Proteomics on porcine haptoglobin and IgG/IgA show protein species distribution and glycosylation pattern to remain similar in PCV2-SD infection

Haptoglobin (Hp) and immunoglobulins are plasma glycoproteins involved in the immune reaction of the organism after infection and/or inflammation. Porcine circovirus type 2-systemic disease (PCV2-SD), formerly known as postweaning multisystemic wasting syndrome (PMWS), is a globally spread pig disease of great economic impact. PCV2-SD affects the immunological system of pigs causing immunosuppression. The aim of this work was to characterize the Hp protein species of healthy and PCV2-SD affected pigs, as well as the protein backbone and the glycan chain composition of porcine Hp. PCV2-SD affected pigs had an increased overall Hp level, but it did not affect the ratio between Hp species. Glycoproteomic analysis of the Hp β subunits confirmed that porcine Hp is N-glycosylated and, unexpectedly, O-glycosylated, a PTM that is not found on Hp from healthy humans. The glyco-profile of porcine IgG and IgA heavy chains was also characterized; decreased levels of both proteins were found in the investigated group of PCV2-SD affected pigs. Obtained results indicate that no significant changes in the N- and O-glycosylation patterns of these major porcine plasma glycoproteins were detectable between healthy and PCV2-SD affected animals.

Phosphorylated polyols, pyrophosphates, and derivatives thereof having biological activity

The present invention provides phosphorylated and pyrophosphate derivatives of polyols, and structural derivatives of these compounds, and provides pharmaceutical compositions comprising the same. The compounds and compositions disclosed herein have various biological activities, including for example, as allosteric effectors of hemoglobin and/or as kinase inhibitors. The present invention further provides methods for therapy in human or mammalian patients, and methods for synthesis of biologically active compounds and their intermediates.

Education, gender, and state-level disparities in the health of older Indians: Evidence from biomarker data

Using new biomarker data from the 2010 pilot round of the Longitudinal Aging Study in India (LASI), we investigate education, gender, and state-level disparities in health. We find that hemoglobin level, a marker for anemia, is lower for respondents with no schooling (0.7 g/dL less in the adjusted model) compared to those with some formal education and is also lower for females than for males (2.0 g/dL less in the adjusted model). In addition, we find that about one third of respondents in our sample aged 45 or older have high C-reaction protein (CRP) levels (>3 mg/L), an indicator of inflammation and a risk factor for cardiovascular disease. We find no evidence of educational or gender differences in CRP, but there are significant state-level disparities, with Kerala residents exhibiting the lowest CRP levels (a mean of 1.96 mg/L compared to 3.28 mg/L in Rajasthan, the state with the highest CRP). We use the Blinder–Oaxaca decomposition approach to explain group-level differences, and find that state-level disparities in CRP are mainly due to heterogeneity in the association of the observed characteristics of respondents with CRP, rather than differences in the distribution of endowments across the sampled state populations.

Epidemiology of Dysglycemia in Pregnant Oklahoma American Indian Women

CONTEXT: Minority communities are disproportionately affected by diabetes, and minority women are at an increased risk for glucose intolerance (dysglycemia) during pregnancy.

OBJECTIVES: In pregnant American Indian women, the objectives of the study were to use current criteria to estimate the prevalence of first-trimester (Tr1) dysglycemia and second-trimester (Tr2) incidence of gestational diabetes mellitus (GDM) and to explore new candidate measures and identify associated clinical factors.

DESIGN: This was a prospective cohort study. In Tr1 we performed a 75-g, 2-hour oral glucose tolerance test (OGTT) and glycated hemoglobin (HbA1c) to determine the following: fasting insulin; homeostasis model assessment of insulin resistance; serum 1,5-anhydroglucitol; noninvasive skin autofluorescence (SCOUT). We defined dysglycemia by American Diabetes Association and Endocrine Society criteria and as HbA1c of 5.7% or greater. In Tr2 in an available subset, we performed a repeat OGTT and SCOUT.

PARTICIPANTS: Pregnant American Indian women (n = 244 at Tr1; n = 114 at Tr2) participated in the study.

OUTCOMES: The prevalence of dysglycemia at Tr1 and incidence of GDM at Tr2 were measured.

RESULTS: At Tr1, one woman had overt diabetes; 36 (15%) had impaired glucose tolerance (American Diabetes Association criteria and/or abnormal HbA1c) and 59 (24%) had GDM-Tr1 (Endocrine Society criteria). Overall, 74 (30%) had some form of dysglycemia. Associated factors were body mass index, hypertension, waist/hip circumferences, SCOUT score, fasting insulin, and homeostasis model assessment of insulin resistance. At Tr2, 114 of the Tr1 cohort underwent a repeat OGTT and SCOUT, and 26 (23%) had GDM. GDM-Tr2 was associated with increased SCOUT scores (P = .029) and Tr1 body mass index, waist/hip circumferences, diastolic blood pressure, fasting insulin, and triglyceride levels. Overall, dysglycemia at Tr1 and/or Tr2 affected 38% of the women.

