A Tale of Two Families: Whole Genome and Segmental Duplications Underlie Glutamine Synthetase and Phosphoenolpyruvate Carboxylase Diversity in Narrow-Leafed Lupin (Lupinus angustifolius L.).

Narrow-leafed lupin (Lupinus angustifolius L.) has recently been supplied with advanced genomic resources and, as such, has become a well-known model for molecular evolutionary studies within the legume family-a group of plants able to fix nitrogen from the atmosphere. The phylogenetic position of lupins in Papilionoideae and their evolutionary distance to other higher plants facilitates the use of this model species to improve our knowledge on genes involved in nitrogen assimilation and primary metabolism, providing novel contributions to our understanding of the evolutionary history of legumes. In this study, we present a complex characterization of two narrow-leafed lupin gene families-glutamine synthetase (GS) and phosphoenolpyruvate carboxylase (PEPC). We combine a comparative analysis of gene structures and a synteny-based approach with phylogenetic reconstruction and reconciliation of the gene family and species history in order to examine events underlying the extant diversity of both families. Employing the available evidence, we show the impact of duplications on the initial complement of the analyzed gene families within the genistoid clade and posit that the function of duplicates has been largely retained. In terms of a broader perspective, our results concerning GS and PEPC gene families corroborate earlier findings pointing to key whole genome duplication/triplication event(s) affecting the genistoid lineage.

Integration of Lupinus angustifolius L. (narrow-leafed lupin) genome maps and comparative mapping within legumes.

Narrow-leafed lupin (Lupinus angustifolius L.) has recently been considered a reference genome for the Lupinus genus. In the present work, genetic and cytogenetic maps of L. angustifolius were supplemented with 30 new molecular markers representing lupin genome regions, harboring genes involved in nitrogen fixation during the symbiotic interaction of legumes and soil bacteria (Rhizobiaceae). Our studies resulted in the precise localization of bacterial artificial chromosomes (BACs) carrying sequence variants for early nodulin 40, nodulin 26, nodulin 45, aspartate aminotransferase P2, asparagine synthetase, cytosolic glutamine synthetase, and phosphoenolpyruvate carboxylase. Together with previously mapped chromosomes, the integrated L. angustifolius map encompasses 73 chromosome markers, including 5S ribosomal DNA (rDNA) and 45S rDNA, and anchors 20 L. angustifolius linkage groups to corresponding chromosomes. Chromosomal identification using BAC fluorescence in situ hybridization identified two BAC clones as narrow-leafed lupin centromere-specific markers, which served as templates for preliminary studies of centromere composition within the genus. Bioinformatic analysis of these two BACs revealed that centromeric/pericentromeric regions of narrow-leafed lupin chromosomes consisted of simple sequence repeats ordered into tandem repeats containing the trinucleotide and pentanucleotide simple sequence repeats AGG and GATAC, structured into long arrays. Moreover, cross-genus microsynteny analysis revealed syntenic patterns of 31 single-locus BAC clones among several legume species. The gene and chromosome level findings provide evidence of ancient duplication events that must have occurred very early in the divergence of papilionoid lineages. This work provides a strong foundation for future comparative mapping among legumes and may facilitate understanding of mechanisms involved in shaping legume chromosomes.

Reclassification of Eimeria pogonae Walden (2009) as Choleoeimeria pogonae comb. nov. (Apicomplexa: Eimeriidae).

The presented paper provides a reclassification of Eimeria pogonae from Pogona vitticeps into the correct genus Choleoeimeria. A description of exogenous and endogenous stages of biliary coccidium is given. Sporulation of the oocysts was endogenous. The mature oocysts contained four sporocysts each with two sporozoites. Oocysts were ellipsoidal in shape, with average length/width ratio 1.7 and measured 28.4 (SD1.5) × 16.8 (SD 1.5). The micropyle, residuum, and polar granules were absent from the sporulated oocysts. Ovoidal in shape, sporosysts without Steida bodies contained residuum and two elongated and boat-shaped sporozoites. The endogenous stages of the coccidia were located mainly in the epithelium of bile ducts; however, single-epithelium cells of the gallbladder were also infected.

Remnants of the Legume Ancestral Genome Preserved in Gene-Rich Regions: Insights from Lupinus angustifolius Physical, Genetic, and Comparative Mapping.

