[Eating Habits of People with Type 2 Diabetes Mellitus in Portugal: A Cross-Sectional Study]. / Hábitos Alimentares das Pessoas com Diabetes Mellitus Tipo 2 em Portugal: Um Estudo Transversal.

INTRODUCTION: Nutrition is a cornerstone of diabetes mellitus prevention and management; therefore, it is essential to enable patients to adopt healthy eating habits. Previous studies have not yet documented the main errors in the eating habits of Portuguese people with type 2 diabetes mellitus. This study aims to identify the main errors in the eating habits of people living with type 2 diabetes mellitus in Portugal and to evaluate its associations with sociodemographic variables. METHODS: Cross-sectional multicentric study in a convenience sample of people with type 2 diabetes mellitus in Primary Health Care Units. The UK Diabetes and Diet Questionnaire (UKDDQ) - translated and adapted, was applied from July to October 2022. Descriptive and inferential statistical analyses were conducted. RESULTS: Of the 550 participants, 52.2% were female, 68.3% were 65 years or over, 55.8 % had an education level up to the fourth grade, 24.7% had economic deprivation, and the mean time since diagnosis was 10.60 ± 8.13 years. Only 36.2% of the sample had a healthy UKDDQ score. Less than 50% of the sample had healthy scores for the items "high-fiber rice or pasta", "high-fiber bread", "butter, margarine and vegetable oils" and "vegetables and pulses". Only 8.9% of the sample had a healthy consumption of fiber. About 70.4% reported healthy scores for the consumption of "high-added-sugar foods" and 54.7% for "high-saturated fat". A statistically significant weak positive correlation was found between the UKDDQ score and age (ρ = 0.201, p < 0.001) with a more frequent choice of healthy foods with increasing age. Female respondents reported healthier habits, particularly in the consumption of "high-saturated fat" and "high-fiber foods". CONCLUSION: The majority of our sample did not take advantage of the potential benefits of healthy eating habits. The main food groups whose consumption should be emphasized or discouraged were individualized, particularly the need to encourage the consumption of high-fiber foods. Targeted educational actions must focus especially on younger and/or male patients.

Introdução: A nutrição é uma área de intervenção na prevenção e gestão da diabetes mellitus; por isso, é fulcral promover a capacitação da população para a adoção de hábitos alimentares saudáveis. Ainda que existam alguns estudos nesta área, não se conhecem os principais erros nos hábitos alimentares das pessoas com diabetes em Portugal. Os objetivos deste estudo foram identificar os principais erros nos hábitos alimentares das pessoas com diabetes mellitus tipo 2 em Portugal e avaliar a sua relação com variáveis sociodemográficas. Métodos: Estudo transversal multicêntrico, em amostra de conveniência de pessoas com diabetes mellitus tipo 2 seguidas em Unidades de Cuidados de Saúde Primários. Aplicação do UK Diabetes and Diet Questionnaire (UKDDQ) ­ traduzido e adaptado, de julho a outubro de 2022. Análise estatística descritiva e inferencial. Resultados: Amostra de 550 participantes, 52,2% do sexo feminino, 68,3% com 65 anos ou mais, 55,8% com nível de escolaridade igual ou inferior ao 1.º ciclo do ensino básico, 24,7% com insuficiência económica e tempo desde o diagnóstico médio de 10,60 ± 8,13 anos. Apenas 36,2% da amostra obteve um score UKDDQ considerado saudável. Menos de 50% obteve scores saudáveis para os itens "arroz ou massa ricos em fibras", "pão integral", "manteiga, margarina e óleos vegetais" e "vegetais e leguminosas". Somente 8,9% da amostra obteve score saudável para o consumo de fibras. Cerca de 70,4% obteve score saudável para o consumo de açúcares livres e 54,7% para o consumo de ácidos gordos saturados. Verificou-se a existência de uma correlação com significado estatístico positiva fraca entre o score UKDDQ e a idade (ρ = 0,201, p < 0,001), com escolha mais frequente de alimentos saudáveis com o aumentar da idade. As pessoas do sexo feminino reportaram hábitos alimentares mais saudáveis, particularmente no consumo de fibras e ácidos gordos saturados. Conclusão: A maior parte da nossa amostra não usufruiu do potencial efeito positivo de uma alimentação saudável. Individualizam-se grupos de alimentos cujos consumos devem ser enfatizados ou desencorajados, particularmente, a necessidade de incentivar o consumo de alimentos ricos em fibra. Ações educacionais dirigidas devem ter especial foco em pessoas mais jovens e/ou do sexo masculino.

Kikuchi-Fujimoto Disease: The Unexpected Diagnosis of a Cervical Adenopathy.

