Assessing the viability of cryopreserved coconut zygotic embryos by electrolytic conductivity and potassium leaching

The objective of this work was to adapt the application of electrolytic conductivity and potassium leaching tests to assess the viability of cryopreserved embryos of 'Anão Verde do Brasil de Jiqui' (AVeJBr) coconut. The zygotic embryos were excised, sterilized and subjected to four cryoprotectant treatments combined with three incubation times (12, 16 and 20 hours), totaling 12 treatments. The pre-treatment of mature zygotic embryos of AVeJBr coconut using a cryoprotectant with 1.75 mol L-1 of sucrose + 15% glycerol for 12 and 16 hours promoted lower embryo humidity and increased viability in electrolytic conductivity and potassium leaching tests. Samples with ten embryos are sufficient for electrolytic conductivity analysis in cryopreserved or non-cryopreserved AVeJBr coconut zygotic embryos. The 4 to 8 hour imbibition period of the embryos is promising for the electrolytic conductivity analysis of non-cryopreserved mature zygotic embryos of AVeJBr coconut.

Baseline Results of an Acute Ischemic Stroke Registry Including Acute Multimodal Imaging (ASTRAL)

Background: The DEFUSE (n_74) and EPITHET (n_101) studies have in common that a baseline MRI was obtained prior to treatment (tPA in DEFUSE; tPA or placebo in EPITHET) in the 3-6 hour time-window. There were however important methodological differences between the studies. A standardized reanalysis of pooled data was undertaken to determine the effect of these differences on baseline characteristics and study outcomes. Methods: To standardize the studies 1) the DWI and PWI source images were reprocessed and segmented using automated image processing software (RAPID); 2) patients were categorized according to their baseline MRI profile as either Target Mismatch (PWITmax_6/DWI ratio_ 1.8 and an absolute mismatch _15mL), Malignant (DWI or PWITmax_10 lesion _ 100 mL), or No Mismatch. 3) favorable clinical response was defined as NIHSS score of 0-1 or a _8 points improvement on the NIHSSS at day 90. Results: Prior to standardization there was no difference in the proportion of Target Mismatch patients between EPITHET and DEFUSE (54% vs 49%, p_0.6), but the EPITHET study had more patients with the Malignant profile than DEFUSE (35% vs 9%, p_0.01) and fewer patients that had No Mismatch (11% vs 42%, p_0.01). These differences in baseline MRI profiles between EPITHET and DEFUSE were largely eliminated by standardized processing of PWI and DWI images with RAPID software (Target Mismatch 49% vs 48%; Malignant 15% vs 8%; No Mismatch 36% vs 25%; p_NS for all comparisons) Reperfusion was strongly associated with a favorable clinical response in mismatch patients (figure). This relationship was not affected by the standardization procedures (pooled odds ratio of 8.8 based on original data and 6.6 based on standardized data). Conclusion: Standardization of image analyses procedures in acute stroke is important as non-standardized techniques introduce significant variability in DWI and PWI imaging characteristics. Despite methodological differences, the DEFUSE and EPITHET studies show a consistent and robust association between reperfusion and favorable clinical response in Target Mismatch patients regardless of standardization. These data support an RCT of iv tPA in the 3-6 hour time-window for Target Mismatch patients identified using RAPID.

Effect of RasGAP N2 fragment-derived peptide on tumor growth in mice.

Peptides that interfere with the natural resistance of cancer cells to genotoxin-induced apoptosis may improve the efficacy of anticancer regimens. We have previously reported that a cell-permeable RasGAP-derived peptide (TAT-RasGAP(317-326)) specifically sensitizes tumor cells to genotoxin-induced apoptosis in vitro. Here, we examined the in vivo stability of a protease-resistant D-form of the peptide, RI.TAT-RasGAP(317-326), and its effect on tumor growth in nude mice bearing subcutaneous human colon cancer HCT116 xenograft tumors. After intraperitoneal injection, RI.TAT-RasGAP(317-326) persisted in the blood of nude mice for more than 1 hour and was detectable in various tissues and subcutaneous tumors. Tumor-bearing mice treated daily for 7 days with RI.TAT-RasGAP(317-326) (1.65 mg/kg body weight) and cisplatin (0.5 mg/kg body weight) or doxorubicin (0.25 mg/kg body weight) displayed reduced tumor growth compared with those treated with either genotoxin alone (n = 5-7 mice per group; P = .004 and P = .005, respectively; repeated measures analysis of variance [ANOVA, two-sided]). This ability of the RI.TAT-RasGAP(317-326) peptide to enhance the tumor growth inhibitory effect of cisplatin was still observed at peptide doses that were at least 150-fold lower than the dose lethal to 50% of mice. These findings provide the proof of principle that RI.TAT-RasGAP(317-326) may be useful for improving the efficacy of chemotherapy in patients.

