Health Effects of Carotenoids during Pregnancy and Lactation.

Adequate nutrition is particularly important during pregnancy since it is needed not only for maintaining the health of the mother, but also determines the course of pregnancy and its outcome, fetus development as well as the child's health after birth and during the later period of life. Data coming from epidemiological and interventions studies support the observation that carotenoids intake provide positive health effects in adults and the elderly population. These health effects are the result of their antioxidant and anti-inflammatory properties. Recent studies have also demonstrated the significant role of carotenoids during pregnancy and infancy. Some studies indicate a correlation between carotenoid status and lower risk of pregnancy pathologies induced by intensified oxidative stress, but results of these investigations are equivocal. Carotenoids have been well studied in relation to their beneficial role in the prevention of preeclampsia. It is currently hypothesized that carotenoids can play an important role in the prevention of preterm birth and intrauterine growth restriction. Carotenoid status in the newborn depends on the nutritional status of the mother, but little is known about the transfer of carotenoids from the mother to the fetus. Carotenoids are among the few nutrients found in breast milk, in which the levels are determined by the mother's diet. Nutritional status of the newborn directly depends on its diet. Both mix feeding and artificial feeding may cause depletion of carotenoids since infant formulas contain only trace amounts of these compounds. Carotenoids, particularly lutein and zeaxanthin play a significant role in the development of vision and nervous system (among others, they are important for the development of retina as well as energy metabolism and brain electrical activity). Furthermore, more scientific evidence is emerging on the role of carotenoids in the prevention of disorders affecting preterm infants, who are susceptible to oxidative stress, particularly retinopathy of prematurity.

Assessment of individual carotenoid and vitamin A dietary intake in overweight and obese Dominican subjects.

INTRODUCTION: Carotenoids are plant pigment with important biological activities in humans, such as provitamin-A among others. At present, there are no individual carotenoid intake data in the Dominican population, which is at risk of vitamin A deficiency and has an important percentage of overweight and obese individuals . OBJECTIVE: To assess the individual components of vitamin A intake (retinol, α-carotene, ß-carotene and ß-cryptoxanthin) and that of other relevant dietary carotenoids like lutein, zeaxanthin and lycopene of Dominican daily food intake. METHODS: Fifty overweigth and obese subjects (22-69 y). Individual carotenoid intake, from whole diet and from the ingestion of fruits and vegetables, was determined using three 24 h diet recalls and a specific carotenoid database. Retinol, macronutrient and energy intake were calculated using DIAL® software. RESULTS: The total carotenoid intake was 6363.2 µg/day, 56. 1% corresponding to provitamin A carotenoids (74.3% ß-carotene). Vitamin A intake was supplied by retinol (40%) and by provitamin A carotenoids (60%); vegetables contributed more than fruits (39.2% and 19.2%, respectively). Non-provitamin A carotenoid intake represents 43.9% of the total intake and is supplied by lycopene and lutein plus zeaxanthin in similar percentages (52.3% and 47.7%, respectively). CONCLUSIONS: The diet of these Dominican subjets met the recommended vitamin A intake, when expressed as retinol equivalents, 59% of which was supplied by provitamin-A carotenoids from plant sources, mainly by red/orange and white/yellow foods. Individual carotenoid intake is an aspect of great interest for issuing dietary recommendations in the public health setting.

Thiamin, Pyridoxine, Vitamin D, and Carotene Deficiency in a Malnourished Patient Following Billroth II Gastrectomy.

We describe the case of a malnourished 48-year-old man who had previously undergone a Billroth II procedure for severe peptic ulcer disease. He was found to have a severely stenotic gastrojejunal anastomosis with inflamed mucosa that prevented him from tolerating solid food. Laboratory assessment revealed deficiencies in thiamin, pyridoxine, vitamin D, and carotene. This case demonstrates potential vital micronutrient complications following a partial gastrectomy.

Low intakes of carotene, vitamin B2 , pantothenate and calcium predict cognitive decline among elderly patients with diabetes mellitus: The Japanese Elderly Diabetes Intervention Trial.

