Dairy Effluent-Saturated Biochar's Short-Term Effects on Vigna unguiculata and Cynodon dactylon Performance and Soil Properties.

We compared the effects of wood-, manure-, and blend-derived biochar (BC) saturated/unsaturated with dairy effluents on Vigna unguiculata and Cynodon dactylon performance and soil characteristics in a greenhouse pot study. Plant samples were assayed for herbage and root dry weight and N and C percentages. Soil samples were assayed for nutrients, pH, and conductivity. Variance analysis, Tukey's tests, Pearson's correlations, and multiple regression analysis were performed. The performance of C. dactylon was not affected. V. unguiculata's herbage and root production responded negatively to manure BC and 2% of any BC, respectively, which is mainly explained by the conductivity and soil P increase, respectively. When V. unguiculata was grown, BC inclusion decreased NO3-N and increased the soil P content. When C. dactylon was grown, only P was altered (increased) when manure or the blend BC were applied. The soil total C increased as the BC loading rate increased. The application of high BC rates was detrimental for V. unguiculata, but showed a neutral effect for C. dactylon. To improve dairy waste recycling, saturated 1% blend BC and saturated 2% blend or manure BC could be applied to V. unguiculata and C. dactylon, respectively, with no short-term negative impacts. Only wood BC avoided soil P build-up. BC application increased the soil total C, showing potential for C sequestration.

Biochar and Dairy Manure Amendment Effects on Cynodon dactylon Performance and Soil Properties.

Studies have determined the separate effects of biochar (BC) and manure application on forage species and soil, but few examined the effects of BCs made from different feedstock applied along with dairy manure. We compared the effect of wood- and manure-derived feedstock BC as well as dairy manure amendment application on Cynodon dactylon performance and soil properties in sandy loam and clay loam soils in a greenhouse pot study. Plant samples were assayed for herbage and root dry weight as well as herbage and root N and C percent and yield. Soil samples were assayed for macronutrients, micronutrients, metals, pH and conductivity. Data analyses involved variance analysis and Tukey's tests using R in RStudio (the IDE). In general, C. dactylon yields or mineral content were not affected by either manure or BC. However, an increase in the total herbage dry weight (30%) and in herbage N% (55%) was observed for clay loam and sandy loam soil, respectively, due to manure amendment application. There were no alterations in clay loam NO3-N and P due to any treatment; however, in sandy loam, these nutrients were not altered only when wood BC was applied. In sandy loam soil, NO3-N and P increased when manure BC along with dairy manure and when manure BC alone were applied, respectively. Thus, wood BC application should be considered to avoid these nutrient buildups when dairy manure is used as a soil amendment. This research shows a neutral (BC) or positive (dairy manure amendment) impact on C. dactylon performance. BC incorporation increases soil total C, showing potential for C sequestration. Long-term field trials could corroborate plant performance and soil parameters.

Extracellular matrix complexity in biomarker studies: a novel assay detecting total serum tenascin-C reveals different distribution to isoform-specific assays.

Serum biomarkers are the gold standard in non-invasive disease diagnosis and have tremendous potential as prognostic and theranostic tools for patient stratification. Circulating levels of extracellular matrix molecules are gaining traction as an easily accessible means to assess tissue pathology. However, matrix molecules are large, multimodular proteins that are subject to a vast array of post-transcriptional and post-translational modifications. These modifications often occur in a tissue- and/or disease-specific manner, generating hundreds of different variants, each with distinct biological roles. Whilst this complexity can offer unique insight into disease processes, it also has the potential to confound biomarker studies. Tenascin-C is a pro-inflammatory matrix protein expressed at low levels in most healthy tissues but elevated in, and associated with the pathogenesis of, a wide range of autoimmune diseases, fibrosis, and cancer. Analysis of circulating tenascin-C has been widely explored as a disease biomarker. Hundreds of different tenascin-C isoforms can be generated by alternative splicing, and this protein is also modified by glycosylation and citrullination. Current enzyme-linked immunosorbent assays (ELISA) are used to measure serum tenascin-C using antibodies, recognising sites within domains that are alternatively spliced. These studies, therefore, report only levels of specific isoforms that contain these domains, and studies on the detection of total tenascin-C are lacking. As such, circulating tenascin-C levels may be underestimated and/or biologically relevant isoforms overlooked. We developed a highly specific and sensitive ELISA measuring total tenascin-C down to 0.78ng/ml, using antibodies that recognise sites in constitutively expressed domains. In cohorts of people with different inflammatory and musculoskeletal diseases, levels of splice-specific tenascin-C variants were lower than and distributed differently from total tenascin-C. Neither total nor splice-specific tenascin-C levels correlated with the presence of autoantibodies to citrullinated tenascin-C in rheumatoid arthritis (RA) patients. Elevated tenascin-C was not restricted to any one disease and levels were heterogeneous amongst patients with the same disease. These data confirm that its upregulation is not disease-specific, instead suggest that different molecular endotypes or disease stages exist in which pathology is associated with, or independent of, tenascin-C. This immunoassay provides a novel tool for the detection of total tenascin-C that is critical for further biomarker studies. Differences between the distribution of tenascin-C variants and total tenascin-C have implications for the interpretation of studies using isoform-targeted assays. These data highlight the importance of assay design for the detection of multimodular matrix molecules and reveal that there is still much to learn about the intriguingly complex biological roles of distinct matrix proteoforms.