The narrow-leafed lupin (Lupinus angustifolius) was recently considered as a legume reference species. Genetic resources have been developed, including a draft genome sequence, linkage maps, nuclear DNA libraries, and cytogenetic chromosome-specific landmarks. Here, we used a complex approach, involving DNA fingerprinting, sequencing, genetic mapping, and molecular cytogenetics, to localize and analyze L. angustifolius gene-rich regions (GRRs). A L. angustifolius genomic bacterial artificial chromosome (BAC) library was screened with short sequence repeat (SSR)-based probes. Selected BACs were fingerprinted and assembled into contigs. BAC-end sequence (BES) annotation allowed us to choose clones for sequencing, targeting GRRs. Additionally, BESs were aligned to the scaffolds of the genome sequence. The genetic map was supplemented with 35 BES-derived markers, distributed in 14 linkage groups and tagging 37 scaffolds. The identified GRRs had an average gene density of 19.6 genes/100 kb and physical-to-genetic distance ratios of 11 to 109 kb/cM. Physical and genetic mapping was supported by multi-BAC-fluorescence in situ hybridization (FISH), and five new linkage groups were assigned to the chromosomes. Syntenic links to the genome sequences of five legume species (Medicago truncatula, Glycine max, Lotus japonicus, Phaseolus vulgaris, and Cajanus cajan) were identified. The comparative mapping of the two largest lupin GRRs provides novel evidence for ancient duplications in all of the studied species. These regions are conserved among representatives of the main clades of Papilionoideae. Furthermore, despite the complex evolution of legumes, some segments of the nuclear genome were not substantially modified and retained their quasi-ancestral structures. Cytogenetic markers anchored in these regions constitute a platform for heterologous mapping of legume genomes.

Comparative genomics of Lupinus angustifolius gene-rich regions: BAC library exploration, genetic mapping and cytogenetics.

BACKGROUND: The narrow-leafed lupin, Lupinus angustifolius L., is a grain legume species with a relatively compact genome. The species has 2n = 40 chromosomes and its genome size is 960 Mbp/1C. During the last decade, L. angustifolius genomic studies have achieved several milestones, such as molecular-marker development, linkage maps, and bacterial artificial chromosome (BAC) libraries. Here, these resources were integratively used to identify and sequence two gene-rich regions (GRRs) of the genome. RESULTS: The genome was screened with a probe representing the sequence of a microsatellite fragment length polymorphism (MFLP) marker linked to Phomopsis stem blight resistance. BAC clones selected by hybridization were subjected to restriction fingerprinting and contig assembly, and 232 BAC-ends were sequenced and annotated. BAC fluorescence in situ hybridization (BAC-FISH) identified eight single-locus clones. Based on physical mapping, cytogenetic localization, and BAC-end annotation, five clones were chosen for sequencing. Within the sequences of clones that hybridized in FISH to a single-locus, two large GRRs were identified. The GRRs showed strong and conserved synteny to Glycine max duplicated genome regions, illustrated by both identical gene order and parallel orientation. In contrast, in the clones with dispersed FISH signals, more than one-third of sequences were transposable elements. Sequenced, single-locus clones were used to develop 12 genetic markers, increasing the number of L. angustifolius chromosomes linked to appropriate linkage groups by five pairs. CONCLUSIONS: In general, probes originating from MFLP sequences can assist genome screening and gene discovery. However, such probes are not useful for positional cloning, because they tend to hybridize to numerous loci. GRRs identified in L. angustifolius contained a low number of interspersed repeats and had a high level of synteny to the genome of the model legume G. max. Our results showed that not only was the gene nucleotide sequence conserved between soybean and lupin GRRs, but the order and orientation of particular genes in syntenic blocks was homologous, as well. These findings will be valuable to the forthcoming sequencing of the lupin genome.

[The evaluation of the stress coping styles and emotional intelligence in psychiatrically treated adolescent patients with deliberate self-harm in relation to chosen clinical features]. / Ocena stylów radzenia sobie ze stresem i inteligencji emocjonalnej u samouszkadzajacej sie mlodziezy leczonej psychiatrycznie w zaleznosci od wybranych czynników klinicznych.

