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mRNA Copy Numbers (mrna + copy_number)

Distribution by Scientific Domains

Medical Sciences	75%

Selected Abstracts

Quantitative PSA RT-PCR for preoperative staging of prostate cancer

THE PROSTATE, Issue 4 2003
Ralf Kurek
Abstract BACKGROUND The clinical value of detecting prostate specific antigen (PSA) mRNA in the peripheral blood mononuclear cell fraction of patients (pts) by standard RT-PCR assays with localized prostate cancer remains controversial. We used a quantitative RT-PCR assay to measure the PSA mRNA copy number in addition to the qualitative PSA RT-PCR and correlated the results with clinical parameters. METHODS Total RNA was extracted from the peripheral blood mononuclear cell fraction of 115 prostate cancer pts prior to radical retropubic prostatectomy (RP) who received 3 months of neoadjuvant androgen deprivation. For quantitative RT-PCR, a PSA-like internal standard (IS) was added to each sample prior to reverse transcription and the PCR products for PSA and IS were selectively detected with fluorescent europium chelates after hybridization. Corresponding qualitative PSA,RT-PCR was performed for all samples. RESULTS The median PSA copy number was 126 (range: 0,37988). There were no significant correlations established between qualitative or quantitative RT-PCR results and given clinical parameters. Corresponding quantitative and qualitative RT-PCR results were significantly associated (P,=,0.01). CONCLUSIONS We were unable to show any additional value of quantitative as well as qualitative PSA RT-PCR for preoperative staging of prostate cancer so far. Nevertheless, the long-term follow up of the patients has to be awaited. Prostate 56: 263,269, 2003. © 2003 Wiley-Liss, Inc. [source]

Matrix metalloproteinase-9 and tissue inhibitor of matrix metalloproteinase-1 in the respiratory tracts of human infants following paramyxovirus infection

JOURNAL OF MEDICAL VIROLOGY, Issue 4 2007
Matthew B. Elliott
Abstract Respiratory syncytial (RSV) and parainfluenza (PIV) viruses are primary causes of acute bronchiolitis and wheezing illnesses in infants and young children. To further understand inflammation in the airways following infection, we tested for the presence of matrix metalloproteinases (MMP) and natural tissue inhibitors of MMP (TIMP) in primary and established human cell lines, and in the nasopharyngeal secretions (NPS) of human infants infected with RSV or PIV. Using ELISA and multiplex-based assays, MMP-9 and TIMP-1 proteins were, respectively, detected in 66/67 and 67/67 NPS. During PIV or RSV infection TIMP-1 concentrations were associated with hypoxic bronchiolitis. TIMP-1 amounts were also negatively correlated with O2 saturation, and positively correlated with IL-6, MIP-1,, and G-CSF amounts following RSV infection. IL-6, MIP-1,, and G-CSF were negatively correlated with O2 saturation during RSV infection. Acute respiratory tract disease was not associated with MMP-9 protein/protease activity. Additional studies using real-time quantitative PCR suggested that MMP-9 mRNA copy numbers were elevated in normal human bronchial epithelial (NHBE) cells infected with RSV, while TIMP-1 and TIMP-2 were not increased. However, ELISA did not reveal MMP-9 protein in the NHBE cell culture supernatants. Hence, the data implied that airway epithelial cells were not the primary source of MMP or TIMP following paramyxovirus infection. Taken together, the data suggested that paramyxovirus infection perturbs MMP-9/TIMP-1 homeostasis that in turn may contribute to the severity of respiratory tract disease. J. Med. Virol. 79:447,456, 2007. © 2007 Wiley-Liss, Inc. [source]

Abnormal basement membrane type IV collagen ,-chain composition in labial salivary glands in Sjögren's syndrome

ARTHRITIS & RHEUMATISM, Issue 4 2009
P. Poduval
Objective Sjögren's syndrome (SS) is characterized by atrophy and malfunction of the acinar cells. The aim of this study was to investigate whether type IV collagen ,-chain composition of acinar cell compartments could be abnormal in diseased glands. Methods Messenger RNA (mRNA) from human submandibular gland (HSG) cells, cultured with or without growth factor,depleted Matrigel, was analyzed using quantitative reverse transcription,polymerase chain reaction (RT-PCR). Labial salivary glands were analyzed using quantitative RT-PCR and immunohistochemistry. Results HSG cells of both the ductal and acinar phenotypes synthesized all ,-chain mRNA, in particular those of the ,1 and ,2 chains. Labial salivary glands (LSGs) contained ,1/2 chains but also contained mRNA of all the other ,-chains, although the mRNA copy numbers for the ,3 and ,4 chains were low, and the corresponding proteins were absent. Type IV collagen ,1/2-chains were observed in all tubuloalveolar basement membranes. In healthy glands, ,5 and ,6 chains were continuous around ducts but discontinuous around acini. In SS glands, these chains were absent or patchy around the ducts and absent around the acini. Conclusion Ductal and acinar epithelial cells are able to locally produce mRNA for all 6 different ,-chains. Type IV collagen ,1/2-chains seem to form the backbone in the tubuloalveolar basement membrane in salivary glands. Type IV collagen ,3 and ,4 chain mRNA were found in cultured salivary epithelial cells and LSG explants but were not translated to the corresponding ,-chains in LSGs. Both ,5 and ,6 mRNA were observed in salivary epithelial cells and glands. In healthy glands, immunolabeling always disclosed corresponding ,-chains around ducts, but their synthesis and/or degradation seemed to be locally regulated around acinar cells. [source]

Expression of LIF and LIF receptor beta in Alzheimer's and Parkinson's diseases

ACTA NEUROLOGICA SCANDINAVICA, Issue 1 2010
M. Soilu-Hänninen
Background,,, Signaling through the leukemia inhibitory factor (LIF) receptor (LIFR) is crucial for nervous system development. There are few studies concerning the expression of LIF and LIFR in normal and degenerating adult human brain. Objectives,,, To study the expression of LIF and LIFR in Alzheimer's disease (AD), Parkinson's disease (PD), and control brains. Patients and methods,,, LIF and LIFR mRNA copy numbers were determined by quantitative real-time RT-PCR from four brain regions of 34 patients with AD, 40 patients with PD, and 40 controls. Immunohistochemistry was performed in seven PD and in four AD patients and in seven normal controls. Results,,, In general, the LIF copy numbers were 1 log higher than the LIFR copy numbers. In the AD brains, LIF expression was higher than in the controls in the hippocampus and in the temporal cortex, and in the PD brains in the hippocampus and in the anterior cingulated cortex. Expressions of LIF and LIFR in different brain regions were opposite except for the AD hippocampus and PD anterior cingulated cortex, where the expression patterns were parallel. Conclusions,,, Co-operative expression of LIF and LIFR in AD hippocampus and PD anterior cingulated cortex may indicate a role for LIF in neuronal damage or repair in these sites. [source]