Current challenges in designer cellulosome engineering.

Designer cellulosomes (DCs) are engineered multi-enzyme complexes, comprising carbohydrate-active enzymes attached to a common backbone, the scaffoldin, via high-affinity cohesin-dockerin interactions. The use of DCs in the degradation of renewable biomass polymers is a promising approach for biorefineries. Indeed, DCs have shown significant hydrolytic activities due to the enhanced enzyme-substrate proximity and inter-enzyme synergies, but technical hurdles in DC engineering have hindered further progress towards industrial application. The challenge in DC engineering lies in the large diversity of possible building blocks and architectures, resulting in a multivariate and immense design space. Simultaneously, the precise DC composition affects many relevant parameters such as activity, stability, and manufacturability. Since protein engineers face a lack of high-throughput approaches to explore this vast design space, DC engineering may result in an unsatisfying outcome. This review provides a roadmap to guide researchers through the process of DC engineering. Each step, starting from concept to evaluation, is described and provided with its challenges, along with possible solutions, both for DCs that are assembled in vitro or are displayed on the yeast cell surface. KEY POINTS: • Construction of designer cellulosomes is a multi-step process. • Designer cellulosome research deals with multivariate construction challenges. • Boosting designer cellulosome efficiency requires exploring a vast design space.

Conversion of the free Cellvibrio japonicus xyloglucan degradation system to the cellulosomal mode.

Cellulosomes are multi-enzyme complexes produced by specialised micro-organisms. The spatial proximity of synergistically acting enzymes incorporated in these naturally occurring complexes supports the efficient hydrolysis of lignocellulosic biomass. Several functional designer cellulosomes, incorporating naturally non-cellulosomal cellulases, have been constructed and can be used for cellulose saccharification. However, in lignocellulosic biomass, cellulose is tightly intertwined with several hemicelluloses and lignin. One of the most abundant hemicelluloses interacting with cellulose microfibrils is xyloglucan, and degradation of these polymers is crucial for complete saccharification. Yet, designer cellulosome studies focusing on the incorporation of hemicellulases have been limited. Here, we report the conversion of the free Cellvibrio japonicus xyloglucan degradation system to the cellulosomal mode. Therefore, we constructed multiple docking enzyme variants of C. japonicus endoxyloglucanase, ß-1,2-galactosidase, α-1,6 xylosidase and ß-1,4-glucosidase, using the combinatorial VersaTile technique dedicated to the design and optimisation of modular proteins. We individually optimised the docking enzymes to degrade the xyloglucan backbone and side chains. Finally, we show that a purified designer xyloglucanosome comprising these docking enzymes was able to release xyloglucan oligosaccharides, galactose, xylose and glucose from tamarind xyloglucan. KEY POINTS: • Construction of xyloglucan-degrading designer cellulosome. • Conversion of free Cellvibrio japonicus enzymes to cellulosomal mode. • Type of linker inserted between dockerin and enzyme module affects docking enzyme activity.

High-Throughput Generation of Product Profiles for Arabinoxylan-Active Enzymes from Metagenomes.