Computational analysis of the genetic and environmental contributions to disease-related human phenotypes: From predicting adverse lipid response of HIV patients receiving antiretroviral therapy to genome-wide association studies of cardiovascular and lipid related disorders

Genetic variants influence the risk to develop certain diseases or give rise to differences in drug response. Recent progresses in cost-effective, high-throughput genome-wide techniques, such as microarrays measuring Single Nucleotide Polymorphisms (SNPs), have facilitated genotyping of large clinical and population cohorts. Combining the massive genotypic data with measurements of phenotypic traits allows for the determination of genetic differences that explain, at least in part, the phenotypic variations within a population. So far, models combining the most significant variants can only explain a small fraction of the variance, indicating the limitations of current models. In particular, researchers have only begun to address the possibility of interactions between genotypes and the environment. Elucidating the contributions of such interactions is a difficult task because of the large number of genetic as well as possible environmental factors.In this thesis, I worked on several projects within this context. My first and main project was the identification of possible SNP-environment interactions, where the phenotypes were serum lipid levels of patients from the Swiss HIV Cohort Study (SHCS) treated with antiretroviral therapy. Here the genotypes consisted of a limited set of SNPs in candidate genes relevant for lipid transport and metabolism. The environmental variables were the specific combinations of drugs given to each patient over the treatment period. My work explored bioinformatic and statistical approaches to relate patients' lipid responses to these SNPs, drugs and, importantly, their interactions. The goal of this project was to improve our understanding and to explore the possibility of predicting dyslipidemia, a well-known adverse drug reaction of antiretroviral therapy. Specifically, I quantified how much of the variance in lipid profiles could be explained by the host genetic variants, the administered drugs and SNP-drug interactions and assessed the predictive power of these features on lipid responses. Using cross-validation stratified by patients, we could not validate our hypothesis that models that select a subset of SNP-drug interactions in a principled way have better predictive power than the control models using "random" subsets. Nevertheless, all models tested containing SNP and/or drug terms, exhibited significant predictive power (as compared to a random predictor) and explained a sizable proportion of variance, in the patient stratified cross-validation context. Importantly, the model containing stepwise selected SNP terms showed higher capacity to predict triglyceride levels than a model containing randomly selected SNPs. Dyslipidemia is a complex trait for which many factors remain to be discovered, thus missing from the data, and possibly explaining the limitations of our analysis. In particular, the interactions of drugs with SNPs selected from the set of candidate genes likely have small effect sizes which we were unable to detect in a sample of the present size (<800 patients).In the second part of my thesis, I performed genome-wide association studies within the Cohorte Lausannoise (CoLaus). I have been involved in several international projects to identify SNPs that are associated with various traits, such as serum calcium, body mass index, two-hour glucose levels, as well as metabolic syndrome and its components. These phenotypes are all related to major human health issues, such as cardiovascular disease. I applied statistical methods to detect new variants associated with these phenotypes, contributing to the identification of new genetic loci that may lead to new insights into the genetic basis of these traits. This kind of research will lead to a better understanding of the mechanisms underlying these pathologies, a better evaluation of disease risk, the identification of new therapeutic leads and may ultimately lead to the realization of "personalized" medicine.

Continuous assessment of electrical epileptic activity in acute stroke.

OBJECTIVE: To determine the incidence and risk factors of electrical seizures and other electrical epileptic activity using continuous EEG (cEEG) in patients with acute stroke. METHODS: One hundred consecutive patients with acute stroke admitted to our stroke unit underwent cEEG using 10 electrodes. In addition to electrical seizures, repetitive focal sharp waves (RSHWs), repetitive focal spikes (RSPs), and periodic lateralized epileptic discharges (PLEDs) were recorded. RESULTS: In the 100 patients, cEEG was recorded for a mean duration of 17 hours 34 minutes (range 1 hour 12 minutes to 37 hours 10 minutes). Epileptic activity occurred in 17 patients and consisted of RSHWs in seven, RSPs in seven, and PLEDs in three. Electrical seizures occurred in two patients. On univariate Cox regression analysis, predictors for electrical epileptic activity were stroke severity (high score on the National Institutes of Health Stroke Scale) (hazard ratio [HR] 1.12; p = 0.002), cortical involvement (HR 5.71; p = 0.021), and thrombolysis (HR 3.27; p = 0.040). Age, sex, stroke type, use of EEG-modifying medication, and cardiovascular risk factors were not predictors of electrical epileptic activity. On multivariate analysis, stroke severity was the only independent predictor (HR 1.09; p = 0.016). CONCLUSION: In patients with acute stroke, electrical epileptic activity occurs more frequently than previously suspected.

