Development of Novel Assays for Measuring Different Molecular Forms of Prostate Specific Antigen

Prostate cancer (PCa) is the most commonly diagnosed non-skin cancer and second leading cause of cancer-related death of men in developed countries. Measurement of prostate specific antigen (PSA) is a very sensitive method for diagnosing and monitoring of prostate cancer (PCa), but the specificity needs improvement. Measurements of different molecular forms of PSA have been shown to improve differentiation between PCa and benign prostatic diseases. However, accurate measurement of some isoforms has not been achieved in previous assays. The aim of the present study was to develop new assays that reliably measure enzymatically active PSA, PSA-α1-chymotryposin (PSA-ACT) and PSA-α1-protease inhibitor (PSA-API), and to evaluate their diagnostic value. Double-label immunofluorometric assays using a novel monoclonal antibody (MAb) and another antibody to either free PSA (fPSA) or total PSA (tPSA) were developed and used to measure PSA-ACT and fPSA or tPSA at the same time. These assays provide enough sensitivity for measurement of PSA-ACT in sera with low PSA levels. The results obtained confirmed that proportion of PSA-ACT to tPSA (%PSA-ACT) was as useful as proportion of fPSA to tPSA (%fPSA) for discrimination between PCa and benign prostatic hyperplasia (BPH). We developed an immunoassay for detection of PSA-API based on proximity ligation, which improved assay sensitivity 10-fold compared with conventional assays. Our results confirmed previous findings that the PSA-API level is somewhat lower in men with than without PCa, and the combination of %fPSA and proportion of PSA-API to tPSA (%PSA-API) provides diagnostic improvement compared with either method alone. Assays based on this principle should be applicable to other immunoassays in which the nonspecific background is a problem. An immunopeptidometric sandwich assay (IPMA) was developed to measure the enzymatically active PSA. This assay showed high specificity, but sensitivity was not good enough for measurement of PSA concentrations in the gray zone, 2-10 µg/L, in which tPSA does not efficiently differentiate between PCa and BPH. We further developed a solid-phase proximity ligation immunoassay, which provided a 10-fold improvement in sensitivity. This proof of concept study shows that peptides reacting with proteins are potentially useful for sensitive and specific measurement of protein variants for which specific MAbs cannot be obtained.Prostataspesifisen antigeenin (PSA) määrittäminen seerumista on herkkä menetelmä eturauhassyövän diagnostiikkaan, mutta se ei sovellu hyvin eturauhassyövän ja eturauhasen hyvänlaatuisen liikakasvun erottamiseen. Spesifisyyttä on pystytty lisäämään erilaisten PSA-muotojen mittaamisella. Kuitenkaan useiden PSA-muotojen luotettava mittaaminen ei ole ollut mahdollista aikaisemmilla menetelmillä. Tutkimukseni tarkoituksena on ollut kehittää uusia määritysmenetelmiä näiden PSA-muotojen määrittämiseksi [entsymaattisesti aktiivinen PSA, sekä PSA-α1-kymotrypsiini (PSA-ACT) ja PSA-α1-proteaasi inhibiittori (PSA-API) kompleksit]. Lisäksi tutkin näiden menetelmien diagnostista merkitystä. Tuotimme uusia vasta-aineita ja pystytin herkkiä määritysmenetelmiä PSA-ACT ja PSA-API kompleksien mittaamiseksi. Kehittämilläni menetelmillä voidaan PSA-ACT:n määrityksen kanssa samanaikaisesti mitata vapaata PSA:ta tai kokonais PSA:ta. Tästä on etua mm. määritettäessä PSA-ACT:n suhdetta kokonais PSA:han, minkä havaittiin myös tässä tutkimuksessa olevan eturauhassyövän diagnostiikassa yhtä hyödyllinen kuin vapaan PSA:n suhde kokonais PSA:han ja huomattavasti parempi kuin pelkkä kokonais PSA:n mittaus. PSA-API kompleksin mittaamiseksi käytettiin ns. läheisyys-ligaatioon perustuvaa immunomääritystä, jossa vasta-aineiden tulee sitoutua toistensa lähelle ja signaali saadaan aikaan DNA:n monistamiseen perustuvalla menetelmällä. Tällä tavoin saavutettiin 10 kertaa parempi herkkyys kuin perinteisillä menetelmillä ja päästiin eroon aiempia menetelmiä vaivanneesta korkeasta taustasta. Kliiniset tuloksemme vahvistavat aikaisemmat löydökset, joiden mukaan PSA-API tasot ovat jonkin verran korkeammat miehillä, joilla on eturauhassyöpä. Menetelmä soveltunee myös muiden immunomääritysten parantamiseksi. Kehitin myös kaksi immunopeptidometristä määritysmenetelmää, joissa PSA:n tunnistamiseen käytettiin aktiiviseen PSA:han sitoutuvaa peptidiä. Ensimmäinen menetelmä oli spesifinen, muttei tarpeeksi herkkä. Parantaakseni herkkyyttä kehitin läheisyys-ligaatio menetelmän, jossa peptidi ja kaksi vasta-ainetta tunnistavat PSA:n. Tämä menetelmä paransi herkkyyttä 10-kertaisesti. Tämä uuteen periaatteeseen perustuva menetelmä osoittaa että peptidejä voidaan käyttää herkkiin ja spesifeihin määritysmenetelmiin, myös mitattaessa sellaisia proteiini-variantteja, joita vastaan on vaikea kehittää vasta-aineita

Official Register of the United States, containing a list of the officers and employes in the Civil, Military, and Naval Service on the first of July, 1893; together with a list of vessels belonging to the United States. Volume II, Pt. 2 of 2.

