Detection of runs of homozygosity in conserved and commercial pig breeds in Poland

Runs of homozygosity (ROH) are continuous segments of the genome that arose as a result of inbreeding, resulting in the inheritance of identical haplotypes from both parents who shared a common ancestor. In the present study, we performed a detailed characterization and comparison of ROH in four pig breeds, including intensively selected Polish Landrace as well as native unselected animals of Puławska and two Złotnicka breeds (White and Spotted). We used a medium-density PorcineSNP60 BeadChip assay (Illumina) and cgaTOH software to detect ROH covering a minimum of 30 adjacent SNPs and maintaining a size over 1 Mb. By analysing ROH distribution and frequency across the genome, we also identified genomic regions with high ROH frequency (so-called “ROH hotspots”). The obtained results showed that the analysed conserved breeds were characterized by a higher ROH span and higher ROH-based inbreeding coefficients (F_ROH), which likely result from past population bottlenecks, increasing the overall inbreeding level within these populations. The analysis of ROH distribution across the genomes revealed the presence of both shared and breed-specific ROH hotspots. These hotspots, presumably representing genome regions under selection, overlapped with a variety of genes associated with processes connected with immune system functioning, reproduction, glucose homeostasis and metabolism. The genome regions with ROH hotspots overlapping in all analysed populations, located on SSC4 (51.9–55.9 Mb) and 13 (92.6–97.8 Mb), covered thirty-one different genes, including MMP16, SLC7A13, ATP6V0D2, CNGB3, WWiP1, RiMDN1 and CPNE3. These genes are primarily associated with biological regulation and metabolism, processes that could be responsible for the variety of the selected production and functional features.     

Coat protein-mediated protection against plum pox virus in herbaceous model plants and transformation of apricot and plum

Summary The expression of the viral coat protein gene in transgenic plants has been shown to induce tolerance against virus infection (Beachy et al., 1990). Transgenic plants ofNicotiana clevelandii andNicotiana benthamiana- herbaceous host plants for PPV - transformed withAgrobacterium strain LBA 4404 containing the plasmid pBinPPVm, regenerated on selection media containing kanamycin were tested for the expression of the PPV coat protein gene by ELISA and immuno western blot. After rooting and acclimatisation plants were tested for the protection against PPV Following the inoculation plants were investigated for symptom development and virus accumulation. Different lines were identified, according to the different reaction to the mechanical inoculation, ranging from a complete absence to a strong reduction of symptoms. There have not been many reports on transformation of trees in general, and in fruit trees particularly. It is obvious that the major obstacle is the regeneration of transformed plantlets. Attempts to improve crop plants by genetic engineering techniques will always depend very strongly on the availability of reliable protocols for transformation, selection and regeneration (Laimer et al., 1989, 1990). Different systems involving juvenile and adult plant material have been developed allowing the transfer of foreign genes into apricot and plum cultivars. We report the transformation and regeneration ofPrunus armeniaca andPrunus domestica plants withAgrobacterium tumefaciens strain LBA 4404 containing various binary plasmids, pBinGUSint, carrying the marker geneβ-glucuronidase (GUS) and pBinPPVm, carrying the coat protein gene of Plum Pox Virus (PPV), the causal agent of Sharka disease. The marker geneGUS was used for the optical evaluation of the efficiency of different transformation systems involving cotyledons of immature embryos as well as somatic embryos and leaf discs. The coat protein gene of PPV was used to introduce the coat protein mediated resistance against one of the most important pathogens of stone fruit trees in Europe and the whole Mediterranean area.     

Npro of classical swine fever virus contributes to pathogenicity in pigs by preventing type I interferon induction at local replication sites

Classical swine fever (CSF) caused by CSF virus (CSFV) is a highly contagious disease of pigs. The viral protein N^pro of CSFV interferes with alpha- and beta-interferon (IFN-α/β) induction by promoting the degradation of interferon regulatory factor 3 (IRF3). During the establishment of the live attenuated CSF vaccine strain GPE^-, N^pro acquired a mutation that abolished its capacity to bind and degrade IRF3, rendering it unable to prevent IFN-α/β induction. In a previous study, we showed that the GPE^- vaccine virus became pathogenic after forced serial passages in pigs, which was attributed to the amino acid substitutions T830A in the viral proteins E2 and V2475A and A2563V in NS4B. Interestingly, during the re-adaptation of the GPE^- vaccine virus in pigs, the IRF3-degrading function of N^pro was not recovered. Therefore, we examined whether restoring the ability of N^pro to block IFN-α/β induction of both the avirulent and moderately virulent GPE^--derived virus would enhance pathogenicity in pigs. Viruses carrying the N136D substitution in N^pro regained the ability to degrade IRF3 and suppress IFN-α/β induction in vitro. In pigs, functional N^pro significantly reduced the local IFN-α mRNA expression in lymphoid organs while it increased quantities of IFN-α/β in the circulation, and enhanced pathogenicity of the moderately virulent virus. In conclusion, the present study demonstrates that functional N^pro influences the innate immune response at local sites of virus replication in pigs and contributes to pathogenicity of CSFV in synergy with viral replication.     

Diversity and pathogenicity of pestalotioid fungi associated with blueberry plants in Portugal,with description of three novel species of Neopestalotiopsis

European Journal of Plant Pathology - Blueberries (Vaccinium corymbosum) have been increasingly cultivated in Portugal. Although blueberry is known to be susceptible to twig blight and dieback...     

Antibacterial and brine shrimp lethality tests of biflavonoids and derivatives of Rheedia gardneriana

The hydroalcoholic extract of the leaves from Rheedia gardneriana yielded volkensiflavone (1), fukugetin (2), fukugiside (3), GB2a-I-7-O-glucoside (4) and epicatechin (5). Compounds 1-5, and some derivatives of 1 and 2 were evaluated for lethality to brine shrimp larvae and for antibacterial activity.     