Comparative demography of the exotic Harmonia axyridis with other aphidophagous coccinellids reared on artificial diet

The objective of this work was to compare biological aspects and life table parameters of the coccinellids Harmonia axyridis, Cycloneda sanguineaand Hippodamia convergens. Insects were fed eggs of Anagasta kuehniella, and reared at 24.5±1ºC, 70±10% relative humidity, with a 12 hour photophase. Hippodamia convergenstook about 1.6 day to complete development, longer than H. axyridis, and 2.4 day longer than C. sanguinea.At immature stages, H. axyridisexhibited the highest survival percentage (49.2%), in comparison to the other coccinellids. For mean adult longevity, H. convergenswas deficient, in comparison with the other species. Mean period of pre oviposition was the longest in C. sanguinea; the longest oviposition time occurred for H. axyridis; and the post oviposition period was similar between the coccinellids. Considering the reproductive parameters, H. axyridisshowed the best performance in all aspects. For life table, the values of H. convergenswere higher than, although close, to those of H. axyridis. Nevertheless, the high net reproductive rate of H. axyridis showed this species potential to increase population size. The biological characteristics of the exotic H. axyridis favors its invasion and establishment in Brazil, corroborating results noticed in other countries.

Plasminogen activator inhibitor 1 and plasminogen activator inhibitor 2 in various disease states.

The association of increased PA-inhibitor (PAI) activity and of PAI-1 and PAI-2 antigen levels with different pathological conditions was studied in a collective of over 300 patients. PAI-1 and PAI-2 levels were measured by specific radioimmunoassays. A good correlation was observed of PAI activity with PAI-1 antigen (r = 0.718; p less than 0.0001) but not with PAI-2 (r = 0.070; n.s.). Both in the controls and in the patients, PAI activity and PAI-1 antigen showed an extremely large range of values. PAI activity ranged from 0.5 to 68 U/ml and PAI-1 antigen from 6 to 600 ng/ml. Increased PAI activity and PAI-1 antigen was observed in patients with malignant tumors, cardiovascular or thromboembolic disease, in the postoperative phase, with hepatic insufficiency, after trauma and after extracorporeal circulation. The large spectrum of disease states with increased PAI activity and PAI-1 antigen reinforces previous suggestions that PAI-1 is an acute phase reactant. After extracorporeal circulation, PAI activity and PAI-1 concentrations strongly increased within one hour, remained elevated for at least one week and returned to preoperation values within 7 days. PAI-2 values ranged from below detection limit (15 ng/ml), observed in half of the plasmas, to 485 ng/ml in a pregnant woman. High values of PAI-2 were only observed in pregnancy.

Selection of bee species for environmental risk assessment of GM cotton in the Brazilian Cerrado

The objective of this work was to list potential candidate bee species for environmental risk assessment (ERA) of genetically modified (GM) cotton and to identify the most suited bee species for this task, according to their abundance and geographical distribution. Field inventories of bee on cotton flowers were performed in the states of Bahia and Mato Grosso, and in Distrito Federal, Brazil. During a 344 hour sampling, 3,470 bees from 74 species were recovered, at eight sites. Apis mellifera dominated the bee assemblages at all sites. Sampling at two sites that received no insecticide application was sufficient to identify the three most common and geographically widespread wild species: Paratrigona lineata, Melissoptila cnecomola, and Trigona spinipes, which could be useful indicators of pollination services in the ERA. Indirect ordination of common wild species revealed that insecticides reduced the number of native bee species and that interannual variation in bee assemblages may be low. Accumulation curves of rare bee species did not saturate, as expected in tropical and megadiverse regions. Species-based approaches are limited to analyze negative impacts of GM cotton on pollinator biological diversity. The accumulation rate of rare bee species, however, may be useful for evaluating possible negative effects of GM cotton on bee diversity.

