Discrimination of GutCheck(NEC): a clinical risk index for necrotizing enterocolitis.

OBJECTIVE: Better measures are needed to identify infants at risk for developing necrotizing enterocolitis (NEC) and facilitate communication about risk across transitions. Although NEC is multi-factorial, quantification of composite risk for NEC in an individual infant is not clearly defined. The objective of this study was to describe the derivation, validation and calibration testing of a novel clinical NEC risk index, GutCheck(NEC). Individual risk factors were weighted to assess composite odds of developing NEC. GutCheck(NEC) is designed to improve communication about NEC risk and coordination of care among clinicians across an infant's clinical course. STUDY DESIGN: On the basis of a synthesis of research evidence about NEC risk and an e-Delphi study including 35 neonatal experts, we identified NEC risk factors believed by the experts to be most relevant for a NEC risk index, then applied a logistic model building process to derive and validate GutCheck(NEC). De-identified data from the Pediatrix BabySteps Clinical Data Warehouse (discharge date 2007 to 2011) were split into three samples for derivation, validation and calibration. By comparing infants with medical NEC, surgical NEC and those who died to infants without NEC, we derived the logistic model using the un-matched derivation set. Discrimination was then tested in a case-control matched validation set and an un-matched calibration set using receiver operating characteristic curves. RESULT: Sampled from a cohort of 58 820 infants, the randomly selected derivation set (n=35 013) revealed nine independent risk factors (gestational age, history of packed red blood cell transfusion, unit NEC rate, late-onset sepsis, multiple infections, hypotension treated with inotropic medications, Black or Hispanic race, outborn status and metabolic acidosis) and two risk reducers (human milk feeding on both days 7 and 14 of life, and probiotics). Unit NEC rate carried the most weight in the summed score. Validation using a 2:1 matched case-control sample (n=360) demonstrated fair to good discrimination. In the calibration set (n=23 447), GutCheck(NEC) scores (range 0 to 58) discriminated those infants who developed surgical NEC (area under the curve (AUC)=0.84, 95% confidence interval (CI) 0.82 to 0.84) and NEC leading to death (AUC=0.83, 95% CI 0.81 to 0.85), more accurately than medical NEC (AUC= 0.72, 95% CI 0.70 to 0.74). CONCLUSION: GutCheck(NEC) represents weighted composite risk for NEC and discriminated infants who developed NEC from those who did not with very good accuracy. We speculate that targeting modifiable NEC risk factors could reduce national NEC prevalence.

Forest restoration potentials of coal-mined lands in the eastern United States.

The Appalachian region in the eastern United Sates is home to the Earth's most extensive temperate deciduous forests, but coal mining has caused forest loss and fragmentation. More than 6000 km in Appalachia have been mined for coal since 1980 under the Surface Mining Control and Reclamation Act (SMCRA). We assessed Appalachian areas mined under SMCRA for forest restoration potentials. Our objectives were to characterize soils and vegetation, to compare soil properties with those of pre-SMCRA mined lands that were reforested successfully, and to determine the effects of site age on measured properties. Soils were sampled and dominant vegetation characterized at up to 10 points on each of 25 post-SMCRA mines. Herbaceous species were dominant on 56%, native trees on 24%, and invasive exotics on 16% of assessed areas. Mean values for soil pH (5.8), electrical conductivity (0.07 dS m(-1)), base saturation (89%), and coarse fragment content (50% by mass) were not significantly different from measured levels on the pre-SMCRA forested sites, but silt+clay soil fraction (61%) was higher, bicarbonate-extractable P (4 mg kg(-1)) was lower, and bulk density (1.20 g cm(-1)) was more variable and often unfavorable. Pedogenic N and bicarbonate-extractable P in surface soils increased with site age and with the presence of weathered rocks among coarse fragments. Our results indicate a potential for many of these soils to support productive forest vegetation if replanted and if cultural practices, including temporary control of existing vegetation, soil density mitigation, and fertilization, are applied to mitigate limitations and aid forest tree reestablishment and growth.

The Perfect Stage of Powdery Mildew (Erysiphe polygoni) of Beta vulgaris Found in Michigan.