Many cases of adenopathies, whose differential diagnosis includes a wide spectrum of pathologies (including some malignant conditions like lymphoproliferative diseases, e.g., lymphomas), resort to primary healthcare. Kikuchi-Fujimoto disease is a rare, benign, self-limiting entity characterized by adenopathies, mainly in the cervical region, which may be associated with constitutional symptoms. This specific pathology is very rare in primary care and is often overlooked. That is why it is essential to promote medical literacy and provide support in managing these cases, which we want to emphasize through this case presentation. This case report presents a 24-year-old female patient who sought a consultation at the Family Health Unit due to a painful swelling in the right cervical region that lasted two weeks. She denied a history of recent infections or constitutional symptoms. A painful and hard right submaxillary mass, measuring 2 cm in diameter, was identified upon palpation. An analytical study and ultrasound of the soft tissues of the cervical region were initially required. Analytically, there were no relevant changes; however, the ultrasound revealed "hypoechoic ganglion formations in the right laterocervical chains, from the retroauricular region to the lower region of the neck, the largest measuring 19x7mm". The patient was reassessed one month later, due to an increase in the number of adenopathies, and a new ultrasound was performed that revealed "supraclavicular adenopathy". After that, she was referred to Secondary Healthcare (Central Hospital), where a lymph node biopsy was performed, with histological results of Kikuchi-Fujimoto disease. The patient maintains a follow-up in a hemato-oncology consultation, with painless adenopathies that, according to her, get worse with anxiety symptoms. Currently, the patient is being treated symptomatically, with stabilization of adenopathies and anxious manifestations. These patients need long-term follow-up due to the possibility of disease recurrence or the development of autoimmune processes. Although it is a diagnosis of exclusion, this disease must always be considered, since it can be mistaken with other serious pathologies that require aggressive treatments. Regarding the relationship between anxiety disorder and the worsening of adenopathies, although no conclusive evidence was found in the literature, there are some studies that have established a connection between inflammation and the deterioration of certain depressive symptoms.

Injection of kaolin/carrageenan in the rat knee joint induces progressive experimental knee osteoarthritis.

ABSTRACT: Osteoarthritis (OA), the most common joint disorder worldwide, is characterized by progressive degeneration of articular and periarticular structures, leading to physical and emotional impairments that greatly affect the quality of life of patients. Unfortunately, no therapy has been able to halt the progression of the disease. Owing to the complexity of OA, most animal models are only able to mimic a specific stage or feature of the human disorder. In this work, we demonstrate the intraarticular injection of kaolin or carrageenan leads to the progressive degeneration of the rat's knee joint, accompanied by mechanical hyperalgesia and allodynia, gait impairments (reduced contact area of the affected limb), and radiological and histopathological findings concomitant with the development of human grade 4 OA. In addition, animals also display emotional impairments 4 weeks after induction, namely, anxious and depressive-like behaviour, important and common comorbidities of human OA patients. Overall, prolonging kaolin or carrageenan-induced monoarthritis mimics several important physical and psychological features of human OA in both male and female rodents and could be further applied in long-term studies of OA-associated chronic pain.

Nutritional intake and food sources in an adult urban Kenyan population.

Urbanisation is hastening the transition from traditional food habits to less healthy diets, which are becoming more common among Kenyans. No up-to-date studies on usual dietary intake and the main food sources of adult Kenyans are available. The aim of the present study was to identify the main food sources of nutrients in the diet of urban adult Kenyans and explore potential associations with demographic variables including age, sex, level of education, occupation and body mass index. The study adopted a cross-sectional design. The dietary intake of 486 adult Kenyans from Nairobi was assessed using a validated, culture-sensitive, semi-quantitative food frequency questionnaire. Binary logistic regression models were used to evaluate associations between food sources and demographic variables. Macronutrient intakes as a proportion of total energy intake (TEI) were within international dietary guidelines. Cereals and grain products (34.0%), sugar, syrups, sweets and snacks (9.8%), fruits (9.7%) and meat and eggs (8.8%) were the major contributors to TEI. Cereals and grain products contributed 42.5% to carbohydrates, followed by fruits (12.4%) and sugar, syrups, sweets and snacks (10.6%). The most important sources of protein and total fat were cereals and grain products (23.3% and 19.7%, respectively) and meat and eggs (22.0% and 18.7%, respectively). Sex, age and level of education were associated with the choice of food groups. Although macronutrient intakes were within guidelines, the Kenyan diet was revealed to be high in sugars, salt and fibre, with differences in food sources according to demographic variables. These results can act as an incentive to national authorities to implement nutritional strategies aiming to raise awareness of healthier dietary patterns among Kenyans.

Comparison of the effects of shock waves versus radiofrequency on abdominal lipolysis: A randomized clinical trial.

