Genomic ancestry and metabolic syndrome in individuals with type 1 diabetes from an admixed population: a multicentre, cross-sectional study in Brazil.

AIMS: To evaluate the relationship between self-reported colour-race, genomic ancestry, and metabolic syndrome in an admixed Brazilian population with type 1 diabetes. METHODS: We included 1640 participants with type 1 diabetes. The proportions of European, African and Amerindian genomic ancestries were determined by 46 ancestry informative markers of insertion deletion. Two different sets of analyses were performed to determine whether self-reported colour-race and genomic ancestry were predictors of metabolic syndrome. RESULTS: Metabolic syndrome was identified in 29.8% of participants. In the first model, the factors associated with metabolic syndrome were: female gender (odds ratio 1.95, P < 0.001); diabetes duration (odds ratio 1.04, P < 0.001); family history of type 2 diabetes (odds ratio 1.36, P = 0.019); and acanthosis nigricans (odds ratio 5.93, P < 0.001). Colour-race was not a predictive factor for metabolic syndrome. In the second model, colour-race was replaced by European genomic ancestry. The associated factors were: female gender (odds ratio 1.95, P < 0.001); diabetes duration (odds ratio 1.04, P < 0.001); family history of type 2 diabetes (odds ratio 1.39, P = 0.011); and acanthosis nigricans (odds ratio 6.12, P < 0.001). Physical exercise (≥3 times a week) was a protective factor (odds ratio 0.77, P = 0.041), and European genomic ancestry was not associated with metabolic syndrome but showed an odds ratio of 1.77 (P = 0.05). CONCLUSIONS: Although a higher level of European genomic ancestry was observed among participants with metabolic syndrome in the univariate analysis, this association did not persist after multivariable adjustments. Further prospective studies in other highly admixed populations remain necessary to better evaluate whether the European ancestral component modulates the development of metabolic syndrome in type 1 diabetes.

Effect of cane use on bone marrow lesion volume in people with medial tibiofemoral knee osteoarthritis: randomized clinical trial.

OBJECTIVE: To evaluate effects of daily cane use for 3 months on medial tibiofemoral bone marrow lesion (BML) volumes in people with medial tibiofemoral osteoarthritis (OA). DESIGN: In this randomized controlled trial (RCT), 79 participants with medial tibiofemoral OA were randomized to either a cane group (using a cane whenever walking) or control group (not using any gait aid) for 3 months. The cane group received a single training session by a physiotherapist, using a biofeedback cane to teach optimal technique and body weight support and motor learning principles to facilitate retention of learning. The primary outcome was change in total medial tibiofemoral BML volume (per unit bone volume) measured from magnetic resonance imaging (MRI) at 3 months. Secondary outcomes were BML volumes (per unit bone volume) of the medial tibia and femur, and patient-reported outcomes of overall knee pain, knee pain on walking, physical function, perceived global symptom changes and health-related quality of life. MRI analyses were performed by a blinded assessor. RESULTS: Seventy-eight participants (99%) completed the primary outcome. Mean (standard deviation) daily cane use was 2.3 (1.7) hours over 3 months. No evidence of between-group differences was found for change in total medial tibiofemoral BML volume (mean difference: -0.0010 (95% confidence intervals: -0.0022, 0.0003)). Most secondary outcomes showed minimal differences between groups. CONCLUSION: Daily use of a cane during walking for 3 months aiming to reduce knee joint loading did not change medial tibiofemoral BML volumes compared to no use of gait aids. CLINICAL TRIAL REGISTRATION: Australian New Zealand Clinical Trial Registry (ACTRN12614000909628).

Effect of repeated intravenous lipopolysaccharide infusions on systemic inflammatory response and endometrium gene expression in Holstein heifers.

