Amplification of Chromosome 17 Centromere (CEP17) in Breast Cancer Patients with a Result of HER2 2± by Immunohistochemistry.

Background: Amplification of the centromeric region of chromosome 17 (CEP17) as measured by In Situ Hybridization (ISH) of the CEP17 probe is used clinically as part of the ISH assay for HER2 status determination in breast cancer. The value of amplification of CEP17 beyond its use in the HER2 ISH test has not been fully explored.Methods: A retrospective review of patients with breast cancer that had a dual probe HER2/CEP17 FISH test during an eight-year period was performed. Data on demographic and cancer-specific characteristics of the included patients were extracted. The group of patients with an amplified CEP17 defined as mean copy number of ≥3 per nucleus was compared with the group without amplification.Results: Two hundred and twelve patients were eligible and included in the analysis. Amplification of CEP17 was observed in 39 patients (18.4%). All patients in the amplified group had a concomitant amplification of HER2 (mean copy number ≥3 per nucleus). In the CEP17 non-amplified group 82 of 172 patients (47.7%) had an amplified HER2 status. More patients in the amplified group had a clinical HER2+ status according to the 3-protein classifier (30.8% versus 12.3% in the non-amplified group) and fewer patients in the amplified group had a clinical ER+/HER2- status (66.7% versus 81.3% in the non-amplified group, x2 p = .01). Other significant differences between the amplified and CEP17 non-amplified groups were observed in their lymph node (LN) status (56.4% of patients in the amplified group versus 38.8% in the non-amplified group were lymph node positive, p = .04) and in the nuclear heterogeneity component of grade (91.2% of patients in the amplified group were nuclear grade 3 versus 67.1% in the non-amplified group, p = .005). There were no statistically significant differences between the groups in overall stage, grade, menopause status or histology. Recurrence Free Survival (RFS) was shorter in stage I to III patients with an amplified CEP17 compared with the non-amplified group.Conclusion: Patients with amplification of CEP17 had a co-amplified HER2 and were more commonly HER2+, LN positive and grade 3 in the nuclear component of grade.

Applying a food processing-based classification system to a food guide: a qualitative analysis of the Brazilian experience.

OBJECTIVE: The present paper aimed to identify the stakeholders, as well as their arguments and recommendations, in the debate on the application of a food processing-based classification system to the new Brazilian Food Guide. DESIGN: Qualitative approach; an analysis was made of documents resulting from the consultation conducted for the development of the new Brazilian Food Guide, which uses the NOVA classification for its dietary recommendations. A thematic matrix was constructed and the resulting themes represented the main points for discussion raised during the consultation. SETTING: Brazil. SUBJECTS: Actors from academia, government and associations/unions/professional bodies/organizations related to the area of nutrition and food security; non-profit institutions linked to consumer interests and civil society organizations; organizations, associations and food unions linked to the food industry; and individuals. RESULTS: Four themes were identified: (i) conflicting paradigms; (ii) different perceptions about the role and need of individuals; (iii) we want more from the new food guide; and (iv) a sustainable guide. CONCLUSIONS: There was extensive participation from different sectors of society. The debate generated by the consultation revealed two main conflicting opinions: a view aligned with the interests of the food industry and a view of healthy eating which serves the interests of the population. The first group was against the adoption of a food processing-based classification system in a public policy such as the new Brazilian Food Guide. The second group, although mostly agreeing with the new food guide, argued that it failed to address some important issues related to the food and nutrition agenda in Brazil.

