical condition, substance use or drug use.
For the diagnosis of Tourette Syndrome, at least two motor tics and one vocal tic are required. The persistence and severity of these tics can vary from person to person. During the diagnostic process, doctors take a detailed medical and family history and perform a physical and neurological examination. Some laboratory tests and neuroimaging methods may be used when deemed necessary. However, usually these tests are not specific for the diagnosis of Tourette Syndrome and are used to rule out other causes.
How Is Tourette Syndrome (tic Disorders) Treated?
The treatment for Tourette’s Syndrome varies depending on the severity of the tics and the impact on the quality of life. If the tics seriously affect daily life or cause problems in social, educational or work life, treatment options are considered. The methods used for the treatment of Tourette Syndrome are as follows;
- Drug treatment,
- Behavioral therapy,
- Training and support,
Does Tourette Syndrome (tic Disorders) Go away?
Tourette Syndrome is usually a lifelong condition. However, the severity and frequency of tics can change over time. For many people, tics peak during adolescence and decrease during adulthood. In some cases, the tics may go away completely. However, it may progress differently in each individual and may persist into adulthood in some individuals.
With treatment, the severity and frequency of tics can be reduced and the quality of life can be improved. However, there is currently no definitive cure for Tourette Syndrome. Therefore, it is important to normalize one’s life as much as possible with early diagnosis and appropriate treatment methods.
Tourette Syndrome (tic Disorders) Treatment Prices in Turkey
Turkey has succeeded in making its name known to the world with its investments and Especially the latest technological devices used in diagnosis and treatment procedures have been a beacon of hope for many diseases. However, there has been an increase in health tourism in Türkiye.
- Hospitals are large, clean, spacious and fully equipped in terms of technological equipment.
- Turkish doctors are specialized, successful, and skilled in their fields.
- Nurses and carers are friendly and compassionate.
- Finding answers to the questions asked quickly and accurately.
- Patience and understanding of all staff, including the intermediary company dealing with the patient.
- Turkey offers holiday opportunities with its natural and historical beauties.
- Easy transportation.
- Diagnosis, treatment, accommodation, eating, drinking, dressing, and holiday needs can be met at affordable prices.
Such situations are shown among the reasons for preference. We can see that patients and their relatives who want to come to Turkey are doing research on Tourette Syndrome (Tic Disorders) Treatment Prices in Turkey. However, it would not be correct to give clear price information at this stage. Many factors such as the type of disease, stage, diagnosis process, treatment process, and stay in Türkiye affect the price issue. If you want to get more detailed price information, you can contact us. In addition, if you come to Turkey for treatment through us, we can facilitate your visa application process with the invitation letter sent by us to the consulate.
Prof.Dr. Ergün Dağlıoğlu (Eğitim Görevlisi), Beyin Cerrahı, Beyin Ve Sinir Cerrahisi