This study aimed to evaluate the effect of repeated intravenous lipopolysaccharide (LPS) infusions in nonlactating heifers on (1) the systemic proinflammatory state as measured by biomarkers in blood and plasma, and (2) endometrial gene expression of candidate transcripts on d 15 of gestation. Our hypothesis was that target transcripts related to a major functional group would be negatively modified in the preimplantation endometrium by the LPS treatments. In the first experiment (n = 13), a systemic proinflammatory state [defined as increased plasma concentrations of tumor necrosis factor (TNF)-α and haptoglobin for 2 wk] was established using 2 different sequential LPS infusion protocols. In the second experiment, heifers (n = 22; 11 mo of age) had their time of ovulation synchronized by a modified Ovsynch protocol and were enrolled in 1 of 2 treatments: control (CON; n = 11), which received sterile saline solution i.v., and LPS treatment (LPS; n = 11), submitted to repeated i.v. LPS injections (0.10, 0.25, 0.50, 0.75, 1.00, and 1.25 µg/kg) starting 2 d after artificial insemination (AI; d 0) and then every other day until d 15 after AI. At each LPS injection, rectal temperatures were measured hourly for 6 h. Blood samples were collected from d -1 to d 13 for analyses of progesterone, TNF-α, and haptoglobin in plasma, along with white blood cell (WBC) count and differential analysis. On d 15, endometrium tissue biopsies were taken and kept at -80°C until quantitative real-time PCR analysis of 30 target transcripts related to the immune system, adhesion molecules, and endometrium receptivity. Data were checked for normality and analyzed by repeated-measures ANOVA using PROC UNIVARIATE and PROC MIXED of SAS (SAS Institute Inc., Cary, NC). After each LPS injection, temperature was greater in the first 4 h in the LPS group compared with CON. Both TNF-α and haptoglobin increased in the LPS treatment with a significant treatment by day interaction. Total leukocyte count did not differ between treatments, but the differential count increased for neutrophils, band cells, and monocytes, and decreased for lymphocytes and eosinophils in LPS compared with CON. Progesterone concentrations in plasma did not differ between treatments during the experimental period. Out of 30 target genes analyzed, 3 transcripts were differentially expressed: indoleamine 2,3-dioxygenase (IDO; fold-change = 0.48) and pentraxin-3 (PTX3; fold-change = 0.38) were downregulated, whereas myxovirus-resistance protein (MX1; fold-change = 2.85) was upregulated in the LPS group. Sequential LPS injections were able to induce a prolonged systemic proinflammatory state, but effects on gene expression were limited to transcripts associated with the immune system. These results suggest that a mechanism for subfertility is linked to a proinflammatory state in dairy heifers.

Effect of dose and timing of prostaglandin F2α treatments during a Resynch protocol on luteal regression and fertility to timed artificial insemination in lactating Holstein cows.

Our objective was to evaluate the effect of a second PGF2α treatment (25 mg of dinoprost) or a double dose of PGF2α (50 mg of dinoprost) during a Resynch protocol on luteal regression and pregnancies per artificial insemination (P/AI) in lactating dairy cows. Lactating Holstein cows (n = 1,100) were randomly assigned at a nonpregnancy diagnosis to receive (1) Ovsynch (control: 100 µg of GnRH; 7 d, 25 mg of PGF2α; 56 h, 100 µg of GnRH), (2) Ovsynch with a second PGF2α treatment (GPPG: 100 µg of GnRH; 7 d, 25 mg of PGF2α; 24 h, 25 mg of PGF2α; 32 h, 100 µg of GnRH), or (3) Ovsynch with a double dose of PGF2α (GDDP: 100 µg of GnRH; 7 d, 50 mg of PGF2α; 56 h, 100 µg of GnRH). All cows received timed artificial insemination (TAI) approximately 16 h after the second GnRH treatment (G2). Pregnancy diagnosis was performed by transrectal palpation 39 ± 3 d after TAI, and pregnancy status was reconfirmed 66 d after TAI. Blood samples collected from a subset of cows in each treatment at the first PGF2α treatment (n = 394) and at G2 (n = 367) were assayed for progesterone (P4). Data were analyzed by logistic regression using the GLIMMIX procedure of SAS (SAS Institute Inc., Cary, NC). At 39 d after TAI, GPPG cows tended to have more P/AI than control cows [35% (137/387) vs. 31% (107/349)], whereas P/AI for GDDP cows [32% (118/364)] did not differ from that for control cows. Pregnancy loss from 38 to 66 d did not differ among treatments and was 8% (30/362). The percentage of cows with complete luteal regression (P4 <0.4 ng/mL at G2) tended to differ among treatments and was greater for GPPG cows than for GDDP and control cows (94% vs. 88% vs. 88%, respectively). Overall, cows with P4 <1 ng/mL at the first PGF2α treatment had fewer P/AI than cows with P4 ≥1 ng/mL (27% vs. 38%), whereas cows with P4 ≥0.4 ng/mL at G2 had fewer P/AI than cows with P4 <0.4 ng/mL (15% vs. 38%). We conclude that adding a second PGF2α treatment 24 h after the first within a Resynch protocol tended to increase the proportion of cows undergoing complete luteal regression and P/AI, whereas treatment with a double dose of PGF2α at a single time did not.