CONCLUSIONS: Dysglycemia at some point during pregnancy was common among American Indian women. It was associated with features of insulin resistance and may confer long-term health risks for mother and child.

Cord blood adipokines, neonatal anthropometrics and postnatal growth in offspring of Hispanic and Native American women with diabetes mellitus

BACKGROUND: Offspring of women with diabetes mellitus (DM) during pregnancy have a risk of developing metabolic disease in adulthood greater than that conferred by genetics alone. The mechanisms responsible are unknown, but likely involve fetal exposure to the in utero milieu, including glucose and circulating adipokines. The purpose of this study was to assess the impact of maternal DM on fetal adipokines and anthropometry in infants of Hispanic and Native American women.

METHODS: We conducted a prospective study of offspring of mothers with normoglycemia (Con-O; n = 79) or type 2 or gestational DM (DM-O; n = 45) pregnancies. Infant anthropometrics were measured at birth and 1-month of age. Cord leptin, high-molecular-weight adiponectin (HMWA), pigment epithelium-derived factor (PEDF) and C-peptide were measured by ELISA. Differences between groups were assessed using the Generalized Linear Model framework. Correlations were calculated as standardized regression coefficients and adjusted for significant covariates.

RESULTS: DM-O were heavier at birth than Con-O (3.7 ± 0.6 vs. 3.4 ± 0.4 kg, p = 0.024), but sum of skinfolds (SSF) were not different. At 1-month, there was no difference in weight, SSF or % body fat or postnatal growth between groups. Leptin was higher in DM-O (20.1 ± 14.9 vs. 9.5 ± 9.9 ng/ml in Con-O, p < 0.0001). Leptin was positively associated with birth weight (p = 0.0007) and SSF (p = 0.002) in Con-O and with maternal hemoglobin A1c in both groups (Con-O, p = 0.023; DM-O, p = 0.006). PEDF was positively associated with birth weight in all infants (p = 0.004). Leptin was positively associated with PEDF in both groups, with a stronger correlation in DM-O (p = 0.009). At 1-month, HMWA was positively associated with body weight (p = 0.004), SSF (p = 0.025) and % body fat (p = 0.004) across the cohort.

CONCLUSIONS: Maternal DM results in fetal hyperleptinemia independent of adiposity. HMWA appears to influence postnatal growth. Thus, in utero exposure to DM imparts hormonal differences on infants even without aberrant growth.

Studying N-linked glycosylation of receptor tyrosine kinases

Metabolic alterations have been identified as a frequent event in cancer. This is often associated with increased flux through glycolysis, and also a secondary pathway to glycolysis, hexosamine biosynthetic pathway (HBP). HBP provides substrate for N-linked glycosylation, which occurs in the endoplasmic reticulum and the Golgi apparatus. N-linked glycosylation supports protein folding and correct sorting of proteins to plasma membrane and secretion. This process generates complex glycoforms, which can be recognized by other proteins and glycosylation of receptor tyrosine kinases (RTK) can also regulate their plasma-membrane retention time. Of special interest for experimental biologists, plants produce proteins, termed lectins, which bind with high specificity to glyco-conjugates. For the purposes of molecular biology, plant lectins can be conjugated to different moieties, such as agarose beads, which enable precipitation of specifically glycosylated proteins. In this chapter, we describe in detail how to perform pull-down experiments with commercially available lectins to identify changes in the glycosylation of RTKs.

The use of erythropoiesis-stimulating agents with ruxolitinib in patients with myelofibrosis in COMFORT-II: an open-label, phase 3 study assessing efficacy and safety of ruxolitinib versus best available therapy in the treatment of myelofibrosis

BACKGROUND: Anemia is considered a negative prognostic risk factor for survival in patients with myelofibrosis. Most patients with myelofibrosis are anemic, and 35-54 % present with anemia at diagnosis. Ruxolitinib, a potent inhibitor of Janus kinase (JAK) 1 and JAK2, was associated with an overall survival benefit and improvements in splenomegaly and patient-reported outcomes in patients with myelofibrosis in the two phase 3 COMFORT studies. Consistent with the ruxolitinib mechanism of action, anemia was a frequently reported adverse event. In clinical practice, anemia is sometimes managed with erythropoiesis-stimulating agents (ESAs). This post hoc analysis evaluated the safety and efficacy of concomitant ruxolitinib and ESA administration in patients enrolled in COMFORT-II, an open-label, phase 3 study comparing the efficacy and safety of ruxolitinib with best available therapy for treatment of myelofibrosis. Patients were randomized (2:1) to receive ruxolitinib 15 or 20 mg twice daily or best available therapy. Spleen volume was assessed by magnetic resonance imaging or computed tomography scan.