domain: medical


text:
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domain: pet


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posted on Sep, 26 2011 @ 05:25 PM
I can't shake the fact of what this world has quickly become, an overpopulated wasteland full of vice and iniquity. I blame parenting, more and more
adults are becoming diagnosed with something called RAD Reactive Attachment Disorder. An attachment disorder is usually seen in children who have been
neglected in some fasion by their parental figures, where the detached individual becomes emotionless and empathetic. This carries on into youth and
adulthood, with an increasing likeliness of a young unintended pregnancy, increasing the chance that the unintended child will suffer from the same
disorder. In adults it can ruin relationships through a lack of emotional values, either through infidelity or through excessive paranoia regarding
the later. This disorder can only spread in these emotionally difficult times, and I see a very cold, untrustworthy future for humankind, the
atrocities of todays and yesterdays do not settle without some impact on the future. I think we are at the pinacle of history, and although we have
seen the devastation caused by our conflicts and malicous intenders throughout history leading to this point, I do believe we reside in the purest of
As fallacious as technology may seem to those of us capable of faithing reality, it does provide us with the greatest advantage had to oppose the
discord running rampant throughout society. It allows for injustices and degregation throughout the world to be brought to the attention of those who
can and would make a difference for those oppressed. It begins in the home, these days when gender lines are evaporating, along with the newfound
freedoms and rights for women comes an increase in responsibility and higher expectations for those women to become selfsufficient. These added
stresses have overwhelmed the female population, by disintegrating the very function and values that have been naturally instilled into the female
psyche and physique.
Women do not react or think like men therefore their interpretation of existence would not be equal to a mans' contextually speaking. Their views
would not be less equal on any account either, instead it should be viewed as just the divergence in society.
We should push towards a partition of sorts in matters involving both sexes, as in that true gender equality can be described as the complete lack of
oppression to an unbiased representation of the opposite gender as a whole, and not the misconception that opposing sexes are equal to and parallel
each other in every manifestation possible. To assume the later is approaching gender unequally, with that one could state that if a women is equal to
a man than a women should be able to compete with a man, and a man should be able to compete with a women on any occasion and both will retain the
same statistical odds of accomplishing the the task at hand. If this is true, then stating that a man can and should give birth to children, is also
true, but we can all conclude that it is, in reality, false.
Another example using childbirth is the notion that married women who have children are expected to provide a portion of finance for the household,
especially as of lately, if they do not, they are viewed as betrayers of the feminist movement, or as lazy and inconsiderate to her spouses
providence. This has added responsibility to the woman in this context, and while erasing crucial nurturing time, it also seperates children from that
ever so important maternal figure. It also leads to stresses that can cause infidelity, or may emasculate the faternal figure in the family setting by
instilling this need to provide more so that his spouse may leave the workforce to reunite with her life at home.
Gender equality and women's rights are a great motivating force towards true equality, but I feel that the fundementals in these movements are
flawed, and that feminism instead of promoting equality of women WITH men in a manner of respect, admiration, and morality, it mistakenly promotes
equality of women TO men in a manner of power, responsibility, and dominance. For equality to exist there must be a loss to both groups and a gain for
both groups. Women will never be fully integrated with me,n instead the sexes will have to rest on compromise in settling disputes, but with a firm
understanding that both parties have availible to themselves specific rights that ensure that their opinions are heard and decided upon with unbiased
If this is not understood we will continue down a tormenting path of blatant disrespect for any value the opposite sex has, men will view women as the
mistress and temptress, whereas the women will view the man as the en-captor and abuser. These critiques will then continue to be evident in society
and all the more prevalent, the mistreatment of men by women and women by men based solely on gender must stop.
We must difine the roles of each sex in the family unit, because the family unit is dying and with it goes the ability to truly love and trust one
another, our kids are growing up abused and neglected without choice, brought into the condition without choice, and often react to the mistreatment
in a negative manner leaving them troubled, angry, and alone, to which they have no choice.
But you as an adult, as a spouse in a relationship, as a father, or as a mother, and both as parental beings, you do have a choice to love, love
eachother and to love your creation, because your impact on the world cannot be determined until your children begin making the choices you have
prepared them for, and how well you prepare them determines whether you suceeded or failed. For those who have no children of their own, it is up to
you to provide where so many have neglected, it is your responsibility to the future of our race to mentor and care, because the children have no
choice, the choice has been passed to all of you to make. Its as easy as love, help save love before its extinction and eradication.
You can make easy decisions your whole life, they are hastened, and so cheap are the rewards, but the decisions you value and invest time debating,
the hard decisions to make, they are the ones that reap rewards of unimaginable wealth. Trust me, ask any heroin addict if it's easy to quit, it's
easier to stay addicted, and the reward is a quick death and an uneventfully short life, but beating the addiction fills you with the confidence to
accomplish your wildest dreams, and restores health and life, but it isn't easy...
I found the love, it's here in the truth, it does not make you feeble, the truth makes you stronger, the truth is discovering yourself and molding
who you want to be in this world, and not giving in to who your expected to be.

