RNA, 40 mg dsRNA or 2.5 mg siRNA. A square-wave protocol was applied with a single 20 ms impulse at 125 V and at room temperature. After electroporation, the worms were transferred to complete M199 medium and incubated for 5 days; 48 hours after electroporation the medium was refreshed. In case of the SmDia RNAi approach, the medium was changes at day 2 and 5 after electroporation and dsRNAs were added to the medium for an additional soaking CJ-023423 effect. For transcript detection the following primer pairs were used for RT-PCRs: a-Int1-RTPCR-59 +a-Int1-RTPCR-39, b-Int1-RTPCR-59 +b-Int1-RTPCR-39, and SmDia-5.1 +SmDia-3.1. For normalization, the transcription of the housekeeping gene SmPDI was monitored using the same cDNAs as template and the following primer combination: SmPDI-59 and SmPDI-39. All PCRs were performed in a final volume of 25 ml. PCR products were separated on 1.5% agarose gels stained with ethidium bromide. The relative intensities of the amplification products were determined densitometrically 
using the program ImageJ. For relative quantification of the Platyhelminth a/b-Integrin Receptors integrin RT-PCR products, the SmPDI amplicons were used as endogenous standard. ~~ The risk for developing chronic trapezius myalgia is increased in several occupations with work tasks such as high exposure to awkward working positions, sustained static and repetitive movements of the arms and movements with high precision demands. Various but often ineffective treatment interventions are frequently implemented. For some of the patients the pain will spread and a chronic widespread pain condition can develop. The pathophysiological mechanisms behind TM and CWP are poorly understood. Microdialysis mimics the function of a capillary blood 17660385 vessel by perfusing a thin dialysis tube implanted in the tissue with a physiological saline solution. Substances can pass across the dialysis membrane along the concentration gradient. The dialysate is analyzed and reflects the composition of the extra cellular fluid where nociceptor free nerve endings terminate. Metabolic, algesic and potential nociception/pain inhibitory substances have been studied in chronic myalgia using MD. Significant increases e.g., in serotonin, glutamate, lactate, pyruvate, N-stearoylethanolamine and palmitoylethanolamide have been reported; for a review see. In the majority of hitherto performed studies one or a few small biochemical substances have been exclusively in focus. There are a few attempts to analyze larger molecules, e.g., proteins, with various results. Proteins can act as signal substances, activate the formation of algesics and control nociceptive processes. Proteomics, defined as the science and the methodology of investigating the proteome has been suggested as a useful technique in order to identify biomarkers Proteomics of Trapezius Muscle Microdialysate of pain conditions. Two-dimensional gel electrophoresis is a widely used technique 19535597 to separate organic substances such as proteins in different tissues. Proteomics have been applied to pain research to identify protein changes in different pain conditions. The proteome has been investigated in animals comparing inflammatory and neuropathic pain. The proteome of the cerebrospinal fluid in healthy subjects, in patients with herniated disc and idiopathic back pain have been studied. Also, nerve samples from patients with complex regional pain syndrome have been analyzed using proteomics in order to detect up/down