RESULTS: Thirteen of 146 ruxolitinib-treated patients had concomitant ESA administration (+ESA). The median exposure to ruxolitinib was 114 weeks in the +ESA group and 111 weeks in the overall ruxolitinib arm; the median ruxolitinib dose intensity was 33 mg/day for each group. Six weeks before the first ESA administration, 10 of the 13 patients had grade 3/4 hemoglobin abnormalities. These had improved to grade 2 in 7 of the 13 patients by 6 weeks after the first ESA administration. The rate of packed red blood cell transfusions per month within 12 weeks before and after first ESA administration remained the same in 1 patient, decreased in 2 patients, and increased in 3 patients; 7 patients remained transfusion independent. Reductions in splenomegaly were observed in 69 % of evaluable patients (9/13) following first ESA administration.

CONCLUSIONS: Concomitant use of an ESA with ruxolitinib was well tolerated and did not affect the efficacy of ruxolitinib. Further investigations evaluating the effects of ESAs to alleviate anemia in ruxolitinib-treated patients are warranted (ClinicalTrials.gov identifier, NCT00934544; July 6, 2009).

Blood: Tests used to assess the physiological and immunological properties of blood.

The properties of blood and the relative ease of access to which it can be retrieved make it an ideal source to gauge different aspects of homeostasis within an individual, form an accurate diagnosis, and formulate an appropriate treatment regime. Tests used to determine blood parameters such as the erythrocyte sedimentation rate, hemoglobin concentration, hematocrit, bleeding and clotting times, mean corpuscular hemoglobin, mean corpuscular hemoglobin concentration, mean cell volume, and determination of blood groups are routinely used clinically, and deviations outside the normal range can indicate a range of conditions such as anemia, pregnancy, dehydration, overhydration, infectious disease, cancer, thyroid disease, and autoimmune conditions, to mention a few. As these tests can be performed relatively inexpensively and do not require high levels of technical expertise, they are ideally suited for use in the teaching laboratory, enabling undergraduate students to link theory to practice. The practicals described here permit students to examine their own blood and that of their peers and compare these with clinically accepted normal ranges. At the end of the practicals, students are required to answer a number of questions about their findings and to link abnormal values to possible pathological conditions by answering a series of questions based on their findings.

A randomized controlled trial on the effects of combined aerobic-resistance exercise on muscle strength and fatigue, glycemic control and health-related quality of life of type 2 diabetes patients

Aim: to evaluate the effects of a 12-weeks combined aerobic-resistance exercise therapy on fatigue and isokinetic muscle strength, glycemic control and health-related quality of life (HRQoL) in moderately affected type 2 diabetes (T2DM) patients. Methods: a randomized controlled trial design was employed. Forty-three T2DM patients were assigned to an exercise group (n = 22), performing 3 weekly sessions of 60 minutes of combined aerobic-resistance exercise for 12-weeks; or a no exercise control group (n = 21). Both groups were evaluated at a baseline and after 12-weeks of exercise therapy for: 1) muscle strength and fatigue by isokinetic dynamometry; 2) plasma glycated hemoglobin A1C (HbA1C); and 3) HRQoL utilizing the SF-36 questionnaire. Results: the exercise therapy led to improvements in muscle fatigue in knee extensors (-55%) and increased muscle strength in knee flexors and extensors (+15 to +30%), while HbA1C decreased (-18%). In addition, the exercising patients showed sizeable improvements in HRQoL: physical function (+53%), vitality (+21%) and mental health (+40%). Conclusion: 12-weeks of combined aerobic-resistance exercise was highly effective to improve muscle strength and fatigue, glycemic control and several aspects of HRQoL in T2DM patients. These data encourage the use of aerobic and resistance exercise in the good clinical care of T2DM.

