[Risk prediction and function evaluation by T-cell epitope model and expression model of HLA-DPB1 mismatching in unrelated-donor hematopoietic stem cell transplantations].

Objective: To evaluate the risk prediction and assessment function of HLA-DPB1 T-cell epitope (TCE) model and expression model in human leukocyte antigen (HLA)-matched unrelated hematopoietic stem cell transplantation (MUD-HSCT) with HLA-DPB1 mismatching. Methods: A total of 364 (182 pairs) potential MUD-HSCT donors and recipients confirmed by HLA high-resolution typing in Shaanxi Blood Center from 2016 to 2019 were analyzed retrospectively. Of the 182 recipients, there were 121 males and 61 females with an average age of (26.3±14.2) years. Of the 182 donors, there were 148 males and 34 females with an average age of (33.7±7.5) years. Polymerase chain reaction-sequence-based typing (PCR-SBT), next-generation sequencing (NGS) and polymerase chain reaction-sequence specific oligonucleotide probe (PCR-SSO) based on LABScan®3D platform were used for high-resolution typing of HLA-A, B, C, DRB1, DQB1, DPB1 gene, and PCR-SBT was used for single nucleotide polymorphism (SNP) typing. TCE model and expression model were used to predict and evaluate the HLA-DPB1 mismatch pattern and acute graft-versus-host-disease (aGVHD) risk. Results: A total of 26 HLA-DPB1 alleles and their 3'-UTR rs9277534 SNP genotypes were detected in this study population, and two new alleles HLA-DPB1*1052∶01 and HLA-DPB1*1119∶01 were found and officially named. The overall mismatch rate of HLA-DPB1 in MUD-HSCT donors and recipients was 90.66% (165/182). In TCE model, the HLA-DPB1 mismatch rates of permissible mismatch (PM) and non-permissible mismatch (non-PM) were 47.80% (87/182) and 42.86% (78/182), respectively. The non-PM in GvH direction was 13.73% (25/182), and which in HvG direction was 29.12% (53/182). A total of 73 pairs of donors and recipients in TCE model met the evaluation criteria of expression model. Among of TCE PM group, recipient DP5 mismatches accounted for 34.25% (25/73) were predicted as aGVHD high risk according to expression model. For the TCE non-PM group, both the recipient DP2 mismatches of 6.85% (5/73) and recipient DP5 mismatches of 10.86% (8/73) were predicted to be at high risk for aGVHD. Risk prediction by TCE model and expression model was 27.27% concordant and 16.97% unconcordant. Conclusions: TCE model and expression model are effective tools to predict aGVHD risk of MUD-HSCT. Comprehensive application of the two models is helpful to the hierarchical assessment of HSCT risk.

[Risk factors analysis of acute kidney injury in patients with exertional heatstroke].

Objective: To explore the risk factors of acute renal injury (AKI) in exertional heat radiation disease (EHS) . Methods: In november 2019, the clinical data of 69 EHS patients admitted from July 2015 to September 2019 were reviewed. The general data, laboratory indexes, Glasgow score (GCS) at admission, 24-hour acute physiology and chronic health score Ⅱ (APACHE Ⅱ) , exposure time rate and physical labor intensity were collected. According to the occurrence of AKI, the patients were divided into AKI group and non-AKI group, 31 and 38 in each group. The differences of general data and laboratory indexes between the two groups were compared, and the t and Mann-Whitney U test were used to compare the two groups. The enumeration data are expressed by examples and constituent ratio (%) . Independent sample χ(2) test is used for inter-group comparison, and multiple test is used for multi-sample comparison. The correlation was analyzed by linear regression. Risk factors were analyzed by Logistic regression analysis. Results: At discharge, 31 of 69 EHS patients developed AKI. Compared with the non-AKI group, the heart rate, white blood cell count, lactic acid, D-dimer and myoglobin were higher; MAP, platelet count and PH were lower in the AKI group. The difference was statistically significant (P<0.05) . APACHE Ⅱ score, core temperature, time to drop to 38.5 ℃, contact time rate, platelet count, pH, lactic acid, D-dimer and myoglobin were all correlated with creatinine (r=0.57, 0.42, 0.80, 0.78, 0.57, 0.43, 0.51, 0.55, 0.79) . APACHE Ⅱ score, time to drop to 38.5C, Lac and MYO are the risk factors of AKI in EHS patients. Multivariate Logistic regression analysis showed that the time required to drop to 38.5C was an independent risk factor for the occurrence of AKI. Conclusion: AKI is a serious complication of EHS. EHS complicated with AKI, should be identified early and effective intervention measures should be taken.

[Hereditary protein S deficiency: survey results from a Chinese pedigree].