Metabolism of oral glucose in children born small for gestational age : evidence for an impaired whole body glucose oxidation

Résumé Les études épidémiologiques indiquent que la restriction intra-utérine confère un risque accru de développement de diabète de type 2 au cours de la vie. Certaines études ont documenté la présence d'une résistance à l'insuline chez les jeunes adultes ou les adolescents nés petits pour l'âge gestationnel. Comme la plupart des études ont impliqués des individus post-pubères et comme la puberté influence de manière marquée le métabolisme énergétique, nous avons évalué le devenir du glucose administré oralement dans un groupe incluant essentiellement des enfants pré-pubères ou en début de puberté avec restriction intra-utérine, et chez des enfants matchés pour l'âge et pour le poids. Tous les enfants ont eu une évaluation de leur composition corporelle par mesure des plis cutanés. Ils ont ensuite été étudiés dans des conditions standardisées et ont reçu 4 charges consécutives orales de glucose à raison de 180 mg/kg de poids corporel jusqu'à atteindre un état d'équilibre relatif. La dépense énergétique et l'oxydation des substrats ont été évaluées durant la quatrième heure par calorimétrie indirecte. Comparativement avec les enfants matchés pour l'âge et le poids, les enfants nés petits pour l'âge gestationnel avaient une plus petite stature. Leur dépense énergétique n'était pas significativement abaissée, mais leur oxydation du glucose était plus basse. Ces résultats indiquent que des altérations métaboliques sont présentes précocement chez les enfants nés petits pour l'âge gestationnel, et qu'elles sont possiblement reliées à des altérations de la composition corporelle. Abstract: Epidemiological studies indicate that intrauterine growth restriction confers an increased risk of developing type 2 diabetes mellitus in subsequent life. Several studies have further documented the presence of insulin resistance in young adults or adolescent children born small for gestational age. Since most studies addressed postpubertal individuals, and since puberty markedly affects energy metabolism, we evaluated the disposal of oral glucose in a group including mainly prepubertal and early pubertal children with intrauterine growth restriction and in healthy age- and weight-matched control children. All children had an evaluation of their body composition by skinfold thickness measurements. They were then studied in standardized conditions and received 4 consecutive hourly loads of 180 mg glucose/kg body weight to reach a near steady state. Energy expenditure and substrate oxidation were evaluated during the fourth hour by indirect calorimetry. Compared to both age- and weight-matched children, children born small for gestational age had lower stature. Their energy expenditure was not significantly decreased, but they had lower glucose oxidation rates. These results indicate that metabolic alterations are present early in children born small for gestational age, and are possibly related to alterations of body composition.

Biases on the administered parenteral doses of an experimental drug during phase I clinical trials

Introduction: The pharmaceutical aspects of drug administration in clinical trials receive poor consideration compared with the important attention devoted to the analytical and mathematical aspects of biological sample exploitation. During PK calculations, many researchers merely use for dose the nominal amount declared, overlooking the noticeable biases that may result in the assessment of PK parameters. The aim of this work was to evaluate the biases related to doses injected of a biosimilar drug in 2 Phase I clinical trials. Patients (or Materials) and Methods: In trial A, 12 healthy volunteers received different doses of a biosimilar of interferon beta-1a by either subcutaneous (SC) or intravenous (IV) injection. The doses were prepared by partially emptying 0.5-mL syringes supplied by the manufacturer (drop count procedure). In trial B, 12 healthy volunteers received 3 different formulations of the drug by IV injection (biosimilar without albumin [HSA], biosimilar with HSA and original brand [Rebif®]) and 2 different formulations as multiple SC injections (biosimilar HSA-free and original brand). In both trials, the actual dose administered was calculated as: D = C·V - losses. The product titer C was assessed by ELISA. The volume administered IV was assessed by weighting. Losses were evaluated by in vitro experiments. Finally, the binding of 125I-interferon to HSA was evaluated by counting the free and HSA complexed molecule fractions separated by gel filtration. Results: Interferon was not significantly adsorbed onto the lines used for its IV administration. In trial A, the titer was very close to the one declared (96 ± 7%). In trial B, it differed significantly (156 ± 10% for biosimilar with/without HSA and 123 ± 5% for original formulation). In trial A, the dose actually administered showed a large variability. The real injected volume could be biased up to 75% compared with the theoretical volume (for the lower dose administered [ie, 0.03 mL]). This was mainly attributed to a partial re-aspiration of the drug solution before withdrawing the syringe needle. A strict procedure was therefore applied in trial B to avoid these inaccuracies. Finally, in trial B, 125I-Interferon beta-1a binding to HSA appeared time dependent and slow, reaching 50% after 16-hour incubation, which is close to steady state reported for the comparator Rebif®. Conclusion: These practical examples (especially biases on actual titer and volume injected) illustrate that actual dose assessment deserves attention to ensure accuracy for estimates of clearance and distribution volume in the scientific literature and for registration purposes, especially for bioequivalence studies.

