Cloning and mapping multiple S-locus F-box genes in European pear (Pyrus communis L.).

European pear, as well as its close relatives Japanese pear and apple, exhibits S-RNase-based gametophytic self-incompatibility. The male determinant of this self-incompatibility mechanism is a pollen-expressed protein containing an F-box domain; in the genera Petunia (Solanaceae), Antirrhinum (Plantaginaceae), and Prunus (Rosaceae), a single F-box gene determines the pollen S. In apple and Japanese pear, however, multiple S-locus F-box genes were recently identified as candidates for the pollen S, and they were named S-locus F-Box Brothers. These genes were considered good candidates for the pollen S determinant since they exhibit S-haplotype-specific polymorphisms, pollen-specific expression, and linkage to the S-RNase. In the present study, S-locus F-Box Brothers homologs have been cloned from two of the most agronomically important European pear varieties, "Abbé Fétel" (S104-2/S105) and“Max Red Bartlett" (S101/S102), and they have been mapped on a genetic linkage map developed on their progeny. Our results suggest that the number of Fbox genes linked to the S-locus of the European pear is higher than expected according with previous reports for apple and Japanese pear, since up to five genes were found to be linked to a single S-haplotype. Moreover, two of these genes exhibited an incomplete linkage to the S-RNase, allowing the identification of low-frequency recombinant haplotypes, generated by a crossing-over event between the two genes. These F-box genes are most likely placed in close proximity of the S-locus but do not belong to it, and they can thus be excluded from being responsible for the determination of pollen S function

Topical Analgesia with Lidocaine Plus Diclofenac Decreases Pain in Benign Anorectal Surgery: Randomized, Double-blind, and Controlled Clinical Trial

Objective: The aim of this study is to evaluate the efficacy and safety of a topical formulation containing lidocaine plus diclofenac (CLIFE1) compared to CLIFE2 (lidocaine), to decrease pain in benign anorectal surgery (BARS) to date not evaluated. Background: More than 50% of patients undergoing BARS, especially hemorrhoidectomy, suffer from moderate and severe postoperative pain. This remains an unresolved problem that could be addressed with the new CLIFE1 topical treatment. Methods: A multicenter, randomized double-blind, active-controlled parallel-group superiority trial, was conducted in two Spanish hospitals. Patients undergoing BARS (hemorrhoids, anal fistula and anal fissure) were randomized at the end of surgery at a 1:1 ratio to receive first dose either CLIFE1 (n=60) or CLIFE2 (n=60) anorectal topical treatment, and after every 12 hours for the first three postoperative days and once a day from the fourth to sixth. The primary outcome was average of pain decrease after topical treatment, measured with visual analogue scale (VAS) by the patients themselves, the evening in the surgery day and four times daily for the first three postoperative days. Results: The results of 120 patients included out of 150 selected undergoing BARS show a decrease in pain after CLIFE1 topical treatment (7.47±13.2) greater than with CLIFE2 (4.38±6.75), difference -3.21 (95% CI) -5.75; -0.676; p=0.008), decreasing significantly postoperative pain (≥ 9 mm, VAS) in 35% of patients undergoing benign anorectal surgery, compared to 18.33 % treated with lidocaine. Conclusions: The CLIFE1 topical treatment shows better analgesic efficacy than CLIFE2 in BARS

Molecular bases and evolutionary dynamics of self-incompatibility in the Pyrinae (Rosaceae).

The molecular bases of the gametophytic self-incompatibility (GSI) system of species of the subtribe Pyrinae (Rosaceae), such as apple and pear, have been widely studied in the last two decades. The characterization of S-locus genes and of the mechanisms underlying pollen acceptance or rejection have been topics of major interest. Besides the single pistil-side S determinant, the S-RNase, multiple related S-locus F-box genes seem to be involved in the determination of pollen S specificity. Here, we collect and review the state of the art of GSI in the Pyrinae. We emphasize recent genomic data that have contributed to unveiling the S-locus structure of the Pyrinae, and discuss their consistency with the models of self-recognition that have been proposed for Prunus and the Solanaceae. Experimental data suggest that the mechanism controlling pollen\u2013pistil recognition specificity of the Pyrinae might fit well with the collaborative \u2018non-self\u2019 recognition system proposed for Petunia (Solanaceae), whereas it presents relevant differences with the mechanism exhibited by the species of the closely related genus Prunus, which uses a single evolutionarily divergent F-box gene as the pollen S determinant. The possible involvement of multiple pollen S genes in the GSI system of Pyrinae, still awaiting experimental confirmation, opens up new perspectives to our understanding of the evolution of S haplotypes, and of the evolution of S-RNase-based GSI within the Rosaceae family. Whereas S-locus genes encode the players determining self-recognition, pollen rejection in the Pyrinae seems to involve a complex cascade of downstream cellular events with significant similarities to programmed cell death