BACKGROUND: High levels of abdominal adiposity mean higher risk of developing cardiovascular diseases. Aerobic exercise per si reduces the risk of developing this type of diseases. Radiofrequency and shockwave therapy showed to be effective in the reduction of localized abdominal fat. AIM (S): To compare the effects of prescribed moderate aerobic exercise associated with 6 sessions of shockwave therapy or radiofrequency, in the subcutaneous abdominal adipose tissue, in overweight female individuals in fertile age. METHODS: Randomized clinical trial, with 30 volunteers in fertile age with overweight and/or obese, randomly and equitably allocated in two experimental groups (1-shockwave therapy and 2-radiofrequency) and one control group. Aerobic physical exercise was prescribed to all groups. The values of anthropometric measurements were measured in two moments. The intervention protocol was performed in a clinic for six weeks, with one session per week. The ANOVA test, paired samples t test, Kruskal-Wallis test, and Wilcoxon test were used to compare the results, for a significance level of 0.05. RESULTS: Of 28 participants completed the study. There were significant differences in waist circumference reduction between the experimental group 1 and 2 when compared to the control group (p = 0.005 and p = 0.014, respectively). There was a statistically significant reduction in navel level circumference in experimental group 1 when compared to experimental group 2 (p = 0.024) and with the control group (p = 0.016). CONCLUSION: Both resources were effective in reducing abdominal measurements when compared to the control group. However, SWT was superior in reducing the navel level circumference.

Integrated bioprocess for structured lipids, emulsifiers and biodiesel production using crude acidic olive pomace oils.

Olive pomace oil (OPO), a by-product of olive oil industry, is directly consumed after refining. The novelty of this study consists of the direct use of crude high acidic OPO (3.4-20% acidity) to produce added-value compounds, using sn-1,3-regioselective lipases: (i) low-calorie dietetic structured lipids (SL) containing caprylic (C8:0) or capric (C10:0) acids by acidolysis or interesterification with their ethyl esters, (ii) fatty acid methyl esters (FAME) for biodiesel, and (iii) sn-2 monoacylglycerols (emulsifiers), as by-product of FAME production by methanolysis. Immobilized Rhizomucor miehei lipase showed similar activity in acidolysis and interesterification for SL production (yields: 47.8-53.4%, 7 h, 50℃) and was not affected by OPO acidity. Batch operational stability decreased with OPO acidity, but it was at least three-fold in interesterification that in acidolysis. Complete conversion of OPO into FAME and sn-2 monoacylglycerols was observed after 3 h-transesterification (glycerol stepwise addition) and lipase deactivation was negligeable after 11 cycles.

Comunicação com a família no serviço de urgência durante a pandemia da COVID-19 / Communication with the family in the emergency service during the pandemic of COVID-19