Objective: To investigate the clinical characteristics and gene mutation, and analyze the association between genotype and phenotype of hereditary protein S deficiency in a Chinese pedigree. Methods: Hereditary protein S deficiency was diagnosed in January 2016 in our hospital. A total of 26 family members were surveyed in this study. Blood samples and clinical data were collected from them, and mutations were identified by Sanger sequencing. Pathogenicity of gene mutations was predicted by protein function prediction software including SIFT, PolyPhen_2, nsSNPAnalyzer and MutPred2. Swiss Model (https://swissmodel.expasy.org/) was used to perform homology modeling of the tertiary structure of the protein S wild-type and mutant-type, and observe the impact of gene mutation on the tertiary structure of the protein. Results: Four out of 26 family members of 4 generations were clinically diagnosed with hereditary protein S deficiency. The proband presented with recurrent pulmonary embolism and venous thromboembolism of the lower extremities, and her uncle and mother had a history of venous thromboembolism. Sequencing revealed a mutation in the c.200A>C gene in the second exon of the PROS1 gene of proband and part of her families (Ⅱ2, Ⅱ6, Ⅲ4, Ⅳ2). The prediction results of this gene mutation performed by SIFT, PolyPhen_2, nsSNPAnalyzer, MutPred2 were all harmful. The results of Swiss-Model homology modeling showed that the 67th amino acid was mutated from glutamic acid to alanine because of this gene mutation. Conclusion: A gene mutation cDNA (c. 200A>T) is identified in a Chinese pedigree with hereditary protein S deficiency. This gene mutation may reduce protein S activity, which may cause recurrent pulmonary embolism and venous thromboembolism of the patients.

[Comparison of different critical care scoring systems in prognosis evaluation of heat stroke].

Objective: To evaluate the prognostic value of different critical care scoring systems in 28-day survival rate of patients with heat stroke. Methods: A retrospective analysis was conducted on the clinical data of 71 patients with heat stroke admitted to the department of emergency medicine of Beijing Luhe Hospital. Capital Medical University from July 2015 to September 2018. The general information and the worst values of vital signs and related pathophysiological indicators within 24 hours were collected and the sequential organ failure assessment (SOFA) , multiple organ dysfunction (MODS) , simplified acute physiological scoreⅡ (SAPS Ⅱ) and acute physiology and chronic health evaluationⅡ (APACHE Ⅱ) were calculated. The patients were divided into the survival group (n=45) and the non-survival group (n=26) according to 28-day prognosis, and the clinical data and scores of the two groups were compared.The ROC curve was drawn to analyze the evaluation value of each scoring system on the survival rate of patients at 28-day. Kaplan-Meier method was used to plot the survival curve of patients. Results: There were no significant differences in age, sex, vital signs and laboratory parameters between two groups (P>0.05) . In non-survival patients, SOFA, SAPS Ⅱ, APACHE Ⅱ scores were significantly elevated in the survival group (P<0.05) . ROC curve analysis showed that the area under ROC curve (AUC) of SOFA score for predicting 28-day survival rate was the highest, which was significantly higher than the APACHE Ⅱ, SAPS Ⅱ, MODS score. When the best cut-off value of SOFA score was 9.0, the sensitivity was 84.6%, and the specificity was 71.1%. Kaplan-Meier survival analysis showed that 28-day survival rate after hospital discharge in patients with SOFA score<9 (n=27) was significantly higher than that in patients with SOFA score ≥9.0 (χ(2)=1.0, P<0.01) . Conclusion: SOFA, APACHE Ⅱ, SAPS Ⅱ on admission have been proved to have good prognostic ability to predict 28-day prognosis in heat stroke patients. Among them, SOFA score system has more accurate prediction value.

A compressible anisotropic hyperelastic model with I5 and I7 strain invariants.

It is obvious that the mechanical properties of arterial tissue include compressibility, anisotropy, and the fact that the out-of-plane shear modulus is smaller than the shear modulus in the plane of the fibers. However, the last point is rarely considered when it comes to compressible anisotropic hyperelastic models. In order to acquire different shear moduli, we propose a modified hyperelastic model including the influence of strain invariants I5 and I7. The convergence and correctness of this model are verified through the hydrostatic tension test, uniaxial tension test, and shear deformation test. It turns out that our model correctly predicts an anisotropic response and volume change to hydrostatic tensile test and the fact that the out-of-plane shear modulus is always smaller than the shear modulus in the plane of the fibers in shear deformation test. We conclude that the influence of strain invariants I5 and I7 is great, especially in the shear deformation, so that it is necessary to include I5 and I7 in the compressible anisotropic hyperelastic model.

[Subcellular localization of GTPase of immunity-associated protein 2].