Toimintasuunnitelman laatiminen ohutlevytuotteita ja hitsauskokoonpanoja valmistavalle yritykselle

Tämän diplomityön tavoitteena oli luoda ohutlevytuotteita ja hitsauskokoonpanoja valmistavalle yritykselle kokonaisvaltainen toimintasuunnitelma ja esitellä erilaisia tuotantomalleja case-esimerkkien avulla. Työn aikana kartoitettiin yrityksennykytilannetta ja ongelmakohtia, pohdittiin nykyistä konekantaa ja löytyneitä pullonkauloja. Lisäksi suunniteltiin korjaustoimenpiteitä ja mitä tulisi tehdä, kun tulevaisuudessa yrityksen liikevaihto tulee kasvamaan ja yritystä tullaan laajentamaan. Pääpaino oli myös laskelmilla, joissa selvitettiinmahdollisten investointien tuntihintaa. Tuntihinnan avulla laadittiin taulukoita, joiden avulla ulosmyyntihinta on helppo määrittää. Tuntihinta kytkettiin sijoitetulle pääomalle haluttuun tuottoprosenttiin.

Sähkömittareiden kaukoluennan kannattavuus ja käyttöönotto sähköverkkoyhtiössä

Sähkömittareiden kaukoluenta (AMR) on tällä hetkellä ajankohtainen aihe, koska useat verkkoyhtiöt ovat tehneet jo päätöksen mittaustoiminnan automatisoinnista ja monissa yhtiöissä ovat selvitykset parhaillaan menossa tai suunnitteilla. Erilaisten ennusteiden mukaan kaikki Suomen sähkömittarit tulevat ajan myötä kaukoluettaviksi, mutta aikataulun arviot vaihtelevat viidestä viiteentoista vuoteen. Diplomityössä on tutkittu Lahti Energia Oy:lle soveltuvaa sähkömittareiden kaukoluentajärjestelmää ja siihen investoimisen kannattavuutta. Tarkastelun kohteena on ollut yhtiölle kaukoluennasta kertyvät kustannukset ja sen avulla saavutettavat säästöt. Koska mittareiden kaukoluennasta on yhtiössä vain vähän kokemusta, lasketut säästöt perustuvat olettamukseen ja arvioituihin lukemiin. Lisäksi työssä on käsitelty yhtiön jakelualueen alueittaista jaottelua kaukoluennan kannattavuuden suhteen ja tutkittu kaukoluennan vaikutuksia yhtiön toimintoihin sekä sen tuomia mahdollisuuksia. Kaukoluennan myötä saadaan täsmällistä tietoa energiankulutuksesta ja asiakkaita voidaan laskuttaa todelliseen mittarinlukemaan perustuen. Sen avulla tulevat mahdollisiksi myös useat lisäpalvelut tai toiminnot, kuten tuntimittaus, sähkön laadun seuranta, sähkökatkojen rekisteröinti ja sähköjen etäkatkaisu. Kaikkia kyseisiä palveluita ei tarvitsevälttämättä hankkia kerralla vaan ne voidaan ottaa myös optiona.