Enquadramento: O presente relatório incorporado no plano curricular do Curso de Mestrado em Enfermagem da Pessoa em Situação Crítica, da Escola Superior de Enfermagem da Universidade do Minho, em consórcio com a Escola Superior de Saúde da Universidade de Trás-os-Montes e Alto Douro, reflete a componente prática, académica e profissional que permitiu o avigoramento de um olhar díspar sobre a comunicação com a família no serviço de urgência durante a atual pandemia da COVID-19. Neste sentido apresenta-se uma revisão integrativa da literatura sobre a comunicação com a família durante a pandemia. Objetivos: Apresentar uma reflexão crítica sobre as experiências vividas ao longo do estágio; relacionar as atividades concretizadas no decorrer do estágio com as competências desenvolvidas, quer as competências comuns de enfermeiro especialista, quer as de enfermeiro especialista em enfermagem médico-cirúrgica, na área de enfermagem à pessoa em situação crítica e, apresentar um estudo de investigação concretizado no decorrer do estágio. Métodos: A metodologia descritiva, analítica e reflexiva, garantiu a concretização dos dois primeiros objetivos. No que se refere ao último objetivo, optou-se pela Metodologia de Projeto, tendo sido desenvolvido um Projeto de Intervenção no âmbito da comunicação com a família no serviço de urgência durante a atual pandemia, destinado aos profissionais de enfermagem que trabalham naquele contexto. Utilizou-se a estratégia PICO e as recomendações PRISMA apresentadas pelo Joanna Briggs Institute, partindo da questão: "Como foram as experiências de comunicação dos enfermeiros com as famílias no serviço de urgência durante a pandemia da COVID-19?". O processo de pesquisa teve início nas bases de dados b-on, Medline, Scopus, PubMed, Web of Science, no Repositório Científico de Acesso Aberto de Portugal e Repositório da Universidade do Minho, com um número total de trezentos e um (N=301) artigos selecionados, com uma filtração cronológica dos últimos dois anos, 2020 e 2021 (anos da pandemia) e com ativação das opções de texto integral na língua inglesa, portuguesa e espanhola. Posteriormente, e fundamentada nesta mesma revisão integrativa da literatura, desenvolveu-se a norma de procedimento de enfermagem. Por forma a dar resposta aos objetivos foi realizada uma atividade formativa sobre a temática, onde os enfermeiros após a sessão procederam à avaliação da mesma e do formador. Resultados: No âmbito deste relatório destacam-se as atividades concretizadas no decorrer do estágio, relacionando-as com o desenvolvimento de competências. A revisão integrativa da literatura releva a necessidade da consciencialização regular dos enfermeiros sobre a comunicação com a família. Também, tendo em conta o período pandémico, há a necessidade da criação de alternativas de comunicação com a pessoa em situação crítica, de modo a facilitar a participação e interação familiar, mediante o recurso à tecnologia virtual, entre outras. Conclusão: No decurso desta especialização houve a necessidade de afluir a visão do cuidar à pessoa em situação crítica em áreas afetas à intervenção de enfermagem. Neste sentido, esta aquisição de competências valoriza o percurso profissional e pessoal, contribuindo para a qualidade dos cuidados. O estágio no serviço de urgência proporcionou uma série de experiências enriquecedoras e permitiu a aquisição de competências especializadas na área da enfermagem médico-cirúrgica, na área de enfermagem à pessoa em situação crítica. O estudo da comunicação no relacionamento entre o enfermeiro e a família assumiu-se como fundamental naquele contexto, justificado pela integração dos familiares no processo terapêutico. A família, como sublinhámos, é uma parte ativa e parceira nos cuidados da pessoa em situação crítica. Como tal, torna-se importante o reforço deste tema, e o treino de competências de comunicação integrado na formação contínua dos enfermeiros. Este será o caminho no sentido de estes profissionais se encontrarem mais capacitados para o desenvolvimento de uma relação interpessoal benéfica e se apresentarem como cuidadores mais disponíveis, dotados de maior sensibilidade e passíveis de atribuir uma atenção personalizada, numa vertente holístico-terapêutica.

Background: This report incorporated in the curriculum of the Master's Course in Nursing of the Person in Critical Situation, of the Higher School of Nursing of the University of Minho, in consortium with the School of Health of the University of Trás-os-Montes and Alto Douro, reflects the practical, academic and professional component which has allowed the invigoration of a different look at communication with the family in the emergency service during the current pandemic of COVID-19. In this sense, an integrative review of the literature on communication with the family during the pandemic is presented. Purposes: To present a critical reflection on the experiences lived along the internship; to relate the activities implemented during the internship with the competences developed, both the common competences of specialist nurses and those of specialist nurses in medical-surgical nursing, in the area of nursing of the person in critical situation and to present a research study implemented during the internship. Methods: The descriptive, analytical and reflective methodology ensured the achievement of the first two objectives. With regard to the last objective, it was chosen the Project Methodology, having been developed an intervention project in the context of communication with the family in the emergency service during the current pandemic, aimed at nursing professionals working in that context. The PICO strategy and the PRISMA recommendations presented by the Joanna Briggs Institute were used, starting from the question: "How were the experiences of communication of nurses with families in the emergency service during the pandemic of COVID-19?". The research process started in the databases b-on, Medline, Scopus, PubMed, Web of Science, in the Scientific Repository of Open Access of Portugal and Repository of the University of Minho, with a total number of three hundred and one (N=301) selected articles, with a chronological filtration of the last two years, 2020 and 2021 (years of the pandemic) and with activation of the options of integral text in English, Portuguese and Spanish. Later, and based on this integrative review of the literature, the nursing procedure standard has been developed. In order to respond to the purposes, a formative activity was carried out on the theme and after the session the nurses have evaluated both the session itself and the trainer. Results: In the context of this report, the activities carried out during the internship, which are related to the development of competences, stand out. The literature review recalls the need for regular awareness of nurses about communication with the family. Besides, taking into account the pandemic period, there is a need to create alternatives for communication with the person in critical situation, in order to facilitate participation and family interaction, through the use of virtual technology, among others. Conclusion: In the course of this specialization, there was the need to relate the vision of caring for the person in critical situation to areas of nursing intervention. In this sense, this acquisition of skills values the professional and personal path, contributing to the quality of caring. The internship in the emergency service provided a series of enriching experiences and allowed the acquisition of specialized competences in the area of medical-surgical nursing nurses, in the area of nursing of the person in critical situation. The study of communication in the relationship between the nurse and the family was fundamental in that context, justified by the integration of family members in the therapeutic process. The family, as we have stressed, is an active part and a partner in the caring of the person in critical situation. As such, it is important to reinforce this theme and integrate the training of communication skills in the continuous instruction of nurses. This will be the way for these professionals to feel themselves more capable of developing a beneficial interpersonal relationship and present themselves as more available caretakers, with greater sensitivity and skilled to give personalized attention, in a holistic-therapeutic approach.

