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Verification of clonal identity of hop ( Humulus lupulus L.) cultivars within breeding programs and germplasm collections is vital to conserving genetic resources. Accurate and economic DNA-based tools are needed in dioecious hop to confirm identity and parentage, neither of which can be reliably determined from morphological observations. In this study, we developed two fingerprinting sets for hop: a 9-SSR fingerprinting set containing high-core repeats that can be run in a single PCR reaction and a kompetitive allele specific PCR (KASP) assay of 25 single nucleotide polymorphisms (SNPs). The SSR set contains a sex-linked primer pair, HI-AGA7, that was used to genotype 629 hop accessions from the US Department of Agriculture (USDA) National Clonal Germplasm Repository (NCGR), the USDA Forage Seed and Cereal Research (FSCR), and the University of Nebraska-Lincoln (UNL) collections. The SSR set identified unique genotypes except for 89 sets of synonymous samples. These synonyms included: cultivars with different designations, the same cultivars from different sources, heat-treated clones, and clonal variants. Population structure analysis clustered accessions into wild North American (WNA) and cultivated groups. Diversity was slightly higher in the cultivated samples due to larger sample size. Parentage and sib-ship analyses were used to identify true-to-type cultivars. The HI-AGA7 marker generated two male- and nine female-specific alleles among the cultivated and WNA samples. The SSR and KASP fingerprinting sets were compared in 190 samples consisting of cultivated and WNA accession for their ability to confirm identity and assess diversity and population structure. The SSR fingerprinting set distinguished cultivars, selections and WNA accessions while the KASP assays were unable to distinguish the WNA samples and had lower diversity estimates than the SSR set. Both fingerprinting sets are valuable tools for identity confirmation and parentage analysis in hop for different purposes. The 9-SSR assay is cost efficient when genotyping a small number of wild and cultivated hop samples (<96) while the KASP assay is easy to interpret and cost efficient for genotyping a large number of cultivated samples (multiples of 96).

Verification of clonal identity of hop (Humulus lupulus L.) cultivars within breeding programs and germplasm collections is vital to conserving genetic resources. Accurate and economic DNA-based tools are needed in dioecious hop to confirm identity and parentage, neither of which can be reliably determined from morphological observations. In this study, we developed two fingerprinting sets for hop: a 9-SSR fingerprinting set containing high-core repeats that can be run in a single PCR reaction and a kompetitive allele specific PCR (KASP) assay of 25 single nucleotide polymorphisms (SNPs). The SSR set contains a sex-linked primer pair, HI-AGA7, that was used to genotype 629 hop accessions from the US Department of Agriculture (USDA) National Clonal Germplasm Repository (NCGR), the USDA Forage Seed and Cereal Research (FSCR), and the University of Nebraska-Lincoln (UNL) collections. The SSR set identified unique genotypes except for 89 sets of synonymous samples. These synonyms included: cultivars with different designations, the same cultivars from different sources, heat-treated clones, and clonal variants. Population structure analysis clustered accessions into wild North American (WNA) and cultivated groups. Diversity was slightly higher in the cultivated samples due to larger sample size. Parentage and sib-ship analyses were used to identify true-to-type cultivars. The HI-AGA7 marker generated two male- and nine female-specific alleles among the cultivated and WNA samples. The SSR and KASP fingerprinting sets were compared in 190 samples consisting of cultivated and WNA accession for their ability to confirm identity and assess diversity and population structure. The SSR fingerprinting set distinguished cultivars, selections and WNA accessions while the KASP assays were unable to distinguish the WNA samples and had lower diversity estimates than the SSR set. Both fingerprinting sets are valuable tools for identity confirmation and parentage analysis in hop for different purposes. The 9-SSR assay is cost efficient when genotyping a small number of wild and cultivated hop samples (<96) while the KASP assay is easy to interpret and cost efficient for genotyping a large number of cultivated samples (multiples of 96).