Genome Sequence of Fowlpox Virus-Integrated Reticuloendotheliosis Virus from a Rio Grande Wild Turkey (Meleagris gallopavo intermedia).

We report the genome sequence of a nearly intact reticuloendotheliosis virus (REV) insertion within a field strain of fowlpox virus from a Rio Grande wild turkey in Gillespie County, TX. The proviral REV genome comprises 7,943 bp and contains partial long terminal repeats.

Near-Complete Proviral Genome Sequence of Reticuloendotheliosis Virus Isolated from an Attwater's Prairie Chicken (Tympanuchus cupido attwateri).

We report the near-complete proviral genome sequence of a reticuloendotheliosis virus isolated and propagated from an endangered Attwater's prairie chicken (Tympanuchus cupido attwateri) during a 2016-2017 outbreak at a captive breeding facility.

A new male sling for minimal to moderate incontinence and climacturia.

Erectile dysfunction (ED) and stress urinary incontinence (SUI) are known bothersome sequelae to radical prostatectomy. In recent years, additional attention has been placed on another less commonly described and reported side effect to this surgery, climacturia. While various noninvasive and surgical interventions have been described for the management of climacturia, until recently, none has provided reliable and meaningful results. In the past few years, the Mini-Jupette sling has gained popularity as an adjunct to inflatable penile prosthesis placement in patients with concomitant ED and climacturia. Recent data have also suggested its feasibility in patients with mild SUI. While the original technique described by Pr. Andrianne has shown long-term, reproducible, and safe results, innovative modifications such as the use of autologous rectus fascia, the Male Urethral Mini-Sling and the Mayo Clinic modified Mini-Jupette sling have also been suggested, proof of widespread interest amongst clinicians toward achieving the optimum surgical technique.

Transporting and Exercising Unconditioned Horses: Effects on Microflora Populations.

The objective of this study was to determine if transportation and exercise stress in horses affect the microflora populations in the equine hindgut. Four horses were subjected to three transport periods (0, 3, and 6 hours) with a 7-d rest period between each transport. Horses were fed 0.91 kg/day of Purina Impact All Stages 12% and had ad libitum access to Cynodon dactylon (Coastal Bermudagrass) hay. Fecal samples were collected before (0 hours) and after (48 hours) transport. In addition, three horses underwent a different standardized exercise test with a 7-d rest period between each exercise. Standardized exercise test intensity was determined by heart rate to validate if the horse was in aerobic or anaerobic work. The protocol for fecal sample collection after exercise was the same as for transport. Prokaryotic community profiling was conducted by 16S metagenomic analysis. After DNA evaluation, differences were found in the microbiome at transport 0 hours and grouped transport 3 hours time 48 and transport 6 hours time 48 (PERMANOVA P = .037) where Bacteroidetes increased 48 hours after transport and Firmicutes decreased 48 hours after transport. Exercise microbial communities showed no difference in either alpha or beta diversity when compared with controls (0 hours). In the present study, difference in microflora may have resulted from stress duration of transport rather than stress duration of exercise.

An observational study of matrix metalloproteinase (MMP)-9 in cystic fibrosis.

