High throughput flow cytometry-based drug sensitivity testing for melflufen in multiple myeloma

Multiple myeloma (MM) is a hematological plasma cell malignancy in the bone marrow. Lately, increased knowledge of MM pathogenesis and advances in therapy have improved the survival of MM patients. However, due to the unique and complex genome of each patient, some patients are resistant to standard therapies while others achieve durable response but eventually experience relapse. Therefore, new strategies especially for relapsed and refractory and high-risk multiple myeloma (RRMM, HRMM) patients, who have poor response to current therapies, are required. Melflufen, a novel prodrug of the alkylating agent melphalan, has shown significantly decreased resistance effects and more selective cytotoxicity compared to melphalan in vitro and in vivo, but the molecular markers identifying the sensitive subgroups of MM patients have not yet been discovered. The aim of this study was to identify a melflufen-sensitive subgroup of MM patients by utilizing a high throughput flow cytometry-based drug sensitivity testing platform. For the assessment, mononuclear cells from bone marrow of MM patients were derived and melflufen sensitivity of different cell populations tested. As a result, malignant plasma cells were significantly more sensitive to melflufen than to melphalan. Importantly, all MM patient samples including RRMM and HRMM patients were sensitive to melflufen although to varying degrees suggesting these patients may benefit from melflufen treatment. Further integration of the results with additional molecular information may lead to discovery of new biomarkers and thus the prediction of melflufen responses can result in more effective disease management and save patients from ineffective therapies

The design and realization of a high temperature printed circuit test board

Tässä työssä suunnitellaan ja toteutetaan korkean lämpötilan testipiirilevy porauksenaikaisessa mittaussovelluksessa käytettävää kiihtyvyysanturituotetta varten. Tavoitteena on valita korkeaan lämpötilaan sopivat piirilevymateriaali ja komponentit, ja koostaa niistä toimiva testipiirilevy. Lisäksi tavoitteena on koekäyttää valmista testipiirilevyä loppukäytönomaisessa vanhennustestissä sen toiminnan varmistamiseksi. Sopivat komponentit ja piirilevymateriaali testipiirilevyllä käytettäviksi valitaan niiden materiaali-ominaisuuksien perusteella. Lasitransformaatiolämpötilaa ja homologista lämpötilaa tutkimalla voidaan ennustaa, miten eri materiaalit käyttäytyvät eri lämpötiloissa. Valmista testipiirilevyä paistetaan 200 °C lämpötilassa 24 tuntia kerrallaan yhteensä kymmenen kertaa. Vanhennuksien välissä todetaan piirilevyn toiminta mittaamalla sen linjojen resistansseja sekä tutkimalla juoteliitoksia mikroskoopin avulla. Työssä tehtyjen mittausten perusteella suunniteltu testipiirilevy säilyy toimintakykyisenä kymmenen loppukäyttöä simuloivan vanhennustestin jälkeen. Levylle suoritettavat vanhennukset eivät vaikuta linjojen resistansseihin merkittävästi, ja ainoa havaittava muutos on piirilevyn ja juoksutejäänteiden tummuminen. Tuloksista päätellään, että lasitransformaatiolämpötilan ja homologisen lämpötilan perusteella tehdyt materiaalivalinnat ovat onnistuneita. Tulosten valossa tässä työssä suunniteltu testipiirilevy on sopiva porauksenaikaisessa mittaussovelluksessa käytettävän kiihtyvyysanturin testaamiseen.In this thesis a printed circuit test board, for an accelerometer for measurement while drilling application, is designed and realized. The goal of this thesis is to select appropriate high temperature compatible PCB laminate materials and components, and to build a working printed circuit test board from them. In addition, the goal is to conduct a test for the designed test PCB similar to the end application to proof its functionality. The suitable components and PCB laminate materials are selected based on their material properties. By studying the glass transition temperature and homologous temperature, the behavior of materials in different temperatures can be predicted. The finished printed circuit test board undergoes a series of ten 24 hour long burn-in tests with a burn-in temperature of 200 °C. In between the burn-ins, the functionality of the test PCB is verified by measuring the resistance of its signal routes and by studying the solder connections with a microscope. According to the measurements made in this thesis, the designed test PCB maintains its functionality after ten burn-in tests, which are similar to the end application of the test PCB. The burn-ins performed do not have an influence on the resistances of the signal routes of the test PCB. The only detectable change is the darkening of the PCB laminate and flux residues. Based on the results, it can be concluded that the material choices made in this thesis, based on the glass transition temperature and homologous temperature, are successful. In the light of the results the printed circuit test board is compatible to be used as a test PCB for an accelerometer for measurement while drilling application