The Centers for Medicare & Medicaid Services (CMS) metric for reporting ultrafiltration (UF) recommends limiting UF at a range of 10 to 13 mL/Kg/Hr to avoid organ stunning. All organ stunning has an impact on patient quality of life (QoL). As this new standard of UF is implemented, the interdisciplinary team (IDT) and the patient must work together toward achieving a less than 13 mL/Kg/Hr UF buy-in. Understanding the CMS goals and target date of January 1, 2018, for data collection purposes was key to early initiation of staff and patient education. Dialysis treatment centers must rise to implement mandated fluid removal objectives. Education and knowledge are the best facilitators of success when implementing change. The IDT must work together to continually reinforce the standards set by CMS.

In order to determine the extent to which care delivered to children is appropriate (in line with evidence-based care and/or clinical practice guidelines (CPGs)) in Australia, we developed a set of clinical indicators for 21 common paediatric medical conditions for use across a range of primary, secondary and tertiary healthcare practice facilities. Clinical indicators were extracted from recommendations found through systematic searches of national and international guidelines, and formatted with explicit criteria for inclusion, exclusion, time frame and setting. Experts reviewed the indicators using a multi-round modified Delphi process and collaborative online wiki to develop consensus on what constituted appropriate care. From 121 clinical practice guidelines, 1098 recommendations were used to draft 451 proposed appropriateness indicators. In total, 61 experts (n = 24 internal reviewers, n = 37 external reviewers) reviewed these indicators over 40 weeks. A final set of 234 indicators resulted, from which 597 indicator items were derived suitable for medical record audit. Most indicator items were geared towards capturing information about under-use in healthcare (n = 551, 92%) across emergency department (n = 457, 77%), hospital (n = 450, 75%) and general practice (n = 434, 73%) healthcare facilities, and based on consensus level recommendations (n = 451, 76%). The main reason for rejecting indicators was 'feasibility' (likely to be able to be used for determining compliance with 'appropriate care' from medical record audit). A set of indicators was developed for the appropriateness of care for 21 paediatric conditions. We describe the processes (methods), provenance (origins and evolution of indicators) and products (indicator characteristics) of creating clinical indicators within the context of Australian healthcare settings. Developing consensus on clinical appropriateness indicators using a Delphi approach and collaborative online wiki has methodological utility. The final indicator set can be used by clinicians and organisations to measure and reflect on their own practice.

ObjectiveTo identify the risk factors associated with complaints, malpractice claims and impaired performance in medical practitioners.DesignSystematic review.Data sourcesOvid-Medline, Ovid Embase, Scopus and Cochrane Central Register of Controlled Trials were searched from 2011 until March 2020. Reference lists and Google were also handsearched.ResultsSixty-seven peer-reviewed papers and three grey literature publications from 2011 to March 2020 were reviewed by pairs of independent reviewers. Twenty-three key factors identified, which were categorised as demographic or workplace related. Gender, age, years spent in practice and greater number of patient lists were associated with higher risk of malpractice claim or complaint. Risk factors associated with physician impaired performance included substance abuse and burn-out.ConclusionsIt is likely that risk factors are interdependent with no single factor as a strong predictor of a doctor’s risk to the public. Risk factors for malpractice claim or complaint are likely to be country specific due to differences in governance structures, processes and funding. Risk factors for impaired performance are likely to be specialty specific due to differences in work culture and access to substances. New ways of supporting doctors might be developed, using risk factor data to reduce adverse events and patient harm.PROSPERO registration numberPROSPERO registration number: CRD42020182045.