AIM: The primary goal of the study was an evaluation of the dominating stress coping styles in adolescent patients with self-harm records, who were psychiatrically treated, taking into account the level of their emotional intelligence vs. the psychiatric diagnosis, the type of motives and decision involved in self-harming and the presence of suicidal attempts (SA) in the past. The secondary goal included an analysis of the correlations between particular stress coping skills and the level of emotional intelligence. METHOD: The reported studies involved self-harming patients aged of 13-18 years during their psychiatric hospitalisation (n=31). The applied tools included the Coping Inventory for Stressful Situations (CISS) and the Two-Dimensional Inventory of Emotional Intelligence (DINEMO). RESULTS: An evaluation of the correlation between stress coping styles and the levels of emotional intelligence in the studied group and the types of mental disorders did not reveal any significant differences between the evaluated subgroups. Patients, who confirmed an instrumental motive, obtained statistically significantly higher scores on the task-oriented scale vs. those who performed the acts of DSH for reactive or pathological reasons. Taking into consideration the type of decision, involved in self-harming acts, did not show any differences in the stress coping styles of the patients, however, those patients, who had planned an act of DSH, achieved statistically significantly higher scores in the OTHERS scale of the DINEMO. Patients with DSH and with SA in the past (77% studied group), achieved similar results in CISS and DINEMO vs. the self-harming patients without SA in the past. In the study group, one statistically significant correlation was demonstrated between CISS--the avoidance-oriented style--and the I in DINEMO. CONCLUSIONS: 1. Patients with DSH records and without SA constitute a fairly uniform group with regards to stress coping styles, taking into account the type of psychic disorders and the urgency of self-harm decision. 2. Their use of the task-oriented style may be associated with instrumental auto-aggression. 3. DSH may be a non-verbal form of communicating other problems to others. 4. Adolescents, who perform the acts of self-harm, are endangered by suicidal attempts, regardless of their stress coping styles or the level of their emotional intelligence. 5. A development of constructive stress coping skills against strong emotional reactions is a recommended method in the therapy of patients with DSH.

Influence of the Tool Cutting Edge Helix Angle on the Surface Roughness after Finish Milling of Magnesium Alloys.

This paper shows the surface quality results after finishing milling of AZ91D and AZ31 magnesium alloys. The study was performed for variable technological parameters: cutting speed, feed per tooth, axial depth of cut and radial depth of cut. The tools used in the study were two carbide cutters with a different tool cutting edge helix angle. The measurement of the research results presented the surface roughness parameters was made on the lateral faces and the end faces of the specimens. Statistical analysis and simulations using artificial neural networks were carried out with the Statistica software. The normality of the distribution was examined, and the hypotheses of the equality of mean values and variance were verified. For the AZ91D magnesium alloy on the lateral and the end faces (Ra, Rz parameters), simulations were carried out. Two types of ANN were used: MLP (Multi-layered perceptron) and RBF (Radial Basis Function). To increase the machining stability and to obtain a high surface finish, the more suitable tool for finishing milling is the tool with a helix angle of λs = 20°. Artificial neural networks have been shown to be a good tool for predicting surface roughness parameters of magnesium alloys after finishing milling.

Retrosternal (Morgagni) diaphragmatic hernia.

This study presents the case of a shih tzu puppy, in which a rare congenital Morgagni diaphragmatic hernia was diagnosed. The diagnosis was based on abdominal and thoracic radiographs, including a contrast study of the gastrointestinal tract, which revealed a co-existing umbilical hernia. Both hernias were repaired by surgery.

Legume Cytosolic and Plastid Acetyl-Coenzyme-A Carboxylase Genes Differ by Evolutionary Patterns and Selection Pressure Schemes Acting before and after Whole-Genome Duplications.