Phosphatidylethanolamines glycoxidation: products and significance

As fosfatidiletanolaminas constituem a segunda classe de fosfolípidos mais abundantes nos organismos. Elas estão presentes nas membranas biológicas e nas lipoproteínas. As alterações estruturais dos fosfolípidos, ocorrem devido ao stress oxidativo e podem manisfestar-se em alterações das suas propriedades e funções. Já são conhecidas algumas condições fisiopatológicas, nas quais os fosfolípidos oxidados estão envolvidos, por exemplo sinalização celular, resposta imunitária, apoptose e doenças relacionadas com o envelhecimento. Por esse motivo, o interesse no estudo dos fosfolípidos oxidados e suas funções tem crescido nos últimos anos. Contudo, a maioria dos estudos realizados, focam a oxidação das fosfatidilcolinas, tendo sido dedicada pouca atenção a outras classes de fosfolipídos, como as fosfatidiletanolaminas. As fosfatidiletanolaminas, podem ainda sofrer outras modificações, devido ao grupo amina livre presente na cabeça polar, como por exemplo a glicação. As fosfatidiletanolaminas glicadas já foram detectadas em condições de hiperglicemia, em pacientes diabéticos, e tem correlação com a hemoglobina glicada. Sabe-se que a glicação de biomoléculas, pode aumentar as modificações oxidativas, que por sua vez, podem ser responsáveis pelo estado inflamatório, existente na diabetes mellitus. Tanto o stress oxidativo, como a inflamação estão relacionados com a diabetes e as suas complicações. A espectrometria de massa tem sido utilizada como uma importante tecnologia na detecção e caracterização de modificações oxidativas de fosfolípidos. Assim, neste trabalho pretendeu-se estudar as modificações oxidativas induzidas em fosfatidiletanolaminas glicadas, e os seus efeitos biológicos nos monócitos e células dendríticas do sangue periférico. Pretendeu-se ainda, estudar as alterações que ocorreram nas espécies de fosfatidiletanolaminas do fígado de ratos diabéticos. Os resultados obtidos permitiram identificar vários produtos de oxidação de fosfatidiletanolaminas glicadas, nomeadamente novos produtos formados pela oxidação da cabeça polar glicada. A oxidação na cabeça polar glicada foi, ainda, confirmada pela realização de experiências com spin traps combinadas com espetrometria de massa. Posteriormente, as fosfatidiletanolaminas oxidadas, glicadas e glicoxidadas demonstraram ter efeitos pró-inflamatórios, confirmados pelo aumento da estimulação monócitos e de células dendríticas, expresso no aumento do número de células produtoras de citocinas em comparação com o estado basal. As diferentes modificações de fosfatidiletanolaminas induziram estímulos distintos nos dois tipos de células. Sendo as fosfatidiletanolaminas glicadas e as glicoxidadas, os compostos que induziram um maior estímulo. Estes resultados sugeriram que as fosfatidiletanolaminas glicadas e as glicoxidadas podem estar associadas com o estado inflamatório que decorre da hiperglicemia crónica. Ainda, a avaliação do perfil lipídico de extratos de fígado de ratos diabéticos demonstrou que a hiperglicemia induz inúmeras alterações das espécies de fosfatidiletanolaminas e das espécies de outras classes de fosfolípidos, em simultâneo com sinais de lesão hepática. Em conclusão, este trabalho demonstra a relação existente entre, a hiperglicémia, o stress oxidativo, a glicação e oxidação de fosfatidiletanolaminas e ainda a inflamação e compliações diabéticas. Portanto a contribuição da lipidómica é importante para compreender os efeitos prejudiciais da hiperglicemia não controlada, e por isso, merece ser mais explorado.

Tradução científica de The thalassaemia syndromes: reflexões sobre questões terminológicas e estratégias de tradução

Trabalho de projecto de mestrado, Tradução, Universidade de Lisboa, Faculdade de Letras, 2012

Malaria and tuberculosis co-infection:role for hemozoin immunosuppression?

Tese de doutoramento, Ciências Biomédicas (Microbiologia e Parasitologia), Universidade de Lisboa, Faculdade de Medicina, 2014

Molecularly imprinted polymer based on MWCNTs-QDs as fluorescent biomimetic sensor for specific recognition of target protein

A novel molecularly imprinted optosensing material based on multi-walled carbon nanotube-quantum dots (MWCNT-QDs) has been designed and synthesized for its high selectivity, sensitivity and specificity in the recognition of a target protein bovine serum albumin (BSA). Molecularly imprinted polymer coated MWCNT-QDs using BSA as the template (BMIP-coated MWCNT-QDs) exhibits a fast mass-transfer speed with a response time of 25 min. It is found that the BSA as a target protein can significantly quench the luminescence of BMIP-coated MWCNT-QDs in a concentration-dependent manner that is best described by a Stem-Volmer equation. The K-SV for BSA is much higher than bovine hemoglobin and lysozyme, implying a highly selective recognition of the BMIP-coated MWCNT-QDs to BSA. Under optimal conditions, the relative fluorescence intensity of BMIP-coated MWCNT-QDs decreases linearly with the increasing target protein BSA in the concentration range of 5.0 x 10(-7)-35.0 x 10(-7) M with a detection limit of 80 nM.