Mandatory food labeling of trans fat acids: qualitative analysis of the public consultation / Lei de rotulagem de alimentos em gordura trans: análise qualitativa da consulta pública / Ley de rotulación de alimentos con grasa transgénica: análisis cualitativa de consulta pública

The objective of this paper is to identify the stakeholders and their contributions to the public consultation carried out with regard to the mandatory trans fat acids food labeling proposal in Brazil. Methodology thematic analysis of the documents. Results 25 stakeholders made suggestions regarding the proposed law, and the majority belonged to the food industry sector. The following issues were raised: lack of knowledge among the consumers about trans fat acids; lack of information about trans fat acids in food composition tables and infrastructure to analyse the trans fat content of foods; voluntary trans fat labelling. Conclusions The stakeholders addressed important issues, such as improving nutritional knowledge among the population. However, few stakeholders linked to consumers took part in the public consultation, which might lead to disproportionate influence of the food industry on the development of future food policies.

Objetivo identificar os atores sociais e as suas contribuições na consulta pública realizada sobre a proposta obrigatória de rotulagem de alimentos no Brasil com gordura trans. Metodologia análise temática dos documentos. Resultados 25 atores apresentaram sugestões para a proposta da lei, e a maioria pertencia ao setor da indústria de alimentos. Temas encontrados: falta de conhecimento entre os consumidores sobre a gordura trans; falta de informação sobre gorduras trans em tabelas de composição de alimentos e de infraestrutura para analisar o teor de gordura trans nos alimentos; rotulagem voluntária da gordura trans. Conclusões Os participantes abordaram questões importantes, como a necessidade de aumentar o conhecimento dos consumidores sobre gordura trans. Entretanto, houve poucos atores sociais ligados aos interesses dos consumidores que participaram na consulta pública, e isso pode levar a um desequiibrio de poderes nas discussões sobre nutrição no Brasil.

Objetivo Identificar actores sociales y sus contribuciones en la consulta pública realizada sobre la propuesta obligatoria de rotulación de alimentos en Brasil con grasas transgénicas. Metodología Análisis temático de documentos. Resultados 25 actores presentaron sugerencias para la propuesta de ley, y la mayoría pertenecía al sector de la industria de alimentos. Temas encontrados Falta de conocimiento entre consumidores sobre grasas transgénicas; falta de información sobre grasas transgénicas en tablas de composición de alimentos e infraestructura para analizar grado de grasas transgénicas en alimentos; rotulación voluntaria de grasas transgénicas. Conclusiones Los participantes abordaron cuestiones importantes, como la necesidad de aumentar el conocimiento de los consumidores sobre grasas transgénicas. No obstante, hubo pocos actores sociales ligados a los intereses de los consumidores que participaron en la consulta y eso puede llevar a un desequilibrio de poderes en discusiones sobre nutrición en Brasil.

Qualitative Analysis of Cognitive Interviews With School Children: A Web-Based Food Intake Questionnaire.

BACKGROUND: The use of computers to administer dietary assessment questionnaires has shown potential, particularly due to the variety of interactive features that can attract and sustain children's attention. Cognitive interviews can help researchers to gain insights into how children understand and elaborate their response processes in this type of questionnaire. OBJECTIVE: To present the cognitive interview results of children who answered the WebCAAFE, a Web-based questionnaire, to obtain an in-depth understanding of children's response processes. METHODS: Cognitive interviews were conducted with children (using a pretested interview script). Analyses were carried out using thematic analysis within a grounded theory framework of inductive coding. RESULTS: A total of 40 children participated in the study, and 4 themes were identified: (1) the meaning of words, (2) understanding instructions, (3) ways to resolve possible problems, and (4) suggestions for improving the questionnaire. Most children understood questions that assessed nutritional intake over the past 24 hours, although the structure of the questionnaire designed to facilitate recall of dietary intake was not always fully understood. Younger children (7 and 8 years old) had more difficulty relating the food images to mixed dishes and foods eaten with bread (eg, jam, cheese). Children were able to provide suggestions for improving future versions of the questionnaire. CONCLUSIONS: More attention should be paid to children aged 8 years or below, as they had the greatest difficulty completing the WebCAAFE.

Children's Cognitive and Affective Responses About a Narrative Versus a Non-Narrative Cartoon Designed for an Active Videogame.