AIM: The present study aimed to examine whether nutrient intakes predicted cognitive decline among elderly patients with diabetes mellitus. METHODS: This study evaluated data from a 6-year prospective follow up of 237 elderly patients (aged ≥65 years) with diabetes mellitus, and the associations of baseline nutrient intakes with cognitive decline. Cognitive decline was defined as a ≥2-point decrease in the Mini-Mental State Examination (MMSE) score. Intakes of food and nutrients were assessed using a validated food frequency questionnaire, and were compared between patients with cognitive decline and intact cognition. Analysis of covariance and logistic regression analysis were used to compare the changes in the MMSE score during the follow up among intake tertile groups for each nutrient. RESULTS: Compared with men with intact cognition, the men with cognitive decline had lower baseline intakes of calcium, vitamin A, vitamin B2 , pantothenate, soluble fiber, green vegetables and milk. However, no significant associations between cognitive decline and nutrient intakes were observed among women. After adjusting for age, body mass index, glycated hemoglobin levels, history of severe hypoglycemia, previous stroke and baseline MMSE score, we found that cognitive decline was significantly associated with low intakes of carotene, vitamin B2 , pantothenate, calcium and green vegetables. Multiple logistic regression analysis showed that intakes of nutrients and green vegetables predicted cognitive decline after adjusting for age, body mass index, glycated hemoglobin levels, baseline MMSE score, and incident stroke during the follow up. CONCLUSIONS: These findings suggest that sufficient intakes of carotene, vitamin B2 , pantothenate, calcium and vegetables could help prevent cognitive decline among elderly men with diabetes mellitus. Geriatr Gerontol Int 2017; 17: 1168-1175.

Lycopene Deficiency in Ageing and Cardiovascular Disease.

Lycopene is a hydrocarbon phytochemical belonging to the tetraterpene carotenoid family and is found in red fruit and vegetables. Eleven conjugated double bonds predetermine the antioxidant properties of lycopene and its ability to scavenge lipid peroxyl radicals, reactive oxygen species, and nitric oxide. Lycopene has a low bioavailability rate and appears in the blood circulation incorporated into chylomicrons and other apo-B containing lipoproteins. The recent body of evidence suggests that plasma concentration of lycopene is not only a function of intestinal absorption rate but also lycopene breakdown via enzymatic and oxidative pathways in blood and tissues. Oxidative stress and the accumulation of reactive oxygen species and nitric oxide may represent a major cause of lycopene depletion in ageing, cardiovascular disease, and type 2 diabetes mellitus. It has been shown recently that low carotenoid levels, and especially decreased serum lycopene levels, are strongly predictive of all-cause mortality and poor outcomes of cardiovascular disease. However, there is a poor statistical association between dietary and serum lycopene levels which occurs due to limited bioavailability of lycopene from dietary sources. Hence, it is very unlikely that nutritional intervention alone could be instrumental in the correction of lycopene and carotenoid deficiency. Therefore, new nutraceutical formulations of carotenoids with enhanced bioavailability are urgently needed.

Maternal Antioxidant Levels in Pregnancy and Risk of Preeclampsia and Small for Gestational Age Birth: A Systematic Review and Meta-Analysis.