domain: sociology


text:
BMW automobile enthusiasts have become used to a certain level of a reputation for performance when driving their 640i xDrive Gran Coupe around town. The vehicle in your driveway is head and shoulders above the rest compared to all the other cars, trucks, and SUVs currently being driven today. While it's not hard to locate replacement parts for BMW vehicles, these days only the absolute best are required to maintain your car or truck. Auto enthusiasts appreciate the luxury built into each BMW, and know that only top quality repair 640i xDrive Gran Coupe parts will suffice when repairs are needed.
If turning didn?t ever exist, then every road would be completely straight. That would be quite boring for BMW 640i xDrive Gran Coupe owners, and it would also mean that tie rods would likely never have been invented. Your vehicle?s tie rod ends connect the center link or the steering rack to the .10, allowing the wheels to pivot and turn. If your BMW 640i xDrive Gran Coupe tie rod is broken, then your ability to steer is reduced and you might not be able to dodge obstacles in the road.
Trust in the value of the parts you get from carpartsdiscount.com and realize incredible savings on all your upcoming repairs. Buying new BMW parts from www.carpartsdiscount.com means you are paying the lowest prices for the most superior original and aftermarket replacement 640i xDrive Gran Coupe parts on the internet. Don't let a broken Tie Rod spell the end of your car, truck, or SUV, and don't risk lowering its resale value by buying anything but the best repair parts. If your car or truck was built in , then you will find the correct BMW 640i xDrive Gran Coupe Tie Rod parts right here.

domain: hobby


text:
Is the moon all it's cracked up to be? Yes — and then some. New analysis of the lunar surface reveals that it's far more fractured than once thought.
Since the moon formed 4.3 billion years ago, asteroid impacts have scarred its face with pits and craters. But the damage goes far deeper than that, with cracks extending to depths of 12 miles (20 kilometers), researchers recently reported.
Though the moon's craters have been well-documented, scientists previously knew little about the upper region of the moon's crust, the megaregolith, which sustained the bulk of the damage from space rock bombardment. In the new study, computer simulations revealed that impacts from single objects could fragment the lunar crust into blocks about 3 feet (1 meter) wide, opening surface cracks that extend for hundreds of kilometers. This suggests that much of the fracturing in the megaregolith could have come from single, high-speed impacts, leaving the crust "thoroughly fractured" early in the moon's history. [When Space Attacks: 6 Craziest Meteor Impacts]
These findings helped to address questions raised by NASA's Gravity Recovery and Interior Laboratory (GRAIL), a mission that sent twin spacecraft to the moon in 2011 to create the most detailed lunar gravity map to date.
Data gathered by GRAIL showed that the moon's crust was far less dense than expected, Sean Wiggins, lead author of the new study and a doctoral candidate with the Earth, Environmental and Planetary Sciences Department at Brown University in Rhode Island, told Live Science.
Wiggins and his colleagues suspected that ancient impacts could have substantially fractured the lunar surface, "adding porosity and therefore lowering the density," he said.
Using simulations, the study authors found that an impact from an object measuring just 0.6 miles (1 km) in diameter could have opened cracks reaching depths of 12 miles (20 km) in the lunar surface. After impacts from objects measuring 6 miles (10 km) in diameter, cracks yawned to similar depths, but also extended laterally to distances up to 186 miles (300 km) from the impact crater.
"There's quite a lot of damage outside of the main crater area," Wiggins said. "Material is still very broken up, farther away than we would have predicted." Over time, networks of cracks grew and connected, creating a fragmented lunar crust, the researchers reported.
The researchers also used the simulations to explore how similar impacts could affect Earth, which has also been pummeled by asteroids, and they found that gravity played an important role in the quantity and severity of the fractures.
Under conditions with higher gravity — such as on Earth — the surface in simulations suffered less damage from impacts, while lower gravity meant that the surface experienced more damage, the simulations showed. This explains why impacts on the moon created surface cracks that penetrated deeper than cracks from asteroid impacts on Earth.
Piecing together a more detailed picture of the megaregolith will help scientists to better understand how that region conducts heat; this could reveal important clues about the formation of other moons and even planets, Wiggins said.
"It definitely opens doors for further investigation into lots of different processes — not just on the moon, but on other bodies as well, like Mars or Earth," he added.
The findings were published online March 12 in the Journal of Geophysical Research: Planets.
- In Photos: The Impact Craters of North America
- Crash! 10 Biggest Impact Craters on Earth
- 10 Things You Didn't Know About the Moon
Originally published on Live Science.
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Mindy Weisberger is an editor at Scholastic and a former Live Science channel editor and senior writer. She has reported on general science, covering climate change, paleontology, biology, and space. Mindy studied film at Columbia University; prior to Live Science she produced, wrote and directed media for the American Museum of Natural History in New York City. Her videos about dinosaurs, astrophysics, biodiversity and evolution appear in museums and science centers worldwide, earning awards such as the CINE Golden Eagle and the Communicator Award of Excellence. Her writing has also appeared in Scientific American, The Washington Post and How It Works Magazine.