Powdery mildew (Erysiphe polygoni DC [synonym E. betae {Vanha} Weltzien]) affects several different crops of Beta vulgaris, including sugar beet, Swiss chard, and table beet. The disease has been prevalent in many sugar beet-growing areas of the United States since the first major epidemic in beet in 1974 (3). Powdery mildew in the United States was primarily associated with the asexual stage of the pathogen until the perfect stage was found, first in western states such as Idaho and Colorado (2), then in more Midwestern states such as Nebraska, and most recently in North Dakota (1). Similar to North Dakota, powdery mildew has not been a major problem in the Michigan growing area. It does appear sporadically, particularly on sugar beets that have not been sprayed to control other foliar diseases. In 2010, powdery mildew infection on sugar beet was observed in late August in a field in the Saginaw Valley of Michigan. Plants were inspected periodically for the presence of the sexual stage. In early October, sugar beet and Swiss chard plants with heavy powdery mildew infection also were observed at the Michigan State University (MSU) Horticultural Demonstration Gardens in East Lansing and on sugar beet at the MSU Plant Pathology and Botany research farms. On both the Saginaw Valley sugar beet and Swiss chard on the MSU campus, ascomata were observed on a few leaves in mid-October. No ascomata were found on sugar beet at the other two locations. The majority of ascomata were dark brown to black when located, although a few light tan ascomata were observed on the Swiss chard. Ascomata varied from 70 to 100 µm in diameter. Asci contained one to four hyaline or golden yellow ascospores similar to those described in other growing regions on sugar beet (1,2). No ascomata had been detected on powdery mildew-infected sugar beet from either the Saginaw Valley or the MSU research farms the previous two years. These results appear to indicate a spread of the ability to form the perfect stage eastward from the western United States. This may be due to movement of one mating type because E. polygoni has been reported to be heterothallic on some crops (4). The presence of the perfect stage indicates that sexual recombination could occur in E. polygoni on Beta species in Michigan, creating the potential for more rapid development of new strains that might vary in fungicide sensitivity and response to host resistance. References: (1) C. A. Bradley et al. Plant Dis. 91:470, 2007 (2) J. J. Gallian and L. E. Hanson. Plant Dis. 87:200, 2003. (3) E. G. Ruppel. Page 13 in: Compendium of Beet Disease and Insects. E. D. Whitney and J. E. Duffus, eds. The American Phytopathological Society, St. Paul, MN, 1986. (4) C. G. Smith. Trans. Br. Mycol. Soc. 55:355, 1970.

Development and characterization of potato-Solanum brevidens chromosomal addition/substitution lines.

Solanum brevidens is a wild diploid potato species possessing high levels of resistances to several major potato diseases. We previously developed fertile somatic hybrids between S. brevidens and the cultivated potato (Solanum tuberosum) in order to introgress disease resistances from this wild species into potato. A series of backcross progenies was developed from a hexaploid somatic hybrid A206. Using a combination of S. brevidens-specific randomly amplified polymorphic DNA (RAPD) markers and a sequential genomic in situ hybridization (GISH) and fluorescence in situ hybridization (FISH) technique, we identified all 12 S. brevidens chromosomes in the backcross progenies. Seven potato-S. brevidens monosomic chromosome addition lines (chromosomes 1, 3, 4, 5, 8, 9 and 10) and one monosomic substitution line (chromosome 6) were identified, and the remaining four S. brevidens chromosomes (2, 7, 11, and 12) were included in two other lines. These chromosomal addition/substitution stocks provide valuable tools for potato cytogenetic research, and can be used to introgress disease resistances from S. brevidens into potato.

Surface runoff losses of phosphorus from Virginia soils amended with turkey manure using phytase and high available phosphorus corn diets.