BACKGROUND: In cystic fibrosis (CF), cross-sectional studies have reported sputum matrix metalloproteinase (MMP)-9 to be elevated and negatively correlated with FEV1. This longitudinal study examined the association between MMP-9 and tissue inhibitors of metalloproteinases (TIMPs) to prognostic parameters in CF. METHOD: A cross-sectional survey of CF and control subjects; CF patients were followed up for a median of 49 months. MMP-9 and TIMP-1 and TIMP-2 were quantified in sputum and plasma. RESULTS: Seventy-three patients with CF, median age 22 years, and 40 controls were recruited. Fifty-three of these CF patients were followed up. Prospectively, in CF subjects, plasma MMP-9 activity was adversely associated with FEV1 (ß -1.15 (95% CI -2.10, -0.20), p = 0.019) and rate of FEV1 decline, and plasma TIMP-1 was adversely associated with mortality: hazard ratio 3.66 (1.91-7.04), p < 0.001. CONCLUSIONS: These associations further justify investigation of MMP-9 and TIMP-1 as biomarkers for short- to medium-term FEV1 decline and mortality in patients with CF.

Quantitative assessment of Naegleria fowleri and Escherichia coli concentrations within a Texas reservoir.

Previous presence/absence studies have indicated a correlation between the presence of the pathogenic amoeba Naegleria fowleri and the presence of bacteria, such as the fecal indicator Escherichia coli, in environmental surface waters. The objective of this study was to use quantitative real-time polymerase chain reaction (qPCR) methodologies to measure N. fowleri and E. coli concentrations within a Texas reservoir in late summer, and to determine if concentrations of N. fowleri and E. coli were statistically correlated. N. fowleri was detected in water samples from 67% of the reservoir sites tested, with concentrations ranging up to an estimated 26 CE (cell equivalents)/100 mL. E. coli was detected in water samples from 60% of the reservoir sites tested, with concentrations ranging up to 427 CE/100 mL. In this study, E. coli concentrations were not indicative of N. fowleri concentrations.

The pursuit of better diagnostic performance: a human factors perspective.

Despite the relatively slow start in treating diagnostic error as an amenable research topic at the beginning of the patient safety movement, interest has steadily increased over the past few years in the form of solicitations for research, regularly scheduled conferences, an expanding literature and even a new professional society. Yet improving diagnostic performance increasingly is recognised as a multifaceted challenge. With the aid of a human factors perspective, this paper addresses a few of these challenges, including questions that focus on who owns the problem, treating cognitive and system shortcomings as separate issues, why knowledge in the head is not enough, and what we are learning from health information technology (IT) and the use of checklists. To encourage empirical testing of interventions that aim to improve diagnostic performance, a systems engineering approach making use of rapid-cycle prototyping and simulation is proposed. To gain a fuller understanding of the complexity of the sociotechnical space where diagnostic work is performed, a final note calls for the formation of substantive partnerships with those in disciplines beyond the clinical domain.

Probe-based real-time PCR method for multilocus melt typing of Xylella fastidiosa strains.

Epidemiological studies of Pierce's disease (PD) can be confounded by a lack of taxonomic detail on the bacterial causative agent, Xylella fastidiosa (Xf). PD in grape is caused by strains of Xylella fastidiosa subsp. fastidiosa, but is not caused by other subspecies of Xf that typically colonize plants other than grape. Detection assays using ELISA and qPCR are effective at detecting and quantifying Xf presence or absence, but offer no information on Xf subspecies or strain identity. Surveying insects or host plants for Xf by current ELISA or qPCR methods provides only presence/absence and quantity information for any and all Xf subspecies, potentially leading to false assessments of disease threat. This study uses a series of adjacent-hybridizing DNA melt analysis probes that are capable of efficiently discriminating Xf subspecies and strain relationships in rapid real-time PCR reactions.

Coincident light and clock regulation of pseudoresponse regulator protein 37 (PRR37) controls photoperiodic flowering in sorghum.

