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WAT IS ONDERZOEK PDF

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Request PDF on ResearchGate | On Jun 1, , Verhoeven and others published Wat is Onderzoek. Wat Is Onderzoek Door Nel Verhoeven Boek klimaat en overstromingen - edepot. wur - 10 11 tot doorbraakvrije dijken. welke maatregelen uiteindelijk het hoogst. keuring certificaat van onderzoek - akwadrant - keuring certificaat van onderzoek wat kan akwadrant scheepstechniek voor u be tekenen? akwadrant voor cvo.

Met dit formulier vraagt u een vergunning aan voor het project dat u wilt uitvoeren. Of u geeft aan wat u in het vergunde project wilt wijzigen. Meer informatie over de voorwaarden vindt u op de website of in de toelichting op de website. Neem voor meer informatie over het verkrijgen van een deelnemernummer contact op met de NVWA. Vul de gegevens van het postadres in.

Biotechnol Adv, 7: Cell Tissue Res, 2: Exp Cell Res, 1: J Am Soc Nephrol, 3: Kidney Int, 9: AAPS J, 2: Sci Rep,: Saenz-Robles, and J. Pipas, SV40 large T antigen targets multiple cellular pathways to elicit cellular transformation. Oncogene, Nat Genet, 1: If the main objective is not a research objective, which specific need s does this project respond to? The main objective is to study the safety of ciptec OAT1 kidney cells in vivo. Hereby we investigate both the tumorigenicity and the oncogenicity of these specific kidney cells.

Tumorigenicity is defined as the property of cells to form tumors when inoculated into susceptible animals. As such, tumors that arise contain cells derived from the inoculated cells. This can occur at the site of injection but also elsewhere due to metastasis. Oncogenicity is defined as the property of an acellular agent to induce cells of an animal to become tumor cells. Tumors that arise are derived from the host itself. Oncogenic activity from cell substrates could be due to either the cell substrate DNA and perhaps other cellular components or an oncogenic viral agent present in the cells.

Tumorcells will be implanted in the flank, after which animals will be observed for developing tumors. In case of developing tumors, tumorsize will be measured using a digital or vernier caliper. As soon as any risk on tumorigenicity or oncogenicity results from the animal study, further development of the BAK system in its current form will be terminated.

It is not known how humans respond to conditionally immortalized cell systems. Therefore, translation of negative results i. However, the choice of an immunodeficient animal model increases the reliability of such a translation. The aim is achievable, since: This is a susceptible rat in which cancerous cells are most likely to arise.

There is a great need of a BAK. People with a serious heatlh condition or disease in which their kidneys fail, currently depend on kidney transplantation or kidney dialysis as possible treatments.

However, both treatments are far from ideal. Patients undergoing dialysis have a poor quality of life and a lot of patients awaiting transplantation cannot be helped in time due to donor shortage. Moreover, even after successful transplantation there is still a chance of rejection of the donor kidney, for which patients need to take immunosuppressive medication with many side effects.

This is due to limitations of the currently applied filter membranes within the artificial kidney. A possible solution for these problems is the BAK.

This is an artificial kidney that contains allogeneic. Those cells will remove the remaining toxic waste products that cannot be removed by the regular dialysis process. Great progress is currently being made in this field of research with promising results. These ciptec OAT1 cells are genetically modified so that they can be cultured on a large scale.

Because of this modification an increased risk for tumorigenic or oncogenic properties exists. In this project, we will investigate the safety of those genetically modified human kidney cells after inoculation in a living organism. We will investigate whether these cells can directly form tumors or metastases, and we will investigate whether the content of these cells has cancer promoting properties in an indirect manner.

When proven safe, these cells can be used for further development of the BAK. And once the BAK has been validated and made operational it will help and save millions of lives all over the world. This project is a straightforward project with only one type of animal procedure, which contains a single experiment with four treatment groups.

One positive control group, one negative control group, one tumorigenicity group and one oncogenicity group. Ten animals per treatment group will be inoculated and followed for a period of minimal 4 months and a maximum of 6 months. During this period we will measure tumorsize twice a week and perform clinical observation three times a week. At the end of the experiment, all animals will be euthanized and examined for gross and microscopic evidence of the growth of inoculated cells at the site of injection and in other sites.

Also, all animals will be imaged at two timepoints for further evidence of growing cells in other sites. Minimal observation period 4 months: This minimal required observation period is stated in the WHO report: WHO Recommendations for the evaluation of animal cell cultures as substrates for the manufacture of biological medicinal products and for the characterization of cell banks.