Many states have passed legislation that regulates agricultural P applications based on soil P levels and crop P uptake in an attempt to protect surface waters from nonpoint P inputs. Phytase enzyme and high available phosphorus (HAP) corn supplements to poultry feed are considered potential remedies to this problem because they can reduce total P concentrations in manure. However, less is known about their water solubility of P and potential nonpoint-source P losses when land-applied. This study was conducted to determine the effects of phytase enzyme and HAP corn supplemented diets on runoff P concentrations from pasture soils receiving surface applications of turkey manure. Manure from five poultry diets consisting of various combinations of phytase enzyme, HAP corn, and normal phytic acid (NPA) corn were surface-applied at 60 kg P ha(-1) to runoff boxes containing tall fescue (Festuca arundinacea Schreb.) and placed under a rainfall simulator for runoff collection. The alternative diets caused a decrease in manure total P and water soluble phosphorus (WSP) compared with the standard diet. Runoff dissolved reactive phosphorus (DRP) concentrations were significantly higher from HAP manure-amended soils while DRP losses from other manure treatments were not significantly different from each other. The DRP concentrations in runoff were not directly related to manure WSP. Instead, because the mass of manure applied varied for each treatment causing different amounts of manure particles lost in runoff, the runoff DRP concentrations were influenced by a combination of runoff sediment concentrations and manure WSP.

Differential induction of glyoxylate cycle enzymes by stress as a marker for seedling vigor in sugar beet (Beta vulgaris).

Significant differences in seedling vigor exist among sugar beet (Beta vulgaris) hybrids; however, traditional approaches to breeding enhanced vigor have not proven effective. Seedling vigor is a complex character, but presumably includes efficient mobilization of seed storage reserves during germination and efficient seedling growth in diverse environments. The involvement of lipid metabolism during germination of sugar beet under stress conditions was suggested by the isolation at high frequency of Expressed Sequence Tags (ESTs) with similarity to isocitrate lyase (EC 4.1.3.1). High-level expression of this glyoxylate cycle enzyme during germination and seedling emergence was also suggested by nucleotide sequencing of cDNA libraries obtained from a well emerging sugar beet hybrid during germination under stress. Genes involved in carbohydrate and lipid catabolism were differentially expressed in a strongly emerging hybrid, relative to a weakly emerging hybrid, during stress germination. Stress markedly reduced the levels of alpha-amylase transcripts in the weakly emerging hybrid. In contrast, the strongly emerging hybrid exhibited only a moderate reduction in alpha-amylase transcript levels under the same conditions, and showed large increases in the expression of genes involved in lipid metabolism, suggesting compensation by lipid for carbohydrate metabolism in the better emerging hybrid. Differential activity of the glyoxylate cycle thus appears to be a physiological marker that distinguishes between high- and low-vigor sugar beet cultivars. This finding suggests, for the first time, a biochemical target for selection for enhanced germination and improved emergence in sugar beet.

A snapshot of the low temperature stress transcriptome of developing rice seedlings (Oryza sativa L.) via ESTs from subtracted cDNA library.

Rice (Oryza sativa L.) is sensitive to chilling particularly during early seedling development. Given the biochemical complexity of tolerance mechanisms, genetic potential for this trait depends on highly coordinated expression of many genes. We used a simple cDNA subtraction strategy to develop Expressed Sequence Tags (ESTs) that represent an important subset of cold stress-upregulated genes. The 3,084 subtracted cDNA clones represent a total of 1,967 unigenes from 1,354 singletons and 613 contigs. As expected in the developing seedlings, genes involved in basic cellular processes, i.e., metabolism, growth and development, protein synthesis, folding and destination, cellular transport, cell division and DNA replication were widely represented. Genes with stress-related and regulatory functions comprised 23.17% of the total ESTs. These categories included proteins with known function in cellular defenses against abiotic (drought, cold and salinity) and biotic (pathogen) stresses, and proteins involved in developmental and stress response signalling and transcription. Based on the types of genes represented, tolerance mechanisms rely on precise integration of developmental processes with stress-related responses. A large fraction of the ESTs (38.7%) represents unknown proteins. This EST library is a rich source of cold stress-related genes, and supplements for other publicly available libraries for comprehensive analysis of the stress-response transcriptome.

Risk factors as the basis for triage in low-resource cervical cancer screening programs.

