Ex vivo animal-model assessment of a non-invasive system for loss of resistance detection during epidural blockade.

During recent decades epidural analgesia has gained widespread recognition in many applications. In this complex procedure, anaesthetist uses a specific needle to inject anesthetic into the epidural space. It is crucial the appropriate insertion of the needle through inhomogeneous tissues placed between the skin and the epidural space to minimize anesthetic-related complications (e.g., nausea, headache, and dural puncture). Usually, anaesthetists perform the procedure without any supporting tools, and stop pushing the syringe when they sense a loss of resistance (LOR). This phenomenon is caused by the physical properties of the epidural space: the needle breaks the ligamentum flavum and reaches the epidural space, in this stage the anaesthetist perceives a LOR because the epidural space is much softer than the ligamentum flavum. To support the clinician in this maneuver we designed a non-invasive system able to detect the LOR by measuring the pressure exerted on the syringe plunger to push the needle up to the epidural space. In a previous work we described the system and its assessment during in vitro tests. This work aims at assessing the feasibility of the system for LOR detection on a more realistic model (ex vivo pig model). The system was assessed by analyzing: its ability to hold a constant value (saturation condition) during the insertion of the needle, and its ability to detect the entrance within the epidural space by a decrease of the system's output. Lastly, the anaesthetist was asked to assess how the ex vivo procedure mimics a clinical scenario. The system reached the saturation condition during the needle insertion; this feature is critical to avoid false positive during the procedure. However, it was not easy to detect the entrance within the epidural space due to its small volume in the animal model. Lastly, the practitioner found real the model, and performed the procedures in a conventional manner because the system did not influence his actions.

A cost-effective, non-invasive system for pressure monitoring during epidural needle insertion: Design, development and bench tests.

Epidural blockade procedures have gained large acceptance during last decades. However, the insertion of the needle during epidural blockade procedures is challenging, and there is an increasing alarming risk in accidental dural puncture. One of the most popular approaches to minimize the mentioned risk is to detect the epidural space on the base of the loss of resistance (LOR) during the epidural needle insertion. The aim of this paper is to illustrate an innovative and non-invasive system able to monitor the pressure exerted during the epidural blockade procedure in order to detect the LOR. The system is based on a Force Sensing Resistor (FSR) sensor arranged on the top of the syringe's plunger. Such a sensor is able to register the resistance opposed to the needle by the different tissues transducing the pressure exerted on the plunger into a change of an electrical resistance. Hence, on the base of a peculiar algorithm, the system automatically detects LOR providing visual and acoustic feedbacks to the operator improving the safety of the procedure. Experiments have been performed to characterize the measurement device and to validate the whole system. Notice that the proposed solution is able to perform an effective detection of the LOR.

Microsatellite and AFLP markers in the Prunus persica [L. (Batsch)]xP. ferganensis BC(1)linkage map: saturation and coverage improvement.

A set of 146 single sequence repeats (SSRs) and 14 amplified fragment length polymorphism (AFLP) primer combinations were used to enrich a previously developed linkage map obtained from a (Prunus persicaxP. ferganensis)xP. persica BC(1) progeny. Forty-one SSR primer pairs gave polymorphic patterns detecting 42 loci. The restriction/selective primer AFLP combinations produced a total of 79 segregating fragments. The resulting map is composed of 216 loci covering 665 cM with an average distance of 3.1 cM. Novel regions were covered by the newly mapped loci for a total of 159 cM. Eight linkage groups were assembled instead of the earlier 10 as two small groups (G1a and G8b), previously independent, were joined to their respective major groups (G1b and G8a). Several gaps were also reduced resulting in an improved saturation of the map. Twelve gaps >or=10 cm are still present. A comparative analysis against the Prunus reference map (71 anchor loci) pointed out an almost complete synteny and colinearity. Six loci were not syntenic and only two were not colinear. Genetic distances were significantly longer in our map than in the reference one.

A peach linkage map integrating RFLPs, SSRs, RAPDs, and morphological markers.

A linkage map was obtained using a BC1 progeny (Prunus persica x (P. persica x P ferganensis)). The map is composed of 109 loci (74 RFLPs, 17 SSRs, 16 RAPDs, and two morphological traits) distributed in 10 linkage groups. Loci, segregating in five different ratios, were integrated in the map with JoinMap 2.0 software. The map covers 521 cM of the peach genome. The average distance between adjacent loci is 4.8 cM. Two monogenic traits, flesh adhesion (F/f) and leaf glands (E/e), were placed on the map. Thirty-two loci in common with a saturated linkage map of Prunus allowed a comparative analysis to be made between the two maps. Homologies were found among the respective linkage groups. No relevant differences were observed in the linear order of the common loci.

Microsatellite DNA in peach (Prunus persica L. Batsch) and its use in fingerprinting and testing the genetic origin of cultivars.

We isolated and sequenced 26 microsatellites from two genomic libraries of peach cultivar 'Redhaven', enriched for AC/GT and AG/CT repeats, respectively. For 17 of these microsatellites, it was possible to demonstrate Mendelian inheritance. Microsatellite polymorphism was assayed in 50 peach and nectarine cultivars. Of the 1300 PCRs carried out, all but two produced amplified products of the expected size. All microsatellites were polymorphic, showing 2-8 alleles per locus. Heterozygosity ranged from 0.04-0.74 (mean 0.47); the discrimination power (PD) ranged from 0.04-0.84 (mean 0.60). Cultivar heterozygosity varied greatly, with one cultivar ('Independence') being homozygous at all loci. The set of microsatellites discriminated all cultivars investigated, except several sport mutations, i.e., 'Dixitime' vs. 'Springcrest', 'Compact Redhaven' vs. 'Redhaven', and two pairs of cultivars, 'Venus' vs. 'Orion' and 'Elegant Lady' vs. 'Rome Star', whose pedigrees are controversial. We were able to analyze the paternity of several cultivars. In most cases, the parenthood was confirmed. The comparison of three long-living 'Redhaven' accessions supplied by different repositories did not provide any evidence of somatic instability of microsatellites. Hence, microsatellites, ranked according to their information content, are recommended as markers of choice for peach fingerprinting and suggestions are provided for interpreting band profiles and the correct sizing of alleles.