Exposição profissional a chumbo : utilização da protoporfirina-zinco (PPZ) na vigilância de saúde de trabalhadores ex

Resumo Uma estratégia de avaliação e prevenção de riscos na exposição a agentes químicos deve ter sempre em conta que a vigilância do ambiente de trabalho e a da saúde dos trabalhadores são aspectos complementares de uma mesma realidade – os riscos resultantes da interacção entre um agente químico e os trabalhadores a ele expostos. Se à Vigilância Ambiental compete apreciar o risco, pela caracterização do agente no ambiente de trabalho, a Vigilância Biológica pronuncia-se sobre a interacção entre o tóxico e o organismo, avaliando a resposta à agressão química e a evolução das reacções de adaptação ou de desajuste face à absorção do tóxico. Os Indicadores Biológicos, deste modo, assumem um estatuto de instrumento privilegiado na vigilância da saúde dos trabalhadores expostos, na medida em que medem a quantidade de tóxico que efectivamente penetrou e foi absorvido, ou o resultado (efeito) determinado por essa mesma dose. O presente estudo procura contribuir para a definição de um quadro metodológico de utilização dos Indicadores Biológicos na avaliação/gestão da exposição profissional ao chumbo, designadamente apreciando a variação da protoporfirina-zinco (PPZ), indicador até ao presente ainda não utilizado em Portugal. O chumbo é um metal de ocorrência natural, cujos níveis nos diversos ecossistemas resultam, principalmente, das actividades antropogénicas de natureza doméstica e industrial. A sua capacidade poluente é assinalável, representando uma fonte de exposição permanente para o homem, demonstrável pela sua constante presença no organismo apesar de não desempenhar qualquer tipo de função fisiológica. São actualmente inúmeras as suas aplicações, tornando a exposição profissional ao chumbo uma realidade vasta: indústrias de acumuladores eléctricos, de vidros, de plásticos e de munições, construção civil, manutenção e reparação automóvel e de navios, fabrico de tintas, indústrias electrónicas, fundições e actividades de soldadura são, entre outras, situações onde é uma realidade a ter em conta. A penetração dos compostos inorgânicos de chumbo no organismo efectua-se principalmente por via respiratória, não sendo, no entanto, desprezável, a sua penetração por via digestiva. As partículas absorvidas são transportadas pelo sangue principalmente ligadas aos eritrocitos (95%), distribuem-se pelos tecidos moles e depositam-se essencialmente no tecido ósseo, onde representam mais de 90% da carga corporal do total absorvido e tendo aí um elevado tempo de semi-vida (mais de 20 anos). Não é metabolizado no organismo e a sua eliminação efectua-se essencialmente por via renal,sendo igualmente excretado, em menor escala, através das fezes, do suor, da saliva, das faneras e do leite materno. O conhecimento científico evidencia que concentrações sanguíneas de chumbo entre 20 e 50 mg/dL são susceptíveis de determinar efeitos adversos no homem, podendo ser afectados o sistema hematopoiético, o sistema nervoso, o sistema cardiovascular, o sistema reprodutor e o sistema imunitário. Contudo, ainda muito há a clarificar no âmbito da toxicidade do chumbo. Os níveis de exposição a que correspondem as alterações nos diversos órgãos e sistemas continuam a ser motivo de alguma controvérsia. As características carcinogénicas e mutagénicas do chumbo são, ainda, um campo de vasta exigência de investigação. A intoxicação por chumbo e seus sais (Saturnismo) de origem ocupacional é reconhecida em Portugal como doença profissional (grupo 1 - Doenças Provocadas por Agentes Químicos, da Lista das Doenças Profissionais). É uma intoxicação do tipo crónico, fruto da absorção contínua de doses relativamente pequenas durante longo período, evidenciando-se no seu início por sinais e sintomas vagos e difusos de grande inespecificidade, que podem incluir, nomeadamente, perda de apetite, sabor metálico na boca, palidez, mal-estar e fadiga, cefaleias, mialgias e artralgias, irritabilidade, tremores finos, obstipação, cólicas abdominais, insónias, déficit da memória de curto prazo e da capacidade de concentração. Um importante conjunto de indicadores biológicos pode ser utilizado na vigilância periódica da saúde de trabalhadores nestas condições de exposição. Tais indicadores (de dose ou de efeito), encerram diferentes significados e comportam distintas exigências, competindo ao Médico do Trabalho, no âmbito dos programas de prevenção dos efeitos adversos relacionados com a exposição profissional a chumbo, seleccionar a sua utilização e interpretar a sua informação, de modo a avaliar a interacção do tóxico com o organismo numa fase de reversibilidade. O presente estudo envolveu 180 trabalhadores dos quais 110 apresentavam plumbémias (Pb-S) iguais ou superiores a 40 mg/dL. Além da Pb-S, a todos foi doseada a protoporfirina-zinco (PPZ) e efectuado o Hemograma e a cerca de 25% foi determinada a concentração do ácido d-aminolevulínico urinário (ALA-U). Os doseamentos da PPZ efectuados em amostra de sangue capilar através de um hematofluorímetro portátil revelaram-se de total fiabilidade, dando significado a uma técnica de fácil execução e baixo custo. A avaliação do tipo de colheita urinária para doseamento do ALA-U concluiu pela necessidade de recurso a urinas de 24 horas.Os resultados do estudo evidenciaram uma elevada associação entre a PPZ e a Pb-S, com uma maior magnitude e de início mais precoce do que o que registado na associação da Pb-S com o ALA-U. Revelaram, ainda, fracos níveis de associação da hemoglobina (e outros parâmetros hematológicos) com a Pb-S. E demonstraram para um cut-off de 100 mg/ dL de PPZ, taxa de falsos negativos e falsos positivos, para plumbémias a partir de 70 mg/dL, inferiores a 20%. Assim, concluiu-se que, nos protocolos de vigilância de saúde de trabalhadores expostos a chumbo, o doseamento da PPZ por hematofluorímetro, em sangue de colheita capilar, é adequado, fiável e de realização preferencial em relação ao do ALA-U. Concluiu-se, também, que a realização do hemograma apenas se justifica em situações individuais que clinicamente o tornem