Optimal flowering time is critical to the success of modern agriculture. Sorghum is a short-day tropical species that exhibits substantial photoperiod sensitivity and delayed flowering in long days. Genotypes with reduced photoperiod sensitivity enabled sorghum's utilization as a grain crop in temperate zones worldwide. In the present study, Ma(1), the major repressor of sorghum flowering in long days, was identified as the pseudoresponse regulator protein 37 (PRR37) through positional cloning and analysis of SbPRR37 alleles that modulate flowering time in grain and energy sorghum. Several allelic variants of SbPRR37 were identified in early flowering grain sorghum germplasm that contain unique loss-of-function mutations. We show that in long days SbPRR37 activates expression of the floral inhibitor CONSTANS and represses expression of the floral activators Early Heading Date 1, FLOWERING LOCUS T, Zea mays CENTRORADIALIS 8, and floral induction. Expression of SbPRR37 is light dependent and regulated by the circadian clock, with peaks of RNA abundance in the morning and evening in long days. In short days, the evening-phase expression of SbPRR37 does not occur due to darkness, allowing sorghum to flower in this photoperiod. This study provides insight into an external coincidence mechanism of photoperiodic regulation of flowering time mediated by PRR37 in the short-day grass sorghum and identifies important alleles of SbPRR37 that are critical for the utilization of this tropical grass in temperate zone grain and bioenergy production.

High-throughput DNA isolation method for detection of Xylella fastidiosa in plant and insect samples.

We report an inexpensive, high-throughput method for isolating DNA from insect and plant samples for the purpose of detecting Xylella fastidiosa infection. Existing methods often copurify inhibitors of DNA polymerases, limiting their usefulness for PCR-based detection assays. When compared to commercially available kits, the method provides enhanced pathogen detection at a fraction of the cost.

Detection and analysis of the bacterium, Xylella fastidiosa, in glassy-winged sharpshooter, Homalodisca vitripennis, populations in Texas.

The glassy-winged sharpshooter, Homalodisca vitripeninis Germar (Hemiptera: Cicadellidae), is a xylophagous insect that is an endemic pest of several economically important plants in Texas. H. vitripennis is the main vector of Xylella fastidiosa Wells (Xanthomonadales: Xanthomonadaceae), the bacterium that causes Pierce's disease of grapevine and can travel long distances putting much of Texas grape production at risk. Understanding the movement of H. vitripennis populations capable of transmitting X. fastidiosa into Pierce's-disease-free areas is critical for developing a management program for Pierce's disease. To that end, the USDA-APHIS has developed a program to sample vineyards across Texas to monitor populations of H. vitripennis. From this sampling, H vitripennis collected during 2005 and 2006 over the months of May, June, and July from eight vineyards in different regions of Texas were recovered from yellow sticky traps and tested for the presence of X. fastidiosa. The foregut contents were vacuum extracted and analyzed using RT-PCR to determine the percentage of H. vitripennis within each population that harbor X. fastidiosa and have the potential to transmit this pathogen. H. vitripennis from vineyards known to have Pierce's disease routinely tested positive for the presence of X. fastidiosa. While almost all H. vitripennis collected from vineyards with no history of Pierce's disease tested negative for the presence of the pathogen, three individual insects tested positive. Furthermore, all three insects were determined, by DNA sequencing, to be carrying a strain of X. fastidiosa homologous to known Pierce's disease strains, signifying them as a risk factor for new X. fastidiosa infections.

Optimization of total protein and activity assays for the detection of MMP-12 in induced human sputum.

BACKGROUND: Proteolysis of matrix components, in particular elastin, is a major contributing factor to the development of lung diseases such as emphysema and chronic obstructive pulmonary disease (COPD). MMP-12 (macrophage elastase) is a protease known to be involved in the progression of lung disease. The relatively low abundance of MMP-12 has precluded the development of quantitative assays that can accurately measure MMP-12 protein levels and activity across cohorts of healthy and diseased individuals. METHODS: Commercial antibodies were screened for performance in sandwich ELISA and capture FRET activity assay formats. Precision, accuracy, sensitivity, dilution linearity, and spike recovery were evaluated using sputum samples. RESULTS: Total protein and capture FRET activity assays were developed that were sensitive enough to detect MMP-12 in 37 of 38 donor sputum samples. A comparison of results between the two assays shows that a majority of sputum MMP-12 is in the active form. No differences were seen between normal, asthmatic, and COPD donors. CONCLUSION: Sensitive and quantitative assays for both MMP-12 activity and total protein in human induced sputum have been developed. These assays can be used to evaluate MMP-12 as a biomarker for lung disease, and to monitor efficacy of potential therapeutic compounds.