BACKGROUND: Oxidative stress in preeclampsia and small for gestational age (SGA) birth suggests antioxidant supplementation could prevent these conditions. However, it remains unclear whether maternal antioxidant levels are systematically lower in these pregnancies. OBJECTIVE: To conduct a systematic review of the association between maternal antioxidant levels during pregnancy and preeclampsia or SGA. METHODS: We searched PubMed, Embase, and several other databases from 1970-2013 for observational studies that measured maternal blood levels of non-enzymatic antioxidants (vitamins A, C, E, and carotenoids) during pregnancy or within 72 hours of delivery. The entire review process was done in duplicate. Study quality was assessed using the Newcastle-Ottawa Scale and additional questions. We pooled the standardized mean difference (SMD) across studies, stratified by outcome and pregnancy trimester, and investigated heterogeneity using meta-regression. RESULTS: We reviewed 1,882 unique citations and 64 studies were included. Most studies were small with important risk of bias. Among studies that addressed preeclampsia (n = 58) and SGA (n = 9), 16% and 66%, respectively, measured levels prior to diagnosis. The SMDs for vitamins A, C, and E were significantly negative for overall preeclampsia, but not for mild or severe preeclampsia subtypes. Significant heterogeneity was observed in all meta-analyses and most could not be explained. Evidence for lower carotenoid antioxidants in preeclampsia and SGA was limited and inconclusive. Publication bias appears likely. CONCLUSIONS: Small, low-quality studies limit conclusions that can be drawn from the available literature. Observational studies inconsistently show that vitamins C and E or other antioxidants are lower in women who develop preeclampsia or SGA. Reverse causality remains a possible explanation for associations observed. New clinical trials are not warranted in light of this evidence; however, additional rigorous observational studies measuring antioxidant levels before clinical detection of preeclampsia and SGA may clarify whether levels are altered at a causally-relevant time of pregnancy.

Relationship of serum carotenoid concentrations with allostatic load as a measure of chronic stress among middle-aged adults in the USA.

OBJECTIVE: Chronic stress and repeated physiological attempts at stress adaptation may result in 'fatigue' and suboptimal performance of multiple physiological systems, i.e. allostatic load (AL). Although carotenoids have been linked with individual cardiovascular, metabolic and inflammatory biomarkers, little is known about the relationship of carotenoids with the multi-system biomarker measure of stress, AL. The present study examined the association of serum concentrations of carotenoids with AL among middle-aged adults. DESIGN: Cross-sectional. AL score was calculated based on nine risk-rated indicators (systolic and diastolic blood pressure, pulse rate, total and HDL-cholesterol, glycosylated Hb, sex-specific waist-to-hip ratio, albumin and C-reactive protein). SUBJECTS: Middle-aged (45-64 years, n 3387) men and women participants in the Third National Health and Nutrition Examination Survey, NHANES III (1988-1994). RESULTS: Serum ß-carotene concentration was inversely associated with high AL after adjusting for age, education, race/ethnicity, serum cotinine, alcohol consumption, physical activity and other carotenoids (α-carotene, ß-cryptoxanthin, lycopene, lutein/zeaxanthin). Females in the lowest ß-carotene quartile were 2·94 (95 % CI 1·74, 4·94) times and males 2·90 (95 % CI 1·43, 5·89) times as likely to have high AL, compared with peers in the highest quartile (P for linear trend 0·001 and 0·018 for females and males, respectively). Mean serum ß-carotene concentrations were also inversely associated with the number of 'high-risk' AL components (P for linear trend <0·001 and 0·004 for females and males, respectively). CONCLUSIONS: Our study adds to evidence linking low ß-carotene levels with unfavourable health outcomes.

Patterns of dietary intake and serum carotenoid and tocopherol status are associated with biomarkers of chronic low-grade systemic inflammation and cardiovascular risk.

Dietary modification may affect inflammatory processes and protect against chronic disease. In the present study, we examined the relationship between dietary patterns, circulating carotenoid and tocopherol concentrations, and biomarkers of chronic low-grade systemic inflammation in a 10-year longitudinal study of Scottish postmenopausal women. Diet was assessed by FFQ during 1997-2000 (n 3237, mean age 54·8 (SD 2·2) years). Participants (n 2130, mean age 66·0 (SD 2·2) years) returned during 2007-11 for follow-up. Diet was assessed by FFQ (n 1682) and blood was collected for the analysis of serum high-sensitivity C-reactive protein (hs-CRP), IL-6, serum amyloid A, E-selectin, lipid profile and dietary biomarkers (carotenoids, tocopherols and retinol). Dietary pattern and dietary biomarker (serum carotenoid) components were generated by principal components analysis. A past 'prudent' dietary pattern predicted serum concentrations of hs-CRP and IL-6 (which decreased across the quintiles of the dietary pattern; P= 0·002 and P= 0·001, respectively; ANCOVA). Contemporary dietary patterns were also associated with inflammatory biomarkers. The concentrations of hs-CRP and IL-6 decreased across the quintiles of the 'prudent' dietary pattern (P= 0·030 and P= 0·006, respectively). hs-CRP concentration increased across the quintiles of a 'meat-dominated' dietary pattern (P= 0·001). Inflammatory biomarker concentrations decreased markedly across the quintiles of carotenoid component score (P< 0·001 for hs-CRP and IL-6, and P= 0·016 for E-selectin; ANCOVA). Prudent dietary pattern and carotenoid component scores were negatively associated with serum hs-CRP concentration (unstandardised ß for prudent component: -0·053, 95% CI -0·102, -0·003; carotenoid component: -0·183, 95% CI -0·233, -0·134) independent of study covariates. A prudent dietary pattern (which reflects a diet high in the intakes of fish, yogurt, pulses, rice, pasta and wine, in addition to fruit and vegetable consumption) and a serum carotenoid profile characteristic of a fruit and vegetable-rich diet are associated with lower concentrations of intermediary markers that are indicative of CVD risk reduction.