OBJECTIVE: To analyze the subcellular localization of GTPase of immunity-associated protein 2 (GIMAP2) for the further functional study. METHODS: In the study, we first obtained the protein sequences of GTPase of immunity-associated protein 2 (GIMAP2) from National Center for Biotechnology Information (NCBI) database, and then performed a prediction analysis of its transmembrane structure, nuclear localization signal (NLS), nuclear export signal (NES) and subcellular localization through bioinformatics online tools. GIMAP2 gene amplified by PCR was inserted into the expression vector pQCXIP-mCherry-N1 and positive clones were selected by ampicillin resistance. After using methods to extract and purify, the sequenced recombinant plasmid pQCXIP-GIMAP2-mCherry, together with the retroviral packaging plasmids VSVG and Gag/pol, was transferred into HEK293FT cells by liposomes for virus packaging. The virus supernatant was collected 48 h after transfection and directly infected the human breast cancer cell line MDA-MB-436. Immunofluorescence staining was constructed to detect the localization of endogenous and exogenous GIMAP2 in MDA-MB-436 cells. Meanwhile, green fluorescent chemical dyes were used to label mitochondria, endoplasmic reticulum, and lipid droplets in living MDA-MB-436 cells stably expressing the GIMAP2-mCherry fusion protein. Images for the three dye-labeled organelles and GIMAP2-mCherry fusion protein were captured by super-resolution microscope N-SIM. RESULTS: Bioinformatics analysis data showed that GIMAP2 protein composed of 337 amino acids might contain two transmembrane helix (TM) structures at the carboxyl terminus, of which TMs were estimated to contain 40-41 expected amino acids, followed by the residual protein structures toward the cytoplasmic side. NES was located at the 279-281 amino acids of the carboxyl terminus whereas NLS was not found. GIMAP2 might locate in the lumen of the endoplasmic reticulum. Sequencing results indicated that the expression vector pQCXIP-GIMAP2-mCherry was successfully constructed. Fluorescent staining confirmed that GIMAP2-mCherry fusion protein, co-localized well with endogenous GIMAP2, expressed successfully in the endoplasmic reticulum and on the surface of lipid droplets in MDA-MB-436 cells. CONCLUSION: GIMAP2 localizes in the endoplasmic reticulum and on the surface of LDs, suggesting potential involvement of GIMAP2 in lipid metabolism.

QTL analysis of durable stripe rust resistance in the North American winter wheat cultivar Skiles.

KEY MESSAGE: This study determined the effects of growth stage and temperature on expression of high-temperature adult-plant resistance to stripe rust, mapped six QTL for durable resistance in winter wheat Skiles using a doubled haploid population, and selected breeding lines with different combinations of the QTL using marker-assisted selection. The winter wheat cultivar Skiles has a high level of high-temperature adult-plant (HTAP) resistance to stripe rust caused by Puccinia striiformis f. sp. tritici (Pst). The Skiles HTAP resistance was highly effective at the adult-plant stage even under low temperatures, but high temperatures induced earlier expression and increased levels of resistance. To map resistance genes, Skiles was crossed with the susceptible cultivar Avocet S and a doubled haploid (DH) population was developed. The DH population was tested in fields at Pullman, WA, in 2016, 2017 and 2018, Mount Vernon, WA, in 2017 and 2018 under natural infection, and an environmentally controlled greenhouse at the adult-plant stage with the currently predominant race PSTv-37. The population was genotyped using the 90 K Illumina iSelect wheat SNP chip and selected SSR markers on specific chromosomes. In total, 2526 polymorphic markers were used for QTL mapping and six QTL were detected. Two of the six QTL had major effects across all environments, with one mapped on chromosome 3BS, explaining up to 28.2% of the phenotypic variation and the other on chromosome 4BL, explaining up to 41.8%. Minor QTL were mapped on chromosomes 1BL, 5AL, 6B and 7DL. Genotyping 140 wheat cultivars from the US Pacific Northwest revealed high polymorphism of markers for five of the QTL, and five highly resistant lines with the five QTL were selected from Skiles-derived breeding lines using the markers. This study demonstrated that multiple QTL with mostly additive effects contributed to the high-level HTAP resistance in Skiles.

Combination of all-stage and high-temperature adult-plant resistance QTL confers high-level, durable resistance to stripe rust in winter wheat cultivar Madsen.

