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Two time-resolved fluorescence-based methods for nucleic acid quantification are described and their results are compared. Both methods use an exogenous internal standard to eliminate errors arising from different steps of the assay. The first method is a competitive end-point assay, where the standard competes for the same primers with the actual target sequence, prostate-specific antigen (PSA) c
Objectives. To design protocols for specific and quantitative immunohistochemical detection of human kallikrein 2 (hK2) using lanthanide chelate-labeled monoclonal antibodies (Mabs) and time-resolved fluorescence imaging. Methods. Anti-prostate-specific antigen (PSA) Mabs were tested in microtiterplate assays for their ability to prevent PSA from cross-reacting with the anti-hK2 Mab 6H10. Europium
BACKGROUND. In semen, prostate-specific antigen (PSA or hK3) digests the gel proteins semenogelin I and II, resulting in liquefaction and the release of motile spermatozoa. We characterized the substrate specificity and zinc-mediated inhibition of PSA. METHODS. The proteolysis of human semenogelin I (SgI) and II (SgII) by PSA was characterized by purification of generated SgI and SgII fragments, N
The 2006 World Health Organization (WHO) growth standard—based on the Multi Growth Reference Study (MGRS)—is a universal standard for healthy height and weight of young children. The share of children that deviate substantially from this growth standard (i.e., fall below −2 standard deviations) defines three important indicators: stunting (low height-for-age), wasting (low weight-for-height), and
Prostate specific antigen (PSA, hK3) in serum is predominantly complexed to α-1-antichymotrypsin (ACT), but a minor fraction remains in a free form despite the very large excess of serine protease inhibitors and α-2- macroglobulin. The fraction of free to total PSA is significantly lower in prostate cancer (CAP) compared to benign prostatic hyperplasia (BPH) which provides improved discrimination
The impact on the performance of GaN high electron mobility transistors (HEMTs) of in situ ammonia (NH3) pre-treatment prior to the deposition of silicon nitride (SiN) passivation with low-pressure chemical vapor deposition (LPCVD ) is investigated. Three different NH3 pre-treatment durations (0, 3, and 10 min) were compared in terms of interface properties and device performance. A reduction o
Human glandular kallikrein 2 (hK2) is a serine protease expressed by the prostate gland with 80% identity in primary structure to prostate-specific antigen (PSA). Recently, hK2 was shown to activate the zymogen form of PSA (proPSA) in vitro and is likely to be the physiological activator of PSA in the prostate, hK2 is also able to activate urokinase and effectively cleave fibronectin. We studied t
BACKGROUND. Protein gene product 9.5 (PGP 9.5) has been considered to be a neuronal marker, but it is also present in extraneuronal tissues, e.g., the human mammary gland and rat epididymis. Its presence and distribution in the developing and adult male human genital tract have been unknown. METHODS. Immunohistochemical reactions were performed on human embryonic and postnatal specimens of the mal
Chelates with fluorescent lanthanides such as europium and terbium are widely used in immunofluorometric assays, e.g. for the measurement of different molecular forms of prostate-specific antigen (PSA) in serum for detection and monitoring of prostate cancer. These chelates have also been introduced as non-radioactive labels in immunocytochemistry and in situ hybridization. In the present study, s
Objectives. The subcellular localization of the breast cancer susceptibility gene product BRCA1 has been controversial. Discrepant results have been reported during the past 3 years, partially because of the unavailability of highly specific reagents for BRCA1 protein. Our objective was to characterize the BRCA1-like immunoreactivity that is detected in human seminal plasma by using monoclonal and
We have developed a direct immunofluorescence technique utilising chelates of the lanthanide ions europium and terbium conjugated to monoclonal IgGs (Mabs) against prostate-specific antigen (PSA) and alpha-1- antichymotrypsin (ACT) for the detection and quantification on the same tissue section. Strong signals without disturbance from tissue autofluorescence were demonstrated in paraffin sections
Seventy-nine monoclonal antibodies submitted to the ISOBM TD-3 PSA Workshop were tested for their reactivity with recombinant human kallikrein-2 (rhK2). A sandwich immunofluorometric assay using polyclonal antiprostate- specific antigen (PSA) antiserum-coated plates was used to capture rhK2 and subsequently the test antibody. The response of each test antibody was compared with 3 reference antibod
The concept of measuring the proportions of various forms of PSA in serum, particularly the proportion of free to total PSA, represents a new and exciting method of detecting early curable prostate cancers and avoiding unnecessary prostate biopsies in men who have BPH only. Compared with other methods of improving diagnostic specificity, it does not require transrectal ultrasound for determination
Objectives. Nearly half of men with clinically localized prostate cancer are understaged. We evaluated whether knowledge of preoperative free prostate-specific antigen (f-PSA), complexed (c-PSA), and total (t-PSA) concentrations or the ratios thereof (f-PSA/t-PSA, c-PSA/t PSA, and f PSA/c- PSA) could improve upon the staging of prostate cancer when compared with standard PSA testing (t-PSA). In ad
At ejaculation, the epididymal spermatozoa mix with the secretions produced by the seminal vesicles and the prostate. The glandular secretions immediately turns into a gel-like structure which entraps the spermatozoa. The ejaculated spermatozoa become progressively motile as the gel dissolves. The discovery, structure, and interactions of proteins involved in these post-ejaculatory phase reactions
Stringent factors orchestrate bacterial cell reprogramming through increasing the level of the alarmones (p)ppGpp. In Beta- and Gammaproteobacteria, SpoT hydrolyzes (p)ppGpp to counteract the synthetase activity of RelA. However, structural information about how SpoT controls the levels of (p)ppGpp is missing. Here we present the crystal structure of the hydrolase-only SpoT from Acinetobacter baum