aconselhável. E que estes protocolos devem incluir a realização da Pb-S e da PPZ, podendo, em situações de controlo rigoroso (ambiental, biológico e clínico), basear-se apenas na determinação da PPZ reservando os outros indicadores para aprofundar a investigação médica nos casos de taxas elevadas desta ou de situações limitantes. ■ Résumée Une stratégie d’évaluation et de prévention des risques d’exposition aux agents chimiques doit toujours tenir en considération que la vigilance du lieu de travail et de la santé des travailleurs sont des aspects complémentaires d’une même réalité – les risques résultant d’une interaction entre l’agent chimique et les travailleurs exposés. Si c’est à la Vigilance Ambiantale de juger le risque, par la caractérisation de l’agent dans le lieu de travail, la Vigilance Biologique, elle, se prononce sur l’interaction entre le toxique et l’organisme, évaluant la réponse à l’agression chimique et l’évolution des réactions d’adaptation ou de rupture face à l’absorption du toxique. Les Indicateurs Biologiques assument ainsi un statut d’instrument privilégié de vigilance de la santé des travailleurs exposés, dans la mesure où ils déterminent la quantité de toxique qui a effectivement été pénétré et absorbé, ou le résultat (effet) déterminé par cette dose. Cette étude-ci cherche à contribuer à la définition d’un cadre méthodologique d’utilisation des Indicateurs Biologiques dans l’évaluation/ gestion de l’exposition professionnelle au plomb inorganique, évaluant spécialement le comportement de la protoporphirine-zinc (PPZ), indicateur pas encore utilisé au Portugal.Le plomb est un métal d’occurrence naturelle dont les niveaux dans les différents écosystèmes en résultent, principalement, des activités anthropogéniques de nature domestique et industrielle. Sa capacité polluante peut être signalée, représentant une source d’exposition permanente pour l’homme, celle-ci démontrable par sa présence continue dans l’organisme, même si elle n’y accomplit aucune fonction physiologique. Actuellement ses applications sont innombrables, faisant de l’exposition professionnelle au plomb une réalité de grande ampleur : industries d’accumulateurs électriques, de verre, de plastique et de munitions, bâtiments, manutention et réparation automobile et navale, fabrication d’encres, industries électroniques, fontes et activités de soudure sont, entre autres, des situations réelles a en tenir compte. La pénétration du plomb inorganique dans l’organisme se fait principalement par voie respiratoire, pouvant se faire également par voie digestive. Les particules absorbées sont transportées par le sang, surtout liées aux érythrocytes (95%), se repartent à travers les tissus mous et se déposent essentiellement dans le tissu osseux, où elles représentent plus de 90% de la charge corporelle de ce qui a été absorbé et ont un temps de demi-vie élevé (plus de 20 ans). Le plomb n’est pas métabolisé dans l’organisme et son élimination se fait essentiellement par voie rénale, pouvant tout de même, à une moindre échelle, être excrété dans les fèces, de la sueur, de la salive, des ongles, des cheveux et du lait maternel. La connaissance scientifique met en évidence que des concentrations sanguines de plomb entre 20 et 50 mg/dL sont susceptibles de déterminer des effets adverses dans l’homme, pouvant les systèmes hématopoïétique, nerveux, cardiovasculaire, reproducteur et immunitaire en être affectés. Cependant, il en reste beaucoup à éclaircir dans le domaine de la toxicité du plomb. Les niveaux d’exposition auxquels correspondent les modifications des divers organes et systèmes, demeurent toujours sujet de quelque controverse. Les caractéristiques carcinogèniques et mutagèniques du plomb restent toujours un champ d’investigation d’une grande exigence. L’intoxication par le plomb et ses sels (Saturnisme) d’origine occupationnelle est reconnue, au Portugal, comme une maladie professionnelle (groupe 1- Maladies Provoquées par des Agents Chimiques, de la Liste des Maladies Professionnelles). C’est une intoxication du tipe chronique, due à l’absorption continue de doses relativement petites pendant une longue période, mise en évidence à travers des signes et des symptômes vagues et diffus sans grande spécificité, lesquels peuvent inclure, particulièrement, le manque d’appétit, goût métallique dans la bouche, pâleur, malaise et fatigue, céphalées, myalgies et arthralgies, irritabilité, tremblements fins, constipation, coliques abdominales, insomnies, déficit de la mémoire à court terme et de la capacité de concentration.Un ensemble important d’indicateurs biologiques peut être employé dans la vigilance périodique de la santé des travailleurs dans ces conditions d’exposition. Ces indicateurs (de dose ou d’effet) renferment différentes significations et comportent diverses exigences, devant le Médecin de Travail, dans le domaine des programmes de prévention des effets adverses qui sont en relation avec l’exposition professionnelle au plomb, sélectionner son utilisation et interpréter son information de façon à évaluer l’interaction de l’élément toxique avec l’organisme à un stade de réversibilité. L’étude ci-présent engloba 180 travailleurs desquels 110 présentaient des plombémies (Pb-S) égales ou supérieures à 40 mg/dL. À part la Pb-S, la protoporphyrine-zinc (PPZ) leur a été prise en dosage et un Hémogramme fut effectué et fut déterminé l’acide d- aminolévulinique urinaire (ALA-U) sur environ 25% des travailleurs. Le dosage de la PPZ efectué en échantillon de sang capillaire par un fluorimètre portable, s’est accomplit d’une fiabilité total, donnant du sgnificat à une téchnique de facile execution et bas prix. L’évaluation de la prise urinaire par dosage du ALA-U conclut au besoin d’un recours aux urines de 24 heures Les résultats de l’étude ont mis en évidence une association élevée entre la PPZ et la Pb- S, avec une intensité majeure et de début plus précoce par rapport à celui qui fut registré lors de l’association de la Pb-S avec la ALA-U. Ces résultats ont également montré de faibles niveaux d’association entre l’hémoglobine (et autres paramètres hématologiques) et