KEY MESSAGE: Wheat cultivar Madsen has a new gene on the short arm of chromosome 1A and two QTL for all-stage resistance and three QTL for high-temperature adult-plant resistance that in combination confer high-level, durable resistance to stripe rust. Wheat cultivar Madsen has maintained a high-level resistance to stripe rust over 30 years. To map quantitative trait loci (QTL) underlying the high-level, durable resistance, 156 recombinant inbred lines (RILs) developed from cross Avocet S × Madsen were phenotyped with selected races of Puccinia striiformis f. sp. tritici in the greenhouse seedling tests, and in naturally infected fields during 2015-2017. The RILs were genotyped by SSR and SNP markers from genotyping by sequencing and the 90 K wheat SNP chip. Three QTL for all-stage resistance were mapped on chromosomes 1AS, 1BS and 2AS, and two QTL for high-temperature adult-plant (HTAP) resistance were mapped on 3BS and 6BS. The most effective QTL on 2AS, explaining 8.97-23.10% of the phenotypic variation in seedling tests and 8.60-71.23% in field tests, contained Yr17 for all-stage resistance and an additional gene for HTAP resistance. The 6BS QTL, detected in all field tests, was identified as Yr78. The 1AS QTL, conferring all-stage resistance, was identified as a new gene, which explained 20.45 and 30.23% of variation in resistance to races PSTv-37 and PSTv-40, respectively, and contributed significantly to field resistance at Pullman in 2015-2017, but was not detected at Mount Vernon. The interactions among QTL were mostly additive, and RILs with all five QTL had the highest level of resistance in the field, similar to Madsen. Genotyping 148 US Pacific Northwest wheat cultivars with markers for the 1AS, 2AS and 6BS QTL validated the genes and markers, and indicated their usefulness for marker-assisted selection.

Gout as a risk factor for osteoporosis: epidemiologic evidence from a population-based longitudinal study involving 108,060 individuals.

Is gout a risk factor for future osteoporosis? This large population-based study comprising two matched groups of individuals with and without gout demonstrates that patients with gout have a 20% increase in the risk of developing osteoporosis in future through an 8-year follow-up. INTRODUCTION: To examine if gout is associated with an increased risk of osteoporosis. METHODS: We conducted a nationwide population-based retrospective matched-cohort study. Two matched cohorts (n = 36,458 with gout and 71,602 without gout) assembled and recruited from the Longitudinal Health Insurance Dataset containing 1 million subjects. Exclusion criteria were missing data, age < 20 years, short follow-up period, and pre-existing osteoporosis. Both cohorts were followed up until incident osteoporosis, death, or the end of the study. Person-year data and incidence rates were evaluated. A multivariable Cox model was used to derive an adjusted hazard ratio (aHR) after controlling for socioeconomic proxy, geographical difference, glucocorticoid and allopurinol exposure, various prespecified medical conditions, and comorbidities. RESULTS: Men comprised 72.8% of the cohorts. With a follow-up of 183,729 and 359,900 person-years for the gout and non-gout cohorts, 517 and 811 incidents of osteoporosis occurred, respectively, after excluding osteoporosis incidents in the first 3 years of follow-up. The cumulative incidence of osteoporosis was statistically higher in the gout cohort than in the non-gout cohort, at 3.3 versus 2.1% (P = 0.0036, log-rank). Our Cox model showed a 1.2-fold increase in the incidence of osteoporosis in the gout cohort, with an aHR of 1.2 (95% confidence interval, 1.06-1.35). CONCLUSIONS: This first population-based epidemiologic study supports the hypothesis that compared with individuals without gout; those with gout have a modest increase in the risk of developing osteoporosis in future.

Synthesis and Characterization of SiO2@Y2MoO6:Eu3+ Core-Shell Structured Spherical Phosphors by Sol-Gel Process.

SiO2@Y2MoO6:Eu3+ core-shell phosphors were prepared by the sol-gel process. X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FT-IR), field emission scanning electron microscopy (FESEM), energy-dispersive X-ray spectra (EDS), transmission electron microscopy (TEM), photoluminescence (PL) spectra as well as kinetic decays were used to characterize the resulting SiO2@Y2MoO6:Eu3+ core-shell phosphors. The XRD results demonstrated that the Y2MoO6:Eu3+ layers on the SiO2 spheres crystallized after being annealed at 700 °C and the crystallinity increased with raising the annealing temperature. The obtained core-shell phosphors have spherical shape with narrow size distribution (average size ca. 640 nm), non-agglomeration, and smooth surface. The thickness of the Y2MoO6:Eu3+ shells on the SiO2 cores could be easily tailored by varying the number of deposition cycles (70 nm for four deposition cycles). The Eul+ shows a strong PL emission (dominated by 5D0-7F2 red emission at 614 nm) under the excitation of 347 nm UV light. The PL intensity of Eu3+ increases with increasing the annealing temperature and the number of coating cycles.

Molecular Mapping of Stripe Rust Resistance Gene Yr76 in Winter Club Wheat Cultivar Tyee.