OBJECTIVE: This article presents the results of interviews conducted with children regarding their cognitive and affective responses toward a narrative and a non-narrative cartoon. The findings will be used to further explore the role of a narrative in motivating continued active videogame play. MATERIALS AND METHODS: Twenty children (8-11 years old of mixed gender) watched two cartoons (narrative and non-narrative) and were subsequently interviewed. A thematic matrix was used to analyze the interviews. RESULTS: The narrative cartoon (n = 11) was only slightly preferred compared with the non-narrative one (n = 9), with little difference among the participants. The theme categories identified during the analyses were plot, characters, and suggestions. The fight scenes were mentioned by the children as a likeable aspect of the narrative cartoon. In the non-narrative cartoon, the vast majority (n = 17) liked the information about physical activity that was provided. The children enjoyed the appearance and personalities of the characters in both cartoons. A discrepancy in the data about the fight scenes (narrative cartoon) and characters (both cartoons) was found among the female participants (i.e., some girls did not like the fight and thought the characters were too aggressive). However, most of the children wanted to see more action in the story, an increase in the number of fight scenes (narrative cartoon), or more information about exercise and examples of exercises they could do (non-narrative cartoon). They also suggested adding a game to the non-narrative cartoon, including more characters, and improving the animation in both cartoons. CONCLUSIONS: The children preferred the narrative cartoon because of the story and the fight. Some gender differences were found, which further studies should investigate.

The Yin and Yang of Formative Research in Designing Serious (Exer-)games.

Despite its relevance, formative research on games may be an undervalued part of the game development process. At the 2014 International Society of Behavioral Nutrition and Physical Activity exergaming preconference satellite meeting, a roundtable discussion was held to assemble experiences and suggestions on enhancing the use of formative research in the development of active videogames (i.e., exergames). This article presents a summary of the concepts discussed. The discussants concluded that, although formative research may slightly expand the project timeline, the potential benefits include a game more in line with preferences of the intended users, with better operationalized theoretical constructs and broader stakeholder support, facilitating implementation and sustainability. It also improves the efficiency of other research parts because of a lower dropout rate of participants. Formative, qualitative research is thus a necessary complement to quantitative measurements of intervention outcomes, in a sort of Yin and Yang dynamic. An adapted version of formative research that casts a wider net may, however, be needed, involving both behavioral scientists and game developers, expanding the topics beyond the game's looks and soliciting the opinions of a larger group of stakeholders, such as implementers, gatekeepers, and funders.

Influence of behavioral theory on fruit and vegetable intervention effectiveness among children: a meta-analysis.

OBJECTIVE: To test the hypotheses that interventions clearly based on theory, multiple theories, or a formal intervention planning process will be more effective in changing fruit and vegetable consumption among children than interventions with no behavioral theoretical foundation. DESIGN: Systematic review and meta-analysis. SETTING: Identification of articles in PubMed, PsycInfo, Medline, Cochrane Collaborative database, and existing literature reviews and meta-analyses. PARTICIPANTS: Children aged 2-18 years. INTERVENTIONS: Change in fruit and/or vegetable consumption in dietary change interventions. METHODS: Meta-analysis, meta-regression analysis, and summary reporting for articles. CONCLUSIONS AND IMPLICATIONS: Predicating an intervention on behavioral theory had a small to moderate enhancement (P < .001) of outcome effectiveness. Differences in mean Hedges' g effect sizes between theory and non-theory interventions were 0.232 for fruit, 0.043 for vegetables, and 0.333 for fruit and vegetables combined. There was mixed support, however, for enhanced dietary change with multiple theories or a formal planning process. After controlling for study quality, theory use was related only to vegetable consumption (ß = 0.373; P < .001). More research is needed on theory's influences on dietary behaviors to guide future interventions among children. More research is also needed to identify what may be effective practical- or experience-based procedures that complement theory, to incorporate into interventions.

Assessment of diet and physical activity of brazilian schoolchildren: usability testing of a web-based questionnaire.