OBJECTIVES: Several studies have shown impressive sensitivity and specificity of visual inspection using acetic acid wash (VIA) in detecting cervical dysplasia. This study examines how risk-factor based triage, using clinical history-taking to limit the testing to those at highest risk of disease, improves the clinical utility of VIA. METHODS: The study population included 2206 woman aged 25-56 from peri-urban primary care clinics in Zimbabwe. Three risk factors variables [age, lifetime number of sexual partners and history of a sexually transmitted infection (STI)] were selected as the basis for computer-simulated patient triage. Criteria for selecting risk factors were biological relevance, programmatic feasibility, historical evidence in the scientific literature as a risk factor for cervical cancer and a significant (P<0.05) predictor of dysplasia in this study population. Predictive values for VIA were calculated on subgroups of women with combinations of the three risk factors and then compared with the predictive value of VIA calculated on the full study population. RESULTS: The positive predictive value (PPV) among women with all three risk factors (30.2%) was 1.6 times higher than that of all study women and 4.5 times higher than women with none of the three risk factors (4.1%). Additionally, the PPV was slightly (30%) higher than that obtained from computer-simulated serial testing using VIA as the primary test followed by HPV testing (27.4%). CONCLUSION: Clinical history-taking could provide the basis for patient triage to increase the clinical utility of VIA. Such an approach could also serve to prioritize who gets tested first in low-resource countries with high disease prevalence that are struggling to strengthen national cervical cancer prevention programs.

The genetic identity of alien chromosomes in potato breeding lines revealed by sequential GISH and FISH analyses using chromosome-specific cytogenetic DNA markers.

Genomic in situ hybridization (GISH) is one of the most popular and effective techniques for detecting alien chromatin introgressed into breeding lines; however, GISH analysis alone does not reveal the genetic identity of the alien chromosomes. We previously isolated a set of bacterial artificial chromosomes (BACs) specific to each of the 12 potato chromosomes. These BAC clones can be used as chromosome-specific cytogenetic DNA markers (CSCDMs) for potato chromosome identification. Here we demonstrate that GISH and fluorescence in situ hybridization (FISH), using CSCDMs, can be performed sequentially on the same chromosome preparations. Somatic metaphase chromosomes prepared using an enzymatic digestion and "flame-drying" procedure allows repeated probing up to five times without significant damage to chromosome morphology. The sequential GISH and FISH analyses reveal the genomic origin and genetic identity of the alien chromosomes in a single experiment and also determine whether an alien chromosome has been added to the genetic background of potato or is substituting for a homoeologous potato chromosome. The sequential GISH and FISH procedures should be widely applicable for germplasm characterization, especially in plant species with small-sized chromosomes.

Analysis of the introgression of Solanum bulbocastanum DNA into potato breeding lines.

Somatic hybrids have been obtained between potato and Solanum bulbocastanum PI 245310, a Mexican diploid (2n=2x=24) species. Through restriction fragment length polymorphism (RFLP) and randomly amplified polymorphic DNA (RAPD) analyses it was found that the somatic hybrids contain each chromosome of the diploid parent and that the synteny of RFLP markers noted with tomato, potato and S. brevidens is largely maintained in S. bulbocastanum. RFLP analyses of BC1 progeny of two different hybrids indicated that a substantial number of markers were either lost or were heterozygous, in marked contrast with results previously noted with S. brevidens. A RAPD map for all 12 chromosomes of S. bulbocastanum was prepared and marker transmission was followed in three BC2 populations. Results with chromosomes 3, 8 and 10 from these populations are compared.

Building relationships with families in the NICU: exploring the guarded alliance.

Extremely low-birthweight infants and their families experience prolonged hospitalization in the newborn intensive care unit (NICU). Prolonged hospitalization is particularly stressful for the family, leading to distress in most aspects of their lives, including separation of parents from each other and from other family members, a need for increased psychological support, and increased economic concerns. Based on these characteristics, these families appear to have needs more similar to that of families caring for chronically ill members. Indeed, this initial hospitalization may be the prelude to coping and surviving with a chronic illness for many of these children and families. This article proposes the adaption of "guarded alliance" as a framework for developing relationships and providing care to families in the NICU. This framework was derived from research with families of the chronically ill. Supportive research is used to justify the adoption of this framework for interactions with families in the NICU. In addition, linkages to family-centered care principles in critical care settings are also discussed.