Tyee, one of the wheat cultivars used to differentiate races of Puccinia striiformis f. sp. tritici in the United States, was identified to have a single gene for all-stage resistance, tentatively named YrTye. To map the gene, Tyee was crossed with 'Avocet Susceptible' (AvS). Genetic analysis of the F1, F2, F2:3, and BC1 progenies confirmed a single dominant gene for resistance to race PSTv-37 that is avirulent to YrTye. A mapping population of 135 F2 plants was phenotyped with PSTv-37 and the derived F2:3 lines were tested with races PSTv-37, PSTv-40, and PSTv-79. The F2 mapping population was genotyped with simple sequence repeat (SSR) markers. A genetic map comprising 13 SSR markers located YrTye in chromosome 3AS flanked distally by SSR marker wmc11 and proximally by wmc532 at 2.6 and 3.4 cM, respectively. Amplification of Chinese Spring 3A deletion lines placed the gene in the distal bin 3AS4-0.45 to 1.00. Because YrTye is different from all formally named Yr genes in chromosomal location, we permanently name the gene Yr76. A near-isogenic line of spring common wheat was developed and selected by testing F3 lines derived from a AvS*4/Tyee cross with Tyee-avirulent and virulent races and the flanking markers. The specific SSR alleles flanking Yr76 were validated using cultivars and breeding lines with and without the gene, and showed high polymorphisms. The specificity of Yr76 is useful in differentiating P. striiformis f. sp. tritici races, and its tightly linked markers will be useful in developing resistant cultivars when combining the gene with other genes for resistance to stripe rust.

Molecular Mapping of YrSP and Its Relationship with Other Genes for Stripe Rust Resistance in Wheat Chromosome 2BL.

Stripe rust, caused by Puccinia striiformis f. sp. tritici, is an important disease of wheat worldwide. Resistance is the best way to control the disease. YrSP, a gene originally from 'Spaldings Prolific' wheat and providing resistance to a broad spectrum of races, is used for differentiating P. striiformis f. sp. tritici races but its chromosomal location is not clear. To map YrSP, a near-isogenic line (AvSYrSPNIL) was backcrossed to the recurrent parent, Avocet S. Genetic analysis of the BC7F1, BC8, BC7F2, and BC7F3 progenies confirmed a single dominant gene for resistance. In total, 182 BC7F2 plants and their derived BC7F3 lines were phenotyped with an avirulent P. striiformis f. sp. tritici race and genotyped with simple-sequence repeat (SSR), single-nucleotide polymorphism (SNP), and sequence-tagged site (STS) markers. A linkage map was constructed with 3 SSR, 17 SNP, and 3 STS markers covering 23.3 centimorgans (cM). Markers IWA638 and dp269 were 0.6 cM proximal and 1.5 cM distal, respectively, to YrSP. The gene was mapped in chromosome bin 2BL-C-0.5, physically within the proximal 50% of the chromosome 2BL arm. Allelism tests based on F2 phenotypes indicated that YrSP is closely linked to but not allelic with genes Yr5, Yr7, Yr43, Yr44, and Yr53. Infection type data from tests with 10 historical and currently predominant P. striiformis f. sp. tritici races in the United States also demonstrated differences in specificity between YrSP and the other genes. The specificity of YrSP is useful in differentiating P. striiformis f. sp. tritici races and studying the plant-pathogen interactions, and the information of chromosomal location of the gene and its tightly linked markers should be useful in developing resistant cultivars when combined with other genes for resistance to stripe rust.

Barberry Does Not Function as an Alternate Host for Puccinia striiformis f. sp. tritici in the U. S. Pacific Northwest Due to Teliospore Degradation and Barberry Phenology.

Sexual reproduction of the stem rust pathogen, Puccinia graminis f. sp. tritici (Pgt), on barberry (Berberis vulgaris) has been shown to provide initial inoculum for the development of the disease on wheat and barley and also generate diverse races of the pathogen. However, in our previous study, the stripe rust pathogen, P. striiformis f. sp. tritici (Pst), was not found on barberry in the U. S. Pacific Northwest. To determine why Pgt is able to infect the alternate host, while Pst cannot under the natural conditions, the viabilities of teliospores of both Pgt and Pst were investigated from 2011 to 2014 by determining the germination rates using telial samples collected periodically from wheat fields. Teliospores of Pst usually produced in July were physically degraded during winter, and their germination rate decreased from 50 to 90% in August to less than 1% in the following March and no germination after May. In contrast, Pgt teliospores usually produced in July and August remained physically intact and physiologically dormant, and could not germinate until February. Germination of Pgt teliospores gradually increased to 90% in May, at which time young leaves of barberry were susceptible to infection. In addition, a time-series experiment was conducted for inoculation of barberry plants with Pst teliospores. The results showed that Pst teliospores need a minimum of 32 h continual dew-forming conditions to infect barberry, and infection reaches a peak after incubation of inoculated plants for 88 h. The lack of a prolonged period of leaf wetness conditions during the season of telial maturity effectively negates Pst infection of barberry plants in the Pacific Northwest.

