Use of a multiplex polymerase chain reaction assay in the antemortem diagnosis of toxoplasmosis and neosporosis in the central nervous system of cats and dogs

2003 ◽  
Vol 64 (12) ◽  
pp. 1507-1513 ◽  
Author(s):  
Scott J. Schatzberg ◽  
Nicholas J. Haley ◽  
Stephen C. Barr ◽  
Alexander deLahunta ◽  
Natasha Olby ◽  
...  
Keyword(s):  
Central Nervous System ◽  
Polymerase Chain Reaction ◽  
Nervous System ◽  
Polymerase Chain Reaction Assay ◽  
Multiplex Polymerase Chain Reaction ◽  
Chain Reaction ◽  
The Central Nervous System ◽  
Polymerase Chain

Expression of the integrin alpha 6 beta 4 in peripheral nerves: localization in Schwann and perineural cells and different variants of the beta 4 subunit

1994 ◽  
Vol 107 (2) ◽  
pp. 543-552 ◽  
Author(s):  
C.M. Niessen ◽  
O. Cremona ◽  
H. Daams ◽  
S. Ferraresi ◽  
A. Sonnenberg ◽  
...  
Keyword(s):  
Central Nervous System ◽  
Polymerase Chain Reaction ◽  
Nervous System ◽  
Peripheral Nerves ◽  
Northern Blot Analysis ◽  
The Other ◽  
Chain Reaction ◽  
The Central Nervous System ◽  
Polymerase Chain ◽  
Integrin Alpha 6

Integrin alpha 6 beta 4 is expressed in human peripheral nerves, but not in the central nervous system. This integrin heterodimer has previously been found in perineural fibroblast-like cells and in Schwann cells (SCs), which both assemble a basement membrane but do not form hemidesmosomes. We show here that in SCs, which had formed a myelin sheath, alpha 6 beta 4 was enriched in the proximity of the nucleus, at Ranvier paranodal areas and at Schmitt-Lanterman clefts; alpha 6 beta 4 was also found at the grooved interface between small axons and non-myelinating SCs. Immunoprecipitation of human peripheral nerves, in combination with Western blotting showed that beta 4 is associated with the alpha 6A subunit. Northern blot analysis of human peripheral nerves showed a single beta 4 transcript of 6 kb. Using the reverse transcriptase polymerase chain reaction, we detected two mRNA species, one for the most common (−70, -53) form of beta 4 and the other encoding the (+53) variant of beta 4. Cultured SCs were devoid of alpha 6 beta 4 but expressed alpha 6 beta 1, indicating that SCs lose beta 4 expression when contact with neurons is lost. Thus, resting SCs in contact with axons express alpha 6A in combination with beta 4, irrespective of myelin formation. We suggest that alpha 6 beta 4 expressed in SCs plays a role in peripheral neurogenesis.

scholarly journals A Single-center, Quasi-experimental Study to Evaluate the Impact of a Multiplex Polymerase Chain Reaction System Combined with Antimicrobial Stewardship Intervention on Time to Targeted Therapy in Patients with Suspected Central Nervous System Infection

2017 ◽  
Vol 4 (suppl_1) ◽  
pp. S8-S8 ◽  
Author(s):  
Kelsey Powell ◽  
Sara Revolinski ◽  
Allison Gibble ◽  
Anne Daniels ◽  
J Njeri Wainaina ◽  
...  
Keyword(s):  
Central Nervous System ◽  
Polymerase Chain Reaction ◽  
Nervous System ◽  
Targeted Therapy ◽  
Multiplex Polymerase Chain Reaction ◽  
Antimicrobial Agents ◽  
Empiric Therapy ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Research Grant

Abstract Background Empiric treatment for central nervous system (CNS) infections consists of coverage with multiple antimicrobial agents that may be continued until a pathogen can be identified. Identification may take significant time to result, leading to extended durations of multiple antimicrobial agents, delays in targeted therapy and subsequent adverse effects, such as nephrotoxicity and Clostridium difficile infection. A multiplex polymerase chain reaction (PCR) system that can identify 14 pathogens responsible for community-acquired CNS infections in 1 hour was recently FDA-approved for cerebrospinal fluid (CSF) analysis. The objective of this study was to determine the effect of this PCR paired with antimicrobial stewardship (AMS) team intervention on the time to targeted therapy. Methods During the intervention (Int) phase (January 25, 2017–April 30, 2017), all PCR results were called to the AMS team, who reviewed clinical data and provided antimicrobial recommendations per pre-determined protocol. Recommendations consisted of de-escalation or addition of therapy. The pre-intervention (PI) group consisted of patients with CSF culture obtained between January 25, 20116 and April 30, 2016. Results A total of 138 patients were evaluated; 46 in the Int group and 92 in the PI. Of the 46 patients in the Int group, 25 had a negative PCR result and were never initiated on antimicrobials. One patient required antimicrobial escalation. Twenty patients were started on empiric therapy and were candidates for de-escalation. In the PI group, there were no patients with CSF cultures that required therapy escalation, while 33 patients were initiated on empiric antimicrobials. Results from the subgroup of patients in whom empiric therapy was started as shown in Table 1. Conclusion Implementation of a multiplex PCR with AMS intervention resulted in decreased time to targeted therapy. This project was funded through a competitive stewardship grant provided by Merck & Co. Disclosures S. Revolinski, Merck: Grant Investigator, Research grant; J. N. Wainaina, Merck: Grant Investigator, Research grant; A. Huang, Merck: Grant Investigator, Research grant

scholarly journals Quantitative RT-PCR Analysis of Uncoupling Protein Isoforms in Mouse Brain Cortex: Methodological Optimization and Comparison of Expression with Brown Adipose Tissue and Skeletal Muscle

2004 ◽  
Vol 24 (7) ◽  
pp. 780-788 ◽  
Author(s):  
Sylvain Lengacher ◽  
Pierre J. Magistretti ◽  
Luc Pellerin
Keyword(s):  
Skeletal Muscle ◽  
Central Nervous System ◽  
Polymerase Chain Reaction ◽  
Nervous System ◽  
Adipose Tissue ◽  
Brain Cortex ◽  
Chain Reaction ◽  
Brown Adipose ◽  
The Central Nervous System ◽  
Polymerase Chain

Uncoupling proteins (UCPs) present in the inner mitochondrial membrane are involved in uncoupling respiration from ATP synthesis. Five UCP isoforms have been identified but information about their presence and level of expression in the central nervous system remains incomplete. To determine the nature and proportion of UCP isoform mRNAs present in brain cortex, we developed and optimized a specific quantitative reverse-transcription polymerase chain reaction procedure. Optimal range of RNA concentrations to be used in the reverse-transcriptase reaction was determined. Primer design and concentration were optimized for each target gene while polymerase chain reaction efficiency was assessed for a range of reverse-transcriptase dilutions. Genomic contribution to the quantitative signal was evaluated for each isoform and minimized. Three reference genes were tested for normalization, and β-actin was found to be the most stable among tissues. Results indicate that brain cortex contains significant amounts of all UCP mRNAs, with UCP5 and UCP4 being the most abundant, as opposed to brown adipose tissue and skeletal muscle, which predominantly express UCP1 and UCP3, respectively. These data provide a first quantitative assessment of UCP mRNA expression in mouse brain, showing the presence of all five isoforms with distinct proportions, thus suggesting specific roles in the central nervous system.

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