Skin and plasma carotenoid response to a provided intervention diet high in vegetables and fruit: uptake and depletion kinetics.

BACKGROUND: Objective biomarkers are needed to assess adherence to vegetable and fruit intervention trials. Blood carotenoids are considered the best biomarker of vegetable and fruit intake, but collecting blood is invasive and the analyses are relatively expensive for population studies. Resonance Raman spectroscopy (RRS) is an innovative method for assessing carotenoids in skin noninvasively. OBJECTIVE: Our objective was to compare blood carotenoid concentrations with skin carotenoid assessments by RRS during a controlled feeding intervention. DESIGN: Twenty-nine participants consumed low-carotenoid diets (6 wk, phases 1 and 3), a provided diet containing 6-cup equivalents (1046 g/d) of vegetables and fruit (8 wk, phase 2), and usual diet (final 8 wk, phase 4). RESULTS: At baseline, skin and plasma total carotenoid values were correlated (r = 0.61, P < 0.001). Skin and plasma carotenoid values decreased (P < 0.001) 36% and 30%, respectively, from baseline to the end of phase 1 and then increased (P < 0.001) by >200% at the end of phase 2. Plasma carotenoids returned to baseline concentrations by the middle of phase 3 and skin carotenoid concentrations by the middle of phase 4. Skin carotenoid status predicted plasma values by using a mixed linear model including all time points (r = 0.72, P < 0.001), which indicates that changes in skin carotenoid status closely follow changes in plasma across a broad range of intakes. At the individual level, skin carotenoids predicted plasma values (r = 0.70, P < 0.001) over all time points. CONCLUSION: Skin carotenoid status assessed by resonance Raman spectroscopy is a noninvasive, objective biomarker of changes in vegetable and fruit intake.

Low serum concentrations of carotenoids and vitamin E are associated with high adiposity in Mexican-American children.

Mexican-American children have a high prevalence of overweight/obesity. Micronutrient deficiencies may be contributing to the development of greater adiposity in these children. This study investigated the relations between adiposity and serum concentrations of carotenoids, retinol, and vitamin E among Mexican-American children 8-15 y of age included in the 2001-2004 U.S. NHANES. Associations of the outcomes of children's body mass index (BMI), truncal fat mass (TrFM), and total body fat mass (TBFM) with serum concentrations of α-carotene, cis-ß-carotene, trans-ß-carotene, retinol, and α-tocopherol were determined by using linear, quantile, and multinomial regression models. BMI was inversely associated with serum concentrations of α-carotene (ß = -0.88, P < 0.05), trans-ß-carotene (ß = -2.21, P < 0.01), cis-ß-carotene (ß = -2.10, P < 0.01), and α-tocopherol adjusted for total cholesterol ratio (ß = -3.66, P < 0.01), respectively. Similar inverse associations were found with TrFM and TBFM. Higher cis-ß-carotene and α-tocopherol serum concentrations were associated with reduced probability of overweight (OR: 0.57; 95% CI: 0.37, 0.89; P < 0.05; and OR: 0.56; 95% CI: 0.37, 0.86; P < 0.05; respectively) and obesity (OR: 0.39; 95% CI: 0.26, 0.58; P < 0.01; and OR: 0.38; 95% CI: 0.24, 0.60; P < 0.01; respectively). Higher retinol serum concentrations were associated with increased probability of overweight and obesity (OR: 2.01; 95% CI: 1.26, 3.22; P < 0.01; and OR: 2.90; 95% CI: 1.65, 5.09; P < 0.01; respectively). Significant inverse associations were found between serum concentrations of carotenoids and vitamin E and adiposity among Mexican-American children, but serum retinol concentrations were positively associated with adiposity. Future research is needed to understand the causes and consequences of micronutrient status on adiposity and comorbidities.