Circulating progesterone concentrations in nonlactating Holstein cows during reuse of intravaginal progesterone implants sanitized by autoclave or chemical disinfection.

The aim of this study was to compare plasma progesterone (P4) concentrations in nonlactating, multiparous Holstein cows (n = 24) treated with 2 types of intravaginal implants containing either 1.0 or 1.9 g of P4 either at the first use or during reuse of the implants after sanitizing the implant by autoclave or chemical disinfection. In a completely randomized design with a 2 × 3 factorial arrangement and 2 replicates, every cow underwent 2 of 6 treatments. Two sources of P4 [controlled internal drug release (1.9 g of P4) from Zoetis (São Paulo, Brazil), and Sincrogest (1.0 g of P4) from Ourofino (Cravinhos, Brazil)] and 3 types of processing, new (N), reused after autoclave (RA), and reused after chemical disinfection (RC), were used. After inducing luteolysis to avoid endogenous circulating P4, the cows were randomized in 1 of 6 treatments (1.9 g of N, 1.9 g of RA, 1.9 g of RC, 1.0 g of N, 1.0 g of RA, and 1.0 g RC). Cows were treated with the implants for 8 d and during this period blood samples were collected at 0, 2, 12, 24, 48, 72, 96, 120, 144, 168, and 192 h. Statistical analyses were performed using Proc-Mixed and the mean ± standard error of the mean P4 concentrations were calculated using the Proc-Means procedures of SAS 9.4 (SAS Institute Inc., Cary, NC). No interaction between treatments was observed. Comparing types of implant, average P4 concentrations during treatments were greater for 1.9 g than 1.0 g (1.46 vs. 1.14 ± 0.04 ng/mL). When types of processing were compared, average P4 concentrations did not differ between autoclaved and new inserts (1.46 vs. 1.37 ± 0.05 ng/mL; respectively), but both were greater than chemically disinfected implants (1.09 ± 0.04 ng/mL). Within 1.9-g P4 inserts, P4 concentrations from autoclaved implants were greater than new, which were greater than chemically disinfected (1.67 ± 0.06 vs. 1.49 ± 0.07 vs. 1.21 ± 0.05 ng/mL; respectively). For 1.0-g P4 implants, P4 concentrations from autoclaved did not differ from new, but both were greater than chemically disinfected (1.20 ± 0.08 vs. 1.24 ± 0.06 vs. 0.97 ± 0.05 ng/mL; respectively). In conclusion, the mean plasma P4 concentration in nonlactating Holstein cows was greater for 1.9 than 1.0 g of P4 and regardless of the type of implant, the autoclaving process provided greater circulating P4 in relation to chemical disinfection, and similar or greater P4 concentrations compared with a new implant.

Follicular dynamics, circulating progesterone, and fertility in Holstein cows synchronized with reused intravaginal progesterone implants that were sanitized by autoclave or chemical disinfection.