BACKGROUND: Information and communication technology (ICT) has been used with increasing frequency for the assessment of diet and physical activity in health surveys. A number of Web-based questionnaires have been developed for children and adolescents. However, their usability characteristics have scarcely been reported, despite their potential importance for improving the feasibility and validity of ICT-based methods. OBJECTIVE: The objective of this study was to describe the usability evaluation of the Consumo Alimentar e Atividade Física de Escolares (CAAFE) questionnaire (Food Consumption and Physical Activity Questionnaire for schoolchildren), a new Web-based survey tool for the self-assessment of diet and physical activity by schoolchildren. METHODS: A total of 114 schoolchildren aged 6 to 12 years took part in questionnaire usability testing carried out in computer classrooms at five elementary schools in the city of Florianopolis, Brazil. Schoolchildren used a personal computer (PC) equipped with software for recording what is on the computer screen and the children's speech during usability testing. Quantitative and qualitative analyses took into account objective usability metrics such as error counts and time to complete a task. Data on the main difficulties in accomplishing the task and the level of satisfaction expressed by the children were assessed by the observers using a standardized form and interviews with the children. Descriptive statistics and content analysis were used to summarize both the quantitative and the qualitative aspects of the data obtained. RESULTS: The mean time for completing the questionnaire was 13.7 minutes (SD 3.68). Compared to the children in 2nd or 3rd grades, those in 4th or 5th grades spent less time completing the questionnaire (median 12.4 vs 13.3 minutes, P=.022), asked for help less frequently (median 0 vs 1.0 count, P=.005), had a lower error count (median 2.0 vs 8.0 count, P<.001), and obtained a higher overall performance score (median 73.0 vs 68.0, P=.005). Children with a PC at home spent less time completing the questionnaire (median 12.3 vs 14.9 minutes, P<.001), had a lower overall error count (median 2.0 vs 9.0 count, P=.03), and had a higher performance score (median 72.0 vs 64.0, P=.005) compared to the children without a PC at home. The most common difficulty in completing the questionnaire was in using the scroll bar. The majority of children reported a positive evaluation (liked a lot or liked) for the four design elements, which were evaluated. CONCLUSIONS: The results of the present study provided feedback to improve the final version of the CAAFE questionnaire. Quantitative data showed minor errors and system failures, while qualitative data indicated that, overall, the children enjoyed the CAAFE questionnaire. Grade levels and PC use must be taken into account in Web-based tools designed for children.

Opa3, a novel regulator of mitochondrial function, controls thermogenesis and abdominal fat mass in a mouse model for Costeff syndrome.

The interrelationship between brown adipose tissue (BAT) and white adipose tissue (WAT) is emerging as an important factor in obesity, but the effect of impairing non-shivering thermogenesis in BAT on lipid storage in WAT remains unclear. To address this, we have characterized the metabolic phenotype of a mouse model for Costeff syndrome, in which a point mutation in the mitochondrial membrane protein Opa3 impairs mitochondrial activity. Opa3(L122P) mice displayed an 80% reduction in insulin-like growth factor 1, postnatal growth retardation and hepatic steatosis. A 90% reduction in uncoupling protein 1 (UCP1) expression in interscapular BAT was accompanied by a marked reduction in surface body temperature, with a 2.5-fold elevation in interscapular BAT mass and lipid storage. The sequestration of circulating lipid into BAT resulted in profound reductions in epididymal and retroperitoneal WAT mass, without affecting subcutaneous WAT. The histological appearance and intense mitochondrial staining in intra-abdominal WAT suggest significant 'browning', but with UCP1 expression in WAT of Opa3(L122P) mice only 62% of that in wild-type littermates, any precursor differentiation does not appear to result in thermogenically active beige adipocytes. Thus, we have identified Opa3 as a novel regulator of lipid metabolism, coupling lipid uptake with lipid processing in liver and with thermogenesis in BAT. These findings indicate that skeletal and metabolic impairment in Costeff syndrome may be more significant than previously thought and that uncoupling lipid uptake from lipid metabolism in BAT may represent a novel approach to controlling WAT mass in obesity.