[Genetic modification of Methylobacterium extorquens G10 producer strain of polyhydroxybutyrate].

The effect of the increased copy number of polyhydroxybutyrate (PHB) biosynthesis genes in pink-pigmented methylobacterium Methylobacterium extorquens G10 on properties of the biopolymer was studied. The activity of poly-3-hydroxybutyril-synthase (PHB-synthase) was shown to increase and the molecular weight of synthesized PHB decreases twofold (150 --> 79 kDa) after insertion of extra copies of phaC and phaCAB genes into cells of the producer strain, whereas the physicochemical properties of the plastic changed insignificantly. White mutant M. extorquens G10-W with disrupted synthesis of the carotenoid pigment (defect by the crtI gene, which codes for phytoene desaturase) was established to have the same rate of growth and level of PHB accumulation as the initial strain G10. The G10-W strain is a promising producer of PHB, with decreased expenses for purification and PHB biosynthesis.

Low vitamin and carotenoid levels are related to cerebral white matter lesions.

PURPOSE: To determine the effects of vitamins and carotenoids on brain white matter lesions (WMLs), we examined the associations between WMLs with vitamin and carotenoid levels in Japanese middle-aged and elderly subjects. SUBJECTS AND METHODS: Four-hundred and sixty-nine healthy participants (male = 317; female = 152) that underwent medical examinations were examined. Deep white matter lesions (DWLs) were detected via magnetic resonance imaging (MRI) in 39 subjects. We evaluated the effects of vitamin and carotenoid levels on DWLs via logistic regression analysis. RESULTS: Lower gamma-tocopherol levels were significantly associated with DWLs in all subjects. While lower gamma-tocopherol and vitamin C levels were significantly associated with DWLs in males, lower delta-tocopherol levels were associated with DWLs in females. The associations between DWLs and lower gamma- and delta-tocopherol and vitamin C levels were independent of age, hypertension, or smoking. However, the associations between DWLs and lower alfa-tocopherol were not significant following adjustments for smoking. CONCLUSION: Lower carotenoid and vitamin levels were independently associated with cerebral DWLs in Japanese subjects.

Fat spectro-colorimetric characteristics of lambs switched from a low to a high dietary carotenoid level for various durations before slaughter.

This study investigated the changes in fat reflectance spectrum characteristics and color in lambs switched from a low to a high dietary carotenoid level for various durations before slaughter. Six treatments, feeding a high dietary carotenoid level for 0, 15, 30, 45, 60 or 75 days before slaughter, were compared in individually indoor penned lambs. Each treatment used 10 Romane lambs; feeding management ensured similar growth pattern and carcass weight for all the treatment groups. There was a change in reflectance spectrum characteristics and yellowness of subcutaneous fat as early as 15 days after the switch. Mean concentration of carotenoid pigments and yellowness of subcutaneous fat increased linearly with the duration of the high dietary carotenoid level. In perirenal fat, the change in reflectance spectrum characteristics was observed as early as 15 days after the switch, but the response to the duration of the high dietary carotenoid level was curvilinear.

Micronutrient deficiency in obese subjects undergoing low calorie diet.