This experiment aimed to compare circulating progesterone (P4), follicular dynamics, and fertility during reuse of intravaginal P4 implants that were sanitized by autoclave or chemical disinfection in lactating Holstein cows submitted to fixed-time artificial insemination (FTAI). For this, 123 primiparous and 226 multiparous cows from 2 farms, averaging (mean ± standard deviation) 163.9 ± 141.9 d in milk, 35.7 ± 11.3 kg of milk/d, and a body condition score of 2.9 ± 0.5, were enrolled in the study. Cows were randomly assigned to 1 of 2 treatments using a completely randomized design and each cow received a reused implant (1.9 g of P4; previously used for 8 d) that was either autoclaved (AUT; n = 177) or chemically disinfected (CHEM; n = 172) on d -10. Also on d -10, cows received 2 mg of estradiol benzoate and 100 µg of GnRH. On d -3, cows received 25 mg of dinoprost (PGF2α). A second PGF2α was given on d -2, along with 1 mg of estradiol cypionate and P4 implant removal. Cows received FTAI on d 0. A subset of cows (n = 143) was evaluated by ultrasound on d -10, -8, -6, -3, -2, 0, and 5 to identify ovarian structures, and blood was sampled on d -10, -3, and -2 for P4 concentrations by RIA. Pregnancy diagnoses were performed at d 32 and 60. Statistical analyses was performed using PROC-MIXED for continuous variables and PROC-GLIMMIX of SAS 9.4 (SAS Institute Inc., Cary, NC) for binomial variables. The treatments did not differ in circulating P4 on d -10 or -3, but P4 was greater on d -2 in CHEM cows. Ovulation to the treatments on d -10 was associated with lower circulating P4 on d -10 (2.0 vs. 3.1 ng/mL) and resulted in greater P4 on d -3 (4.0 vs. 2.4 ng/mL) and more cows with a corpus luteum on d -3 (100 vs. 40%) than nonovulating cows. Cows that ovulated to d -10 treatments were more likely to have a synchronized new follicular wave (97.9 vs. 63.2%) and had an earlier wave emergence (1.9 vs. 2.6 d), resulting in less cows ovulating a persistent follicle (0.0 vs. 35.7%). Type of P4 implant, corpus luteum presence on d -10, and ovulation to d -10 treatments did not affect fertility (pregnancy per AI; P/AI). However, P/AI on farm A was greater than on farm B at 32 (40.8 vs. 27.8%) and 60 d (35.8 vs. 24.3%), independent of treatment. In conclusion, P4 implants with different P4 release patterns did not produce detectable differences in follicular dynamics, synchronization rate, or P/AI. Nevertheless, presence of corpus luteum or ovulation at the beginning of the FTAI protocol affected reproductive variables, such as timing and synchronization of follicular wave emergence, and size of the ovulatory follicle. Beyond that, more overall synchronized cows became pregnant to the FTAI protocol.

A new species of Hemibrycon (Characiformes, Characidae, Stevardiinae) from the upper Magdalena River basin in Colombia.

Hemibrycon iqueima sp. nov., is described from small streams in the Magdalena drainage at the foothills of the western slope of the Eastern Cordillera of the Colombian Andes, Suarez municipality, Tolima Department, Colombia. The new species is distinguished from its congeners in the Magdalena-Cauca River basin by a combination of characters related to snout-anal-fin origin length, head length, dorsal-pectoral fin distance, dorsal-fin-hypural distance, postorbital distance, orbital diameter, snout length, number of total vertebrae, pre-dorsal scales, scale rows between anal-fin origin and lateral line, number of branched rays of the anal fin, maxillary teeth number and number and arrangement of hooks on the branched rays of the pectoral and dorsal fins. In addition, the validity of this species is supported by previous molecular analyses that included specimens of the new species that had been erroneously identified. Phylogenetic relationships between the new species and congeners from Pacific coast basins are discussed.

Knee osteoarthritis phenotypes and their relevance for outcomes: a systematic review.

OBJECTIVE: To systematically review the literature for studies investigating knee osteoarthritis (OA) phenotypes to examine what OA characteristics are relevant for phenotyping. METHODS: A comprehensive search was performed in Medline, EMBASE, Web of Sciences, CINAHL, and Scopus databases from inception to September 2016. Inclusion was limited to observational studies of individuals with symptomatic knee OA that identified phenotypes based on any OA characteristics and assessed their association with clinically important outcomes. A descriptive synthesis of the data was performed. RESULTS: Of the 2777 citations retrieved, 34 studies were included. Clinical phenotypes were investigated most frequently, followed by laboratory, imaging and aetiologic phenotypes. Eight studies defined subgroups based on outcome trajectories (pain, function and radiographic progression trajectories). Most studies used a single patient or disease characteristic to identify patients subgroups while five included characteristics from multiple domains. We found evidence from multiple studies suggesting that pain sensitization, psychological distress, radiographic severity, body mass index (BMI), muscle strength, inflammation and comorbidities are associated with clinically distinct phenotypes. Gender, obesity and other metabolic abnormalities, the pattern of cartilage damage, and inflammation may be implicated in delineating distinct structural phenotypes. Only a few studies investigated the external validity of the phenotypes or their prospective validity using longitudinal outcomes. CONCLUSIONS: There is marked heterogeneity in the data selected by the studies investigating knee OA phenotypes. We identified the phenotypic characteristics that can be considered for a comprehensive phenotype classification in future studies. A framework for the investigation of phenotypes could be useful for future studies. PROTOCOL REGISTRATION: PROSPERO CRD42016036220.

