Corporate strategy development: International distance model for the Brazilian market

A Work Project, presented as part of the requirements for the Award of a Masters Degree in Management from the NOVA – School of Business and EconomicsThis paper aims to understand and examine the critical factors, which might help the companies willing to export to Brazil to enter that market. Those factors can be also defined as distances between Brazil and the other countries (companies) and have been analysed in the International Distance Model. In order to understand the findings of that model, it has been applied to one real world example of the Polish company exporting its product to Brazil

Ultrasonography as a Modern Teaching Support to the Anatomy Course: Is It Beneficial for Medical Students

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The impact of cystocele repair on urge symptoms in women with pelvic organ prolapse

Objectives: The purpose of this study was to evaluate the impact of cystocele repair on urinary urge symptoms and to determine the likelihood that urge symptoms are caused by cystocele and therefore cured by cystocele repair. The secondary aim was to assess the impact of baseline cystocele stage POP on the improvement of urge symptoms following surgical treatment of POP. Material and methods: A total of 321 female patients with cystocele stages II, III or IV (POP), who underwent repair surgery for pelvic organ prolapse, were included. A retrospective analysis was performed to determine the presence of urge symptoms in patients with cystocele and to evaluate how many patients were cured from urge symptoms by the cystocele repair. Postoperative data were obtained by interview during a follow-up examination six weeks after surgery. Results: Preoperatively, 52.02% of all patients diagnosed with cystocele stages II, III or IV POP experienced urge symptoms. Urge symptoms were cured in 88.62% of patients with cystocele stages  II after POP repair (p < 0.005). 88.60% of patients with cystocele stage II POP and 88.68% of patients with cystocele stages III to IV POP reported improvement in urge symptoms (p < 0.005). Despite cystocele repair, 11.4% of patients with preoperative cystocele stage II POP and 11.32% with preoperative cystocele stages III and IV POP reported persistent urge symptoms. 5.84% of the study group who showed no urge symptoms preoperatively, experienced de novo urge symptoms after following surgery (p < 0.005). Conclusions: Cystocele repair cured urge symptoms in the majority of patients. Therefore, repair of bladder prolapse may help to differentiate urge symptoms from other urinary tract dysfunctions and assist in determining a proper diagnosis and treatment. However, the severity of POP had no significant influence on the improvement in urge symptoms following cystocele repair. Risk of de novo urge symptoms after anatomical repair still needs to be explored

Application of Least Squares with Conditional Equations Method for Railway Track Inventory Using GNSS Observations

Satellite geodetic networks are commonly used in surveying tasks, but they can also be used in mobile surveys. Mobile satellite surveys can be used for trackage inventory, diagnostics and design. The combination of modern technological solutions with the adaptation of research methods known in other fields of science offers an opportunity to acquire highly accurate solutions for railway track inventory. This article presents the effects of work carried out using a mobile surveying platform on which Global Navigation Satellite System (GNSS) receivers were mounted. The satellite observations (surveys) obtained were aligned using one of the methods known from classical land surveying. The records obtained during the surveying campaign on a 246th km railway track section were subjected to alignment. This article provides a description of the surveying campaign necessary to obtain measurement data and a theoretical description of the method employed to align observation results as well as their visualisation. Document type: Articl

Cloning and purification of functionally active Fas ligand interfering protein (FIP) expressed in Escherichia coli

This report presents purification and characterization of the extracellular domain of rat Fas protein, called FIP (FasL interfering protein), expressed as inclusion bodies in Escherichia coli. FIP was extracted from the inclusion bodies, solubilized with 8 M urea, purified by a single-step immobilized metal ion (Ni2+) affinity chromatography and refolded. SDS/PAGE and mass spectrometry analysis of the purified protein verified its purity. Fluorescence spectrum analysis showed that the refolding procedure caused structural changes which presumably might have led to oligomerization. The purified FIP has biological activities: it binds specifically soluble Fas ligand and protects human Jurkat lymphocytes against FasL-dependent apoptosis. This efficient procedure of FIP expression in E. coli and renaturation may be useful for production of therapeutically important proteins

