Bioeffects of albumin-encapsulated microbubbles and real-time myocardial contrast echocardiography in an experimental canine model

Myocardial contrast echocardiography has been used for assessing myocardial perfusion. Some concerns regarding its safety still remain, mainly regarding the induction of microvascular alterations. We sought to determine the bioeffects of microbubbles and real-time myocardial contrast echocardiography (RTMCE) in a closed-chest canine model. Eighteen mongrel dogs were randomly assigned to two groups. Nine were submitted to continuous intravenous infusion of perfluorocarbon-exposed sonicated dextrose albumin (PESDA) plus continuous imaging using power pulse inversion RTMCE for 180 min, associated with manually deflagrated high-mechanical index impulses. The control group consisted of 3 dogs submitted to continuous imaging using RTMCE without PESDA, 3 dogs received PESDA alone, and 3 dogs were sham-operated. Hemodynamics and cardiac rhythm were monitored continuously. Histological analysis was performed on cardiac and pulmonary tissues. No hemodynamic changes or cardiac arrhythmias were observed in any group. Normal left ventricular ejection fraction and myocardial perfusion were maintained throughout the protocol. Frequency of mild and focal microhemorrhage areas in myocardial and pulmonary tissue was similar in PESDA plus RTMCE and control groups. The percentages of positive microscopical fields in the myocardium were 0.4 and 0.7% (P = NS) in the PESDA plus RTMCE and control groups, respectively, and in the lungs they were 2.1 and 1.1%, respectively (P = NS). In this canine model, myocardial perfusion imaging obtained with PESDA and RTMCE was safe, with no alteration in cardiac rhythm or left ventricular function. Mild and focal myocardial and pulmonary microhemorrhages were observed in both groups, and may be attributed to surgical tissue manipulation.

Gene expression profiling of clinical stages II and III breast cancer

Clinical stage (CS) is an established indicator of breast cancer outcome. In the present study, a cDNA microarray platform containing 692 genes was used to identify molecular differences between CSII and CSIII disease. Tumor samples were collected from patients with CSII or CSIII breast cancer, and normal breast tissue was collected from women without invasive cancer. Seventy-eight genes were deregulated in CSIII tumors and 22 in CSII tumors when compared to normal tissue, and 20 of them were differentially expressed in both CSII and CSIII tumors. In addition, 58 genes were specifically altered in CSIII and expression of 6 of them was tested by real time RT-PCR in another cohort of patients with CSII or CSIII breast cancer and in women without cancer. Among these genes, MAX, KRT15 and S100A14, but not APOBEC3G or KRT19, were differentially expressed on both CSIII and CSII tumors as compared to normal tissue. Increased HMOX1 levels were detected only in CSIII tumors and may represent a molecular marker of this stage. A clear difference in gene expression pattern occurs at the normal-to-cancer transition; however, most of the differentially expressed genes are deregulated in tumors of both CS (II and III) compared to normal breast tissue.

Johdon mittariston kehittäminen

Tämän diplomityön tarkoituksena on tarjota yritykselle ratkaisu talouden seu-rantaan reaaliaikaisesti. Nykyaikaisessa maailmassa yrityksen on kehitettävä toimintaansa jatkuvasti toimintaansa pysyäkseen kilpailussa mukana. Yrityksen toiminnan kehittämisen taustalla ovat myös muuttuneet omistussuhteet ja yri-tyksen uuden pääomistajan kiinnostus seurata taloutta reaaliaikaisemmin. Diplomityössä yritykselle tarjottavat ratkaisut tuotetaan teorian ja yritykseltä saadun tiedon yhdistämisellä. Teoriaosuudessa käydään erilaiset ratkaisumallit perusteluineen läpi. Empiirisessä osassa sovelletaan teoriaosa käytäntöön. Lop-putuloksena yrityksen johdolle tarjotaan valmiita työkaluja talouden seurantaan. Työn tulosten luotettavuus perustuu kirjoittajan laajaan yrityksen tuntemukseen, sekä avoimiin keskusteluihin yrityksen ylimmän johdon kanssa. Yritysjohto an-toi työssä käyttöön kaiken mahdollisen, hyvin salaisenkin materiaalin käyttöön, työn mahdollisimman hyvän lopputuloksen saamiseksi. Pääasiallisen lopputuloksena luotiin sähköinen tuntiseurantalomake, jota nimetyt henkilöt pystyvät seuraamaan reaaliaikaisesti internetin avulla. Yritykselle mää-ritettiin olennaisimmat taloudenmittarit, joita yrityksen tulee seurata ja päivittää määritellyin aikavälein, lisäksi määrättiin vastuu alueet, kenelle asioiden päivitys kuuluu. Lisäksi luotiin avainlukujen seurantatyökalupohja, jonne on kerätty kaikki yrityksen toiminnan kannalta tärkeimmät tiedot. Työkalut ovat valmiina käyttöönotettavaksi. Lopullinen luodun ohjelman hyödyntäminen ja käyttöönot-to tapahtuu myöhemmin.