Progesterone-based fixed-time artificial insemination protocols for dairy cows: Gonadotropin-releasing hormone versus estradiol benzoate at initiation and estradiol cypionate versus estradiol benzoate at the end.

Our objectives were to evaluate ovarian dynamics and fertility comparing 2 treatments at the start of a progesterone (P4)-based fixed-time artificial insemination (FTAI) protocol and 2 treatments at the end of the protocol. Thus, 1,035 lactating Holstein cows were assigned in a random phase of the estrous cycle to 1 of 4 treatments using a completely randomized design with a 2×2 factorial arrangement. At the beginning of the protocol (d -10), cows received GnRH or estradiol benzoate (EB) and, at the end, EB (d -1) or estradiol cypionate (ECP; d -2), resulting in 4 treatments: GnRH-EB, GnRH-ECP, EB-EB, and EB-ECP. All cows received an intravaginal P4 device on d -10, which was removed on d -2. Cows also received PGF2α on d -3 and -2. The FTAI was performed on d 0. Ovaries were evaluated by ultrasound for corpus luteum (CL) presence and regression (d -10 and -3) and follicle measurements (d -10 and 0), as well as the uterus for percentage pregnant per AI (P/AI; d 32 and 60). Blood samples were collected (d -10 and -3) for P4 measurements. Treatment with GnRH rather than EB tended to increase P/AI on d 32 (38.2 vs. 33.7%) and on d 60 (32.9 vs. 28.9%). More cows treated with GnRH had CL on d -3 compared with EB-treated cows (77.3 vs. 58.3%), due to less CL regression between d -10 and -3 (24.7 vs. 43.8%) and more cows with a new CL on d -3 (35.9 vs. 25.0%). Cows treated with GnRH also had greater P4 concentrations on d -3 than EB cows (3.4 vs. 2.0 ng/mL). Increased circulating P4 at the start of the protocol (d -10) decreased the probability of ovulation to EB or GnRH at that time. Cows from GnRH group also ovulated a larger-diameter follicle at the end of the protocol (15.5 vs. 14.7mm). No difference between EB and ECP in P/AI on d 32 (34.8 vs. 37.0) and 60 (30.8 vs. 31.0%) or in pregnancy loss (11.1 vs. 15.4%) was detected and we found no interaction between treatments for P/AI. Independent of treatment, cows with CL on d -10 and -3 had the greatest P/AI on d 60 (36.9%). In conclusion, treatments at the end of the protocol were similar for ECP or EB and we found no additive effect or interactions on P/AI between treatments. However, cows treated with GnRH rather than EB on d -10 had less luteolysis and tended to have greater P/AI, probably because P4 concentrations were greater during the protocol. Finally, regardless of treatments, cows with CL at the beginning of the protocol as well as at the time of PGF2α had greater fertility.

Genetic potential of black bean genotypes with predictable behaviors in multienvironment trials.

The aim of this study was to evaluate the phenotypic stability and specific and broad adaptability of common black bean genotypes for the Central and Center-South regions of Brazil by using the Annicchiarico and AMMI (weighted average of absolute scores: WAAS, and weighted average of absolute scores and productivity: WAASP) methodologies. We carried out 69 trials, with 43 and 26 trials in the Central and Center-South regions, respectively. Thirteen genotypes were evaluated in a randomized block design with three replications, during the rainy, dry, and winter seasons in 2 years. To obtain estimates of specific adaptation, we analyzed the parameters for each method obtained in the two geographic regions separately. To estimate broad adaptation, we used the average of the parameters obtained from each region. The lines identified with high specific adaptation in each region were not the same based on the Annicchiarico and AMMI (WAAS) methodologies. It was not possible to identify the same genotypes with specific or broad stability by using these methods. By contrast, the Annicchiarico and AMMI (WAASP) methods presented very similar estimates of broad and specific adaptation. Based on these methods, the lines with more specific adaptation were CNFP 8000 and CNFP 7994, in the Central and Center-South regions, respectively, of which the CNFP 8000 line was more widely adapted.

Genetic potential of common bean progenies obtained by different breeding methods evaluated in various environments.