Netflix & Search: Katsottavan hakeminen ja löytäminen Netflixistä

Tässä pro gradu -tutkielmassa tarkastellaan katsottavan hakemista ja löytämistä Netflix-suoratoistopalvelusta. Aihetta lähestytään erityisesti tiedonhakemisen ja informaatioarkkitehtuurin näkökulmasta. Tutkielman päätavoitteena oli kartoittaa ja kuvailla ihmisten eri tapoja hakea ja löytää katsottavaa. Erityisesti selvitettiin Netflix-käyttöliittymän informaatioarkkitehtuurin mahdollistamia tapoja hakea ja löytää katsottavaa, sekä sitä toimintaa, jota ilmenee hakijan ollessa vuorovaikutuksessa Netflix-käyttöliittymän kanssa, mutta myös käyttöliittymän ulkopuolella tapahtuvaa toimintaa huomioitiin. Tarkastelun kohteena oli sekä aktiivinen että passiivinen katsottavan hakemiseen ja löytämiseen liittyvä toiminta. Hakemisen ja löytämisen tapojen puhtaan kuvailun ohella kartoitettiin katsottavan hakemiseen ja löytämiseen mahdollisesti vaikuttavia tekijöitä, ja sitä, miten ne vaikuttavat. Osana tarkastelua tutustuttiin Netflix-käyttöliittymään ja sen informaatioarkkitehtuuriin. Käyttöliittymän tarkastelun lisäksi toteutettiin kyselytutkimus, jonka avulla pyrittiin saamaan kattavampi kuva katsottavan hakemisesta ja löytämisestä ilmiönä. Kyselyllä kerättiin laaja-alaisesti tietoa Netflixin käyttöön liittyvistä tekijöistä kuten tyypillisistä tavoista käyttää Netflixiä ja sen eri toimintoja, kontekstitekijöistä ja käyttökokemuksista. Kyselyn kohderyhmänä olivat yliopisto-opiskelijat, ja kyselyyn vastasi 137 Netflixiä käyttänyttä eri alojen opiskelijaa. Tutkimuksessa tunnistettiin joitakin katsottavan hakemiseen ja löytämiseen liittyviä tyypillisiä piirteitä. Mikäli käyttäjällä on jo palvelun avatessa mielessä, mitä hän haluaa katsoa, on katsottavan hakeminen palvelusta melko suoraviivaista toimintaa. Monipuolisempaa hakemistoimintaa ilmenee erilaisissa tilanteissa, ja hakemiseen vaikuttavat useat niin käyttäjään liittyvät sisäiset tekijät kuin ulkopuoliset kontekstitekijät. Netflixin suositusjärjestelmällä ja ulkoisilla lähteillä kuten sosiaalisella medialla ja muiden ihmisten suosituksilla voidaan nähdä olevan keskeinen rooli katsottavan löytämisessä

Analysis and interpretation of magnetic anomalies observed in north-central California

To assist in the assessment of the geothermal potential of north-central California and to aid in defining the geologic transitions between the physiographic provinces of the Klamath Range, the Cascade Range, the Modoc Plateau, the Great Valley, and the Sierra Nevada Range, personnel from the Geophysics Group in the College of Oceanography at Oregon State University conducted a detailed aeromagnetic survey extending from 40°15' to 42°00'N latitude and from 120°45' to l22°45'W longitude. Two forms of spectral analysis, the energy spectrum and the exponential methods, were used to make source-top and source-bottom depth calculations. The magnetic source-bottom depths were interpreted as Curie-point isotherm depths. Based on the energy spectrum analysis, several regions with elevated Curie-point isotherm depths were mapped: (1) the Secret Spring Mountain-National Lava Beds Monument area, (2) the Mount Shasta area, (3) the Big Valley Mountains area, and (4) an area northeast of Lassen Peak. The elevated Curie-point isotherm depths within these areas, as shallow as 4 to 7 km below sea level (BSL) in the Secret Spring Mountain-National Lava Beds Monument area, the Mount Shasta area, and the area northeast of Lassen Peak, and 4 to 6 km BSL in the Big Valley Mountains area, imply vertical temperature gradients in excess of 70°C/km and heat flow greater than 100 mW/m² when assuming a Curie-point temperature of 580°C. Shallow source-bottom depths of 4 to 5 km BSL were mapped in the Eddys Mountain area and interpreted to be the depth of a lithologic contact. Source-top depths show that the magnetic basement varies from about 3.5 km BSL, beneath the sedimentary assemblages of the Great Valley and the eastern Klamath Range, to near sea level in the Cascade Range. The exponential approximation method yielded source-bottom depths which agreed, in general, with depths determined by the energy spectrum method. However, this method appears less reliable and its depth estimates less accurate compared to the energy spectrum method. A broad negative anomaly, observed on the total field magnetic intensity map and low-pass filtered anomaly maps, suggests the sedimentary rocks of the Klamath Complex underlie Mount Shasta and the Medicine Lake Highlands. Magnetic lineations are oriented mainly NW