Noncovalent Interactions with PAMAM and PPI Dendrimers Promote the Cellular Uptake and Photodynamic Activity of Rose Bengal: The Role of the Dendrimer Structure.

Rose bengal is an anionic dye considered as a potential photosensitizer for anticancer photodynamic therapy. The clinical utility of rose bengal is hampered by its short half-life, limited transmembrane transport, aggregation, and self-quenching; consequently, efficient drug carriers that overcome these obstacles are urgently required. In this study, we performed multilevel in vitro and in silico characterization of interactions between rose bengal and cationic poly(amidoamine) (PAMAM) and poly(propyleneimine) (PPI) dendrimers of the third and fourth generation and assessed the ability of the resultant complexes to modulate the photosensitizing properties of the drug. We focused on explaining the molecular basis of this phenomenon and proved that the generation- and structure-dependent binding of the dye by the dendrimers increases the cellular uptake and production of singlet oxygen and intracellular reactive oxygen species, leading to an increase in phototoxicity. We conclude that the application of dendrimer carriers could enable the design of efficient photodynamic therapies based on rose bengal.

A decade of experience with genetically tailored pig models for diabetes and metabolic research.

The global prevalence of diabetes mellitus and other metabolic diseases is rapidly increasing. Animal models play pivotal roles in unravelling disease mechanisms and developing and testing therapeutic strategies. Rodents are the most widely used animal models but may have limitations in their resemblance to human disease mechanisms and phenotypes. Findings in rodent models are consequently often difficult to extrapolate to human clinical trials. To overcome this 'translational gap', we and other groups are developing porcine disease models. Pigs share many anatomical and physiological traits with humans and thus hold great promise as translational animal models. Importantly, the toolbox for genetic engineering of pigs is rapidly expanding. Human disease mechanisms and targets can therefore be reproduced in pigs on a molecular level, resulting in precise and predictive porcine (PPP) models. In this short review, we summarize our work on the development of genetically (pre)diabetic pig models and how they have been used to study disease mechanisms and test therapeutic strategies. This includes the generation of reporter pigs for studying beta-cell maturation and physiology. Furthermore, genetically engineered pigs are promising donors of pancreatic islets for xenotransplantation. In summary, genetically tailored pig models have become an important link in the chain of translational diabetes and metabolic research.

How Can Biomolecules Improve Mucoadhesion of Oral Insulin? A Comprehensive Insight using Ex-Vivo, In Silico, and In Vivo Models.

Currently, insulin can only be administered through the subcutaneous route. Due to the flaws associated with this route, it is of interest to orally deliver this drug. However, insulin delivered orally has several barriers to overcome as it is degraded by the stomach's low pH, enzymatic content, and poor absorption in the gastrointestinal tract. Polymers with marine source like chitosan are commonly used in nanotechnology and drug delivery due to their biocompatibility and special features. This work focuses on the preparation and characterization of mucoadhesive insulin-loaded polymeric nanoparticles. Results showed a suitable mean size for oral administration (<600 nm by dynamic laser scattering), spherical shape, encapsulation efficiency (59.8%), and high recovery yield (80.6%). Circular dichroism spectroscopy demonstrated that protein retained its secondary structure after encapsulation. Moreover, the mucoadhesive potential of the nanoparticles was assessed in silico and the results, corroborated with ex-vivo experiments, showed that using chitosan strongly increases mucoadhesion. Besides, in vitro and in vivo safety assessment of the final formulation were performed, showing no toxicity. Lastly, the insulin-loaded nanoparticles were effective in reducing diabetic rats' glycemia. Overall, the coating of insulin-loaded nanoparticles with chitosan represents a potentially safe and promising approach to protect insulin and enhance peroral delivery.

Mild maternal hyperglycemia in INSC93S transgenic pigs causes impaired glucose tolerance and metabolic alterations in neonatal offspring.