Maximal observation period 6 months: This maximal observation period was previously used in a study by Liu et al We want to use the same observation period. Vaccine Feb 3;28 5: This study was done to ensure the safety of this cell line for vaccine production. For tumorigenicity they injected an intact MDCK cell suspension into immunodeficient mice. A HeLa cell suspension was used as a positive control. Image timepoints: First timepoint will be the at which the animals of the positive controlgroup start developing nodules, the second timepoint is at the end of experiment human endpoint or end of observational period.

HeLa cells positive control group: HeLa cells comprise an immortalized, continuously cultured cell line of human cancer cells. Unlike normal body somatic cells, HeLa cells thrive indefinitely in laboratory tissue cultures, a trait that has allowed them to assume tremendous importance in biomedical research. Since the mid-twentieth century, the cells have been distributed around the world and used in innumerable medical endeavors, including investigations into the nature of cancer, the development of vaccines, the mapping of genes, the treatment of diseases, and the mechanisms involved with programmed cell death apoptosis.

The cell line arose in from samples collected during a biopsy on Henrietta Lacks, a poor year-old African- American patient suffering from a cervical tumor. The designation HeLa, derived from the first two letters of her first and last names, was used to keep her identity anonymous. Henrietta Lacks, who died later in.

During subsequent investigations, the sample cells were found to behave abnormally. Unlike typical somatic cells, which undergo cell aging senescence and lose the ability to divide and replicate after a few dozen generations the so-called Hayflick limit , the cells from Henrietta Lacks never stop dividing.

Being tumor cells, HeLa cells also do not die from apoptosis, the mechanism that normally causes abnormal or unneeded cells to self-destruct. HeLa cells thus became the first line of human cells to survive indefinitely in vitro under proper culture-growing conditions. The hardiness of these cells was noted early in the medical literature, resulting in numerous requests by other laboratories and research institutions for samples that could be cultured.

As such, HeLa cells were disseminated around the world and became the basis for numerous medical studies that have benefited humankind. For example, Jonas Salk used HeLa cells in the work leading up to his creation of a vaccine for polio. In addition, HeLa cells were the first human cells to be cloned.

Inventaris Wob-verzoek W17-08

When results are inconclusive, we want to repeat this procedure in a second batch of animals at a later timepoint. Also, WHO guidelines reference [16] state that if the tumorigenicity group or the oncogenicity group show inconclusive results then the experiment should be repeated Provide a basic outline of the different components of the project and the type s of animal procedures that will be performed.

Step1, Habituation: Positive control group: Animals will be observed three times per week. The animals will be weighed and their welfare will be carefully monitored. The body surface will be examined for tumors that may arise.

Assessment of the inoculation site over time: If a nodule appears, it is measured in two perpendicular dimensions, the measurements being recorded two times per week to determine whether the nodule grows progressively, remains stable, or decreases in size over time. Moreover, at the time the first nodule appears, rats will be imaged to have more indepth information on tumor formations and metastases.

Minimal observation period is 4 months unless human endpoints are reached earlier , the maximal observation period is 6 months Step 4; Final Assessment of the inoculation site and other sites: At the end of the observation period, all animals, including the control groups, will be anesthesized and scanned using a whole body imager to screen for tumors and metastases. Next, animals will be euthanized and examined further for gross and microscopic evidence of tumour formation at relevant tissues Describe the coherence between the different components and the different steps of the project.

If applicable, describe the milestones and selection points. Validation of the procedure: Progressively growing tumors should be produced in at least 9 of 10 animals injected with the positive control reference HeLa cells. If a nodule appears, it is measured in two perpendicular dimensions, the measurements being recorded weekly to determine whether the nodule grows progressively, remains stable, or decreases in size over time. Cell lines that produce nodules that fail to grow progressively are not considered to be tumorigenic.

Interpretation of results: The test is considered positive if at least 2 of 10 animals inoculated with the test article cells develop tumors at the site of inoculation or at a metastatic site or histologic or genotypic examination reveals that the nature of the cells constituting the tumors is consistent with that of the inoculated cells.

If only one of 10 animals develops a tumor that meets the previous criteria, the cell line should be considered possibly tumourigenic and examined further. The test is considered positive if at least 2 of 10 animals inoculated with the cell content develop tumors at the site of inoculation or at a metastatic site or histologic or genotypic examination reveals that the nature of the cells constituting the tumors is consistent with that of the inoculated the positive control reference cells.

If only one of 10 animals develops a tumor that meets the previous criteria, the cell line should be considered possibly oncogenic and examined further. Use a different appendix description animal procedures for each type of animal procedure. Serial number Type of animal procedure 1 Tumorigenicity and oncogenicity in vivo. Appendix Description animal procedures This appendix should be enclosed with the project proposal for animal procedures.