Nutritive sucking and neurobehavioral development in preterm infants from 34 weeks PCA to term.

PURPOSE: To investigate changes in nutritive sucking patterns, behavioral state, and neurobehavioral development of preterm infants from the 34 weeks postconceptional age (PCA) to term. DESIGN: Nonexperimental descriptive correlational design was used. METHODS: A convenience sample included 66 preterm infants with a gestational age between 24 and 34 weeks at birth. Mean gestational age was 29.7 weeks, with a mean birthweight of 1379 grams. Instruments included the Kron Nutritive Sucking Apparatus, the Anderson Behavioral State Scale (ABSS), and the Brazelton Neonatal Behavioral Assessment Scale (BNBAS). A feeding procedure was administered at 34 and 40 weeks PCA with BNBAS administered at 40 weeks PCA. Data analysis strategies included t-tests, Spearman rho correlations, Wilcoxon matched-pairs signed-ranks test, and a Random Coefficients Model. RESULTS: There were significant differences in number of sucks (p < 0.001), intensity of sucking pressures (p < 0.001), average time between sucks (p < 0.001), and average time between bursts (p < 0.001) from 34 weeks PCA to term. With maturation, it was noted that the preterm infants were significantly more alert during the sucking protocol from 34 weeks to term (p < 0.001). Yet, infant sucking parameters at 34 weeks PCA and term were not significantly related to the BNBAS or to the infant's state. CLINICAL IMPLICATIONS: These results demonstrate that high risk preterm infants mature significantly in their sucking behaviors and in their ability to maintain a more alert behavioral state from 34 weeks PCA to term. However, unlike full term infants--where state is positively correlated to feeding (sucking) success--the preterm infant state of alertness may not enhance sucking skills at 34 weeks PCA. These infants may be too stressed to manage both feeding and stimulation from the environment that is unavoidable when alert. However, this relationship does appear to change by 40 weeks PCA.

Developmentally supportive caregiving and technology in the NICU: isolation or merger of intervention strategies?

Providing nursing care in the highly technical environment of the neonatal intensive care unit is changing as concepts of developmentally supportive caregiving merge with conventional nursing interventions and are integrated into routine practice. Medical knowledge with the use of technology is insufficient in guiding the whole of neonatal nursing care. Integration of other patterns and concepts is essential to expert nursing practice with high risk infants and their families.

Physicians' perspectives on communicating prescription drug information.

Physician-patient communication has been identified as one factor that influences prescription drug compliance rates, even though the nature of such communication has not been well described. This study used a qualitative approach (in-depth interviews with 20 physicians representing a variety of medical areas) to better understand the process by which physicians communicate prescription drug information to their patients. A content analysis of interview transcripts indicated that two-way interactions focusing on the specific needs of the patients should be encouraged over simply disseminating more and more information about prescriptions. Information regarding unlikely outcomes was suggested to cause patient anxiety, which might be tied to decreased compliance rates. Physicians stressed the importance of "patient-directed communication" and expressed their concern over what they considered to be inappropriate sources of drug information. Other specific reasons cited for noncompliance included forgetfulness, ignoring medication when feeling better, and apprehensions about side effects.

Methylation of somatic and sperm DNA in the homosporous fern Ceratopteris richardii.

Plants, in general, have a high proportion of their CpG and CpNpG nucleotide motifs modified with 5-methylcytosine (5mC). Developmental changes in the proportion of 5mC are evident in mammals, particularly during gametogenesis and embryogenesis, but little information is available from flowering plants due to the intimate association of gametes with sporophytic tissues. In ferns, sperm are uninucleate and free-swimming and thus are easily isolated. We have examined 5mC in DNA isolated from fern sperm and other tissues with methylation-sensitive and -insensitive restriction enzyme isoschizomers, Southern blots probed with chloroplast and nuclear ribosomal RNA genes and end-labeled restriction fragments. We conclude that fern sperm DNA is methylated to a similar or greater degree than DNA isolated from either sporophytes or gametophytes.