Grain yield is strongly influenced by the environment, has polygenic and complex inheritance, and is a key trait in the selection and recommendation of cultivars. Breeding programs should efficiently explore the genetic variability resulting from crosses by selecting the most appropriate method for breeding in segregating populations. The goal of this study was to evaluate and compare the genetic potential of common bean progenies of carioca grain for grain yield, obtained by different breeding methods and evaluated in different environments. Progenies originating from crosses between lines and CNFC 7812 and CNFC 7829 were replanted up to the F7 generation using three breeding methods in segregating populations: population (bulk), bulk within F2 progenies, and single-seed descent (SSD). Fifteen F8 progenies per method, two controls (BRS Estilo and Perola), and the parents were evaluated in a 7 x 7 simple lattice design, with plots of two 4-m rows. The tests were conducted in 10 environments in four States of Brazil and in three growing seasons in 2009 and 2010. Genetic parameters including genetic variance, heritability, variance of interaction, and expected selection gain were estimated. Genetic variability among progenies and the effect of progeny-environment interactions were determined for the three methods. The breeding methods differed significantly due to the effects of sampling procedures on the progenies and due to natural selection, which mainly affected the bulk method. The SSD and bulk methods provided populations with better estimates of genetic parameters and more stable progenies that were less affected by interaction with the environment.

Estimation of genetic parameters and selection of high-yielding, upright common bean lines with slow seed-coat darkening.

Slow seed coat darkening is desirable in common bean cultivars and genetic parameters are important to define breeding strategies. The aims of this study were to estimate genetic parameters for plant architecture, grain yield, grain size, and seed-coat darkening in common bean; identify any genetic association among these traits; and select lines that associate desirable phenotypes for these traits. Three experiments were set up in the winter 2012 growing season, in Santo Antônio de Goiás and Brasília, Brazil, including 220 lines obtained from four segregating populations and five parents. A triple lattice 15 x 15 experimental design was used. The traits evaluated were plant architecture, grain yield, grain size, and seed-coat darkening. Analyses of variance were carried out and genetic parameters such as heritability, gain expected from selection, and correlations, were estimated. For selection of superior lines, a "weight-free and parameter-free" index was used. The estimates of genetic variance, heritability, and gain expected from selection were high, indicating good possibility for success in selection of the four traits. The genotype x environment interaction was proportionally more important for yield than for the other traits. There was no strong genetic correlation observed among the four traits, which indicates the possibility of selection of superior lines with many traits. Considering simultaneous selection, it was not possible to join high genetic gains for the four traits. Forty-four lines that combined high yield, more upright plant architecture, slow darkening grains, and commercial grade size were selected.

Genetic parameters and breeding strategies for high levels of iron and zinc in Phaseolus vulgaris L.

One of the current focus of common bean breeding programs in Brazil is to increase iron (FeC) and zinc content (ZnC) in grains. The objectives of this study were to estimate genetic parameters for FeC and ZnC in common bean, verify the need for conducting multi-site evaluation tests, identify elite lines that combine high FeC and ZnC with good adaptability, stability, and agronomic potential, and examine the genetic association between FeC and ZnC. Elite lines (140) were evaluated for important agronomic traits in multiple environments. In one trial, FeC and ZnC were evaluated and genetic parameters were estimated. Based on the high heritability estimates and significant selection gains obtained, the conditions for a successful selection was favorable. Of the 140 evaluated lines, 17 had higher FeC and ZnC, and were included in the validation test (2013, five environments), specifically for the evaluation of FeC and ZnC. The line by environment interaction for FeC and ZnC was detected, but it was predominantly simple. The environmental effect strongly influenced FeC and ZnC . The environment Brasília/rainy season was selected as the best evaluation site for preliminary tests for FeC and ZnC, because it resulted in similar conclusions as the mean of the five environments. The lines CNFP 15701 and CNFC 15865 had higher FeC and ZnC and were highly adaptable and stable, and are recommended for utilization in breeding programs. The lines CNFC 15833, CNFC 15703, and CNFP 15676 showed excellent combined agronomic and nutritional traits, and were selected for the development of biofortified cultivars. Additionally, the genetic association between FeC and ZnC was detected.

Differential interactions between Curtobacterium flaccumfaciens pv. flaccumfaciens and common bean.