domain: news


text:
November 5 2015
"We report that cultured CRC cells harboring KRAS or BRAF mutations are selectively killed when exposed to high levels of vitamin C."
August 5 2015
"With studies proving that vitamin C deficiency is a problem for all people who have a thyroid condition, it’s possible that adrenal fatigue could be the cause. Therefore, it’s imperative to include foods and/or supplements containing vitamin C, especially with extreme or ongoing stress."
April 7 2015
"With the recent price jump in sterilized vitamin C for injection, the Vitamin C Foundation shares that the sharp increase may be better for patients in the long term. Doctors who utilize intravenous vitamin C therapy will be spurred to seek out lower-cost options that may work even better."
March 25 2015
"Based on their results, the researchers concluded that vitamin C and serum ascorbate intake may be inversely related to the risk of cataract and therefore, vitamin C intake should be encouraged for the primary prevention of cataract."


text: Delve 23rd October – 16th November 2013 Leyden Gallery are proud to present the work of the artist Lindsay Moran, sculptor Atsuko Nakamura and photographers Stuart Kerr and Joanna Casey; each of whom contribute to the concept of delving implicit to this show. To delve implies a movement into another space, a desire to immerse elsewhere. The structures and languages of the urban sites we occupy still hold the mythic remnants of this desire – living in an ‘urban jungle’ connotes architectural profusion and confusion. Within this modern mayhem, we still value and implant spaces of green and patches of nature. As cities overcrowd and forests diminish; becoming the reduced sites of production for our overloaded lives, we turn to delving elsewhere. ‘Delve’, in transporting images and remnants of forests and greenery into the heart of the urban landscape, questions, and in doing so, reconfigures these sites imbued with legendary and mythical tales. Here, where art, memory and yearning enter into a bond of visual splendour, the skewed nature of a precious eco balance is also highlighted. Two of the artists in this show literally take their practice into the forests around Britain – ancient, yet ever-becoming sites, filled with fabulous tales and magical light.