Portfolio as an Indicator of Young Learners’ English Proficiency in Mainstream Language Instruction (EFL) and Bilingual Content Instruction (CLIL)

Tämä soveltavan kielitieteen ja kielitaidon arvioinnin toimintatutkimus tarkasteli kieliportfolion ominaisuuksia ja mahdollisuuksia nuorten oppijoiden englannin kielen arvioinnissa kahdessa eri oppimiskontekstissa: englanti oppiaineena (EFL) ja kaksikielinen sisällönopetus (CLIL). Tutkielman itsenäiset, kahteen eri englannin kielen rekisteriin (arkikieli ja akateeminen kieli) kohdistuneet portfoliokokeilut olivat erillisiä tapaustutkimuksia. Molemmat portfoliot perustuivat väljästi Eurooppalaiseen kielisalkkumalliin, ja ne olivat osa tutkielmantekijän luokkaopetusta ja -toimintaa. EFL -portfoliokokeilu 9-10-vuotiaille kolmasluokkalaisille toteutettiin marraskuun 2011 ja toukokuun 2012 välisenä aikana, kun CLIL -portfoliokokeilu n. 7-9-vuotiallle ensimmäisen ja toisen luokan oppilaille kesti kaksi lukuvuotta 2012–2014. Molemmissa kokeiluissa myös oppilaiden vanhemmat kuuluivat tutkimusjoukkoon, samoin CLIL -portfolion toteutuksessa avustaneet ja opettajanäkökulmaa edustaneet opettajaopiskelijat. Portfoliokokeilun aloitti myös kaksi muuta CLIL -opettajaa, mutta kumpikin kokeilu päättyi alkuvaiheeseensa. Tarkemman tarkastelun kohteina olivat tutkimuksen osallistujien kokemukset ja mielipiteet portfoliokokeiluista. Erityisesti tavoitteena oli selvittää, miten informatiivisena englannin kielitaidon indikaattorina kieliportfoliota pidettiin. Myös kehitysehdotuksia kerättiin. Trianguloitu aineisto koottiin sekä puolistrukturoiduin kyselyin että vapaaehtoisin teemahaastatteluin, jotka äänitettiin. EFL -aineisto koostui 18 oppilaskyselystä, 17 huoltajakyselystä ja 7 oppilashaastattelusta. CLIL -aineistoon sisältyi 19 oppilaskyselyä, 18 huoltajakyselyä, 7 oppilashaastattelua ja yksi opettajaopiskelijoiden (N=3) ryhmähaastattelu. Aineisto analysoitiin pääosin kvalitatiivisin menetelmin temaattisen sisältöanalyysin keinoin, mutta myös laskien frekvenssejä ja prosenttisosuuksia. Osallistujien mielipiteet ja kokemukset olivat hyvin samankaltaiset ja positiiviset kummassakin portfoliokokeilussa. Merkittävä enemmistö sekä oppilaista että huoltajista koki, että portfolion avulla on mahdollista osoittaa englannin kielitaitoa ja sen kehittymistä. Oppilaat kuvailivat portfoliotyötä hauskaksi ja kivaksi, ja heidän mielestään portfoliotehtävien pitäisi olla tarpeeksi haastavia, sisältää taiteellisia ja luovia elementtejä sekä kohdistua tuttuihin, mielenkiintoisiin aiheisiin. He totesivat, että portfolion avulla voi oppia lisää kieltä. Vanhempien mielestä portfolio kertoo koulun vieraiksi jääneistä oppisisällöistä, auttaa ymmärtämään lapsen ajatusmaailmaa ja motivaatiotasoa sekä paljastaa heidän kielitaidostaan uusia ulottuvuuksia. Opettajaopiskelijat havaitsivat, että portfolion avulla voi tutustua oppilaiden kieli- ja kulttuuritaustoihin sekä kartoittaa heidän kielellisiä tarpeitaan. Tämän tutkielman teoreettisen tarkastelun ja tulosten mukaan kieliportfolio tukee erinomaisesti uuden Perusopetuksen Opetussuunnitelman (NCC 2014) tavoitteita ja arvioinnin uudistuspyrkimyksiä sekä lainsäädännön arvioinnille asettamia edellytyksiä. Portfolio on erittäin suositeltava nuorten oppijoiden kielitaidon arviointimenetelmä perinteisten rinnalle.

