Immunohistochemistry should undergo robust validation equivalent to that of molecular diagnostics

Immunohistochemistry (IHC) is a widely available and highly utilised tool in diagnostic histopathology and is used to guide treatment options as well as provide prognostic information. IHC is subjected to qualitative and subjective assessment, which has been criticised for a lack of stringency, while PCR-based molecular diagnostic validations by comparison are regarded as very rigorous. It is essential that IHC tests are validated through evidence-based procedures. With the move to ISO15189 (2012), not just of the accuracy, specificity and reproducibility of each test need to be determined and managed, but also the degree of uncertainty and the delivery of such tests. The recent update to ISO 15189 (2012) states that it is appropriate to consider the potential uncertainty of measurement of the value obtained in the laboratory and how that may impact on prognostic or predictive thresholds. In order to highlight the problems surrounding IHC validity, we reviewed the measurement of Ki67and p53 in the literature. Both of these biomarkers have been incorporated into clinical care by pathology laboratories worldwide. The variation seen appears excessive even when measuring centrally stained slides from the same cases. We therefore propose in this paper to establish the basis on which IHC laboratories can bring the same level of robust validation seen in the molecular pathology laboratories and the principles applied to all routine IHC tests.

Quantification of HER2 heterogeneity in breast cancer-implications for identification of sub-dominant clones for personalised treatment

Breast cancer is a heterogeneous disease, at both an inter- and intra-tumoural level. Appreciating heterogeneity through the application of biomarkers and molecular signatures adds complexity to tumour taxonomy but is key to personalising diagnosis, treatment and prognosis. The extent to which heterogeneity exists, and its interpretation remains a challenge to pathologists. Using HER2 as an exemplar, we have developed a simple reproducible heterogeneity index. Cell-to-cell HER2 heterogeneity was extensive in a proportion of both reported 'amplified' and 'non-amplified' cases. The highest levels of heterogeneity objectively identified occurred in borderline categories and higher ratio non-amplified cases. A case with particularly striking heterogeneity was analysed further with an array of biomarkers in order to assign a molecular diagnosis. Broad biological complexity was evident. In essence, interpretation, depending on the area of tumour sampled, could have been one of three distinct phenotypes, each of which would infer different therapeutic interventions. Therefore, we recommend that heterogeneity is assessed and taken into account when determining treatment options.

Evidence for the reliability and validity, but not the practical utility of the two-factor Consideration of Future Consequences Scale-14

Researchers have proposed 1-factor, 2-factor, and bifactor solutions to the 12-item Consideration of Future Consequences Scale (CFCS-12). In order to overcome some measurement problems and to create a robust and conceptually useful two-factor scale the CFCS-12 was recently modified to include two new items and to become the CFCS-14. Using a University sample, we tested four competing models for the CFCS-14: (a) a 12-item unidimensional model, (b) a model fitted for two uncorrelated factors (CFC-Immediate and CFC-Future), (c) a model fitted for two correlated factors (CFC-I and CFC-F), and (d) a bifactor model. Results suggested that the addition of the two new items has strengthened the viability of a two factor solution of the CFCS-14. Results of linear regression models suggest that the CFC-F factor is redundant. Further studies using alcohol and mental health indicators are required to test this redundancy.

Urinary thrombomodulin levels were significantly higher following occupational exposure to chemicals, in the presence of dipstick protein, but not in the presence of dipstick blood