domain: artistic


text:
What Role Did Shakespeare Play In The Original Production Of Hamlet The Tempest did that. original music for the show. As Reich and Company re-imagine this production, audiences can expect some changes. The company cast gender-blind so there are many female.
Jenny and the Cat Club, written and illustrated by Esther Averill, published by the New York Review of Books, £10. Ages 4+ Poor Jenny, bright as a penny, is a little black cat simply too timid to have.
Feb 5, 2018. It's like a book club in your ears and everyone's invited. From nonfiction books about food culture to classic children's books, no genre is off.
This book is Witherspoon’s June book club book, and it’s one that she completely. She came alive while she was reading." One of the books that Witherspoon recalled was Harper Lee’s classic. Set in.
May 6, 2017. Both social and fun, book clubs are the perfect way to take my. also have a monthly book club featuring hard-hitting classic novels that tend to.
Three works of literature that share a common theme (e.g. dystopia, books. At least one classic and one contemporary novel are included in each theme's book.
It’s been nearly 50 years since US author Judy Blume published her groundbreaking and controversial debut novel Are You There God. says she even started her own period club with her friends after.
Poet Who Wrote Of Daedalus A Tale Of Two Cities Charles Dickens Introduction It was the best of times, it was the worst of times. — Charles Dickens Precious metal investors have two distinct mindsets:
The Top 50 greatest fiction books of all time determined by 120 lists and articles. From the preeminent prose satirist in the English language, a great classic. The editors of the Norwegian Book Clubs, with the Norwegian Nobel Institute,
Welcome to the California Classic. A sellout crowd flooded Golden 1 Center to catch. By waiving him, that money is now off.
Classic or vintage books with tea or coffee and perhaps some stationery too. There are three clubs for you to be a part of, choose your favourite and join the fun!
Netflix has announced that it will be releasing a live-action TV series based on classic Young Adult novel series The Baby-Sitters Club. The ten-part series will. The ten-episode series is based on.
Phobos And Deimos Mythology Deimos will suffer the opposite fate. Its orbit is slowly drawing it away from Mars, and eventually the moon will be cast off into space. Deimos has a number of
With a long print history, this instruction classic is popular for its easy. thus producing revealing looks at multiple aspects of the club and its legendary personalities. There were previous.
A different way to do leadership development could be to start a companywide classics reading group. Many of the greatest books ever written address both the rewards and the risks of seeking.
If you need help choosing your next Book Club book to discuss, you'll find plenty of inspiration in this Book Bucket List from. All these hand-picked books are available at unbelievable prices. This is a classic story that just has to be read!
Bokklubben World Library (Norwegian: Verdensbiblioteket) is a series of classical books, mostly novels, published by the Norwegian Book Club since. Books portal · Classic book · Larry McCaffery's list of the 20th Century's Greatest Hits: 100.
Happy Step Fathers Day Poems • Father’s Day eCards for Everyone: Simply choose a card you know they’ll love and send it to them via email or post it to their Facebook wall. • Happy
The Classic Book Club – next meeting (date TBA). Current host Frances Quesnel: “ For books are not absolutely dead things, but do contain a potency of life….
Jul 14, 2016. With that reputation comes a vibrant book club community, but mind you, you don' t. to comic books groups, the city has a book club for every taste. separate book clubs revolving around the topics of classic science fiction,
Solid State Books hosts multiple book clubs, all free and open to the public. Join bookseller Gwen Hunter to read and discuss selections of classic and.
Mar 21, 2019. It wasn't just that they weakened the spines of the books or that some of them were youth-inappropriate cigarette ads. A fair fraction of them.
Online shopping for 100 Books To Read In A Lifetime from a great selection at Books Store. Edition: $11.99. Alice's Adventures in Wonderland & Through the Looking-Glass (Bantam Classics). Breath, Eyes, Memory (Oprah's Book Club).
Dec 31, 2017. Book clubs encourage you to try books you might not have picked. I've picked up a bunch of newer and classic novels and nonfiction reads.
Feb 10, 2015. Last week, celebrity-loved book club expert Julie Goler taught us how to start a book club that doesn't turn into Lord of the Flies. This week, she.
Hosted by John Mullan, professor of English at University College London, the Guardian's Book Club examines a book a month, via a weekly column in the.
Who Was Ajax In Greek Mythology It was said that as a boy Ajax was given into the care of the centaur Chiron for training; Chiron would indeed train many of the great heroes of Greek
Jul 28, 2017. It illustrates how books can bring people together. The Joy Luck Club. "This is an unsurpassed classic filled with stories for the personal.
Join a book club to meet friends, discuss great books, and experience new literature. Discusses classic novels and recent award winners. Meets the first.
Most of our book club books are available through Library2Go at no cost, and. Between The Panels; Classic Re-Reads; Great Reads; Life After Life; Pints &.
But Kevin Kisner did have to turn to playing competitor and tournament host Rickie Fowler Thursday morning after playing his.
Numerous past NFL and NBA stars participated in the 10th annual Andre Reed Celebrity Golf Classic at the Lehigh Country Club.
There are two classic questions beloved by both interviewers. [The tale of the ‘lost’ Gutenberg Bible and other books about..books] Because it tracks at least a dozen figures, “The Club” can’t.