Fuzzy real option analysis in patent related decision making and patent valuation

The shift towards a knowledge-based economy has inevitably prompted the evolution of patent exploitation. Nowadays, patent is more than just a prevention tool for a company to block its competitors from developing rival technologies, but lies at the very heart of its strategy for value creation and is therefore strategically exploited for economic pro t and competitive advantage. Along with the evolution of patent exploitation, the demand for reliable and systematic patent valuation has also reached an unprecedented level. However, most of the quantitative approaches in use to assess patent could arguably fall into four categories and they are based solely on the conventional discounted cash flow analysis, whose usability and reliability in the context of patent valuation are greatly limited by five practical issues: the market illiquidity, the poor data availability, discriminatory cash-flow estimations, and its incapability to account for changing risk and managerial flexibility. This dissertation attempts to overcome these impeding barriers by rationalizing the use of two techniques, namely fuzzy set theory (aiming at the first three issues) and real option analysis (aiming at the last two). It commences with an investigation into the nature of the uncertainties inherent in patent cash flow estimation and claims that two levels of uncertainties must be properly accounted for. Further investigation reveals that both levels of uncertainties fall under the categorization of subjective uncertainty, which differs from objective uncertainty originating from inherent randomness in that uncertainties labelled as subjective are highly related to the behavioural aspects of decision making and are usually witnessed whenever human judgement, evaluation or reasoning is crucial to the system under consideration and there exists a lack of complete knowledge on its variables. Having clarified their nature, the application of fuzzy set theory in modelling patent-related uncertain quantities is effortlessly justified. The application of real option analysis to patent valuation is prompted by the fact that both patent application process and the subsequent patent exploitation (or commercialization) are subject to a wide range of decisions at multiple successive stages. In other words, both patent applicants and patentees are faced with a large variety of courses of action as to how their patent applications and granted patents can be managed. Since they have the right to run their projects actively, this flexibility has value and thus must be properly accounted for. Accordingly, an explicit identification of the types of managerial flexibility inherent in patent-related decision making problems and in patent valuation, and a discussion on how they could be interpreted in terms of real options are provided in this dissertation. Additionally, the use of the proposed techniques in practical applications is demonstrated by three fuzzy real option analysis based models. In particular, the pay-of method and the extended fuzzy Black-Scholes model are employed to investigate the profitability of a patent application project for a new process for the preparation of a gypsum-fibre composite and to justify the subsequent patent commercialization decision, respectively; a fuzzy binomial model is designed to reveal the economic potential of a patent licensing opportunity.

Simultaneous detection of the C282Y, H63D and S65C mutations in the hemochromatosis gene using quenched-FRET real-time PCR