Currently, there are no biomarkers which can identify patients with an increased risk of developing urothelial cancer as a result of occupational chemical exposure. The aim of this study was to evaluate the relationships between final diagnosis and 22 biomarkers measured in urine, serum and plasma collected from 156 hematuric patients. Fourteen of the 80 patients (17.5%) with urothelial cancer and 13/76 (17.1%) of the controls were deemed to have a history of chemical exposure. We applied Fisher's exact tests to explore associations between chemical exposure and final diagnosis, and tumor stage and grade, where applicable; ANOVA and t-test to compare age across patients with and without chemical exposure; and Zelen's exact test to evaluate relationships across final diagnosis, chemical exposure and smoking. Following pre-selection of biomarkers using Lasso, we identified biomarkers with differential levels across patients with and without chemical exposure using Welch's t-test. Using a one-sided t-test and considering multiple testing using FDR, we observed that TM levels in urine were significantly higher in samples from patients with a history of chemical exposure regardless of their diagnosis as control or urothelial cancer (one-sided t-test, p_UC = 0.014 and p_CTL = 0.043); in the presence of dipstick protein and when urinary pH levels ≤ 6 (p = 0.003), but not in the presence of dipstick blood (p = 0.115). Urothelial cancer patients with a history of chemical exposure were significantly younger (64.1 years) than those without chemical exposure (70.2 years) (one-sided t-test p-value = 0.012); and their tumors were higher grade (Fisher's exact test; p = 0.008). There was a strong association between a history of chemical exposure and smoking in urothelial cancer patients (Zelen's exact test; p = 0.025). Elevated urinary thrombomodulin levels could have the potential to identify chemical exposure in hematuric patients at high risk of developing urothelial cancer.

A synopsis of the genus Bursaphelenchus Fuchs, 1937 (Aphelenchida: Parasitaphelenchidae) with keys to species

The 75 valid species of the genus Bursaphelenchus are listed together with their synonyms. Diagnostic characters and their states are discussed and illustrated. Tabular and traditional text keys are provided for the genus. Two new subspecies are proposed to distinguish populations of B. piniperdae and B. poligraphi, as described by Rühm (1956), from the original descriptions of these species published by Fuchs (1937). Known records of Bursaphelenchus species with their associated natural vectors, plants and plant families are given. Dendrograms of species relationships (UPGMA, standard distance: mean character difference) based on combined taxonomic characters and also on spicule characters only, are provided. Discussion as to whether the species groups are natural or artificial (and therefore purely diagnostic) is based on their relationships in the dendrogram and the vector and associated plant ranges of the species. Of the six species groups distinguished, two appear to represent natural assemblages, these being the xylophilus-group (with ten species) and the hunti-group (seven species), of which two, B. cocophilus and B. dongguanensis, form the cocophilus-cluster which is separated on the dendrogram from the main clusters. The remaining four species groups appear to be artificial and purely diagnostic in function, namely the aberrans-group (four species); the eidmanni-group (six species); the borealis-group (five species), and the piniperdae-group (43 species). Two new subspecies, both in the piniperdae-group, viz. B. piniperdae ruehmpiniperdae n. subsp. and B. poligraphi ruehmpoligraphi n. subsp., are proposed and diagnosed from B. piniperdae piniperdae and B. poligraphi poligraphi the respective type subspecies. Bursaphelenchus dongguanensis is regarded as being a valid member of the genus and its transfer to Parasitaphelenchus is rejected.

Chromosome structure and behaviour in Bursaphelenchus xylophilus (Nematoda: Parasitaphelenchidae) germ cells and early embryo

Chromosome structure and behaviour in both meiosis of the germ cells and mitosis of the embryo from fertilisation to the two-cell stage in Bursaphelenchus xylophilus were examined by DAPI staining and three-dimensional reconstruction of serial-section images from confocal laser-scanning microscopy. By this method, each chromosome’s shape and behaviour were clearly visible in early embryogenesis from fertilisation through the formation and fusion of the male and female pronuclei to the first mitotic division. The male pronucleus was bigger than that of the female, although the oocyte is larger and richer in nutrients than the sperm. From the shape of the separating chromosomes at anaphase, the mitotic chromosomes appeared to be polycentric or holocentric rather than monocentric. Each chromosome was clearly distinguishable in the male and female germ cells, pronuclei of the one-cell stage embryo, and the early embryonic nuclei. The haploid number of chromosomes (N) was six (2n = 12), and all chromosomes appeared similar. The chromosome pair containing the ribosomal RNA-coding site was visualised by fluorescence in situ hybridisation. Unlike the sex determination system in Caenorhabditis elegans (XX in hermaphrodite and XO in male), the system for B. xylophilus may consist of an XX female and an XY male.