domain: entertainment


text:
Rising for the third consecutive month and reaching a 10-year high. But what does it mean?
PALM COAST, FL – May 27, 2016 – Pending home sales rose for the third consecutive month in April and reached their highest level in over a decade, according to the National Association of Realtors®. All major regions saw gains in contract activity last month except for the Midwest, which saw a meager decline. What is driving the increase?
The Pending Home Sales Index,* a forward-looking indicator based on contract signings, hiked up 5.1 percent to 116.3 in April from an upwardly revised 110.7 in March and is now 4.6 percent above April 2015 (111.2). After last month's gain, the index has now increased year-over-year for 20 consecutive months.
Lawrence Yun, NAR chief economist, says vast gains in the South and West propelled pending sales in April to their highest level since February 2006 (117.4). "The ability to sign a contract on a home is slightly exceeding expectations this spring even with the affordability stresses and inventory squeezes affecting buyers in a number of markets," he said. "The building momentum from the over 14 million jobs created since 2010 and the prospect of facing higher rents and mortgage rates down the road appear to be bringing more interested buyers into the market."
On the topic of mortgage rates, which have remained below 4 percent in 16 of the past 17 months1, Yun says it remains to be seen how long they will stay this low. Along with rent growth, rising gas prices — and the fading effects of last year's cheap oil on consumer prices — could edge up inflation and push rates higher. For now, he foresees mortgage rates continuing to hover around 4 percent in coming months, but inflation could potentially surprise the market and cause rates to increase suddenly.
Adds Yun, "Even if rates rise soon, sales have legs for further expansion this summer if housing supply increases enough to give buyers an adequate number of affordable choices during their search."
Following the housing market's best first quarter of existing-sales since 2007 (5.66 million)2 and a decent increase (1.7 percent) in April, Yun expects sales this year to climb above earlier estimates and be around 5.41 million, a 3.0 percent boost from 2015. After accelerating to 6.8 percent a year ago, national median existing-home price growth is forecast to slightly moderate to between 4 and 5 percent.
The PHSI in the Northeast climbed 1.2 percent to 98.2 in April, and is now 10.1 percent above a year ago. In the Midwest the index declined slightly (0.6 percent) to 112.9 in April, but is still 2.0 percent above April 2015.
Pending home sales in the South jumped 6.8 percent to an index of 133.9 in April and are 5.1 percent higher than last April. The index in the West soared 11.4 percent in April to 106.2, and is now 2.8 percent above a year ago.
Note: The Pending Home Sales Index is a leading indicator for the housing sector, based on pending sales of existing homes. A sale is listed as pending when the contract has been signed but the transaction has not closed, though the sale usually is finalized within one or two months of signing.

domain: geography


text:
|aMathematics and statistics for financial risk management /|c Michael B. Miller.
|aHoboken, N.J. :|bJohn Wiley & Sons,|cc2012.
|axi, 291 p. :|bill. ;|c24 cm.
|aIncludes bibliographical references and index.
|a"In chapter 1, there is a review three math topics -- logarithms, combinatorics, and geometric series - and one financial topic, discount factors. Emphasis will be given to the specific aspects of these topics that are most relevant to risk management. In chapter 2, the author explores the application of probabilities to risk management. There is also an introduction to basic terminology and notations that will be used throughout the rest of the book. In chapter 3, Miller teaches how to describe a collection of data in precise statistical terms. Many of the concepts will be familiar, but the notation and terminology might be new. This notation and terminology will be used throughout the rest of the book. In chapter 4, some of the most common probability distributions will be pointed out, followed by a chapter on two closely related topics, confidence intervals and hypothesis testing. For risk management, these are possibly the two most important concepts in statistics. Chapter 6 provides a basic introduction to linear regression models. At the end of the chapter, Miller explores two risk management applications, factor analysis and stress testing. The final chapter is on a class of estimators, which has become very popular in finance and risk management for analyzing historical data. These models hint at the limitations of the type of analysis that we have been explores in previous chapters. This book has a lot of charts and equations"--|c Provided by publisher.
Mathematics and Statistics for Financial Risk Management is a practical guide to modern financial risk management for both practitioners and academics. The recent financial crisis and its impact on the broader economy underscore the importance of financial risk management in today's world. At the same time, financial products and investment strategies are becoming increasingly complex. Today, it is more important than ever that risk managers possess a sound understanding of mathematics and statistics. In a concise and easy-to-read style, each chapter of this book introduces a different topic in mathematics or statistics. As different techniques are introduced, sample problems and application sections demonstrate how these techniques can be applied to actual risk management problems. Exercises at the end of each chapter and the accompanying solutions at the end of the book allow readers to practice the techniques they are learning and monitor their progress. A companion website includes interactive Excel spreadsheet examples and templates. This comprehensive resource covers basic statistical concepts from volatility and Bayes' Law to regression analysis and hypothesis testing. Widely used risk models, including Value-at-Risk, factor analysis, Monte Carlo simulations, and stress testing are also explored. A chapter on time series analysis introduces interest rate modeling, GARCH, and jump-diffusion models. Bond pricing, portfolio credit risk, optimal hedging, and many other financial risk topics are covered as well. If you're looking for a book that will help you understand the mathematics and statistics of financial risk management, look no further.