la Pb-S. Ils ont démontré aussi, une valeur de cut-off de 100 mg/dL de PPZ, des taux de faux négatifs et faux positifs, pour des plombémies de 70 mg/dL, inférieurs à 20%. On peut donc conclure que dans les protocoles de vigilance de la santé des travailleurs exposés au plomb, le dosage de la PPZ par fluorimetrie dans le sang capillaire est adéquat, fiable et de réalisation préférentielle par rapport à celui du ALA-U. On peut également conclure que la réalisation de l’hémogramme ne se justifie que dans les cas individuels où, cliniquement, celui-ci est conseillé. De plus, ces protocoles doivent inclure la réalisation de la Pb-S et de la PPZ, pouvant, en cas de contrôle rigoureux (ambiantal, biologique et clinique), s’appuyer que dans la détermination de la PPZ réservant les autres indicateurs pour approfondir l’investigation médicale dans les cas où les taux de celle-ci sont élevés ou dans les cas de situations limitantes. ■ Summary Any strategy to evaluate and prevent the risks of chemical agents exposure must always regard the work environment and workers health as complementary aspects of one reality - the resulting risks from the interaction between the chemical agent and the exposed workers. It is the responsibility of Environmental Monitoring to evaluate the risks of exposure by the characterization of the chemical agent in the work environment. Biological Monitoring, on the other hand, pronounces itself over the toxin and body interaction, evaluating human response to the chemical aggression and the body adaptations to the toxic absorption. Biological Exposure Indices (BEI) assume, therefore, a privileged status among exposed workers' health monitoring instruments, as they measure the actual penetrated and absorbed toxic quantity and the effect it produces. This research study aims to contribute to the definition of a methodological strategy on the utilization of BEI’s in evaluating inorganic lead's occupational exposure, more specifically appreciating the zinc protoporphyrin (ZPP) variation, an index that has never been taken under consideration in Portugal until now. Lead is a natural metal whose ecosystem’s levels are mainly due to domestic and industrial anthropogenic activities. Its pollutant capacity is notable, representing a permanent exposure risk shown by its constant presence in the human body, although it has no physiologic function. Nowadays, lead's applications are countless, turning its professional exposure a huge reality: storage batteries industries, glass industries, plasterers and munitions industries, building construction, ships and motor car maintenance and repairing, ink manufacture, electronics industries, foundries and other soldering activities are, among so many other, realities to attend to. Respiration is the main cause of human body's inorganic lead absorption, although digestive pathway must not to be ignored. The absorbed particles are transported by blood, essentially bounded to erythrocytes (95%). It is distributed by soft tissues and settled mainly on bone tissues, where it represents approximately 90% of the total body charge and has a high half-life time (more than 20 years). It is not metabolized by the organism, its elimination being effectuated by renal activity and, in smaller scale, through lees, sweat, saliva, nails, hair and maternal milk.Scientific knowledge shows that concentrations of lead in blood between 20 e 50 mg/dL are susceptible to determine adverse effects in man and able to affect the hematopoietic system, the nervous system, the cardiovascular system, the reproductive system and the immunological system. Nevertheless, there's still much to be learned and clarified about lead's toxicity. The correlation between exposure levels and human's systems and organs alteration levels continues to be a centre of controversies. Still, lead's carcinogenic and mutagenic characteristics continue to be a high demanding research field. Intoxication by lead and its compounds (saturnism), from occupational origin, is recognized in Portugal as an occupational disease, included in Group 1 - Chemical Agents Caused Diseases, on the Occupational Diseases List. It is a chronic intoxication caused by a continuous absorption of small doses, throughout a long period of time. Its signs and symptoms are diffuse and imprecise, of great unspecificity, such as loss of appetite, metallic flavor in the mouth, paleness, ailment and fatigue, headaches, myalgia and arthralgia, irritability, thin tremors, constipation, abdominal pain, insomnias, short memory loses and inability to concentrate. A considered number of BEI’s can be used in Periodic Health Monitoring of workers in such exposure conditions. Such BEI (dose indices or effect indices) provide different meanings and imply different procedures, being Occupational Doctors responsibility, in the context of lead related adverse effects preventive programmes, to select and interpret its information, in order to evaluate the interaction between toxic and organism in a reversible phase of the toxic action. The present research study involved 180 workers, 110 of which presented blood lead levels (PbB) above or equal to 40 mg/dL. Besides PbB, all workers has been evaluated for zinc protoporphyrin levels (PPZ) and submitted to a haemogram. About 25% of the workers were selected for d-aminolevulinic urinary acid (ALA-U) determination. The evaluation of PPZ, by a portable hematofluorometer using capillary blood samples, turned out to be an easy procedure with low costs and total warrantability. As in regard for ALA- U procedure, it was concluded the necessity of 24 hours urine samples. This research results underlined a strong connection between ZPP and PbB, which was found to to be stringer and to begin earlier than it was registered for PbB and ALA-U association. The same study also revealed a low association level between PbB and hemoglobin or other hematological indices. It was also verified less than 20% of false negatives and false positives cases when admitted a ZPP 100 mg/dL cut off value for PbB³ 70 mg/dL. As in result it was concluded that in Health Monit