Medida da atividade física de crianças: o que professores de educação física têm a nos dizer? / Physical activity assessment in children: what physical education teachers tell us?

Este estudo objetivou obter informações sobre o padrão de atividade física de escolares de sete a dez anos para subsidiar o desenvolvimento de um instrumento baseado em web, destinado a monitorar este comportamento com o uso do computador. Foram realizados seis grupos focais com professores de Educação Física (n = 37). As transcrições foram submetidas à análise qualitativa utilizando uma matriz temática que permitiu a identificação de três categorias: padrões de atividade física dos escolares; limitações do instrumento na versão papel e lápis (Questionário de Atividade Física do Dia Anterior); sugestões de melhorias para o instrumento baseado em web. As principais contribuições para o desenho do instrumento foram: o aumento do número de atividades representadas; inclusão de atividades sedentárias; segmentação do dia para auxiliar a recuperação das informações; melhoria da qualidade gráfica dos desenhos. As informações obtidas permitiram estabelecer as características necessárias para o instrumento proposto, contribuindo para o alcance de maiores níveis de validade de conteúdo e de processo de resposta.

This study aimed to obtain information on the physical activity patterns of schoolchildren aged 7-10 years to support the development of a web-based tool designed to monitor this behavior. Six focal groups were carried out with physical education teachers (n = 37). Thematic analyses were performed based on written transcripts. Three major categories were identified: schoolchildren physical activity patterns; limitations of the pencil and paper instrument format (Previous Day Physical Activity Questionnaire); and suggestions to improve the web based instrument. The main contributions for the instrument design were: increasing the number of physical activities presented in paper and pencil format; inclusion of sedentary activities; segmented day format in order to improve information recall; improvements in graphical design quality. Will be used along with researcher?s expertise as well as usability and validity tests in order to obtain a friendly and intelligible interface for 7 to 10 year old schoolchildren. Information gathered through focus groups allowed to establish the necessary characteristics for the proposed instrument, contributing to the achievement of higher levels of content validity and response process.

OPA1 mutation and late-onset cardiomyopathy: mitochondrial dysfunction and mtDNA instability.

BACKGROUND: Mitochondrial fusion protein mutations are a cause of inherited neuropathies such as Charcot-Marie-Tooth disease and dominant optic atrophy. Previously we reported that the fusion protein optic atrophy 1 (OPA1) is decreased in heart failure. METHODS AND RESULTS: We investigated cardiac function, mitochondrial function, and mtDNA stability in a mouse model of the disease with OPA1 mutation. The homozygous mutation is embryonic lethal. Heterozygous OPA(+/-) mice exhibit reduced mtDNA copy number and decreased expression of nuclear antioxidant genes at 3 to 4 months. Although initial cardiac function was normal, at 12 months the OPA1(+/-) mouse hearts had decreased fractional shortening, cardiac output, and myocyte contraction. This coincided with the onset of blindness. In addition to small fragmented mitochondria, aged OPA1(+/-) mice had impaired cardiac mitochondrial function compared with wild-type littermates. CONCLUSIONS: OPA1 mutation leads to deficiency in antioxidant transcripts, increased reactive oxygen species, mitochondrial dysfunction, and late-onset cardiomyopathy.

Specific deficits in visual electrophysiology in a mouse model of dominant optic atrophy.