Bacterial wilt of common bean caused by Curtobacterium flaccumfaciens pv. flaccumfaciens is an important disease in terms of economic importance. It reduces grain yield by colonizing xylem vessels, subsequently impeding the translocation of water and nutrients to the superior plant parts. The existence of physiological races in C. flaccumfaciens pv. flaccumfaciens has not so far been reported. The objective of the present investigation was to identify physiological races, evaluate differential interaction, and select resistant genotypes of common bean. Initially, 30 genotypes of common bean were inoculated with eight isolates exhibiting different levels of aggressiveness, under controlled greenhouse conditions. Disease was assessed 15 days after inoculation. The existence of differential interactions between C. flaccumfaciens pv. flaccumfaciens isolates and common bean genotypes were identified by utilizing partial diallel analysis. The most aggressive isolates were BRM 14939 and BRM 14942 and the least aggressive isolates were BRM 14941 and BRM 14946. The genotypes IPA 9, Ouro Branco, and Michelite were selected as more resistant among the test isolates. The genotypes IAC Carioca Akytã, BRS Notável, Pérola, IAC Carioca Aruã, and Coquinho contributed more to the isolate x genotype interaction according to the ecovalence method of estimation, and were, therefore, indicated as differentials. Based on these results, it was possible to conclude that physiological races of the pathogen exist, to select resistant genotypes, and to propose a set of differentials.

Phosphorylated H2AX in parthenogenetically activated, in vitro fertilized and cloned bovine embryos.

In vitro embryo production methods induce DNA damage in the embryos. In response to these injuries, histone H2AX is phosphorylated (γH2AX) and forms foci at the sites of DNA breaks to recruit repair proteins. In this work, we quantified the DNA damage in bovine embryos undergoing parthenogenetic activation (PA), in vitro fertilization (IVF) or somatic cell nuclear transfer (SCNT) by measuring γH2AX accumulation at different developmental stages: 1-cell, 2-cell and blastocyst. At the 1-cell stage, IVF embryos exhibited a greater number of γH2AX foci (606.1 ± 103.2) and greater area of γH2AX staining (12923.6 ± 3214.1) than did PA and SCNT embryos. No differences at the 2-cell stage were observed among embryo types. Although PA, IVF and SCNT were associated with different blastocyst formation rates (31.1%, 19.7% and 8.3%, P < 0.05), no differences in the number of γH2AX foci or area were detected among the treatments. γH2AX is detected in bovine preimplantation embryos produced by PA, IVF and SCNT; the amount of DNA damage was comparable among those embryos developing to the blastocyst stage among different methods for in vitro embryo production. While IVF resulted in increased damage at the 1-cell embryo stage, no difference was observed between PA and SCNT embryos at any developmental stage. The decrease in the number of double-stranded breaks at the blastocyst stage seems to indicate that DNA repair mechanisms are functional during embryo development.

Genetic potential of common bean progenies selected for crude fiber content obtained through different breeding methods.

Gastrointestinal health is of great importance due to the increasing consumption of functional foods, especially those concern-ing diets rich in fiber content. The common bean has been valorized as a nutritious food due to its appreciable fiber content and the fact that it is consumed in many countries. The current study aimed to evaluate and compare the genetic potential of common bean progenies of the carioca group, developed through different breeding methods, for crude fiber content. The progenies originated through hybridization of two advanced strains, CNFC 7812 and CNFC 7829, up to the F7 generation using three breeding methods: bulk-population, bulk within F2 families, and single seed descent. Fifteen F8 progenies were evaluated in each method, as well as two check cultivars and both parents, us-ing a 7 x 7 simple lattice design, with experimental plots comprised of two 4-m long rows. Field trials were conducted in eleven environments encompassing four Brazilian states and three different sowing times during 2009 and 2010. Estimates of genetic parameters indicate differences among the breeding methods, which seem to be related to the different processes for sampling the advanced progenies inherent to each method, given that the trait in question is not subject to natural selection. Variability amongst progenies occurred within the three breeding methods and there was also a significant effect of environment on the progeny for all methods. Progenies developed by bulk-population attained the highest estimates of genetic parameters, had less interaction with the environment, and greater variability.

Comparative analysis of laparoscopic and ultrasound-guided biopsy methods for gene expression analysis in transgenic goats.