Integrating family-centered developmental assessment and intervention into routine care in the neonatal intensive care unit.

Incorporating developmental intervention into routine practices requires time available during caregiving for continued infant assessment and intervention and requires time after and between caregiving for relief of infant distress. In an era of health-care reform, reorganization, and restructuring, this added caregiver time appears unavailable. However, nurses need to work together in meeting the developmental challenges of the NICU. Further research needs to be done to validate which interventions are appropriate for which babies and with what medical procedures. In addition, collaboration and sharing of responsibilities and resources with all care providers needs to be investigated. Moreover, research needs to be done that acknowledges that the environment of the NICU is also a world in which many adults work and live a large portion of their daily life. This environment must be supportive of their social needs as well as the needs of the high-risk infant. There are many pieces to the puzzle of providing developmentally supportive caregiving in the NICU. The number of different issues increases the complexity of changing the standard of care: Infant, family, environment. Each has its own challenges. However, with sensitivity, a collaborative approach, and a sincere effort to change, neonatal health-care professionals can integrate developmental practices into the NICU.

Implementing a research-based kangaroo care program in the NICU.

Kangaroo care or skin-to-skin holding of preterm infants requires consistent implementation for best outcomes with infants and families. Successful implementation of a project of this type demands an organized approach. This article describes how a standard of care was developed using research findings and then implemented with a step-by-step approach in the neonatal intensive care unit.

Segregation and recombination of Solanum brevidens synteny groups in progeny of somatic hybrids with S. tuberosum: intragenomic equals or exceeds intergenomic recombination.

The Solanum brevidens genome (2n = 2x = 24) was examined with randomly applied polymorphic DNA (RAPD) markers in a second backcross population derived from a S. brevidens + S. tuberosum somatic hybrid. RAPD markers cosegregated into 12 different S. brevidens synteny groups. Most synteny groups were nonrecombinant. However, nearly 40% of the S. brevidens synteny groups detected in this population were recombinant deletions that carried at least one, but not all, synteny group-specific RAPD markers. All S. brevidens synteny groups (except chromosome 5) were involved in recombination, and recombination occurred within most intervals between markers. About 20% of the recombinant S. brevidens synteny groups involved a single synteny group-specific marker. The inheritance of some single-marker representatives was followed in four BC3 families. At least nine changes in S. brevidens synteny groups had occurred during backcrossing. Six of the nine changes involved translocation of S. brevidens markers between nonhomologous S. brevidens chromosomes, and three S. brevidens markers may have been introgressed into the potato genome.

An evaluation of the effect of anesthetic technique on reproductive success after laparoscopic pronuclear stage transfer. Propofol/nitrous oxide versus isoflurane/nitrous oxide.

BACKGROUND: Laparoscopic pronuclear stage transfer (PROST) is the preferred method of embryo transfer after in vitro fertilization in many infertility programs. There are scant data to recommend the use or avoidance of any particular anesthetic agent for use in women undergoing this procedure. The authors hypothesized that propofol would be an ideal anesthetic for laparoscopic PROST because of its characteristic favorable recovery profile that includes minimal sedation and a low incidence of postoperative nausea and vomiting. The purpose of the study was to compare propofol and isoflurance with respect to postanesthetic recovery and pregnancy outcomes after laparoscopic PROST. METHODS: One hundred twelve women scheduled for laparoscopic PROST were randomized to receive either propofol/nitrous oxide or isoflurane/nitrous oxide for maintenance of anesthesia. RESULTS: Visual analog scale scores for sedation were lower in the propofol group than in the isoflurance group at all measurements between 30 min and 3 h after surgery. More women experienced emesis and were given an antiemetic during recovery in the isoflurance group than in the propofol group. However, the percentage of pregnancies with evidence of fetal cardiac activity was 54% in the isoflurane group compared with only 30% in the propofol group (P = 0.023). Also, the ongoing pregnancy rate was greater in the isoflurane group than in the propofol group (54% vs. 29%, P = 0.014). CONCLUSIONS: Propofol/nitrous oxide anesthesia was associated with lower clinical and ongoing pregnancy rates compared with isoflurane/nitrous oxide anesthesia.