BACKGROUND: The prevalence of micronutrient deficiencies is higher in obese individuals compared to normal-weight people, probably because of inadequate eating habits but also due to increased demands among overweight persons, which are underestimated by dietary reference intakes (DRI) intended for the general population. We therefore evaluated the dietary micronutrient intake in obese individuals compared to a reference population and DRI recommendations. Furthermore, we determined the micronutrient status in obese subjects undergoing a standardized DRI-covering low-calorie formula diet to analyze if the DRI meet the micronutrient requirements of obese individuals. METHODS: In 104 subjects baseline micronutrient intake was determined by dietary record collection. A randomly assigned subgroup of subjects (n = 32) underwent a standardized DRI-covering low-calorie formula diet over a period of three months. Pre- and post-interventional intracellular micronutrient status in buccal mucosa cells (BMC) was analyzed, as well as additional micronutrient serum concentrations in 14 of the subjects. RESULTS: Prior to dietetic intervention, nutrition was calorie-rich and micronutrient-poor. Baseline deficiencies in serum concentrations were observed for 25-hydroxyvitamin-D, vitamin C, selenium, iron, as well as ß-carotene, vitamin C, and lycopene in BMC. After a three-month period of formula diet even more subjects had reduced micronutrient levels of vitamin C (serum, BMC), zinc, and lycopene. There was a significant negative correlation between lipophilic serum vitamin concentrations and body fat, as well as between iron and C-reactive protein. CONCLUSIONS: The present pilot study shows that micronutrient deficiency occurring in obese individuals is not corrected by protein-rich formula diet containing vitamins and minerals according to DRI. In contrast, micronutrient levels remain low or become even lower, which might be explained by insufficient intake, increased demand and unbalanced dispersal of lipophilic compounds in the body. TRIAL REGISTRATION: The study was registered at ClinicalTrials.gov (NCT01344525). The study protocol comprises only a part of the approved trial protocol.

Immune status is more affected by age than by carotenoid depletion-repletion in healthy human subjects.

Prospective studies have indicated an age-related impairment of the immune response. Carotenoids have been hypothesised to enhance immune cell function. The aim of the present study was to describe the age-related effects and the impact of in vivo dietary carotenoid depletion and repletion on specific and non-specific immunity. A total of ninety-eight healthy male subjects (aged 20-75 years) received a carotenoid-depleted diet for 3 weeks and were then supplemented daily for 5 weeks with 30 mg ß-carotene, 15 mg lycopene and 9 mg lutein. Blood samples were collected at study entry, after depletion and supplementation, and biomarkers of immune status were determined. We found that serum IgA levels were positively correlated with ageing. Lymphocyte phenotyping indicated an increase with age in the memory T-helper cell subpopulation (CD4⁺CD45RO⁺) concomitantly with a decrease in naive T-helper cells (CD4⁺CD45RA⁺). A significant increase in the natural killer cells subpopulation and a small decrease in B lymphocytes were also observed, especially for the oldest volunteers. From ex vivo cell function exploration, a positive correlation was observed between age and IL-2 production of phytohaemagglutinin-stimulated lymphocytes. Neutrophils' bactericidal activity was significantly impaired with age (from 50 years) and was modulated by carotenoid status. An age effect was found on neutrophils' spontaneous migration but not on directed migration. Immune response in healthy human subjects is mostly affected by age rather than by dietary carotenoid depletion and repletion. Even in carefully selected healthy volunteers, some age-related immune changes occur predominantly from 50 years onwards. This immunosenescence could generate a loss in the immune system adjustment capacity.

Carotenoid deficiency triggers autophagy in the model green alga Chlamydomonas reinhardtii.

All aerobic organisms have developed sophisticated mechanisms to prevent, detect and respond to cell damage caused by the unavoidable production of reactive oxygen species (ROS). Plants and algae are able to synthesize specific pigments in the chloroplast called carotenoids to prevent photo-oxidative damage caused by highly reactive by-products of photosynthesis. In this study we used the unicellular green alga Chlamydomonas reinhardtii to demonstrate that defects in carotenoid biosynthesis lead to the activation of autophagy, a membrane-trafficking process that participates in the recycling and degradation of damaged or toxic cellular components. Carotenoid depletion caused by either the mutation of phytoene synthase or the inhibition of phytoene desaturase by the herbicide norflurazon, resulted in a strong induction of autophagy. We found that high light transiently activates autophagy in wild-type Chlamydomonas cells as part of an adaptation response to this stress. Our results showed that a Chlamydomonas mutant defective in the synthesis of specific carotenoids that accumulate during high light stress exhibits constitutive autophagy. Moreover, inhibition of the ROS-generating NADPH oxidase partially reduced the autophagy induction associated to carotenoid deficiency, which revealed a link between photo-oxidative damage, ROS accumulation and autophagy activation in Chlamydomonas cells with a reduced carotenoid content.