Characterization of the proteins from Vigna unguiculata seeds

The proteins from Vigna unguiculata (L.) Walp. (cowpea) seeds were investigated. Globulins constitute over 51% of the total seed protein, with albumins composing approximately 45%. The globulins may be fractionated by native electrophoresis or anion exchange chromatography into three main components, which were termed (in decreasing order of anodic mobility) alpha-vignin, beta-vignin, and gamma-vignin. alpha-Vignin, with a sedimentation coefficient of 16.5S, is a major, nonglycosylated globulin, composed of a major 80 kDa subunit, which upon reduction, produces two polypeptides (20 and 60 kDa). beta-Vignin, with a sedimentation coefficient of 13S, is a major, glycosylated globulin, composed of two main polypeptides (55 and 60 kDa) with no disulfide bonds. Finally, gamma-vignin, a minor globulin, is composed by one main type of subunit (22 kDa), which upon reduction, is converted into a single, apparently heavier polypeptide chain (30 kDa) due to the presence of an internal disulfide bond. Immunological analyses revealed structural homology between beta-vignin and beta-conglutin (the vicilin from Lupinus seeds) but not between alpha- or gamma-vignins and their Lupinus counterparts. Haemagglutination activity toward trypsinized rabbit erythrocytes was found exclusively in the albumin fraction and was strongly inhibited by N-acetylglucosamine or chitin.     

Ustekinumab pharmacokinetics after subcutaneous administration in swine model

BackgroundDue to multiple similarities in the structure and physiology of human and pig skin, the pig model is extremely useful for biological drug testing after subcutaneous administration. Knowledge of the differences between subcutaneous injection sites could have a significant impact on the absorption phase and pharmacokinetic profiles of biological drugs.ObjectivesThis study aimed to analyze the impact of administration site on pharmacokinetics and selected biochemical and hematological parameters after a single subcutaneous administration of ustekinumab in pigs. Drug concentrations in blood plasma were analyzed by enzyme-linked immunosorbent assay. Pharmacokinetic analyses were performed based on raw data using Phoenix WinNonlin 8.1 software and ThothPro v 4.1.MethodsThe study included 12 healthy, female, large white piglets. Each group received a single dose of ustekinumab given as a 1 mg/kg subcutaneous injection into the internal part of the inguinal fold or the external part of the inguinal fold.ResultsThe differences in absorption rate between the internal and external parts of the inguinal fold were not significant. However, the time of maximal concentration, clearance, area under the curve calculated between zero and mean residence time and mean residence time between groups were substantially different (p > 0.05). The relative bioavailability after administration of ustekinumab into the external part of the inguinal fold was 40.36% lower than after administration of ustekinumab into the internal part of the inguinal fold.ConclusionsHealthy breeding pigs are a relevant model to study the pharmacokinetic profile of subcutaneously administered ustekinumab.     

Isolation and structural characterization of new anthocyanin-derived yellow pigments in aged red wines

Two newly formed yellow pigments that revealed unique spectral features were detected and isolated from an aged Port red wine by TSK Toyopearl HW-40(s) gel chromatography and characterized by UV-visible spectrophotometry, 1H NMR and 13C NMR, and mass spectrometry (LC-ESI/MS). The UV-vis spectra of these pigments showed maximum absorption at 478 nm that is significantly hypsochromically shifted from those of original grape anthocyanins and other pyranoanthocyanins, exhibiting a more yellow hue in acidic solution. The structures of these pigments correspond to methyl-linked pyranomalvidin 3-glucoside and its respective coumaroyl glucoside derivative. They were shown to arise from the reaction between acetoacetic acid and genuine grape anthocyanins. Isolation and NMR identification using 1D and 2D NMR techniques are reported for the first time for this new family of anthocyanin-derived yellow pigments occurring in red wines.     

A novel cytometric approach to study intestinal mucosa rebuilding in weaned pigs fed with dietary nucleotides

In the weaning period, transition from sow's milk to the post-weaning diet causes the withdrawal of important nutrients as milk nucleotides, which are known to be determinant for the development of the gastrointestinal tract and immune function. The role of dietary nucleotides in the rebuilding of gut epithelium remains unclear. The aim of the study was to evaluate using appropriate markers the rate of mitosis (Ki67) in the crypt epithelial cell, and the rate of apoptosis (active caspase 3), autophagy (MAP I LC3) and DNA damage (p53) in the crypt and villi epithelial cells of the small intestine of weaned pigs fed diets supplemented without/with nucleotides. An in-tissue cytometry method, based on confocal imaging and automated quantitative analysis was implemented. The studies allowed us to understand molecular bases of animal performance which could not be accessed by a routine histometric approach. Namely, the dietary nucleotides provided more uniform small intestine epithelium with considerably less animal-to-animal variation in respect to mitosis (SEM = 1.02 in control vs. 0.62 in supplemented group), autophagy (SEM = 1.22 vs. 0.99) and expression of p53 protein (SEM = 0.72 vs. 0.28). A significant increase in ratio of apoptosis and autophagy, and significantly lower p53 expression was found in the nucleotide-supplemented pigs as compared to control. The mitosis/apoptosis index was lower in the nucleotide-supplemented group suggesting slower rebuilding of gut epithelium in these pigs as compared to the control, respectively 0.454 to 0.191. Finally, in the supplemented group a drop in p53 expression was observed, however, it remains uncertain whether the reduction in DNA damage index is due to the supplementation with nucleotides utilized as a source for repair processes or due to the reduced DNA alterations.