Official Register of the U.S. 1 July. HMD 29 (pts. 1 and 2), 53-2. v2-3. 2779p. [3230-3231] Lists all employees of the Indian service

U.S. GLOBAL CHANGE RESEARCH PROGRAM CLIMATE SCIENCE SPECIAL REPORT (CSSR)

Fifth-Order Draft Table of Contents Front Matter About This Report........................................................................................ 1 Guide to the Report......................................................................................4 Executive Summary ................................................................................... 12 Chapters 1. Our Globally Changing Climate .......................................................... 38 2. Physical Drivers of Climate Change ................................................... 98 3. Detection and Attribution of Climate Change .................................... 160 4. Climate Models, Scenarios, and Projections .................................... 186 5. Large-Scale Circulation and Climate Variability ................................ 228 6. Temperature Changes in the United States ...................................... 267 7. Precipitation Change in the United States ......................................... 301 8. Droughts, Floods, and Hydrology ......................................................... 336 9. Extreme Storms ....................................................................................... 375 10. Changes in Land Cover and Terrestrial Biogeochemistry ............ 405 11. Arctic Changes and their Effects on Alaska and the Rest of the United States..... 443 12. Sea Level Rise ....................................................................................... 493 13. Ocean Acidification and Other Ocean Changes .............................. 540 14. Perspectives on Climate Change Mitigation .................................... 584 15. Potential Surprises: Compound Extremes and Tipping Elements .......... 608 Appendices A. Observational Datasets Used in Climate Studies ............................. 636 B. Weighting Strategy for the Fourth National Climate Assessment ................ 642 C. Detection and Attribution Methodologies Overview ............................ 652 D. Acronyms and Units ................................................................................. 664 E. Glossary ...................................................................................................... 66

U.S. GLOBAL CHANGE RESEARCH PROGRAM CLIMATE SCIENCE SPECIAL REPORT (CSSR)

Fifth-Order Draft Table of Contents Front Matter About This Report........................................................................................ 1 Guide to the Report......................................................................................4 Executive Summary ................................................................................... 12 Chapters 1. Our Globally Changing Climate .......................................................... 38 2. Physical Drivers of Climate Change ................................................... 98 3. Detection and Attribution of Climate Change .................................... 160 4. Climate Models, Scenarios, and Projections .................................... 186 5. Large-Scale Circulation and Climate Variability ................................ 228 6. Temperature Changes in the United States ...................................... 267 7. Precipitation Change in the United States ......................................... 301 8. Droughts, Floods, and Hydrology ......................................................... 336 9. Extreme Storms ....................................................................................... 375 10. Changes in Land Cover and Terrestrial Biogeochemistry ............ 405 11. Arctic Changes and their Effects on Alaska and the Rest of the United States..... 443 12. Sea Level Rise ....................................................................................... 493 13. Ocean Acidification and Other Ocean Changes .............................. 540 14. Perspectives on Climate Change Mitigation .................................... 584 15. Potential Surprises: Compound Extremes and Tipping Elements .......... 608 Appendices A. Observational Datasets Used in Climate Studies ............................. 636 B. Weighting Strategy for the Fourth National Climate Assessment ................ 642 C. Detection and Attribution Methodologies Overview ............................ 652 D. Acronyms and Units ................................................................................. 664 E. Glossary ...................................................................................................... 66

The Pathophysiology of Preeclampsia and Eclampsia

Preeclampsia is a hypertensive disorder of pregnancy, diagnosed after the 20th week of gestation in women experiencing new-onset hypertension along with symptoms affecting the liver, kidneys, or brain. In some cases, women with preeclampsia develop novel seizures or unexplained coma, at which time they are diagnosed with eclampsia. The mechanisms contributing to preeclampsia and eclampsia are not fully elucidated, although the placenta seems to play a critical role. Previous studies suggest that improper placentation stimulates mitochondrial dysfunction and the exaggerated release of placental-derived molecules including inflammatory cytokines, anti-angiogenic factors, reactive oxygen species, and cell-free nucleic acids into the maternal circulation that cause systemic vascular dysfunction. These, along with maternally derived molecules, act in concert, leading to hypertension and target organ damage during pregnancies complicated by preeclampsia and eclampsia.In this reprint, we present the original research articles and review papers published as part of the Special Issue: "The Pathophysiology of Preeclampsia and Eclampsia" in Cells

1995 BRAC Commission

Naval Research Lab: Military Value Data Call. Tabular data, memos, documents. Box 178, L-110

Climate Science Special Report: Fourth National Climate Assessment (NCA4), Volume I

New observations and new research have increased our understanding of past, current, and future climate change since the Third U.S. National Climate Assessment (NCA3) was published in May 2014. This Climate Science Special Report (CSSR) is designed to capture that new information and build on the existing body of science in order to summarize the current state of knowledge and provide the scientific foundation for the Fourth National Climate Assessment (NCA4)

Research Topics in Medicines and How Our Board Members Are Engaged in Them

With the development of analytical instruments, the academic system has become more complicated, producing new journals one after another. Therefore, it became much important to clarify what is original of “Medicines”. As the name Medicines indicates, it includes science and practice of caring for a patient and managing the diagnosis, prognosis, prevention, treatment or palliation of their injury or disease. Therefore, Medicines differs from other journals with similar title in that it covers a wide range of fields, from traditional medicine to rapidly developing molecular-targeted drugs, focusing on their pharmacological effects, structure–activity relationships, metabolic pathways, gene expression, using cultured cells, animals, and clinical trials. Most parts of this Special Issue were written by our editorial board members, who described the research topics and how they are engaged in their field of research. All of these articles are their life-long story. We collected a total number of 22 original works by basic researchers and clinical doctors. We hope that readers can get a chance to know the current status of diverse fields of medicines, and what your most important research themes are after reading these articles