Assessment of nutritional and metabolic profiles of pea shoots: The new ready-to-eat baby-leaf vegetable

Pea-shoots are a new option as ready-to-eat baby-leaf vegetable. However, data about the nutritional composition and the shelf-life stability of these leaves, especially their phytonutrient composition is scarce. In this work, the macronutrient, micronutrient and phytonutrients profile of minimally processed pea shoots were evaluated at the beginning and at the end of a 10-day storage period. Several physicochemical characteristics (color, pH, total soluble solids, and total titratable acidity) were also monitored. Standard AOAC methods were applied in the nutritional value evaluation, while chromatographic methods with UV–vis and mass detection were used to analyze free forms of vitamins (HPLC-DAD-ESI-MS/MS), carotenoids (HPLC-DAD-APCI-MSn) and flavonoid compounds (HPLC-DAD-ESI-MSn). Atomic absorption spectrometry (HR-CS-AAS) was employed to characterize the mineral content of the leaves. As expected, pea leaves had a high water (91.5%) and low fat (0.3%) and carbohydrate (1.9%) contents, being a good source of dietary fiber (2.1%). Pea shoots showed a high content of vitamins C, E and A, potassium and phosphorous compared to other ready-to-eat green leafy vegetables. The carotenoid profile revealed a high content of β-carotene and lutein, typical from green leafy vegetables. The leaves had a mean flavonoid content of 329 mg/100 g of fresh product, mainly composed by glycosylated quercetin and kaempferol derivatives. Pea shoots kept their fresh appearance during the storage being color maintained throughout the shelf-life. The nutritional composition was in general stable during storage, showing some significant (p < 0.05) variation in certain water-soluble vitamins.