Light sheet fluorescence microscopy (LSFM) of optically cleared biological samples represents a powerful tool to analyze the 3-dimensional morphology of tissues and organs. Multimodal combinations of LSFM with additional analyses of the identical sample help to limit the consumption of restricted specimen and reduce inter-sample variation. Here, we demonstrate the proof-of-concept that LSFM of cleared brain tissue samples can be combined with Matrix Assisted Laser Desorption/Ionization-Mass Spectrometry Imaging (MALDI-MSI) for detection and quantification of proteins. Samples of freshly dissected murine brain and of archived formalin-fixed paraffin-embedded (FFPE) human brain tissue were cleared (3DISCO). Tissue regions of interest were defined by LSFM and excised, (re)-embedded in paraffin, and sectioned. Mouse sections were coated with sinapinic acid matrix. Human brain sections were pre-digested with trypsin and coated with α-cyano-4-hydroxycinnamic acid matrix. Subsequently, sections were subjected to MALDI-time-of-flight (TOF)-MSI in mass ranges between 0.8 to 4 kDa (human tissue sections), or 2.5–25 kDa (mouse tissue sections) with a lateral resolution of 50 µm. Protein- and peptide-identities corresponding to acquired MALDI-MSI spectra were confirmed by parallel liquid chromatography tandem mass spectrometry (LC–MS/MS) analysis. The spatial abundance- and intensity-patterns of established marker proteins detected by MALDI-MSI were also confirmed by immunohistochemistry.

Nucleotide sequences of most vertebrate genes will be available in the next few years as ESTs and genomic sequences. This information will allow the deduction of encoded proteins, possibly providing clues to their functions in vivo. To determine unequivocally the physiological roles of all genes and in particular those of medical importance, mutational analyses must be performed. Large-scale random mutagenesis screens, which allow the systematic mutation of all genes, are a valuable approach to this task. To complement the sequencing efforts, we established a consortium generating a reference library of gene-trap sequence tags (GTST) which eventually will represent insertional mutations for most mammalian genes expressed in mouse embryonic stem (ES) cells. Here we report a large-scale gene-trap screen in ES cells using plasmid and retroviral gene-trap (GT) vectors that permit rapid generation and identification of mutant genes in ES cells.

The literature concerning the subcellular location of Y-box binding protein 1 (YB-1), its abundance in normal and cancer tissues, and its prognostic significance is replete with inconsistencies. An explanation for this could be due in part to the use of different antibodies in immunohistochemical and immunofluorescent labeling of cells and tissues. The inconsistencies could also be due to poor resolution of immunohistochemical data. We analyzed two cohorts of breast tumours for both abundance and subcellular location of YB-1 using three different antibodies; two targeting N-terminal epitopes (AB-a and AB-b) and another (AB-c) targeting a C-terminal epitope. We also investigated stress-induced nuclear translocation of YB-1 in cell culture. We report that both AB-a and AB-c detected increased YB-1 in the cytoplasm of high-grade breast cancers, and in those lacking estrogen and progesterone receptors; however the amount of YB-1 detected by AB-a in these cancers is significantly greater than that detected by AB-c. We confirm our previously published findings that AB-b is also detecting hnRNP A1, and cannot therefore be used to reliably detect YB-1 by immunohistochemistry. We also report that AB-a detected nuclear YB-1 in some tumour tissues and stress treated cells, whereas AB-c did not. To understand this, cancer cell lines were analyzed using native gel electrophoresis, which revealed that the antibodies detect different complexes in which YB-1 is a component. Our data suggest that different YB-1 antibodies show different staining patterns that are determined by the accessibility of epitopes, and this depends on the nature of the YB-1 complexes. It is important therefore to standardize the protocols if YB-1 is to be used reproducibly as a prognostic guide for different cancers.

A good understanding of nutrition concepts enables a person to convert basic knowledge into the ability to take action. At the time of this study a game testing nutrition education concepts in English among isiZulu speaking learners had not been investigated. The objective of the study was to determine the effectiveness of an English Food-Based Dietary Guideline (FBDG) nutrition education game on the retention of nutrition knowledge among isiZulu speaking learners. An intervention using a pre-test post-test design. A five part questionnaire was administered to determine levels of nutrition knowledge before and six weeks after a nutrition education board game intervention. A total of 169 Grade 5 learners from two schools in Sweetwaters, KwaZulu-Natal, participated in this study. Baseline results showed that the learners had very poor knowledge of the basic FBDG. The question regarding the food fortification logo showed most improvement overall, with statistical significance ( p = 0.000). The pictorial representation of information in the game helped the learners improve their knowledge retention for certain questions. While there was very little improvement in the retention of knowledge as a whole, the control group experienced a significant increase in their post-test knowledge score (p = 0.011). Poor English proficiency may have influenced the effectiveness of the intervention. Nutrition education games have the potential to improve nutrition knowledge. Priority should be given to increasing English language proficiency among isiZulu speaking learners to improve their ability to retain nutrition information taught at school.