Acetyl-coenzyme A carboxylase (ACCase, E.C.6.4.1.2) catalyzes acetyl-coenzyme A carboxylation to malonyl coenzyme A. Plants possess two distinct ACCases differing by cellular compartment and function. Plastid ACCase contributes to de novo fatty acid synthesis, whereas cytosolic enzyme to the synthesis of very long chain fatty acids, phytoalexins, flavonoids, and anthocyanins. The narrow leafed lupin (Lupinus angustifolius L.) represents legumes, a plant family which evolved by whole-genome duplications (WGDs). The study aimed on the contribution of these WGDs to the multiplication of ACCase genes and their further evolutionary patterns. The molecular approach involved bacterial artificial chromosome (BAC) library screening, fluorescent in situ hybridization, linkage mapping, and BAC sequencing. In silico analysis encompassed sequence annotation, comparative mapping, selection pressure calculation, phylogenetic inference, and gene expression profiling. Among sequenced legumes, the highest number of ACCase genes was identified in lupin and soybean. The most abundant plastid ACCase subunit genes were accB. ACCase genes in legumes evolved by WGDs, evidenced by shared synteny and Bayesian phylogenetic inference. Transcriptional activity of almost all copies was confirmed. Gene duplicates were conserved by strong purifying selection, however, positive selection occurred in Arachis (accB2) and Lupinus (accC) lineages, putatively predating the WGD event(s). Early duplicated accA and accB genes underwent transcriptional sub-functionalization.

Determination of reference values and frequency of occurrence of patella alta in German shepherd dogs: a retrospective study.

BACKGROUND: Patella alta and patella baja are important conditions underlying a predisposition to many joint diseases, including patellar luxation and patellar chondromalacia of the articular cartilage. The frequencies of patella alta and patella baja have not yet been determined. The objectives of this study were to determine the frequency of patella alta and to determine reference values to the position of the vertical patella according to two modified techniques of the Insall-Salvati method in a group of 65 German shepherd dogs (115 stifle joints). RESULTS: The upper limits of reference values for the normal vertical position of the patella were 1.79 and 2.13, depending on the method of measurement. A high prevalence of patella alta was observed in the group of German shepherd dogs. A correlation was demonstrated between the classification of dogs' joints in the patella alta group and the multiplied risk of canine hip dysplasia (CHD) through the estimation of odds ratios. CONCLUSIONS: Dogs with patella alta were healthy dogs that did not exhibit orthopaedic problems in the stifle joints. The results revealed that the risk of CHD is twice as high in dogs with higher patellar ligament length to patella length ratio.

Canine Trichomonas tenax mandibular gland infestation.

BACKGROUND: Several species of trichomonads are intestinal or urogenital parasites of humans and animals, with only a few species typically being located in the oral cavity. The prevalence of oral trichomoniasis in dogs is approximately 15-25 %, although the prevalence varies among different populations and depends on age, sex and the health of the oral cavity. CASE PRESENTATION: A case of mandibular gland infestation by trichomonads in a 13-year-old female Dachshund with advanced periodontal disease and oral trichomoniasis is reported. The dog was referred due to a history of a painless swelling over the left submandibular region that increased in size over time. Based on physical and ultrasound examinations, a final diagnosis of mandibular gland cyst was established and transcutaneous needle aspiration was carried out. Numerous mobile trophozoites of trichomonads were found by microscopy and culturing for trichomonas was performed. The species was finally characterized as Trichomonas tenax by polymerase chain reaction and sequencing. CONCLUSIONS: Studies have shown that T. tenax can be found in humans in atypical locations such as the salivary glands and upper and lower respiratory tracts. According to our knowledge this is the first report of T. tenax being present in the salivary glands of a dog. Because of the relatively high prevalence of trichomoniasis in dogs with periodontal diseases, these parasites should be considered together with bacterial and viral agents in salivary gland infections, especially in individuals with compromised oral health.

Lumbosacral transitional vertebrae, canine hip dysplasia, and sacroiliac joint degenerative changes on ventrodorsal radiographs of the pelvis in police working German shepherd dogs.

Lumbosacral transitional vertebrae (LTV) frequently occur in German shepherd dogs. The aim of the study was to evaluate the prevalence and interdependence between LTV and canine hip dysplasia (CHD) as well as sacroiliac joint degenerative changes visualized on ventrodorsal radiographs of the pelvis in both working and companion German shepherd dogs. The presence of LTV was found in 12% of working dogs and in 33% of companion dogs. Similar incidence of hip dysplasia in both the groups was found. It has been shown that dogs with LTV have a higher frequency of severe CHD. A higher percentage of sacroiliac joint degenerative changes was observed in dogs with no signs of LTV and in working dogs.