Biosensorsüsteem mastiiti põhjustavate bakterite kiireks ja samaaegseks määramiseks piimas

Väitekirja elektrooniline versioon ei sisalda publikatsiooneMastiit on udarapõletik, mis enamasti tekib patogeensete mikroorganismide sattumisel läbi nisajuha udaraveerandisse, olles peamine lüpsilehmade nakkushaigus. Mastiidi poolt põhjustatud kahju kogu maailma 271 miljoni piimalehma kohta on hinnanguliselt 16-26 miljardit eurot aastas. Traditsiooniliste meetoditena mastiiti tekitavate bakterite identifitseerimiseks on tänapäeval kasutusel mikrobioloogilised analüüsid, mis võtavad aega 1-2 päeva ja laboratoorsetes tingimustes tehtavad patogeenide geenianalüüsid, mille tegemiseks kulub 6 tundi. Kuna ravi edukuse tagamiseks on selle täpsuse kõrval väga oluline ka selle võimalikult operatiivne alustamine, siis on vajalik välja töötada sellised analüüsimeetodid, mis võimaldavad patogeenide identifitseerimist oluliselt kiiremini kui praegu ning mida on võimalik kasutada farmides kohapeal. Tänasel päeval sellised meetodid puuduvad. Doktoritöö käigus töötati välja biosensorsüsteem ja mõõtemetoodika kolme peamise mastiiti tekitava patogeeni – Staphylococcus aureus’e Escherichia coli ja Streptococcus uberis’e määramiseks nii eraldi kui ühtlasi ka kõikide nimetatud patogeenide koos määramiseks. Biosensorsüsteemi konstrueerimisel uuriti selle tundlikkust, tööpiirkonda, selektiivsust ja sobivust rakendamiseks keerulistes maatriksites nagu piim. Biosensori selektiivsus kindla patogeeni määramiseks teiste bakterite olemasolul oli väga hea ning seda on võimalik kasutada mitme bakteri samaaegseks määramiseks. Väljapakutud biosensorsüsteemil põhinevat mõõtesüsteemi on võimalik kasutada loomade tervise automatiseeritud kontrolliks farmis kohapeal ja seeläbi kiiresti identifitseerida juba varajases staadiumis potentsiaalne haigus. Varajane haiguse avastamine aitab alustada koheselt kiiret ja sobivat ravi, parandades seeläbi looma heaolu ja piima kvaliteeti ning vähendada tootmiskulusid ja majanduslikku kahju.Mastitis, mostly caused by bacterial infection of the mammary gland, is a major health problem of dairy cows. The resulting decrease of milk production and reduction of its quality along with medication costs and probable premature culling of animals cause essential economic burden. The total mastitis caused losses in dairy industry are estimated to be 16–26 billion € annually in view of a global population of 271 million dairy cows. For the identification of mastitis-causing pathogens, the gold standard is microbiological culturing of bacteria, which in recent years has been partially replaced by polymerase chain reaction analyses of bacterial DNA. Although reliable, these methods require hours to obtain results, not allowing effective treatment of animals and optimal milk processing. Therefore to assure timely and correct treatment of animals, there is a great need for a method applicable for rapid automatic detection of mastitis causing pathogens in milk. A rapid method and an immuno-biosensing system have been proposed and applied for the rapid multiplex detection of major mastitis causing pathogens (Staphylococcus aureus, Escherichia coli and Streptococcus uberis) in milk. The key features of the biosensor development were its selectivity, sensitivity, working range and applicability for rapid automated analyses in complex biological sample matrix like milk. The selectivity of the biosensing system in the presence of several pathogens was very good and it was applicable for the multiplex detection of different pathogens. Based on the results of the present study, the proposed biosensing system has a great potential to serve as a tool for in-line automatic monitoring of milk and animal health in dairy farms

A Novel Monoclonal Antibody to Human Laminin alpha 5 Chain Strongly Inhibits Integrin-Mediated Cell Adhesion and Migration on Laminins 511 and 521

Peer reviewe

Ubiquitin Carboxyl-Terminal Esterase LI (UCHLI) SI8Y Polymorphism in Patients with Cataracts

Background: Cataract is characterized by light-scattering protein aggregates. The ubiquitin-proteasome system has been proposed a role in proteolytic removal of these protein aggregates. Ubiquitin carboxyl-terminal esterase L1 (UCHL1) is a de-ubiquitinating enzyme wit

Gene Expression and Functional Studies of the Optic Nerve Head Astrocyte Transcriptome from Normal African Americans and Caucasian Americans Donors

To determine whether optic nerve head (ONH) astrocytes, a key cellular component of glaucomatous neuropathy, exhibit differential gene expression in primary cultures of astrocytes from normal African American (AA) donors compared to astrocytes from normal Caucasian American (CA) donors.We used oligonucleotide Affymetrix microarray (HG U133A & HG U133A 2.0 chips) to compare gene expression levels in cultured ONH astrocytes from twelve CA and twelve AA normal age matched donor eyes. Chips were normalized with Robust Microarray Analysis (RMA) in R using Bioconductor. Significant differential gene expression levels were detected using mixed effects modeling and Statistical Analysis of Microarray (SAM). Functional analysis and Gene Ontology were used to classify differentially expressed genes. Differential gene expression was validated by quantitative real time RT-PCR. Protein levels were detected by Western blots and ELISA. Cell adhesion and migration assays tested physiological responses. Glutathione (GSH) assay detected levels of intracellular GSH.Multiple analyses selected 87 genes differentially expressed between normal AA and CA (P<0.01). The most relevant genes expressed in AA were categorized by function, including: signal transduction, response to stress, ECM genes, migration and cell adhesion.These data show that normal astrocytes from AA and CA normal donors display distinct expression profiles that impact astrocyte functions in the ONH. Our data suggests that differences in gene expression in ONH astrocytes may be specific to the development and/or progression of glaucoma in AA

The Presumed Principle of Confidentiality in Arbitration : A Challenging Balance Between the Presumed Principle of Confidentiality and Transparency in International Arbitration

International arbitration is one of the most important alternative dispute resolution methods in international law. It is valued especially for the reason that dispute resolution can be executed in a confidential and private manner. The principle of confidentiality has been thought to be automatically part of the arbitration since the very first arbitration cases. Hence, in this thesis, it was examined whether the principle of confidentiality can be assumed to be included in arbitration and whether it can be automatically part of arbitral proceedings. To addition, in this thesis were presented other potential advantages that the principle is bringing to arbitration and discussed the reasons of why this principle is really important. Different measures and matters that should be taken into account in order for the parties to maximise those mentioned benefits were also presented in this thesis. To the contrast, the negative aspects of the principle and the problems that it may bring to arbitration and other important matters that in specific circumstances outweigh the importance of the principle and override it were also discussed. In this thesis, such conclusion was reached that the mentioned principle cannot be absolutely taken for granted and assumed that the principle is applied to every case without limitations. The target audience of the research consists of different legal practitioners, legal scholars and possible students. The questions were approached by utilising different legal written material and source material in this thesis primarily consisted of different articles and literature that deals with the subject. The research method of legal dogmatics was applied to this work to approach the subject