Barberry as Alternate Host Is Important for Puccinia graminis f. sp. tritici But Not for Puccinia striiformis f. sp. tritici in the U.S. Pacific Northwest.

Common barberry (Berberis vulgaris) is the alternate host of the wheat stem rust pathogen, Puccinia graminis f. sp. tritici, under natural conditions in the U.S. Pacific Northwest. Barberry was recently shown to be infected by basidiospores of the wheat stripe rust pathogen, Puccinia striiformis f. sp. tritici, under controlled conditions, but it is unclear if barberry plays any role in stripe rust epidemics under natural conditions. Aecial samples of Puccinia spp. collected from barberry plants in the Pacific Northwest from 2010 to 2013 were characterized to species by inoculation on wheat plants under controlled conditions and by molecular markers and sequences of the internal transcribed spacer (ITS) region of nuclear ribosomal DNA. Inoculation of wheat plants with bulked aecia-bearing barberry samples resulted in most P. graminis f. sp. tritici uredia and some P. striiformis f. sp. tritici uredinia. Virulence tests demonstrated that the P. graminis f. sp. tritici isolates were sexually produced, whereas the P. striiformis f. sp. tritici isolates were clonal based on both virulence and simple sequence repeat marker tests, indicating urediniospores from wheat fields landing on barberry leaves as the possible source of P. striiformis f. sp. tritici inoculum. A method for simultaneously testing individual aecia for identifying of P. graminis f. sp. tritici and P. striiformis f. sp. tritici by pathogenicity and ITS markers. Using the method together with ITS sequencing, tested individual aecia were mostly P. graminis f. sp. tritici and occasionally some other formae speciales of P. graminis, but not P. striiformis. The results imply that barberry is essential for stem rust epidemics, but not for stripe rust under the natural conditions in the U.S. Pacific Northwest.

Molecular mapping of genes Yr64 and Yr65 for stripe rust resistance in hexaploid derivatives of durum wheat accessions PI 331260 and PI 480016.

KEY MESSAGE: This manuscript reports two new genes ( Yr64 and Yr65 ) for effective resistance to stripe rust and usefulness of their flanking SSR markers for marker-assisted selection. Stripe rust, caused by Puccinia striiformis f. sp. tritici (Pst), is one of the most important diseases of wheat worldwide and resistance is the best control strategy. Durum wheat accessions PI 331260 and PI 480016 were resistant to all tested Pst races. To transfer the resistance genes to common wheat and map them to wheat chromosomes, both accessions were crossed with the stripe rust-susceptible spring wheat 'Avocet S'. Resistant F3 plants with 42 chromosomes were selected cytologically and by rust phenotype. A single dominant gene for resistance was identified in segregating F4 lines from each cross. F6 populations for each cross were developed from single F5 plants and used for genetic mapping. Different genes from PI 331260 and PI 480016 were mapped to different loci in chromosome 1BS using simple sequence repeat markers. The gene from PI 331260 was flanked by Xgwm413 and Xgdm33 in bin 1BS9-0.84-1.06 at genetic distances of 3.5 and 2.0 cM; and the gene from PI 480016 was flanked by Xgwm18 and Xgwm11 in chromosome bin C-1BS10-0.50 at 1.2 and 2.1 cM, respectively. Chromosomal locations and race and allelism tests indicated that the two genes are different from previously reported stripe rust resistance genes, and therefore are named as Yr64 from PI 331260 and Yr65 from PI 480016. These genes and their flanking markers, and selected common wheat lines with the genes should be valuable for diversifying resistance genes used in breeding wheat cultivars with stripe rust resistance.

Identification of Yr59 conferring high-temperature adult-plant resistance to stripe rust in wheat germplasm PI 178759.

KEY MESSAGE: This manuscript reports a new gene for non-race-specific resistance to stripe rust and molecular markers for incorporating it into wheat cultivars for control of the disease with durable resistance. Stripe rust, caused by Puccinia striiformis f. sp. tritici, is one of the most destructive wheat diseases worldwide. The spring wheat germplasm 'PI 178759' originating from Iraq showed effective resistance to stripe rust in field evaluations over 8 years in Washington state, USA. To map the resistance gene(s), PI 178759 was crossed with 'Avocet Susceptible', and the parents and 176 F2:3 lines were phenotyped in the fields under natural infection and in a greenhouse with selected races of P. striiformis f. sp. tritici. PI 178759 was identified to have high-temperature adult-plant (HTAP) resistance. Resistance gene analog polymorphism and simple sequence repeat techniques were used to identify molecular markers linked to the resistance gene and a chromosome region was mapped using a quantitative trait locus approach. One major gene was mapped to the long arm of chromosome 7B. Flanked by Xwgp5175 and Xbarc32 in a 2.1 cM region, the gene explained 31.8 and 54.7 % of the phenotypic variation in rAUDPC and IT, respectively. Based on genetic distances among markers and allelism tests, the HTAP resistance gene in PI 178759 is different from the previously reported Yr39, Yr52, YrZH84, and YrC591, also located on chromosome 7BL, and is therefore designated as Yr59. The gene and its flanking markers should be useful for developing wheat cultivars with durable resistance.