Bursaphelenchus antoniae sp. n. (Nematoda: Parasitaphelenchidae) associated with Hylobius sp. from Pinus pinaster in Portugal

Bursaphelenchus antoniae sp. n. is described and illustrated. Dauer juveniles were isolated from the body of the large pine weevil, Hylobius sp., collected from maritime pine (Pinus pinaster) stumps, in Portugal. Bursaphelenchus antoniae sp. n. was reared and maintained in P. pinaster wood segments and on Petri dish cultures of the fungi Botrytis cinerea and Monilinia fructicola. The new species is characterised by a relatively small body length of ca 583 μm (females) and 578 μm (males), a lateral field with two incisures, presence of a small vulval flap and a conoid female tail with a rounded or pointed terminus. Males have stout spicules with a disc-like cucullus and seven caudal papillae arranged as a single midventral precloacal papilla, one precloacal pair and two postcloacal pairs. In the character of the lateral field, B. antoniae sp. n. comes close to B. abietinus, B. rainulfi and B. hylobianum, whilst spicule characters place it within the piniperdae-group sensu Ryss et al. Morphologically, B. antoniae sp. n. is closest to B. hylobianum; the spicules of these two species having flattened, wing-like, alae on the distal third of the lamina. Bursaphelenchus antoniae sp. n. is distinguished from B. hylobianum on the arrangement of the caudal papillae (two vs three pairs). ITS-RFLP profiles and the failure to hybridise support the separation of the two species. Phylogenetic analysis of the new species, based on the 18S rDNA sequence, supports the inclusion of this new species in the B. hylobianum-group sensu Braasch. Sequence analysis of the 28S rDNA D2/D3 domain did not place the new species in a definite group.

On the vulval morphology of some species of Bursaphelenchus (Nematoda: Parasitaphelenchinae)

The vulval pattern of six species of the genus Bursaphelenchus (B. abruptus, B. conicaudatus, B. fraudulentus, B. luxuriosae, B. mucronatus and B. xylophilus) was studied using scanning electron microscopy. A terminology for the vulval region structures observed is proposed herein and illustrated by micrographs and line drawings. It was shown that, of the studied species, only B. mucronatus and B. xylophilus share an identical morphology of the vulval region, all other species differing significantly from each other and from both B. mucronatus and B. xylophilus. This study indicates the diagnostic potential for variation in vulval morphology within Bursaphelenchus and it is recommended that such features are recorded in all future descriptions.

Processos e modelos de raciocínio na tomada de decisão: contributos conceptuais e interrogações