Alongside the obesity epidemic, the prevalence of maternal diabetes is rising worldwide, and adverse effects on fetal development and metabolic disturbances in the offspring's later life have been described. To clarify whether metabolic programming effects are due to mild maternal hyperglycemia without confounding obesity, we investigated wild-type offspring of INSC93S transgenic pigs, which are a novel genetically modified large-animal model expressing mutant insulin (INS) C93S in pancreatic ß-cells. This mutation results in impaired glucose tolerance, mild fasting hyperglycemia and insulin resistance during late pregnancy. Compared with offspring from wild-type sows, piglets from hyperglycemic mothers showed impaired glucose tolerance and insulin resistance (homeostatic model assessment of insulin resistance: +3-fold in males; +4.4-fold in females) prior to colostrum uptake. Targeted metabolomics in the fasting and insulin-stimulated state revealed distinct alterations in the plasma metabolic profile of piglets from hyperglycemic mothers. They showed increased levels of acylcarnitines, gluconeogenic precursors such as alanine, phospholipids (in particular lyso-phosphatidylcholines) and α-aminoadipic acid, a potential biomarker for type 2 diabetes. These observations indicate that mild gestational hyperglycemia can cause impaired glucose tolerance, insulin resistance and associated metabolic alterations in neonatal offspring of a large-animal model born at a developmental maturation status comparable to human babies.

Urine Nuclear Magnetic Resonance (NMR) Metabolomics in Age-Related Macular Degeneration.

Biofluid biomarkers of age-related macular degeneration (AMD) are still lacking, and their identification is challenging. Metabolomics is well-suited to address this need, and urine is a valuable accessible biofluid. This study aimed to characterize the urinary metabolomic signatures of patients with different stages of AMD and a control group (>50 years). It was a prospective, cross-sectional study, where subjects from two cohorts were included: 305 from Coimbra, Portugal (AMD patients n = 252; controls n = 53) and 194 from Boston, United States (AMD patients n = 147; controls n = 47). For all participants, we obtained color fundus photographs (for AMD staging) and fasting urine samples, which were analyzed using 1H nuclear magnetic resonance (NMR) spectroscopy. Our results revealed that in both cohorts, urinary metabolomic profiles differed mostly between controls and late AMD patients, but important differences were also found between controls and subjects with early AMD. Analysis of the metabolites responsible for these separations revealed that, even though distinct features were observed for each cohort, AMD was in general associated with depletion of excreted citrate and selected amino acids at some stage of the disease, suggesting enhanced energy requirements. In conclusion, NMR metabolomics enabled the identification of urinary signals of AMD and its severity stages, which might represent potential metabolomic biomarkers of the disease.

The long non-coding RNA MYCNOS-01 regulates MYCN protein levels and affects growth of MYCN-amplified rhabdomyosarcoma and neuroblastoma cells.

BACKGROUND: MYCN is amplified in small cell lung cancers and several pediatric tumors, including alveolar rhabdomyosarcomas and neuroblastomas. MYCN protein is known to play a key oncogenic role in both alveolar rhabdomyosarcomas and neuroblastomas. MYCN opposite strand (MYCNOS) is a gene located on the antisense strand to MYCN that encodes alternatively spliced transcripts, two of which (MYCNOS-01 and MYCNOS-02) are known to be expressed in neuroblastoma and small cell lung cancer with reciprocal regulation between MYCNOS-02 and MYCN reported for neuroblastomas. We sought to determine a functional role for MYCNOS-01 in alveolar rhabdomyosarcoma and neuroblastoma cells and identify any associated regulatory effects between MYCN and MYCNOS-01. METHODS: MYCNOS-01, MYCNOS-02 and MYCN expression levels were assessed in alveolar rhabdomyosarcoma and neuroblastoma cell lines and tumor samples from patients using Affymetrix microarray data and quantitative RT-PCR. Following MYCNOS-01 or MYCN siRNA knockdown and MYCNOS-01 overexpression, transcript levels were assayed by quantitative RT-PCR and MYCN protein expression assessed by Western blot and immunofluorescence. Additionally, effects on cell growth, apoptosis and cell cycle profiles were determined by a metabolic assay, caspase activity and flow cytometry, respectively. RESULTS: MYCNOS-01 transcript levels were generally higher in NB and RMS tumor samples and cell lines with MYCN genomic amplification. RNA interference of MYCNOS-01 expression did not alter MYCN transcript levels but decreased MYCN protein levels. Conversely, MYCN reduction increased MYCNOS-01 transcript levels, creating a negative feedback loop on MYCN protein levels. Reduction of MYCNOS-01 or MYCN expression decreased cell growth in MYCN-amplified alveolar rhabdomyosarcoma and neuroblastoma cell lines. This is consistent with MYCNOS-01-mediated regulation of MYCN contributing to the phenotype observed. CONCLUSIONS: An alternative transcript of MYCNOS, MYCNOS-01, post-transcriptionally regulates MYCN levels and affects growth in MYCN-amplified rhabdomyosarcoma and neuroblastoma cells.

Hyperglycaemic Hemichorea.