STRUCTURAL AND FUNCTIONAL CONSERVATION OF S-SPECIFICITIES AMONG PYRINAE SPECIES

Gametophytic self-incompatibility (GSI) is the main mechanism that controls fertilization in many rosaceous species, including those belonging to the subtribe Pyrinae (formerly the Maloideae). In natural conditions, S-specificities are subject to frequency-dependent balancing selection; the genetic imprint left by this kind of selection is visible on the S-RNase gene in terms of high sequence diversity, evidence of positive selection, and shared ancestral polymorphisms: thus, some alleles have been maintained almost unaltered during evolution in different but related species, as is the case of Malus and Pyrus species. We have questioned whether the same expected features can be extended to whole S haplotypes, thus including not only the female S determinant (the S-RNase) but also its male counterpart, most likely provided by S-locus F-box Brothers (SFBB) genes - even though direct functional evidence is still needed. On the one side, coevolution between female and male S genes has been postulated several times, given that the generation of any new Sspecificity requires a coordinated change on both sides in order to maintain the full S-haplotypes functionality. But on the other side, recent models for S-RNase-based GSI suggest a key difference between the control of female and male S functions: while the first entirely depends on the single S-RNase gene, the second might likely be provided by multiple SFBB genes acting in a collaborative mode. Even though this hypothesis makes it necessary to re-discuss the mode of coevolution between the female and male S genes, it might provide a suitable explanation for the complex phylogenetic profiles of SFBB genes, which are only in part in agreement with that of the S-RNase. Phylogenetic and segregation analyses of S-locus genes, together with the increasing amount of genomic information available for apple and pear, provide a valuable tool for understanding both the molecular mechanism of SRNase-based GSI, and the complex evolutionary pattern of S-haplotypes

Structural and functional conservation of S-specificities among Pyrinae species.

Gametophytic self-incompatibility (GSI) is the main mechanism that controls fertilization in many rosaceous species, including those belonging to the subtribe Pyrinae (formerly the Maloideae). In natural conditions, S-specificities are subject to frequency-dependent balancing selection; the genetic imprint left by this kind of selection is visible on the S-RNase gene in terms of high sequence diversity, evidence of positive selection, and shared ancestral polymorphisms: thus, some alleles have been maintained almost unaltered during evolution in different but related species, as is the case of Malus and Pyrus species. We have questioned whether the same expected features can be extended to whole S haplotypes, thus including not only the female S determinant (the S-RNase) but also its male counterpart, most likely provided by S-locus F-box Brothers (SFBB) genes - even though direct functional evidence is still needed. On the one side, coevolution between female and male S genes has been postulated several times, given that the generation of any new Sspecificity requires a coordinated change on both sides in order to maintain the full S-haplotypes functionality. But on the other side, recent models for S-RNase-based GSI suggest a key difference between the control of female and male S functions: while the first entirely depends on the single S-RNase gene, the second might likely be provided by multiple SFBB genes acting in a collaborative mode. Even though this hypothesis makes it necessary to re-discuss the mode of coevolution between the female and male S genes, it might provide a suitable explanation for the complex phylogenetic profiles of SFBB genes, which are only in part in agreement with that of the S-RNase. Phylogenetic and segregation analyses of S-locus genes, together with the increasing amount of genomic information available for apple and pear, provide a valuable tool for understanding both the molecular mechanism of SRNase-based GSI, and the complex evolutionary pattern of S-haplotypes

Evaluation of candidate F-box genes for the pollen S of gametophytic self-incompatibility in the Pyrinae (Rosaceae) on the basis of their phylogenomic context

The recent analysis of the S-locus region of apple and Japanese pear, two species of Pyrinae (Rosaceae), suggested multiple and different F-box genes (called SFBBs) as candidates for the male determinant (pollen S) of RNase-based gametophytic self-incompatibility in these two species. Here, we followed a phylogenetic approach to take advantage of the pattern of molecular evolution of the S-locus of Pyrinae in characterizing SFBB homologs belonging to S-haplotypes of apple and three species of Pyrus (European, Japanese, and Chinese pears). Our results suggested that the S-locus region of Pyrinae contains no less than six SFBB members and that its structure seems to be rather conserved between apple and pear species. In accordance with the prevailing theory on S-haplotype evolution, the pollen S is expected to have coevolved with the S-RNase and to show some common features derived from the long-term evolution under frequency-dependent balancing selection, i.e., high sequence diversity, evidence of positive selection, and shared ancestral polymorphisms. Using this conceptual framework, we present evidence that some SFBB genes may be better candidates for pollen S in Pyrinae than others. Overall, the SFBB genes analyzed exhibited much lower sequence diversity than their associated S-RNases; likewise, they showed little or no evidence of positive selection. However, evidence of coevolution with the S-RNase clearly emerged for two of them. Altogether our results suggested different evolutionary histories for different SFBBs putatively derived from their distinct involvement in self-incompatibility

Postbiotic yeast fermentation product supplementation to lactating goats increases the efficiency of milk production by enhancing fiber digestibility and ruminal propionate, and reduces energy losses in methane

Although in vitro data with mixed ruminal fluid demonstrated positive effects of posbiotic diet (POS) from lactobacilli on measures of fermentation and microbial profiles, there is a paucity of in vivo data with lactating ruminants. The aim of the study was to evaluate the effects of incorporating POS into diets of lactating goats on energy (E) partitioning, carbon (C) and nitrogen (N) balance, and performance. Ten late-lactation Murciano-Granadina goats were used in a crossover design with 26-d periods. Goats in the control diet (CON) were fed daily at the rate of 1 kg alfalfa hay and 1.5 kg concentrate, and the treatment group (POS) was fed CON with the addition of 3.75 g/d of Probisan Ruminants (PENTABIOL S.L., Navarra, Spain). No differences in DMI were detected. However, ruminal fluid propionate and apparent total tract digestibilities of NDF and ADF were greater (18%, 4.7%, and 5.2%, respectively; P  0.05) in POS compared with the CON diet. Daily amount of CH4 emission did not differ P > 0.05 between diets. However, when expressed relative to unit of edible product, feeding POS reduced (P < 0.05) the amount of CH4 by 46 g/kg of milk fat, 97 g/kg of milk protein, and 3 g/kg of milk compared with CON. Overall, data indicated that feeding a postbiotic in late-lactation increased energy efficiency for milk production partly by reducing CH4 emission.info:eu-repo/semantics/acceptedVersio