Processos e Modelos de Raciocínio na Tomada de Decisão: contributos concetuais e interrogações Dulce Magalhães - Departamento de Enfermagem, Universidade de Évora Manuel Lopes - Departamento de Enfermagem, Universidade de Évora Ana Fonseca - Departamento de Enfermagem, Universidade de Évora Os enfermeiros que integram as equipas de unidades de cuidados hospitalares têm que prevenir, minimizar e corrigir situações clínicas instáveis. Em consequência disso defrontam-se diária e sistematicamente com a obrigatoriedade de tomar decisões no contexto da ação (Thompson & Dowding, 2005) e agir em conformidade. Eles têm que tomar decisões num espaço que não é reservado e que está confinado a uma teia de relações que acontecem entre o doente, a família e os restantes membros da equipa de saúde (Benner et al, 2011; Tanner, 2006, Lopes, 2006). Um espaço onde acontecem múltiplas interrupções que provêm de procedimentos de trabalho e exigências constantes de atenção a cada doente (Hedberg & Larsson, 2004; Potter et al, 2005; Wolf et al, 2006). São decisões processuais e multidimensionais que têm que considerar em simultâneo, o que deve ser feito, como deve ser feito e quando deve ser feito, no respeito da singularidade do doente e do seu contexto circundante (Gillespie, 2009). Esta dinâmica também está interligada com os dados clínicos que recorrentemente emergem, bem como à acessibilidade, multiplicidade e ambiguidade dos dados clínicos (Junnola et al, 2002; Carnevali &Thomas, 1993 Simmons, 2010), ao tempo de decisão e às decisões conflituosas (Thompson, 2004). São ambientes clínicos onde acontecem contingências, algumas delas aleatórias, que exigem que os enfermeiros independentemente dos níveis de proficiência sejam obrigados a tomar decisões complexas de forma rápida e precisa, para optimizar os resultados esperados (Curry & Botti, 2006). O conhecimento e o raciocínio que os enfermeiros usam na tomada de decisão são elementos, entre outros, que definem muito da prática profissional (Cody, 2006). Por isso, conscientes da realidade enunciada através da investigação que tem vindo a ser desenvolvida, foi nossa intenção analisar criticamente os modelos de decisão que têm expressão na literatura de enfermagem e que têm sido desenvolvidos em diferentes áreas do conhecimento (Concoran-Perry et al, 1999), no sentido de reconhecer o seu poder explicativo para a tomada de decisão dos enfermeiros. Alguns deles são apresentados numa estrutura normativa, outros numa descritiva e em função disso, podem dividir-se em duas categorias teóricas, sistemático-positivista e intuitivo-humanista (Thompson, 1999; Aitken, Marshall, Elliott & McKinley, 2007). As abordagens sistemático-positivistas sugerem que a tomada de decisão acontece num processo sequencial previamente definido e explicitado. Assentam no pressuposto que existe alguma atividade de análise ou de resolução de problemas em curso, para o enfermeiro tomar uma decisão. Nas abordagens intuitivo-humanistas a decisão é entendida no seio de um processo como um todo e no contexto de uma abordagem naturalista (Benner, 1984; Thompson, 1999). Elas podem ser diferenciadas, desde logo, a partir dos seus centros de interesse. Enquanto a primeira se interessa pelo número de dados e pelos processos de análise cognitiva para uma melhor decisão. A segunda dá primazia aos contextos relacionais e de comunicação com os doentes e serve-se da performance clínica para eleger as melhores ações.

Antioxidant activity of some Morrocan hydrosols

Essential oils are used in Cosmetic, Perfumery, Food and Pharmaceutical Industries as flavours and/or medicines. However, part of the essential oil components that remains in the distillation water (hydrosol or distillate water) has been less studied both in chemical and biological terms. This research concerns the antioxidant activity, measured through several methods, of Lavandula officinalis L., Origanum majorana L., Rosmarinus officinalis L., Salvia officinalis L. and Thymus vulgaris L., Cinnamomum verum J. Presl. and Syzygium aromaticum (L.) Merrill and Perry hydrosols. The ability of hydrosols to prevent oxidation was checked by two main methods: prevention of lipid peroxidation through the measurement of malonaldehyde produced after degradation of hydroperoxides; and ability for scavenging free radicals including hydroxyl and superoxide anion radicals. The S. aromaticum and T. vulgaris hydrosols, predominantly constituted by eugenol and carvacrol, respectively, were the most effective as antioxidants, except for scavenging superoxide anion radical. In this case, L. officinalis hydrosol in which linalool prevailed was a stronger antioxidant. The worst hydrosol as antioxidant was that of S. officinalis, independent on the method checked.