Autosomal dominant optic atrophy (ADOA) is a slowly progressive optic neuropathy caused by mutations in the OPA1 gene. OPA1 is ubiquitously expressed and plays a key role in mitochondrial fusion. Heterozygous Opa1 mutant mice (B6; C3-Opa1(Q285STOP)), have previously been reported to develop visual defects and optic nerve changes. In this study, in vivo visual electrophysiological testing (ERGs and VEPs) was performed on 11-13 month old B6; C3-Opa1(Q285STOP) mice (n = 5) and age/sex matched wildtype littermate controls. Full intensity series were recorded in response to brief (4 ms) single flash stimuli delivered in a Ganzfeld dome under dark- and light-adapted conditions. The major ERG components (a-wave and b-wave) showed no detectable difference from wildtype in the amplitude or implicit time of dark-adapted ERGs across the full intensity range tested. This was also true for the components of the dark-adapted VEP. However, the light-adapted ERG responses revealed a significant reduction in the photopic negative response (PhNR) amplitude in Opa1(+/-) animals relative to wildtypes at the brighter intensities tested. Elements of the light-adapted VEP were also abnormal in mutant mice. Overall Opa1(+/-) mice display functional deficits in electrophysiology that are consistent with ganglion cell dysfunction. These deficits may correlate with a reduction in the dendritic arborisation of retinal ganglion cells, which has been previously reported to occur at a similar age in the same mutant mouse line (Williams et al., 2010). The functional phenotype we have described in this mouse model may be useful in the robust and accurate assessment of potential treatments for ADOA.

The effects of compression garments on recovery.

The purpose of this study was to investigate whether wearing lower-body compression garments attenuate indices of muscle damage and decrements in performance following drop-jump training. Seven trained female and four trained male subjects undertook blood collection for creatine kinase (CK) and lactate dehydrogenase (LDH), a mid-thigh girth measurement, and reported their perceived muscle soreness (PMS). A series of performance tests were then completed including sprints (5 m, 10 m, and 20 m), a 5-0-5 agility test, and a countermovement jump test. In a randomized crossover experimental design, separated by 1 week, subjects completed 5 x 20 maximal drop-jumps, followed immediately after exercise by either wearing graduated compression tights (CG) or undertook passive recovery as a control (CON) for 48 hours. CK, LDH, mid-thigh girth, and PMS were retested after 24 hours and 48 hours of recovery. The performance tests were repeated after 48 hours of recovery. Analysis of variance for repeated measures indicated that for female subjects, CK values were elevated after 24-hour recovery (p = 0.020) and a greater PMS was observed after 48-hour recovery in the CON condition (p = 0.002) but not for the CG condition. For all the subjects (n = 11), a greater PMS was observed after 48-hour recovery in the CON condition (p = 0.001) but not the CG condition. Significant increases in time were reported for 10-m (p = 0.016, 0.004) and 20-m sprints (p = 0.004, 0.001) in both the CON and CG conditions and for the 5-m sprint (p = 0.014) in the CG condition. All other parameters were unchanged in either condition. Data indicates that CK responses and PMS might be attenuated by wearing compression tights in some participants after drop-jump training; however, no benefit in performance was observed.

Secondary mtDNA defects do not cause optic nerve dysfunction in a mouse model of dominant optic atrophy.

PURPOSE: The majority of patients with autosomal dominant optic atrophy (DOA) harbor pathogenic OPA1 mutations and certain missense mutations, mostly within the GTPase domain, have recently been shown to cause multiple mitochondrial DNA (mtDNA) deletions in skeletal muscle. This raises the possibility that the optic neuropathy could be the result of secondary mtDNA defects accumulating within retinal ganglion cells (RGCs). To explore this hypothesis, the authors looked for evidence of mitochondrial dysfunction in a mouse model of DOA and documented the visual and neurologic progression in aging mutant mice. METHODS: Visual function was assessed with a rotating optokinetic (OKN) drum at ages 13 and 18 months and neurologic phenotyping was performed using the primary SHIRPA screen at age 13 months, comparing mutant Opa1(+/)(-) mice with wild-type C57Bl/6 mice. The presence of cytochrome c oxidase (COX) deficiency and multiple mtDNA deletions was investigated in gastrocnemius muscle and eye specimens harvested from 2- and 11-month-old Opa1(+/+) and Opa1(+/)(-) mice. RESULTS: At age 13 months, Opa1(+/)(-) mice had a statistically significant reduction in OKN responses compared to C57Bl/6 controls with both 2 degrees and 8 degrees gratings (P < 0.001). At age 18 months, the difference between the two groups was significant for the 8 degrees grating (P = 0.003) but not for the 2 degrees grating (P = 0.082). Opa1(+/)(-) mice did not exhibit any significant neuromuscular deficits and no COX deficient areas or secondary mtDNA deletions were identified in skeletal muscle or the RGC layer. There was also no evidence of significant mtDNA depletion or proliferation in skeletal muscle from Opa1(+/)(-) mice. CONCLUSIONS: COX deficiency and mtDNA abnormalities do not contribute to optic nerve dysfunction in pure DOA.