The present study aimed to compare laparoscopic (LP) and ultrasound-guided (US) biopsy methods to obtain either liver or splenic tissue samples for ectopic gene expression analysis in transgenic goats. Tissue samples were collected from human granulocyte colony stimulating factor (hG-CSF)-transgenic bucks and submitted to real-time PCR for the endogenous genes (Sp1, Baff, and Gapdh) and the transgene (hG-CSF). Both LP and US biopsy methods were successful in obtaining liver and splenic samples that could be analyzed by PCR (i.e., sufficient sample sizes and RNA yield were obtained). Although the number of attempts made to obtain the tissue samples was similar (P > 0.05), LP procedures took considerably longer than the US method (P = 0.03). Finally, transgene transcripts were not detected in spleen or liver samples. Thus, for the phenotypic characterization of a transgenic goat line, investigation of ectopic gene expression can be made successfully by LP or US biopsy, avoiding the traditional approach of euthanasia.

Biochar application to a contaminated soil reduces the availability and plant uptake of zinc, lead and cadmium.

Heavy metals in soil are naturally occurring but may be enhanced by anthropogenic activities such as mining. Bio-accumulation of heavy metals in the food chain, following their uptake to plants can increase the ecotoxicological risks associated with remediation of contaminated soils using plants. In the current experiment sugar cane straw-derived biochar (BC), produced at 700 °C, was applied to a heavy metal contaminated mine soil at 1.5%, 3.0% and 5.0% (w/w). Jack bean (Canavalia ensiformis) and Mucuna aterrima were grown in pots containing soil and biochar mixtures, and control pots without biochar. Pore water was sampled from each pot to confirm the effects of biochar on metal solubility, whilst soils were analyzed by DTPA extraction to confirm available metal concentrations. Leaves were sampled for SEM analysis to detect possible morphological and anatomical changes. The application of BC decreased the available concentrations of Cd, Pb and Zn in 56, 50 and 54% respectively, in the mine contaminated soil leading to a consistent reduction in the concentration of Zn in the pore water (1st collect: 99 to 39 µg L(-1), 2nd: 97 to 57 µg L(-1) and 3rd: 71 to 12 µg L(-1)). The application of BC reduced the uptake of Cd, Pb and Zn by plants with the jack bean translocating high proportions of metals (especially Cd) to shoots. Metals were also taken up by Mucuna aterrima but translocation to shoot was more limited than for jack bean. There were no differences in the internal structures of leaves observed by scanning electron microscopy. This study indicates that biochar application during mine soil remediation reduce plant concentrations of potential toxic metals.

Template-oriented genetic algorithm feature selection of analyte wavelets in the Raman spectrum of a complex mixture.

We introduce a fast computational method for feature selection that facilitates the accurate spectral analysis of a chemical species of interest in the presence of overlapping uncorrelated variance. Using a genetic algorithm in a data-driven approach, our method assigns predictors according to a template determined to minimize prediction variance in a calibration space. This template-oriented genetic algorithm (TOGA) efficiently establishes features of greatest significance and determines their optimal combination. We demonstrate the efficacy of TOGA using an elementary model system in which we seek to quantify a target monosaccharide in mixtures containing other sugars added in random amounts. The results establish TOGA as an effective and reliable technique for isolating signature spectra of targeted substances in complex mixtures.

96-well microtiter plates for biofouling simulation in biomedical settings.

Microtiter plates with 96 wells are routinely used in biofilm research mainly because they enable high-throughput assays. These platforms are used in a variety of conditions ranging from static to dynamic operation using different shaking frequencies and orbital diameters. The main goals of this work were to assess the influence of nutrient concentration and flow conditions on biofilm formation by Escherichia coli in microtiter plates and to define the operational conditions to be used in order to simulate relevant biomedical scenarios. Assays were performed in static mode and in incubators with distinct orbital diameters using different concentrations of glucose, peptone and yeast extract. Computational fluid dynamics (CFD) was used to simulate the flow inside the wells for shaking frequencies ranging from 50 to 200 rpm and orbital diameters from 25 to 100 mm. Higher glucose concentrations enhanced adhesion of E. coli in the first 24 h, but variation in peptone and yeast extract concentration had no significant impact on biofilm formation. Numerical simulations indicate that 96-well microtiter plates can be used to simulate a variety of biomedical scenarios if the operating conditions are carefully set.