First Report of Oregon Grape (Mahonia aquifolium) as an Alternate Host for the Wheat Stripe Rust Pathogen (Puccinia striiformis f. sp. tritici) Under Artificial Inoculation.

As the primary host of the stripe rust pathogen, Puccinia striiformis f. sp. tritici (Pst), wheat can be infected by both aeciospores and urediniospores, and later is the host that gives rise to urediniospores and teliospores. Barberry species (e.g., Berberis vulgaris) can be infected by basidiospores, produced from the teliospores of wheat plants, and later gives rise to pycniospores and aeciospores, which has been demonstrated through artificial inoculation (3). Oregon grape (Mahonia aquifolium), closely related to Berberis, is a native evergreen shrub that is also grown as an ornamental plant in the Pacific Northwest. To determine if M. aquifolium can also serve as an alternate host for Pst, we conducted artificial inoculations under controlled conditions. Seeds of M. aquifolium collected from Pullman, WA, were sown in pots filled with soil mixture, and plants were grown in a greenhouse under wheat-growing conditions (1). In the first experiment, conducted in May to June 2011, the inoculum was telia collected from artificially inoculated wheat cv. Avocet S with urediniospores of isolate 09-134 (race PST-127) from the greenhouse. In the second experiment, conducted in July to August 2011, the inoculum was telia collected from naturally infected wheat cv. Nugaines with urediniospores from isolate 11-292 (race PST-127) from an experimental field near Pullman. For each experiment, mature teliospores of 60 telia from a single wheat plant were suspended in 1.0 ml of distilled water and inoculated with a fine paint brush onto the leaves of seven or eight 10- to 15-day-old plants of M. aquifolium. Plants were incubated initially in a dew chamber at 10°C for 72 h in darkness, then transferred to a growth chamber with a diurnal temperature cycle of 10 to 24°C and a 16 h light/8 h dark cycle (1). Reddish pycnia with nectar appeared on adaxial surfaces of inoculated leaves at 12 days post-inoculation (DPI), and reddish aecia were produced on the baxial surface at 16 DPI. All 15 M. aquifolium leaves of the 15 plants inoculated with teliospores produced pycnia and aecia. Seedlings of Nugaines and Avocet S, wheat cultivars that are susceptible to all Pst races (1), were then inoculated with a water suspension of aeciospores of 30 aecia collected from the M. aquifolium plants. Wheat plants were incubated as described above for M. aquifolium. Uredinia appeared at 15 DPI, and telia were produced after an additional 15 days. From these uredinia that formed on inoculated wheat, a total of 30 single-uredinium isolates were obtained using the standard procedure (1). Virulence tests were carried out on 20 wheat differentials for 10 randomly selected urediniospore isolates, revealing six virulence patterns. When tested with four selected Pst SSR markers (PstP001, PstP003, PstP005, PstP029) (2) and compared to other race PST-127 isolates, all 10 progeny isolates were homozygous, as were the parental isolates (09-134, 11-292). The virulence tests and marker genotypes verified that the urediniospore isolates resulted from infection by aecia, produced by parental isolate 09-134 through its sexual cycle on M. aquifolium. The study exhibited the completed sexual lifecycle of Pst through the five spore stages on wheat and M. aquifolium in a controlled setting, and suggests that under appropriate weather conditions, M. aquifolium may serve as an alternate host for Pst. Due to the wide distribution of M. aquifolium, further studies are needed to determine if the species can be infected by Pst under natural conditions. References: (1) X. M. Chen et al. Can. J. Plant Pathol. 32:315, 2010. (2) P. Cheng et al. Mol. Ecol. Resour. 12:779, 2012. (3) Y. Jin et al. Phytopathology 100:432, 2010.

Molecular mapping of Yr53, a new gene for stripe rust resistance in durum wheat accession PI 480148 and its transfer to common wheat.

