We selected introns with nucleotide sequence similarity using the SSEARCH with Smith-Waterman similarity scores (http://fasta.bioch.virginia.edu/fasta_www2/fasta_list2.shtml) using 200 PRSS iterations [22] among the 335 introns used in this study.
The gene expression pattern between B13 cells with and without Ad-PNM was then compared by performing qRT PCR for rat Ins1, Ins2, Iapp, Pax4, Neurod1, Nkx2.2, Arx, endogenous Pdx, endogenous Ngn3, endogenous MafA, Isl1, Mnx1, Hes1, Slc2a2, Kcnj11, Abcc8, Amy, Ptf1a, Cpe, Pcsk2, Pcsk1, Prss, Cpa, Ctrb1, Rbpj1 and Gapdh (glyceraldehyde-3-phosphate dehydrogenase).
No differences were found between the groups at T1. PRSS and PRCS results were analyzed in 25 patients (14 in Bfb group, 11 in Control group) because their questionnaires were invalid.
Untreated pollen from these lines had normal filamentous actin organization, and they retained this actin configuration after addition of compatible combinations of PrsS (AtPpS1 with PrsS8, Figure 3E; AtPpS3 with PrsS1, Figure 3F).
Cognitive variables were assessed using the 32-item Pain-Related Self-Statements Scale (PRSS) [ 31] with the subscales 'active coping' (for example, 'I can handle my pain') and 'catastrophising' (for example, 'I am a hopeless case') shown to have excellent psychometric properties [ 31].
To provide further evidence for PrpS elicitation of a Papaver-like SI response, we investigated whether death was triggered in AtPpS1 and AtPpS3 pollen after addition of PrsS, by assessing viability of pollen using Evans blue at 8 hr.
Quantification showed that filamentous actin is the predominant phenotype, except for the combination of cognate recombinant PrsS with PrpS pollen (AtPpS1 pollen with PrsS1 added, and AtPpS3 pollen with PrsS3).
The PRSS is an 18-item questionnaire that assesses situation-specific aspects of patients' cognitive coping strategies for pain.
We next investigated whether expression of PrpS in A. thaliana pollen was sufficient to induce similar intracellular responses to those elicited in incompatible Papaver pollen [ 7] by adding incompatible recombinant PrsS.
As Papaver SI relies on a DEVDase/caspase-3-like activity [ 14, 15], we assessed whether a similar activity was involved in the death of PrpS-expressing A. thaliana pollen, by adding Ac-DEVD-CHO, a caspase-3 inhibitor before addition of PrsS.
