Impact of reduced-representation sequencing protocols on detecting population structure in a threatened marsupial.

Reduced-representation sequencing methods have wide utility in conservation genetics of non-model species. Several methods are now available that reduce genome complexity to examine a wide range of markers in a large number of individuals. We produced two datasets collected using different laboratory techniques, comprising a common set of samples from the greater bilby (Macrotis lagotis). We examined the impact of differing data filtering thresholds on downstream population inferences. We found that choice of restriction enzyme and data filtering thresholds, especially the rate of allowable missing data, impacted our ability to detect population structure. Estimates of FST were robust to alterations in laboratory and bioinformatic protocols while principal coordinates and STRUCTURE analyses showed variation according to the number of loci and percent missing data. We advise researchers using reduced-representation sequencing in conservation projects to examine a range of data thresholds, and follow these through to downstream population inferences. Multiple measures of population differentiation should be used in order to fully understand how data filtering thresholds influence the final dataset, paying particular attention to the impact of allowable missing data. Our results indicate that failure to follow these checks could impact conclusions drawn, and conservation management decisions made.

Human remyelination promoting antibody inhibits apoptotic signaling and differentiation through Lyn kinase in primary rat oligodendrocytes.

PURPOSE: Human remyelination promoting IgM mAbs target oligodendrocytes (OLs) and function in animal models of multiple sclerosis (MS). However, their mechanism of action is unknown. This study seeks to identify the cellular mechanism of action of a recombinant human IgM on OL survival. METHODS: Binding of rHIgM22 to the surface of rat OLs was studied by co-localization with various markers. RHIgM22-mediated effects on apoptotic signaling in OLs, differentiation markers, and signaling molecules were detected by Western blotting and immunoprecipitation. RESULTS: RHIgM22 co-localized with integrin ß3 but not other integrin ß-chains in OLs. Downstream of integrin ß3 we identified Src family kinase (SFK) Lyn as a key player of rHIgM22-mediated actions in OLs. Lyn immunoprecipitated in a complex together with integrin αvß3 and PDGFαR. Lyn expression was 9-fold up-regulated and Lyn activation was 3-fold higher inrHIgM22-treated OL cultures compared with controls. RHIgM22 inhibited apoptotic signaling by greater than 10-fold reduction of caspase-3 and capsase-9 cleavage and reduced by 4-fold expression of differentiation markers MBP and MOG in OLs. SFK inhibitors PP2 and SU6656 inhibited Lyn activity and restored caspase-cleavage in OLs. A human IgM that did not promote remyelination and medium wereused as controls. CONCLUSIONS: rHIgM22 prevented apoptotic signaling andinhibited OL differentiation by Lyn implying thatIgM-mediated remyelination is due toprotection of OPC and OLs rather than promotion of OPC differentiation.

The effect of Rapid Induction Analgesia on subjective pain ratings and pain tolerance.

The effect of Rapid Induction Analgesia (RIA) on pain tolerance and ratings of mechanically induced pain in the pain-sensitized forearm was investigated in 58 undergraduates. Posthypnotic suggestions of relaxation and analgesia did not influence pain ratings or tolerance, but relaxation ratings increased after RIA. When suggestions for analgesia were made throughout pain testing, ratings of pain unpleasantness at the pain tolerance point decreased more in the RIA group than in the attention control group. However, RIA did not influence pain threshold or tolerance. It was concluded that RIA was more effective in reducing subjective reports of pain (particularly the affective component) than in altering pain tolerance, and that maintenance of hypnotic suggestions was more effective than posthypnotic suggestions of comfort and relaxation in alleviating the affective component of pain.

Rapid induction analgesia for the alleviation of procedural pain during burn care.

Burn patients must often endure intense pain during their regular dressing changes. The aim of the present study was to investigate the therapeutic effect of rapid induction analgesia (RIA) on resting and procedural pain, anticipatory anxiety, relaxation levels and medication consumption in 30 hospitalized burn patients. Patients rated levels of pain and relaxation for four burn care sessions. RIA was conducted twice on 15 patients, whereas dressing changes proceeded as usual in 15 control patients. When asked to recall pain during the dressing changes, patients remembered an experience which was worse in its entirety than the average of spot ratings taken during the burn care procedure. However, self-reported ratings of the sensory and affective components of pain decreased significantly during and after RIA, particularly in patients who became readily absorbed, and relaxation increased during burn care. Anticipatory anxiety decreased before dressing changes in the RIA group, and analgesic intake decreased between treatment sessions. The promising outcome of this study confirms RIA as a viable adjunct to narcotic treatment for pain control during burn care.