Ilmavirtausten laskennallinen mallinnus

Diplomityössä tutkitaan kolmea erilaista virtausongelmaa CFD-mallinnuksella. Yhteistä näille ongelmille on virtaavana aineena oleva ilma. Lisäksi tapausten perinteinen mittaus on erittäin vaikeaa tai mahdotonta. Ensimmäinen tutkimusongelma on tarrapaperirainan kuivain, jonka tuotantomäärä halutaan nostaa kaksinkertaiseksi. Tämä vaatii kuivatustehon kaksinkertaistamista, koska rainan viipymäaika kuivausalueella puolittuu. Laskentayhtälöillä ja CFD-mallinnuksella tutkitaan puhallussuihkun nopeuden ja lämpötilan muutoksien vaikutusta rainan pinnan lämmön- ja massansiirtokertoimiin. Tuloksena saadaan varioitujen suureiden sekä massan- ja lämmönsiirtokertoimien välille riippuvuuskäyrät, joiden perusteella kuivain voidaan säätää parhaallamahdollisella tavalla. Toinen ongelma käsittelee suunnitteilla olevan kuparikonvertterin sekundaarihuuvan sieppausasteen optimointia. Ilman parannustoimenpiteitä käännetyn konvertterin päästöistä suurin osa karkaa ohi sekundaarihuuvan. Tilannetta tutkitaan konvertterissa syntyvän konvektiivisen nostevirtauksen eli päästöpluumin sekä erilaisten puhallussuihkuratkaisujen CFD-mallinnuksella. Tuloksena saadaan puhallussuihkuilla päästöpluumia poikkeuttava ilmaverho. Suurin osa nousevasta päästöpluumista indusoituu ilmaverhoon ja kulkeutuu poistokanavaan. Kolmas tutkittava kohde on suunnitteilla oleva kuparielektrolyysihalli, jossa ilmanvaihtoperiaatteena on luonnollinen ilmanvaihto ja mekaaninen happosumun keräysjärjestelmä. Ilmanvaihtosysteemin tehokkuus ja sisäilman virtaukset halutaan selvittää ennen hallin rakentamista. CFD-mallinnuksella ja laskentayhtälöillä tutkitaan lämpötila- ja virtauskentät sekä hallin läpi virtaava ilmamäärä ja ilmanvaihtoaste. Tulo- ja poistoilma-aukkojen mitoitukseen ja sijoitukseen liittyvät suunnitteluarvot varmennetaan sekä löydetään ilmanvaihdon ongelmakohdat. Ongelmakohtia tutkitaan ja niille esitetään parannusehdotukset.

Identification of in vitro HSC fate regulators by differential lipid raft clustering.

Most hematopoietic stem cells (HSC) in the bone marrow reside in a quiescent state and occasionally enter the cell cycle upon cytokine-induced activation. Although the mechanisms regulating HSC quiescence and activation remain poorly defined, recent studies have revealed a role of lipid raft clustering (LRC) in HSC activation. Here, we tested the hypothesis that changes in lipid raft distribution could serve as an indicator of the quiescent and activated state of HSCs in response to putative niche signals. A semi-automated image analysis tool was developed to map the presence or absence of lipid raft clusters in live HSCs cultured for just one hour in serum-free medium supplemented with stem cell factor (SCF). By screening the ability of 19 protein candidates to alter lipid raft dynamics, we identified six factors that induced either a marked decrease (Wnt5a, Wnt3a and Osteopontin) or increase (IL3, IL6 and VEGF) in LRC. Cell cycle kinetics of single HSCs exposed to these factors revealed a correlation of LRC dynamics and proliferation kinetics: factors that decreased LRC slowed down cell cycle kinetics, while factors that increased LRC led to faster and more synchronous cycling. The possibility of identifying, by LRC analysis at very early time points, whether a stem cell is activated and possibly committed upon exposure to a signaling cue of interest could open up new avenues for large-scale screening efforts.

EkyNet-kunnossapitojärjestelmän arkkitehtuuri ja toteutus

Eky-Verkkoon kuuluu Etelä-Karjalan alueelta noin 60 metsäteollisuudelle palveluita tuottavaa pk-yritystä sekä neljä kemiallisen metsäteollisuuden tehdasta. Verkoston jäsenyrityksille on toteutettu web-pohjaiset -tietojärjestelmät EkyNet jaEKMet. EkyNet on tilaajaosapuolen tietojärjestelmä, jonka välityksellä he voivat tiedottaa toimittajaosapuolille mm. tehtaalla tapahtuvista seisokkitöistä. EKMet on Etelä-Karjalan metallitoimialan yritysrekisteri, jonka avulla toimittajayritykset ylläpitävät tietoja resursseistaan. Yhdessä EkyNet ja EKMet toimivat yritysten välisenä tiedonvälityksen työkaluna ja mahdollistavat tarvittavan tiedon saannin verkoston yritysten välillä ajasta ja paikasta riippumatta. Työssä tarkastellaan EkyNet-järjestelmän arkkitehtuuria, sekä EKMetin integroitumista siihen. EkyNetiä on jatkokehitetty ja siihen on toteutettuominaisuuksia, jotka mahdollistavat seisokkiin liittyvien tilausten tunti-ilmoitusten sähköisen hyväksynnän. Lisäksi kuvataan tunti-ilmoitukseen liittyvää problematiikkaa Eky-Verkostossa. Uusiversio EkyNet-järjestelmästä on toteutettu ja se on käyttöönottovaiheessa.