Hereditary hemochromatosis (HH) is a common autosomal disorder of iron metabolism mainly affecting Caucasian populations. Three recurrent disease-associated mutations have been detected in the hemochromatosis gene (HFE): C282Y, H63D, and S65C. Although HH phenotype has been associated with all three mutations, C282Y is considered the most relevant mutation responsible for hemochromatosis. Clinical complications of HH include cirrhosis of the liver, congestive cardiac failure and cardiac arrhythmias, endocrine pancreatic disease, which can be prevented by early diagnosis and treatment. Therefore, a reliable genotyping method is required for presymptomatic diagnosis. We describe the simultaneous detection of the C282Y, H63D and S65C mutations in the hemochromatosis gene by real-time PCR followed by melting curve analysis using fluorescence resonance energy transfer (FRET) probes. The acceptor fluorophore may be replaced by a quencher, increasing multiplex possibilities. Real-time PCR results were compared to the results of sequencing and conventional PCR followed by restriction digestion and detection by agarose gel electrophoresis (PCR-RFLP). Genotypes from 80 individuals obtained both by the conventional PCR-RFLP method and quenched-FRET real-time PCR were in full agreement. Sequencing also confirmed the results obtained by the new method, which proved to be an accurate, rapid and cost-effective diagnostic assay. Our findings demonstrate the usefulness of real-time PCR for the simultaneous detection of mutations in the HFE gene, which allows a reduction of a significant amount of time in sample processing compared to the PCR-RFLP method, eliminates the use of toxic reagents, reduces the risk of contamination in the laboratory, and enables full process automation.

Proximal tubular dysfunction as an indicator of chronic graft dysfunction

New strategies are being devised to limit the impact of renal sclerosis on graft function. Individualization of immunosuppression, specifically the interruption of calcineurin-inhibitors has been tried in order to promote better graft survival once chronic graft dysfunction has been established. However, the long-term impact of these approaches is still not totally clear. Nevertheless, patients at higher risk for tubular atrophy and interstitial fibrosis (TA/IF) development should be carefully monitored for tubular function as well as glomerular performance. Since tubular-interstitial impairment is an early event in TA/IF pathogenesis and associated with graft function, it seems reasonable that strategies directed at assessing tubular structural integrity and function would yield important functional and prognostic data. The measurement of small proteins in urine such as α-1-microglobulin, N-acetyl-beta-D-glucosaminidase, alpha/pi S-glutathione transferases, β-2 microglobulin, and retinol binding protein is associated with proximal tubular cell dysfunction. Therefore, its straightforward assessment could provide a powerful tool in patient monitoring and ongoing clinical assessment of graft function, ultimately helping to facilitate longer patient and graft survival associated with good graft function.

The performance of semi-quantitative differential PCR is similar to that of real-time PCR for the detection of the MYCN gene in neuroblastomas

Amplification of the MYCN gene in neuroblastomas is a potent biological marker of highly aggressive tumors, which are invariably fatal unless sound clinical management is applied. To determine the usefulness of semi-quantitative differential PCR (SQ-PCR) for accurate quantification of MYCN gene copy number, we evaluated the analytical performance of this method by comparing the results obtained with it for 101 tumor samples of neuroblastoma to that obtained by absolute and relative real-time PCR. Similar results were obtained for 100 (99%) samples, no significant difference was detected between the median log10 MYCN copy number (1.53 by SQ-PCR versus 1.55 by absolute real-time PCR), and the results of the two assays correlated closely (r = 0.8, Pearson correlation; P < 0.001). In the comparison of SQ-PCR and relative real-time PCR, SQ-PCR versus relative real-time PCR concordant results were found in 100 (99%) samples, no significant difference was found in median log10 MYCN copy number (1.53 by SQ-PCR versus 1.27 by relative real-time PCR), and the results of the two assays correlated closely (r = 0.8, Pearson correlation; P < 0.001). These findings indicate that the performance of SQ-PCR was comparable to that of real-time PCR for the amplification and quantification of MYCN copy number. Thus, SQ-PCR can be reliably used as an alternative assay in laboratories without facilities for real-time PCR.

Citations to papers from Brazilian institutions: a more effective indicator to assess productivity and the impact of research in graduate programs

A recent assessment of 4400 postgraduate courses in Brazil by CAPES (a federal government agency dedicated to the improvement of the quality of and research at the postgraduate level) stimulated a large amount of manifestations in the press, scientific journals and scientific congresses. This gigantic effort to classify 16,400 scientific journals in order to provide indicators for assessment proved to be puzzling and methodologically erroneous in terms of gauging the institutions from a metric point of view. A simple algorithm is proposed here to weigh the scientometric indicators that should be considered in the assessment of a scientific institution. I conclude here that the simple gauge of the total number of citations accounts for both the productivity of scientists and the impact of articles. The effort spent in this exercise is relatively small, and the sources of information are fully accessible. As an exercise to estimate the value of the methodology, 12 institutions of physics (10 from Brazil, one from the USA and one from Italy) have been evaluated.