FITS into Practice: Translating Research into Practice in Reducing the Use of Anti-psychotic Medication for People with Dementia Living in Care Homes

Objectives: This paper reports on the acceptability and effectiveness of the FITS (Focussed Intervention Training and Support) into Practice Programme. This intervention was scaled up from an earlier cluster randomised-controlled trial that had proven successful in significantly decreasing antipsychotic prescribing in care homes. Method: An in depth 10-day education course in person-centred care was delivered over a three-month period, followed by six supervision sessions. Participants were care-home staff designated as Dementia Care Coaches (DCCs) responsible for implementing interventions in 1 or 2 care homes. The course and supervision was provided by educators called Dementia Practice Development Coaches (DPDCs). Effectiveness data included monitoring antipsychotic prescriptions, goal attainment, knowledge, attitudes and implementation questionnaires. Qualitative data included case studies and reflective journals to elucidate issues of implementation. Results: Of the 100 DCCs recruited, 66 DCCs completed the programme. Pre-post questionnaires demonstrated increased knowledge and confidence and improved attitudes to dementia. Twenty per cent of residents were prescribed antipsychotics at baseline which reduced to 14% (31% reduction) with additional dose reductions being reported alongside improved personalised goal attainment. Crucial for FITS into Practice to succeed was the allocation and protection of time for the DCC to attend training and supervision and to carry out implementation tasks in addition to their existing job role. Evaluation data showed that this was a substantial barrier to implementation in a small number of homes. Discussion and conclusions: The FITS into practice programme was well evaluated and resulted in reduction in inappropriate anti-psychotic prescribing. Revisions to the intervention are suggested to maximise successful implementation.

Underway O2/Ar-estiment net community production across the Kuroshio Extension and impact on winter pCO2 drawdown

Senior thesis written for Oceanography 445

Affective Instability, Childhood Trauma and Major Affective Disorders

BACKGROUND: Affective instability (AI), childhood trauma, and mental illness are linked, but evidence in affective disorders is limited, despite both AI and childhood trauma being associated with poorer outcomes. Aims were to compare AI levels in bipolar disorder I (BPI) and II (BPII), and major depressive disorder recurrent (MDDR), and to examine the association of AI and childhood trauma within each diagnostic group. METHODS: AI, measured using the Affective Lability Scale (ALS), was compared between people with DSM-IV BPI (n=923), BPII (n=363) and MDDR (n=207) accounting for confounders and current mood. Regression modelling was used to examine the association between AI and childhood traumas in each diagnostic group. RESULTS: ALS scores in descending order were BPII, BPI, MDDR, and differences between groups were significant (p<0.05). Within the BPI group any childhood abuse (p=0.021), childhood physical abuse (p=0.003) and the death of a close friend in childhood (p=0.002) were significantly associated with higher ALS score but no association was found between childhood trauma and AI in BPII and MDDR. LIMITATIONS: The ALS is a self-report scale and is subject to retrospective recall bias. CONCLUSIONS: AI is an important dimension in bipolar disorder independent of current mood state. There is a strong link between childhood traumatic events and AI levels in BPI and this may be one way in which exposure and disorder are linked. Clinical interventions targeting AI in people who have suffered significant childhood trauma could potentially change the clinical course of bipolar disorder.

Energy Management in UK Non-Domestic Buildings - a New Perspective to Provoke Reduced Carbon Emissions

Politicians, industry and the public generally accept the need for energy consumption to be cut to deliver climate change mitigation measures essential for us to avoid climate disaster. For non-domestic fuel users current energy policy has attempted to drive this through rational economic responses to energy cost pressures. This reliance on voluntary action has created an “Energy Inconsistency”, that is a marked difference between energy opportunities that have been proven technically viable, financially rational and retrofit feasible and those actually adopted. Other factors must therefore be involved to influence what appear to be simple carbon and cost saving opportunities. This paper presents a new approach to energy efficiency and consumption in non-domestic buildings, viewing attitudes and behaviours of building owners and users as the key driver of energy consumption. A new framework is proposed as a method to examine the impact of building ownership on the users’ and owners’ abilities to improve energy efficiency and consumption and identify opportunities to overcome the barriers inherent in these ownership structures.