Low-serum carotenoid concentrations and carotenoid interactions predict mortality in US adults: the Third National Health and Nutrition Examination Survey.

Evidence regarding the health benefits of carotenoids is controversial. Effects of serum carotenoids and their interactions on mortality have not been examined in a representative sample of US adults. The objective was to examine whether serum carotenoid concentrations predict mortality among US adults. The study consisted of adults aged ≥20 years enrolled in the Third National Health and Nutrition Examination Survey, 1988 to 1994, with measured serum carotenoids and mortality follow-up through 2006 (N = 13,293). Outcomes were all-cause, cardiovascular disease, and cancer mortality. In adjusted Cox proportional hazards models, participants in the lowest total carotenoid quartile (<1.01 µmol/L) had significantly higher all-cause mortality (mortality rate ratio, 1.38; 95% confidence interval, 1.15-1.65; P = .005) than those in the highest total carotenoid quartile (>1.75 µmol/L). For α-carotene, the highest quartile (>0.11 µmol/L) had the lowest all-cause mortality rates (P < .001). For lycopene, the middle 2 quartiles (0.29-0.58 µmol/L) had the lowest all-cause mortality rates (P = .047). Analyses with continuous carotenoids confirmed associations of serum total carotenoids, α-carotene, and lycopene with all-cause mortality (P < .001). In a random survival forest analysis, very low lycopene was the carotenoid most strongly predictive of all-cause mortality, followed by very low total carotenoids. α-Carotene/ß-cryptoxanthin, α-carotene/lutein+zeaxanthin and lycopene/lutein+zeaxanthin interactions were significantly related to all-cause mortality (P < .05). Low α-carotene was the only carotenoid associated with cardiovascular disease mortality (P = .002). No carotenoids were significantly associated with cancer mortality. Very low serum total carotenoid, α-carotene, and lycopene concentrations may be risk factors for mortality, but carotenoids show interaction effects on mortality. Interventions of balanced carotenoid combinations are needed for confirmation.

Characterization of cell response in Chlamydomonas moewusii cultures exposed to the herbicide paraquat: Induction of chlorosis.

The use of herbicides constitutes the principal method of weed control, but the introduction of these compounds into the aquatic environment can provoke severe consequences for non-target organisms such as microalgae. Effects of the widely used herbicide paraquat were assessed on the green freshwater microalga Chlamydomonas moewusii by means of the analysis of its photosynthetic pigment content, using a traditional spectrophotometric technique that provides population bulk measurements, and by means of flow cytometry, which allowed characterizing the microalgal response at a single-cell level. Results obtained reveal that paraquat concentrations above 50nM induce chlorosis in a percentage of microalgal cells depending on herbicide concentration and exposure time, as reflected by a reduced cell chlorophyll autofluorescence and pigment content of the biomass. Cell viability in these cultures was also reduced in a concentration dependent way. The possibility of analysing chlorotic and non-chlorotic subpopulations separately allowed the study of morphological properties and physiological status of both cell types, leading to the conclusion that chlorotic cells are non-viable cells, based on their reduced size and complexity and their inability to be stained in the fluorescein diacetate assay. In the case of non-chlorotic cells, cell viability was reduced with the increase of paraquat concentration. Non-chlorotic cells in these cultures showed an increased size and complexity in comparison with control cells, probably due to a growth inhibition. Chlorophyll fluorescence was the most sensitive parameter since even cells exposed to the lowest concentration assayed, 50nM, although not chlorotic, showed a significantly reduced chlorophyll fluorescence with respect to control cells, reflected also by a reduced chlorophyll content of the biomass.