OPA1 deficiency associated with increased autophagy in retinal ganglion cells in a murine model of dominant optic atrophy.

PURPOSE: To examine retinal ganglion cell (RGC) and axonal abnormalities in an ENU-induced mutant mouse carrying a protein-truncating nonsense mutation in OPA1. Mutations in the OPA1 gene cause autosomal dominant optic atrophy (ADOA) in which loss of RGCs followed by myelin degeneration in the optic nerve leads to progressive decrease in visual acuity. METHODS: Ultrastructure of the optic nerve was examined in heterozygous mutants and wild-type littermate controls at 6, 9, and 24 months using electron microscopy. The RGC layer was examined at 6 and 24 months. RESULTS: There was an increase in the number of autophagosomes in the RGC layer in heterozygous mutants compared with wild type at 24 months. Signs of optic nerve degeneration were seen as early as 9 months in Opa1(+/-) mice, with more severe degeneration by 24 months. By 24 months, degeneration of axons was also seen in control mice. Numbers of opaque mitochondria in the Opa1(+/-) mice increased at 6 and 24 months, possibly representing an increase in the density of cristae to fulfill the energy requirements of the axon. In addition, mitochondria with vesiculation of the inner membranes, similar to the mutant mitochondria described in a mouse model of Charcot-Marie-Tooth type 2A, were observed. CONCLUSIONS: Mutations in OPA1 cause pathologic changes to optic nerve axons that are similar to, but occur earlier than, age-related degeneration. Increased autophagy is likely to result from an increase in abnormal mitochondria and could be one mechanism contributing to RGC loss and subsequent optic atrophy seen in ADOA.

A missense mutation in the murine Opa3 gene models human Costeff syndrome.

Opa3 mRNA is expressed in all tissues examined to date, but currently the function of the OPA3 protein is unknown. Intriguingly, various mutations in the OPA3 gene lead to two similar diseases in humans: autosomal dominant inherited optic atrophy and cataract (ADOAC) and a metabolic condition; type 3-methylglutaconic aciduria (MGA). Early onset bilateral optic atrophy is a common characteristic of both disorders; retinal ganglion cells are lost and visual acuity is impaired from an early age. In order to investigate the function of the OPA3 protein, we have generated a novel ENU-induced mutant mouse carrying a missense mutation in the OPA3 gene. The heterozygous mutation in exon 2, causes an amino acid change p.L122P (c.365T>C), which is predicted to alter tertiary protein structure. In the heterozygous state, the mice appear uncompromised however; in the homozygous state mice display some of the features of MGA. Visual function is severely reduced, consistent with significant loss of retinal ganglion cells and degeneration of axons in the optic nerve. In the homozygous optic nerve, there was evidence of increased mitochondrial activity, as demonstrated by the increased presence of mitochondrial marker Cytochrome C Oxidase (COX) histochemistry. Mice homozygous for the opa3(L122P) mutation also display a severe multi-systemic disease characterized by reduced lifespan (majority dying before 4 months), decreased weight, dilated cardiomyopathy, extrapyramidal dysfunction and gross neuro-muscular defects. All of these defects are synonymous with the phenotypic characteristics of Type III MGA found in humans. This model will be of major importance for future studies of the specific function of the OPA3 gene.