domain: finance


text:
Oath Keepers founder convicted of seditious conspiracy
A Washington, D.C., jury on Tuesday convicted Stewart Rhodes, the founder of the far-right group Oath Keepers, of seditious conspiracy for his part in the riot and storming of the Capitol on Jan. 6, 2021. A jury found that Rhodes encouraged his followers to be prepared for a civil war and was in contact with them from outside while they stormed the Capitol. This is the first time someone has been convicted of seditious conspiracy since 1995. The charge calls for up to 20 years in prison.
What’s happening to other Oath Keepers members? Four other Oath Keepers members were co-defendants to Rhodes in the trial, which lasted nearly two months. The jury also convicted Florida chapter leader Kelly Meggs of seditious conspiracy and convicted all five defendants of obstruction of an official proceeding. Rhodes is among almost a dozen Oath Keepers charged with seditious conspiracy. The Justice Department has charged more than 700 people in its investigation into the Jan. 6 riot. Many of the higher-profile cases have yet to be adjudicated.
Dig deeper: Read Carolina Lumetta and Esther Eaton’s report in The Stew about judicial and security updates one year after the riots.
If you enjoyed this article and would like to support WORLD's brand of Biblically sound journalism, click here.
An actual newsletter worth subscribing to instead of just a collection of links. —AdamSign up to receive The Sift email newsletter each weekday morning for the latest headlines from WORLD’s breaking news team.


text:
LUPITA AROUND THE PLANET
Musicals Interstitial 3d Animation
Target: Toddlers (0-3)
Status: Available Now
Launch Year: 2019
Looking for: New broadcasters licensing, and financing and co-production for new seasons
Rights Available: All available, Worldwide (all languages)
Languages: Brazilian Portuguese and English
Co-production: Druzina Content e Petit Fabrik
Highlights: It´s unprecedented and new content.
Created independently, unpublished and original, they are musical interstitial programs of Lupita traveling through diverse countries and cultures discovering the world! Come dance and sing with us!


text:
Need a redesigned front page for [login to view URL]
The approximate new design for the desktop is presented in attached file - [login to view URL]
The developer will need to use/modify the html top bar and the site's logo from existing URL --
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Feel free to improve the fonts - but make sure they are consistent.
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Part of the web page elements and an example of how we want the design to change if the screen size changes from desktop to mobile can be found at our URL: [login to view URL]
The developer will not have access to our server but, instead, will present the work from their own URL.
20 pekerja bebas membida secara purata $151 untuk pekerjaan ini
We will redesign from the scratch with unlimited revisions. Our Portfolio. [login to view URL] I would love to discuss your project over chat. Sincerely Regards David
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I will Build a starting web page for desktop and mobile devices. I have 4+ years experience. I will start right now. CSS, Graphic Design, HTML, Website Design NJ
Hi, I have strong experience and skill in PSD to HTML. I'm interested with this job posting and I can help you to build your starting web page in various resolutions. Thanks & Regards

domain: design