Stripe rust, caused by Puccinia striiformis f. sp. tritici (Pst), is one of the most damaging diseases of wheat worldwide. It is essential to identify new genes for effective resistance against the disease. Durum wheat PI 480148, originally from Ethiopia, was resistant in all seedling tests with several predominant Pst races in the US under controlled greenhouse conditions and at multiple locations subject to natural infection for several years. To map the resistance gene(s) and to transfer it to common wheat, a cross was made between PI 480148 and susceptible common wheat genotype Avocet S (AvS). Resistant F(3) plants with 42 chromosomes were selected cytologically and by testing with Pst race PST-100. A total of 157 F(4) plants from a single F(3) plant with 2n = 42 tested with PST-100 segregated in a 3 resistant: 1 susceptible ratio, indicating that a single dominant gene from PI 480148 conferred resistance. Using the F(3:4) population and the resistance gene-analog polymorphism (RGAP) and simple sequence repeat (SSR) markers, the gene was mapped to the long arm of chromosome 2B. SSR marker Xwmc441 and RGAP marker XLRRrev/NLRRrev ( 350 ) flanked the resistance gene by 5.6 and 2.7 cM, respectively. The effective resistance of the gene to an Australian Pst isolate virulent to Yr5, which is also located on 2BL and confers resistance to all US Pst races, together with an allelism test of the two genes, indicated that the gene from PI 480148 is different from Yr5 and should be a new and useful gene for resistance to stripe rust. Resistant common wheat lines with plant types similar to AvS were selected for use in breeding programs.

Higher serum retinol binding protein 4 may be a predictor of weak metabolic control in chinese patients with type 2 diabetes mellitus.

OBJECTIVE: To investigate relationships between serum levels of retinol binding protein 4 (RBP4) and clinical and metabolic variables in Chinese patients with type 2 diabetes mellitus. METHODS: A total of 513 patients (286 males/227 females) provided clinical and lifestyle data and blood and urine samples for analysis. Patients were stratified into four quartile groups according to serum RBP4 concentrations. RESULTS: RBP4 concentration was independently associated with gender, systolic blood pressure, serum triglyceride and creatinine levels, and estimated glomerular filtration rate (eGFR). Hypertension, dyslipidaemia, micro albuminuria and impaired eGFR (<60 ml/min per 1.73 m2) were significantly more prevalent in patients with the highest RBP4 levels than in those with the lowest levels. Increased serum RBP4 was associated with increased risk of hypertension, dyslipidaemia, micro albuminuria and impaired eGFR after adjusting for gender, age, body mass index and duration of diabetes. CONCLUSION: Serum RBP4 may be a useful marker of overall metabolic control in Chinese patients with type 2 diabetes mellitus.

Characterization and molecular mapping of Yr52 for high-temperature adult-plant resistance to stripe rust in spring wheat germplasm PI 183527.

Stripe rust, caused by Puccinia striiformis f. sp. tritici, is one of the most destructive diseases of wheat worldwide. Resistance is the best approach to control the disease. High-temperature adult-plant (HTAP) stripe rust resistance has proven to be race non-specific and durable. However, genes conferring high-levels of HTAP resistance are limited in number and new genes are urgently needed for breeding programs to develop cultivars with durable high-level resistance to stripe rust. Spring wheat germplasm PI 183527 showed a high-level of HTAP resistance against stripe rust in our germplasm evaluations over several years. To elucidate the genetic basis of resistance, we crossed PI 183527 and susceptible wheat line Avocet S. Adult plants of parents, F(1), F(2) and F(2:3) progeny were tested with selected races under the controlled greenhouse conditions and in fields under natural infection. PI 183527 has a single dominant gene conferring HTAP resistance. Resistance gene analog polymorphism (RGAP) and simple sequence repeat (SSR) markers in combination with bulked segregant analysis (BSA) were used to identify markers linked to the resistance gene. A linkage map consisting of 4 RGAP and 7 SSR markers was constructed for the resistance gene using data from 175 F(2) plants and their derived F(2:3) lines. Amplification of nulli-tetrasomic, ditelosomic and deletion lines of Chinese Spring with three RGAP markers mapped the gene to the distal region (0.86-1.0) of chromosome 7BL. The molecular map spanned a genetic distance of 27.3 cM, and the resistance gene was narrowed to a 2.3-cM interval flanked by markers Xbarc182 and Xwgp5258. The polymorphism rates of the flanking markers in 74 wheat lines were 74 and 30 %, respectively; and the two markers in combination could distinguish the alleles at the resistance locus in 82 % of tested genotypes. To determine the genetic relationship between this resistance gene and Yr39, a gene also on 7BL conferring HTAP resistance in Alpowa, a cross was made between PI 183527 and Alpowa. F(2) segregation indicated that the genes were 36.5 ± 6.75 cM apart. The gene in PI 183527 was therefore designed as Yr52. This new gene and flanking markers should be useful in developing wheat cultivars with high-level and possible durable resistance to stripe rust.