Matrix metalloproteinase-12 is a therapeutic target for asthma in children and young adults.

BACKGROUND: Matrix metalloproteinase (MMP)-12-mediated pathologic degradation of the extracellular matrix and the subsequent repair cycles influence the airway changes in patients with asthma and chronic obstructive pulmonary disease (COPD). The common serine variant at codon 357 of the MMP12 gene (rs652438) is associated with clinical manifestations consistent with more aggressive matrix degradation in other tissues. OBJECTIVE: We sought to explore the hypothesis that MMP12 represents a novel therapeutic target in asthma. METHODS: The role of the rs652438 variant on clinical phenotype was explored in young asthmatic patients and patients with COPD. Candidate MMP-12 inhibitors were identified on the basis of potency and selectivity against a panel of other MMPs. The role of MMP-12-specific inhibition was tested in vitro, as well as in animal models of allergic airway inflammation. RESULTS: The odds ratio for having greater asthma severity was 2.00 (95% CI, 1.24-3.24; P = .004) when comparing asthmatic patients with at least 1 copy of the serine variant with those with none. The carrier frequency for the variant increased in line with asthma treatment step (P = .000). The presence of the variant nearly doubled the odds in favor of asthmatic exacerbations (odds ratio, 1.90; 95% CI, 1.19-3.04; P = .008) over the previous 6 months. The serine variant was also associated with increased disease severity in patients with COPD (P = .016). Prior administration of an MMP-12-specific inhibitor attenuated the early airway response and completely blocked the late airway response with subsequent Ascaris suum challenge in sheep. CONCLUSION: Studies on human participants with asthma and COPD show that the risk MMP12 gene variant is associated with disease severity. In allergen-sensitized sheep pharmacologic inhibition of MMP12 downregulates both early and late airway responses in response to allergic stimuli.

Seasonal increase of Xylella fastidiosa in hemiptera collected from central Texas vineyards.

Yellow sticky traps were placed in six vineyards in central Texas from 2003 to 2006 and in locations outside the vineyards in 2004-2006. In total, 72 collections on 55 dates were examined. Xylem fluid-feeding insects were removed and identified to species and then analyzed by polymerase chain reaction to determine the presence or absence of Xylella fastidiosa Wells et al. Of the 1318 insects removed, 13 species were found, dominated by Homalodisca vitripennis (Germar), Clastoptera xanthocepahala Germar, and Graphocephala versuta (Say). Insects testing positive for X. fastidiosa were analyzed further using fluorescence resonance energy transfer probes to determine the genotype of the bacterium, which fell into four groups: subspecies fastidiosa, multiplex, sandyi, and unknown subspecies. Vineyards known to be affected by Pierce's disease had more insects that were contaminated by the bacterium than those that were not as affected. X. fastidiosa subsp. fastidiosa, the causative agent of Pierce's disease, was found more commonly in insects collected from vineyards than from insects collected outside the vineyards. Conversely, the subspecies multiplex and sandyi, which are not known to cause disease in grape, were more commonly found in insects collected outside the vineyard. The percentage of individuals contaminated with the bacterium increased over the course of the growing season, and the data suggest that vector insects acquired X. fastidiosa subsp. fastidiosa from infected grapevines, a necessary precursor for vine to vine transmission to occur. Management options, including the use of systemic insecticides and plant roguing, would be effective for this type of transmission model.

The "male lumpectomy": focal therapy for prostate cancer using cryoablation results in 48 patients with at least 2-year follow-up.