Impaired processing of local geometric features during navigation in a water maze following hippocampal lesions in rats.

Hippocampal damage impairs navigation with respect to information provided by the shape of an arena. Recent evidence has suggested that normal rats use local geometric information, as opposed to a global geometric representation, to navigate to a correct corner. One implication of this pattern of results is that hippocampal lesions may impair processing of 1 or more of the local geometric features of an environment. The authors therefore investigated the effects of hippocampal cell loss in rats on navigation to a hidden goal with respect to a variety of local cues in an environment with a distinctive shape. Rats with lesions of the hippocampus were impaired in discriminating a right-angled corner from its mirror image. However, they were able to use cues provided by an acute-angled corner (Experiment 1) or a local polarizing cue (Experiment 2). In contrast, lesioned rats were impaired in discriminating long versus short walls (Experiment 3). Results indicate that the hippocampus plays a role in disambiguating locations by processing (metric) information related to the distance between corners.

Re-amputation occurrence in the diabetic population in South Wales, UK.

The incidence of re-amputation following lower extremity amputations (LEA) among the diabetic patients referred to the Artificial Limb and Appliance Centers (ALAC) in South Wales, UK, was investigated. Manual and electronic data-gathering systems were used to extract the medical records of 473 people with various causes of LEA referred to the ALAC in South Wales during 2001-2003. The data included demographic information, causes of amputation and occurrence of various levels of re-amputation. Two hundred and five subjects with diabetes underwent 316 amputations, 44 were foot amputations and 272 major amputations on the ipsilateral and contra-lateral sides. Of the diabetic patients, 45.9% with single LEA underwent re-amputations with 22% incidence of contra-lateral LEA within 2 years. In comparison, 15% underwent re-amputations in the non diabetic dysvascular patients. Ipsilateral re-amputations occurred much earlier (average 21 weeks) compared with the contra-lateral amputations which took an average of 82 weeks following the first amputation. Nearly half of the diabetic patients with single LEA referred for rehabilitation underwent re-amputations within 2 years; out of which 22% of the patients underwent contra-lateral LEA. Although the progression of level of amputations does not follow a particular pattern, re-amputation on the contra-lateral side occurred almost four times later than that on the ipsilateral side.

Opa1 deficiency in a mouse model of autosomal dominant optic atrophy impairs mitochondrial morphology, optic nerve structure and visual function.

OPA1 is a ubiquitously expressed, nuclear dynamin-related GTPase, targeted to the inner mitochondrial membrane, which plays a role in mitochondrial fusion. Mutations in the OPA1 gene on chromosome 3q28-qter are associated with autosomal dominant optic atrophy (ADOA), the most common inherited optic neuropathy, in which retinal ganglion cells (RGCs) are lost and visual acuity is impaired from an early age. We have generated a novel ENU-induced mutant mouse carrying a protein-truncating nonsense mutation in opa1 in order to explore the pathophysiology of ADOA. The heterozygous mutation, B6; C3-Opa1(Q285STOP), located in exon 8 immediately before the central dynamin-GTPase, leads to approximately 50% reduction in opa1 protein in retina and all tissues on western analysis. The homozygous mutation is embryonic lethal by 13.5 days post coitum, demonstrating the importance of Opa1 during early development. Fibroblasts taken from adult heterozygous mutant mice show an apparent alteration in morphology, with an increase in mitochondrial fission and fragmentation. Heterozygous mutants show a slow onset of degeneration in the optic nerve electron microscopy. Furthermore, they demonstrate a functional reduction in visual function on testing with the optokinetic drum and the circadian running wheel. These findings indicate that the opa1 GTPase contains crucial information required for the survival of RGCs and that Opa1 is essential for early embryonic survival. The Opa1 +/- mice described here provide a means to directly investigate the cellular pathophysiology of OPA1 ADOA.