SuperSILAC mix for quantitative proteomics of human tumor tissue

SILAC-labeled cell lines as internal standard for comprehensive human tumor tissue proteome quantification. Initially, we SILAC-labeled the breast cancer cell line HCC1599 and mixed the lysate with the lysate of mammary carcinoma tissue from an individual with grade II lobular carcinoma. We digested the resulting protein mixture according to the filter-aided sample preparation protocol 10 and separated peptides into six fractions by anion exchange chromatography in a StageTip format 11 . We analyzed each fraction by online reverse-phase chromatography coupled to high-resolution, quantitative mass spectrometric analysis using the LTQ Orbitrap Adding the labeled mixture, rather than the single cell line, to the tumor achieved the same number of quantified proteins in triplicate analysis but drastically improved quantification accuracy We also shortened the analysis time from one day per replicate to four hours per replicate by measuring the digest without prior separation on the new generation LTQ-Orbitrap Velos instrument. Despite the lower We describe a method to accurately quantify human tumor proteomes by combining a mixture of five stable-isotope labeling by amino acids in cell culture (siLac)-labeled cell lines with human carcinoma tissue. this generated hundreds of thousands of isotopically labeled peptides in appropriate amounts to serve as internal standards for mass spectrometry-based analysis. by decoupling the labeling from the measurement, this super-siLac method broadens the scope of siLac-based proteomics. In recent years, mass spectrometry has made great technological progress and is increasingly broadly applied in cell culture-based studies 1,2 . Nevertheless, accurate quantification of human tissue proteomes by high-resolution mass spectrometry methods is still in its infancy 3,4 . In the stable-isotope labeling by amino acids in cell culture (SILAC) method, cell lines are labeled through the incorporation of stable 'heavy' versions of essential amino acids in the cell populations to be compared A few studies have expanded the use of SILAC to tissue analysis. The Neuro2A cell line has been metabolically labeled and compared to total mouse brain 7 . A total of 602 proteins had been quantified, albeit with up to tenfold quantitative ratios between cell line and tissue. Such high ratios between sample and internal standard make accurate quantification difficult because the signal of the lowerabundance peptide in the SILAC pair may be close to the noise level. We have recently quantified proteins from primary mouse hepatocytes using SILAC-labeled Hepa1-6 cells 8 . Furthermore, entire mice can be SILAC-labeled, but this technology is not applicable to human subjects 9 . When applying proteomics to tumor biology, it is imperative to quantify a representative number of proteins, to obtain reproducible results and to study cancer-relevant proteins of low abundance. Therefore, we explored the use of a mix of multipl

Development and Characterization of a Camelid Single Domain Antibody–Urease Conjugate That Targets Vascular Endothelial Growth Factor Receptor 2

Angiogenesis is the process of new blood vessel formation and is essential for a tumor to grow beyond a certain size. Tumors secrete the pro-angiogenic factor vascular endothelial growth factor, which acts upon local endothelial cells by binding to vascular endothelial growth factor receptors (VEGFRs). In this study, we describe the development and characterization of V21-DOS47, an immunoconjugate that targets VEGFR2. V21-DOS47 is composed of a camelid single domain anti-VEGFR2 antibody (V21) and the enzyme urease. The conjugate specifically binds to VEGFR2 and urease converts endogenous urea into ammonia, which is toxic to tumor cells. Previously, we developed a similar antibody–urease conjugate, L-DOS47, which is currently in clinical trials for non-small cell lung cancer. Although V21-DOS47 was designed from parameters learned from the generation of L-DOS47, additional optimization was required to produce V21-DOS47. In this study, we describe the expression and purification of two versions of the V21 antibody: V21H1 and V21H4. Each was conjugated to urease using a different chemical cross-linker. The conjugates were characterized by a panel of analytical techniques, including SDS-PAGE, size exclusion chromatography, Western blotting, and LC-MSE peptide mapping. Binding characteristics were determined by ELISA and flow cytometry assays. To improve the stability of the conjugates at physiologic pH, the pIs of the V21 antibodies were adjusted by adding several amino acid residues to the C-terminus. For V21H4, a terminal cysteine was also added for use in the conjugation chemistry. The modified V21 antibodies were expressed in the E. coli BL21 (DE3) pT7 system. V21H1 was conjugated to urease using the heterobifunctional cross-linker succinimidyl-[(N-maleimidopropionamido)-diethyleneglycol] ester (SM(PEG)2), which targets lysine resides in the antibody. V21H4 was conjugated to urease using the homobifunctional cross-linker, 1,8-bis(maleimido)diethylene glycol (BM(PEG)2), which targets the cysteine added to the antibody C-terminus. V21H4-DOS47 was determined to be the superior conjugate as the antibody is easily produced and purified at high levels, and the conjugate can be efficiently generated and purified using methods easily transferrable for cGMP production. In addition, V21H4-DOS47 retains higher binding activity than V21H1-DOS47, as the native lysine residues are unmodified