BACKGROUND: Hyperkinetic disorders such as hemichorea can be caused by cerebrovascular, infectious or inflammatory diseases or by metabolic conditions such as hyperglycaemia. Hyperglycaemic hemichorea is a rare movement disorder which is frequently misdiagnosed. It is characterized by involuntary, continuous, non-patterned movements on one side of the body, basal ganglia lesions seen on head CT or MRI, and clinical improvement after blood glucose normalization. We describe the case of a female patient with uncontrolled diabetes who presented with hemichorea. CASE PRESENTATION: We report the case of a 69-year-old woman with type 2 diabetes who presented with abnormal movements of the right upper limb. She had no neurological signs other than hemichorea. Her blood glucose level was 349 mg/dl and her glycosylated haemoglobin level (HbA1c) was 10.5%. Head CT and MRI showed no changes in the basal ganglia or ischaemic lesions. The patient was started on insulin and haloperidol with clinical improvement. CONCLUSION: Larger case series are needed to establish better understanding of the physiopathological mechanisms and diagnostic criteria of hyperglycaemic hemichorea. The most important diagnostic criterion is clinical improvement after glycaemic control. LEARNING POINTS: Hyperglycaemia is a rare cause of hemichorea.Better understanding of the physiopathology and the establishment of diagnostic criteria are required.Correction of the underlying hyperglycaemia will lead to rapid improvement of the movements and is the most important feature for diagnosis.

Human plasma metabolomics in age-related macular degeneration (AMD) using nuclear magnetic resonance spectroscopy.

PURPOSE: To differentiate the plasma metabolomic profile of patients with age related macular degeneration (AMD) from that of controls, by Nuclear Magnetic Resonance (NMR) spectroscopy. METHODS: Two cohorts (total of 396 subjects) representative of central Portugal and Boston, USA phenotypes were studied. For each cohort, subjects were grouped according to AMD stage (early, intermediate and late). Multivariate analysis of plasma NMR spectra was performed, followed by signal integration and univariate analysis. RESULTS: Small changes were detected in the levels of some amino acids, organic acids, dimethyl sulfone and specific lipid moieties, thus providing some biochemical information on the disease. The possible confounding effects of gender, smoking history and age were assessed in each cohort and found to be minimal when compared to that of the disease. A similar observation was noted in relation to age-related comorbidities. Furthermore, partially distinct putative AMD metabolite fingerprints were noted for the two cohorts studied, reflecting the importance of nutritional and other lifestyle habits in determining AMD metabolic response and potential biomarker fingerprints. Notably, some of the metabolite changes detected were noted as potentially differentiating controls from patients diagnosed with early AMD. CONCLUSION: For the first time, this study showed metabolite changes in the plasma of patients with AMD as compared to controls, using NMR. Geographical origins were seen to affect AMD patients´ metabolic profile and some metabolites were found to be valuable in potentially differentiating controls from early stage AMD patients. Metabolomics has the potential of identifying biomarkers for AMD, and further work in this area is warranted.

The Unfolded Protein Response in Homeostasis and Modulation of Mammalian Immune Cells.

The endoplasmic reticulum (ER) plays important roles in eukaryotic protein folding and lipid biosynthesis. Several exogenous and endogenous cellular sources of stress can perturb ER homeostasis leading to the accumulation of unfolded proteins in the lumen. Unfolded protein accumulation triggers a signal-transduction cascade known as the unfolded protein response (UPR), an adaptive mechanism which aims to protect cells from protein aggregates and to restore ER functions. Further to this protective mechanism, in immune cells, UPR molecular effectors have been shown to participate in a wide range of biological processes such as cell differentiation, survival and immunoglobulin and cytokine production. Recent findings also highlight the involvement of the UPR machinery in the maturational program and antigen presentation capacities of dendritic cells. UPR is therefore a key element in immune system homeostasis with direct implications on both adaptive and innate immune responses. The present review summarizes the knowledge on the emerging roles of UPR signaling cascades in mammalian immune cells as well as the consequences of their dysregulation in relation to the pathogenesis of several diseases.

Hyperthermia adds to trabectedin effectiveness and thermal enhancement is associated with BRCA2 degradation and impairment of DNA homologous recombination repair.