Aminopeptidase Expression in Multiple Myeloma Associates with Disease Progression and Sensitivity to Melflufen

Multiple myeloma (MM) is characterized by extensive immunoglobulin production leading to an excessive load on protein homeostasis in tumor cells. Aminopeptidases contribute to proteolysis by catalyzing the hydrolysis of amino acids from proteins or peptides and function downstream of the ubiquitin–proteasome pathway. Notably, aminopeptidases can be utilized in the delivery of antibody and peptide-conjugated drugs, such as melflufen, currently in clinical trials. We analyzed the expression of 39 aminopeptidase genes in MM samples from 122 patients treated at Finnish cancer centers and 892 patients from the CoMMpass database. Based on ranked abundance, LAP3, ERAP2, METAP2, TTP2, and DPP7 were highly expressed in MM. ERAP2, XPNPEP1, DPP3, RNPEP, and CTSV were differentially expressed between relapsed/refractory and newly diagnosed MM samples (p < 0.05). Sensitivity to melflufen was detected ex vivo in 11/15 MM patient samples, and high sensitivity was observed, especially in relapsed/refractory samples. Survival analysis revealed that high expression of XPNPEP1, RNPEP, DPP3, and BLMH (p < 0.05) was associated with shorter overall survival. Hydrolysis analysis demonstrated that melflufen is a substrate for aminopeptidases LAP3, LTA4H, RNPEP, and ANPEP. The sensitivity of MM cell lines to melflufen was reduced by aminopeptidase inhibitors. These results indicate critical roles of aminopeptidases in disease progression and the activity of melflufen in MM

Aminopeptidase Expression in Multiple Myeloma Associates with Disease Progression and Sensitivity to Melflufen

Multiple myeloma (MM) is characterized by extensive immunoglobulin production leading to an excessive load on protein homeostasis in tumor cells. Aminopeptidases contribute to proteolysis by catalyzing the hydrolysis of amino acids from proteins or peptides and function downstream of the ubiquitin–proteasome pathway. Notably, aminopeptidases can be utilized in the delivery of antibody and peptide-conjugated drugs, such as melflufen, currently in clinical trials. We analyzed the expression of 39 aminopeptidase genes in MM samples from 122 patients treated at Finnish cancer centers and 892 patients from the CoMMpass database. Based on ranked abundance, LAP3, ERAP2, METAP2, TTP2, and DPP7 were highly expressed in MM. ERAP2, XPNPEP1, DPP3, RNPEP, and CTSV were differentially expressed between relapsed/refractory and newly diagnosed MM samples (p < 0.05). Sensitivity to melflufen was detected ex vivo in 11/15 MM patient samples, and high sensitivity was observed, especially in relapsed/refractory samples. Survival analysis revealed that high expression of XPNPEP1, RNPEP, DPP3, and BLMH (p < 0.05) was associated with shorter overall survival. Hydrolysis analysis demonstrated that melflufen is a substrate for aminopeptidases LAP3, LTA4H, RNPEP, and ANPEP. The sensitivity of MM cell lines to melflufen was reduced by aminopeptidase inhibitors. These results indicate critical roles of aminopeptidases in disease progression and the activity of melflufen in MM

Esrrg functions in early branch generation of the ureteric bud and is essential for normal development of the renal papilla

Congenital anomalies of the kidney and urinary tract (CAKUTs) are common disorders of human development affecting the renal parechyma, renal pelvis, ureter, bladder and urethra; they show evidence of shared genetic aetiology, although the molecular basis of this remains unknown in the majority of cases. Breakpoint mapping of a de novo, apparently balanced, reciprocal translocation associated with bilateral renal agenesis has implicated the gene encoding the nuclear steroid hormone receptor ESRRG as a candidate gene for CAKUT. Here we show that the Esrrg protein is detected throughout early ureteric ducts as cytoplasmic/sub-membranous staining; with nuclear localization seen in developing nephrons. In 14.5–16.5 dpc (days post-conception) mouse embryos, Esrrg localizes to the subset of ductal tissue within the kidney, liver and lung. The renal ductal expression becomes localized to renal papilla by 18.5 dpc. Perturbation of function was performed in embryonic mouse kidney culture using pooled siRNA to induce knock-down and a specific small-molecule agonist to induce aberrant activation of Esrrg. Both resulted in severe abnormality of early branching events of the ureteric duct. Mouse embryos with a targeted inactivation of Esrrg on both alleles (Esrrg−/−) showed agenesis of the renal papilla but normal development of the cortex and remaining medulla. Taken together, these results suggest that Esrrg is required for early branching events of the ureteric duct that occur prior to the onset of nephrogenesis. These findings confirm ESRRG as a strong candidate gene for CAKUT