’n Goeie begrip van voedingskonsepte stel mens in staat om basiese kennis in optrede om te sit. Met die doen van hierdie studie is daar nog nie onder isiZulu-sprekende leerders ondersoek ingestel na ’n speletjie wat opvoedkundige voedingskonsepte in Engels toets nie. Die doelstelling van hierdie studie was om die doeltreffendheid te bepaal van ’n Engelstalige voedselgebaseerde dieetriglyn-voedingspeletjie (FBDG) oor die vaslegging van voedingskennis by isiZulusprekende leerders. Die ontwerp is in die vorm van intervensie waar ’n metode van vóóren návrae en -antwoorde gebruik is. ’n Vraelys bestaande uit vyf afdelings is opgestel om die vlakke van voedingskennis vóór en ses weke ná ingryping in die vorm van ’n opvoedkundige voedingspeletjie te bepaal. Altesaam 169 Graad 5-leerders van twee skole in Sweetwaters, KwaZulu-Natal, het aan die studie deelgeneem. Voorlopige resultate het getoon dat die leerders oor gebrekkige kennis van die basiese FBDG beskik. Die vraag oor die voedselverrykingshandelsmerk het die grootste algemene verbetering met statistiese beduidendheid (p = 0.000) getoon. Die oordra van inligting met behulp van prentjies in die speletjie het die leerders gehelp om kennis oor bepaalde vrae makliker te behou. Terwyl daar min verbetering was in die behoud van kennis oor die algemeen, het die kontrolegroep ’n beduidende toename in die behoud van kennis in hulle nátoets-kennistelling (p = 0.011) waargeneem. Gebrekkige kennis van die Engelse taal kon die doeltreffendheid van die intervensie beïnvloed het. Opvoedkundige voedingspeletjies het die potensiaal om voedingskennis te verbeter. Voorkeur moet verleen word aan die verhoging van vaardigheid in Engels onder isiZulu-sprekende leerders om hulle vermoë te verbeter om voedingsinligting wat op skool geleer word, vas te lê. The effectiveness of an English nutrition education game on knowledge retention in Grade 5 isiZulu-speaking learners. A good understanding of nutrition concepts enables a person to convert basic knowledge into the ability to take action. At the time of this study a game testing nutrition education concepts in English among isiZulu speaking learners had not been investigated. The objective of the study was to determine the effectiveness of an English Food-Based Dietary Guideline (FBDG) nutrition education game on the retention of nutrition knowledge among isiZulu speaking learners. An intervention using a pre-test post-test design. A five part questionnaire was administered to determine levels of nutrition knowledge before and six weeks after a nutrition education board game intervention. A total of 169 Grade 5 learners from two schools in Sweetwaters, KwaZulu-Natal, participated in this study. Baseline results showed that the learners had very poor knowledge of the basic FBDG. The question regarding the food fortification logo showed most improvement overall, with statistical significance (p = 0.000). The pictorial representation of information in the game helped the learners improve their knowledge retention for certain questions. While there was very little improvement in the retention of knowledge as a whole, the control group experienced a significant increase in their post-test knowledge score (p = 0.011). Poor English proficiency may have influenced the effectiveness of the intervention. Nutrition education games have the potential to improve nutrition knowledge. Priority should be given to increasing English language proficiency among IsiZulu speaking learners to improve their ability to retain nutrition information taught at school.