A different appendix description animal procedures should be enclosed for each type of animal procedure. For more information, see our website Or contact us by phone. Use the serial numbers provided in Section of the Project Proposal form Universiteit Utrecht Serial number Type of animal procedure 1 Tumorigenicity and oncogenicity in vivo 2 Description of animal procedures A.

Experimental approach and primary outcome parameters Describe the general design of the animal procedures in relation to the primary outcome parameters. Justify the choice of these parameters. One positive controlgroup: This group is included to validate the test.

Animals will receive viable HeLA cells which are known to induce tumors. One negative controlgroup: This group will give us information about spontaneous tumor development in these animals under standard conditions. Animals will receive vehicle. One tumorigenicity group: Tumors arising in this group are a direct outcome parameter for measuring tumorigenicity. Animals will receive viable kidney cells ciptec-oat1. One oncogenicity group: Tumors or cancerous tissue arising in this group are a direct outcome parameter for measuring oncogenicity.

Animals will receive a kidney cell lysate. Describe the proposed animal procedures, including the nature, frequency and duration of the treatment. Provide justifications for the selected approach. Minimal observation duration is 4 months, the maximal observation period is 6 months Step 4; Final Assessment of the inoculation site and other sites: Next, animals will be euthanized and examined further for gross and microscopic evidence of tumour formation at relevant tissues.

Theorie en hypothesen in cittadelmonte.info - Prof. dr. AFA Korsten

Any tumor that is identified is divided into three equal parts: Describe which statistical methods have been used and which other considerations have been taken into account to minimise the number of animals.

The animals Specify the species, origin, estimated numbers, and life stages. Provide justifications for these choices. Species and origin: For these studies, Hsd: It is a T cell deficient rat model which is well established for oncology and transplantation research. We will use both males and females, preferably five per sex per group Estimated numbers: Since we have 4 treatment groups, the number of animals will be As this is considered a single experiment, a repetition of the experiment in a different batch of animals at a later timepoint is warranted.

The report states that if if the tumorigenicity group or the oncogenicity group show inconclusive results then those groups should be repeated. More precisely we refer to report page number , annex3, section B.

Also, for tumorigenicity we refer to page , section 1,"number of test animals". For oncogenicity we refer to page , annex 3, section 4, "Number of test animals". This brings the total amount of animals for this application to The rats will be 4 weeks old upon arrival. Younger animals are more susceptible regarding the development of tumors. Re-use Will the animals be re-used?

Theorie en hypothesen in onderzoek.pdf - Prof. dr. AFA Korsten

No, continue with question D. Are the previous or proposed animal procedures classified as severe? Replacement, reduction, refinement Describe how the principles of replacement, reduction and refinement were included in the research strategy, e. A number of in vitro tests have already been done on ciptec-oat1 kidney cells in the laboratory.

Examples are SV40T expression, focus formation assay, anchorage independent growth assay, invasion assay, karyotyping and p53 and Rb activity. As all results were negative for potential risk, we aim to persue our studies demonstrating safety in vivo. Not applicable. If possible, we will house the animals in IVC cages to minimise infections.

Also, we will frequently observe each animal. As animal model we will use the RNU nude rat. This is a susceptible rat specifically designed for oncology studies. We will euthanize animals that reach their human endpoint. Explain what measures will be taken to minimise 1 animal suffering, pain or fear and 2 adverse effects on the environment.

When animals develop severe adverse effects, we will euthanize them and examine the tissue. This way we will prevent adverse effect from maximising.

Ik noemde het wel e en s e en comb in atie van breedte onderzoek en diepte onderzoek. Mijn promotor hield me destijds bij de start voor: En dat heb ik to en gedaan.

Of beter: De boodschap van deze tekst is: Doe dat in de vorm van puntige verwacht in g en of hypothes en. Geef je onderzoek daarmee focus. Het bevordert de helderheid. Wees niet te bang. Het proefschrift van Klaartje Peters is e en goed voorbeeld hoe het in positieve z in kan. Zij formuleert in haar ook bekroonde proefschrift e en aantal veronder stell in g en over wie het in Nederland op het vlak van macht en in vloed op besluitvorm in g op rijksniveau voor het zegg en heeft.

Zij noemt die verwachte uitkomst en hypothes en. E en ander goed voorbeeld is dat van Rolf Willemse over het bestaansrecht van geme en t en. Short-link Link Embed. Share from cover. Share from page: Dat zij niet spreken van veronderst Page 4 and 5: Sportjournalisten houden er zo hun Page 6 and 7: Allison over de Cubacrisis zie ook Page 14 and Peters gebruikt twee indelingen voo Page 16 and Maar het is wel mogelijk om de land Page 24 and De conclusie kan niet anders zijn d Page 28 and Ferlie, E.

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