BACKGROUND: The use of breast sparing surgery, i.e., "lumpectomy", revolutionized management of breast cancer. Lumpectomy confirmed that quality of life issues can successfully be addressed without compromising treatment efficacy. Complications of prostate cancer treatment, including impotence and incontinence, affect the male self image no less than the loss of a breast does a woman. Traditional thinking held that prostate cancer was multifocal and therefore not amenable to a focal treatment approach. Recent pathology literature indicates, however, that up to 25% of prostate cancers are solitary and unilateral. This raises the question of whether these patients can be identified and treated with a limited "lumpectomy" or focal cancer treatment. METHODS: Focal cryoablation was planned to encompass the area of known tumor based on staging biopsies. PSAs were obtained every 3 months for 2 years and then every 6 months thereafter. RESULTS: Forty-eight patients with at least 2-year follow-up had focal cryoablation. Follow-up ranged from 2 years 10 years with a mean of 4.5 years; 45 of 48 patients (94%) have stable PSAs [American Society of Therapeutic Radiology and Oncology (ASTRO) criteria] with no evidence for cancer, despite 25 patients being medium to high risk for recurrence. Of the 24 patients with stable PSAs who were routinely biopsied (n = 24) all were negative. No local recurrences were noted in areas treated. Potency was maintained to the satisfaction of the patient in of 36 of 40 patients who were potent preoperatively. Of the 48, all were continent. CONCLUSION: These preliminary results indicate a "male lumpectomy" in which the prostate tumor region itself is destroyed, appears to preserve potency in a majority of patients and limits other complications (particularly incontinence), without compromising cancer control. If confirmed by further studies and long-term follow-up, this treatment approach could have a profound effect on prostate cancer management.

Active B12: a rapid, automated assay for holotranscobalamin on the Abbott AxSYM analyzer.

BACKGROUND: Conventional tests for vitamin B(12) deficiency measure total serum vitamin B12, whereas only that portion of vitamin B12 carried by transcobalamin (holotranscobalamin) is metabolically active. Measurement of holotranscobalamin (holoTC) may be more diagnostically accurate for detecting B(12) deficiency that requires therapy. We developed an automated assay for holoTC that can be used on the Abbott AxSYM immunoassay analyzer. METHODS: AxSYM Active B12 is a 2-step sandwich microparticle enzyme immunoassay. In step 1, a holoTC-specific antibody immobilized onto latex microparticles captures holoTC in samples of serum or plasma. In step 2, the captured holoTC is detected with a conjugate of alkaline phosphatase and antiTC antibody. RESULTS: Neither apoTC nor haptocorrin exhibited detectable cross-reactivity. The detection limit was < or = 0.1 pmol/L. Within-run and total imprecision (CV ranges) were 3.4%-5.1% and 6.3%-8.5%, respectively. Assay CVs were < 20% from at least 3 pmol/L to 107 pmol/L. With diluted serum samples, measured concentrations were 104%-114% of the expected values in the working range of the assay. No interference from bilirubin, hemoglobin, triglycerides, erythrocytes, rheumatoid factor, or total protein was detected at expected (abnormal) concentrations. A comparison of the AxSYM Active B12 assay with a commercial RIA for holoTC yielded the regression equation: AxSYM = 0.98RIA + 4.7 pmol/L (S(y x), 11.4 pmol/L; n = 204). Assay throughput was 45 tests/h. A 95% reference interval of 19-134 pmol/L holoTC was established with samples from 292 healthy individuals. CONCLUSIONS: The AxSYM Active B12 assay allows rapid, precise, sensitive, specific, and automated measurement of human holoTC in serum and plasma.

Black soldier fly (Diptera: Stratiomyidae) larvae reduce Escherichia coli in dairy manure.

Escherichia coli labeled with a green fluorescent protein was inoculated into sterile dairy manure at 7.0 log cfu/g. Approximately 125 black soldier fly larvae were placed in manure inoculated and homogenized with E. coli. Manure inoculated with E. coli but without black soldier fly larvae served as the control. For the first experiment, larvae were introduced into 50, 75, 100, or 125 g sterilized dairy manure inoculated and homogenized with E. coli and stored 72 h at 27 degrees C. Black soldier fly larvae significantly reduced E. coli counts in all treatments. However, varying the amount of manure provided the black soldier fly larvae significantly affected their weight gain and their ability to reduce E. coli populations present. For the second experiment, larvae were introduced into 50 g manure inoculated with E. coli and stored for 72 h at 23, 27, 31, or 35 degrees C. Minimal bacterial growth was recorded in the control held at 35 degrees C and was excluded from the analysis. Black soldier fly larvae significantly reduced E. coli counts in manure held at remaining temperatures. Accordingly, temperature significantly influenced the ability of black soldier fly larvae to develop and reduce E. coli counts with greatest suppression occurring at 27 degrees C.