The tetrahydroisoquinoline trabectedin is a marine compound with approved activity against human soft-tissue sarcoma. It exerts antiproliferative activity mainly by specific binding to the DNA and inducing DNA double-strand breaks (DSB). As homologous recombination repair (HRR)-deficient tumors are more susceptible to trabectedin, hyperthermia-mediated on-demand induction of HRR deficiency represents a novel and promising strategy to boost trabectedin treatment. For the first time, we demonstrate enhancement of trabectedin effectiveness in human sarcoma cell lines by heat and characterize cellular events and molecular mechanisms related to heat-induced effects. Hyperthermic temperatures (41.8 or 43°C) enhanced significantly trabectedin-related clonogenic cell death and G2/M cell cycle arrest followed by cell type-dependent induction of apoptosis or senescence. Heat combination increased accumulation of γH2AX foci as key marker of DSBs. Expression of BRCA2 protein, an integral protein of the HRR machinery, was significantly decreased by heat. Consequently, recruitment of downstream RAD51 to γH2AX-positive repair foci was almost abolished indicating relevant impairment of HRR by heat. Accordingly, enhancement of trabectedin effectiveness was significantly augmented in BRCA2-proficient cells by hyperthermia and alleviated in BRCA2 knockout or siRNA-transfected BRCA2 knockdown cells. In peripheral blood mononuclear cells isolated from sarcoma patients, increased numbers of nuclear γH2AX foci were detected after systemic treatment with trabectedin and hyperthermia of the tumor region. The findings establish BRCA2 degradation by heat as a key factor for a novel treatment strategy that allows targeted chemosensitization to trabectedin and other DNA damaging antitumor drugs by on-demand induction of HRR deficiency.

Impact of fetal chromosomal disorders on maternal blood metabolome: toward new biomarkers?

OBJECTIVE: This study aimed at determining the relationship between fetal chromosomal disorders (CDs), including trisomy 21 (T21), and on first- and second-trimester maternal blood plasma, to identify the time-course metabolic adaptations to the conditions and the possible new plasma biomarkers. Furthermore, a definition of a joint circulatory (plasma) and excretory (urine) metabolic description of second-trimester CDs was sought. STUDY DESIGN: Plasma was obtained for 119 pregnant women: 74 controls and 45 CD cases, including 22 T21 cases. Plasma and lipid extracts (for T21 only) were analyzed by nuclear magnetic resonance spectroscopy, and data were handled by variable selection and multivariate analysis. Correlation analysis was used on a concatenated plasma/urine matrix descriptive of second-trimester CD, based on previously obtained urine data. RESULTS: CD cases were accompanied by enhanced lipid ß-oxidation (increased ketone bodies) and underutilization of glucose, pyruvate, and citrate. Lower circulating high-density lipoprotein levels were noted, along with changes in the proline and methanol in the first trimester, and also the urea, creatinine, acetate, and low-density lipoprotein plus very low-density lipoprotein in the second trimester and the different urea and creatinine levels, suggesting fetal renal dysfunction. In terms of plasma composition, T21 cases were indistinguishable from other CDs in the first trimester, whereas in the second trimester, increased methanol and albumin may be T21 specific. Furthermore, first-trimester lipid extracts of T21 showed decreased levels of 18:2 fatty acids, whereas in the second trimester, lower levels of 20:4 and 22:6 fatty acids were noted, possibly indicative of inflammation mechanisms. In both trimesters, high classification rates for CDs (88-89%) and T21 (85-92%) generally relied on variable selection of nuclear magnetic resonance data. Plasma/urine correlations confirmed most metabolic deviations and unveiled possible new ones regarding low-density lipoprotein plus very low-density lipoprotein, sugar, and gut-microflora metabolisms. CONCLUSION: This work partially confirmed previously reported data on first-trimester T21 and provided additional information on time-course metabolic changes accompanying CD and T21, in particular regarding plasma lipid composition. These results demonstrate the potential of plasma metabolomics in monitoring and characterizing CD cases; however, validation in larger cohorts is desirable.

Targeting the Insulin-Like Growth Factor 1 Receptor in Ewing's Sarcoma: Reality and Expectations.

Ewing's sarcoma family of tumours comprises a group of very aggressive diseases that are potentially curable with multimodality treatment. Despite the undoubted success of current treatment, approximately 30% of patients will relapse and ultimately die of disease. The insulin-like growth factor 1 receptor (IGF-1R) has been implicated in the genesis, growth, proliferation, and the development of metastatic disease in Ewing's sarcoma. In addition, IGF1-R has been validated, both in vitro and in vivo, as a potential therapeutic target in Ewing's sarcoma. Phase I studies of IGF-1R monoclonal antibodies reported several radiological and clinical responses in Ewing's sarcoma patients, and initial reports of several Phase II studies suggest that about a fourth of the patients would benefit from IGF-1R monoclonal antibodies as single therapy, with approximately 10% of patients achieving objective responses. Furthermore, these therapies are well tolerated, and thus far severe toxicity has been rare. Other studies assessing IGF-1R monoclonal antibodies in combination with traditional cytotoxics or other targeted therapies are expected. Despite, the initial promising results, not all patients benefit from IGF-1R inhibition, and consequently, there is an urgent need for the identification of predictive markers of response.