Leptin receptor (Ob-R) mRNA expression and serum leptin concentration in patients with colorectal and metastatic colorectal cancer

The objective of the present study was to investigate the effect of leptin on the progression of colorectal carcinoma to metastatic disease by analyzing the serum leptin concentration and Ob-R gene expression in colon cancer tissues. Tissue samples were obtained from 31 patients who underwent surgical resection for colon (18 cases) and metastatic colon (13 cases) cancer. Serum leptin concentration was determined by an enzyme-linked immunosorbent assay (ELISA) and Ob-R mRNA expression by real-time polymerase chain reaction (RT-PCR) for both groups. ELISA data were analyzed by the Student t-test and RT-PCR data were analyzed by the Mann-Whitney U-test. RT-PCR results demonstrated that mRNA expression of Ob-R in human metastatic colorectal cancer was higher than in local colorectal cancer tissues. On the other hand, mean serum leptin concentration was significantly higher in local colorectal cancer patients compared to patients with metastatic colorectal cancer. The results of the present study suggest a role for leptin in the progression of colon cancer to metastatic disease without weight loss. In other words, significantly increased Ob-R mRNA expression and decreased serum leptin concentration in patients with metastatic colon cancer indicate that sensitization to leptin activity may be a major indicator of metastasis to the colon tissue and the determination of leptin concentration and leptin gene expression may be used to aid the diagnosis.

Identification of main contamination points by hygiene indicator microorganisms in beef processing plants

The microbiological quality of beef and meat products is strongly influenced by the conditions of hygiene prevailing during their production and handling. Without proper hygienic control, the environment in slaughterhouses and butcher shops can act as an important source of microbiological contamination. To identify the main points of microbiological contamination in the beef processing chain, 443 samples of equipment, installations and products were collected from 11 establishments (1 slaughterhouse and 10 butcher shops) located in the state of Paraná, Brazil. The microbiological quality of all the samples was evaluated using Petri dishes to obtain counts of mesophilic aerobes (AC), total coliforms, Escherichia coli (EC), yeasts and molds (YM). The main contamination points identified in butcher shops, in decreasing order, were stainless steel boxes, beef tenderizers, grinders, knives, mixers, sausage stuffers, plastic boxes, floors and drains. In the slaughterhouse, these points were sausage stuffers, platforms, floors and drains. The most severely contaminated products were fresh sausages and ground beef. This information about the main points of microbiological contamination in the beef processing chain is expected to aid professionals responsible for hygiene in similar establishments to set up proper hygienic procedures to prevent or reduce microbiological contamination of beef and meat products.

Detection and quantification of Roundup Ready® soybean residues in sausage samples by conventional and real-time PCR

The increasing presence of products derived from genetically modified (GM) plants in human and animal diets has led to the development of detection methods to distinguish biotechnology-derived foods from conventional ones. The conventional and real-time PCR have been used, respectively, to detect and quantify GM residues in highly processed foods. DNA extraction is a critical step during the analysis process. Some factors such as DNA degradation, matrix effects, and the presence of PCR inhibitors imply that a detection or quantification limit, established for a given method, is restricted to a matrix used during validation and cannot be projected to any other matrix outside the scope of the method. In Brazil, sausage samples were the main class of processed products in which Roundup Ready® (RR) soybean residues were detected. Thus, the validation of methodologies for the detection and quantification of those residues is absolutely necessary. Sausage samples were submitted to two different methods of DNA extraction: modified Wizard and the CTAB method. The yield and quality were compared for both methods. DNA samples were analyzed by conventional and real-time PCR for the detection and quantification of Roundup Ready® soybean in the samples. At least 200 ng of total sausage DNA was necessary for a reliable quantification. Reactions containing DNA amounts below this value led to large variations on the expected GM percentage value. In conventional PCR, the detection limit varied from 1.0 to 500 ng, depending on the GM soybean content in the sample. The precision, performance, and linearity were relatively high indicating that the method used for analysis was satisfactory.