Metagenomics is an exciting alternative to seek carbohydrate-active enzymes from a range of sources. Typically, metagenomics reveals dozens of putative catalysts that require functional characterization for further application in industrial processes. High-throughput screening methods compatible with adequate natural substrates are crucial for an accurate functional elucidation of substrate preferences. Based on DNA sequencer-aided fluorophore-assisted carbohydrate electrophoresis (DSA-FACE) analysis of enzymatic-reaction products, we generated product profiles to consequently infer substrate cleavage positions, resulting in the generation of enzymatic-degradation maps. Product profiles were produced in high throughput for arabinoxylan (AX)-active enzymes belonging to the glycoside hydrolase families GH43 (subfamilies 2 [MG432], 7 [MG437], and 28 [MG4328]) and GH8 (MG8) starting from 12 (arabino)xylo-oligosaccharides. These enzymes were discovered through functional metagenomic studies of feces from the North American beaver (Castor canadensis). This work shows how enzyme loading alters the product profiles of all enzymes studied and gives insight into AX degradation patterns, revealing sequential substrate preferences of AX-active enzymes.IMPORTANCE Arabinoxylan is mainly found in the hemicellulosic fractions of rice straw, corn cobs, and rice husk. Converting arabinoxylan into (arabino)xylo-oligosaccharides as added-value products that can be applied in food, feed, and cosmetics presents a sustainable and economic alternative for the biorefinery industries. Efficient and profitable AX degradation requires a set of enzymes with particular characteristics. Therefore, enzyme discovery and the study of substrate preferences are of utmost importance. Beavers, as consumers of woody biomass, are a promising source of a repertoire of enzymes able to deconstruct hemicelluloses into soluble oligosaccharides. High-throughput analysis of the oligosaccharide profiles produced by these enzymes will assist in the selection of the most appropriate enzymes for the biorefinery.

A Multistate Model for Analyzing Transitions Between Body Mass Index Categories During Childhood: The Generation XXI Birth Cohort Study.

Prevalences of overweight and obesity in young children have risen dramatically in the last several decades in most developed countries. Childhood overweight and obesity are known to have immediate and long-term health consequences and are now recognized as important public health concerns. We used a Markov 4-state model with states defined by 4 body mass index (BMI; weight (kg)/height (m)2) categories (underweight (<-2 standard deviations (SDs) of BMI z score), normal weight (-2 ≤ SD ≤ 1), overweight (1 < SD ≤ 2), and obese (>2 SDs of BMI z score)) to study the rates of transition to higher or lower BMI categories among children aged 4-10 years. We also used this model to study the relationships between explanatory variables and their transition rates. The participants consisted of 4,887 children from the Generation XXI Birth Cohort Study (Porto, Portugal; 2005-2017) who underwent anthropometric evaluation at age 4 years and in at least 1 of the subsequent follow-up waves (ages 7 and 10 years). Children who were normal weight were more likely to move to higher BMI categories than to lower categories, whereas overweight children had similar rates of transition to the 2 adjacent categories. We evaluated the associations of maternal age and education, type of delivery, sex, and birth weight with childhood overweight and obesity, but we observed statistically significant results only for sex and maternal education with regard to the progressive transitions.

Direct and BMI-mediated effect of birthweight on childhood cardio-metabolic health-a birth cohort study.

BACKGROUND: Different directions of the association of birthweight with cardio-metabolic health have been found, especially in children, which may be explained by the mediating effect of attained adiposity. We aimed to untangle direct and BMI-mediated associations of birthweight with childhood cardio-metabolic indicators. METHODS: Children from Generation XXI birth cohort were included (n = 4881). Birthweight was abstracted from clinical files. At age 4 and 7, children were re-evaluated. Glucose, triglycerides, LDL-cholesterol, systolic (SBP) and diastolic blood pressure (DBP) z-scores were the cardio-metabolic traits analyzed. Regression coefficients and respective 95% confidence intervals [ß (95%CI)] were computed using path analysis. RESULTS: Birthweight had inverse total effect on SBP at age 4 [-0.005 (-0.010; -0.001)] and 7 [-0.011 (-0.017; -0.006)] and DBP at 7 [-0.008 (-0.012; -0.004)]. Direct effects were found for SBP at 4 [-0.013 (-0.018; -0.009)] and 7 [-0.014 (-0.019; -0.009)], and DBP at 7 [-0.010 (-0.015; -0.006)], explaining the inverse total effects. Positive BMI-mediated indirect effects were found for all cardio-metabolic traits: higher birthweight was associated with higher childhood BMI, which in turn was associated with higher levels of cardio-metabolic traits. CONCLUSIONS: Positive BMI-mediated effect of birthweight on all cardio-metabolic traits was found. However, direct effects were in the opposite direction, significant for blood pressure, which may explain the diversity of results observed in the literature. Combining the direct and BMI-mediated effects, higher birthweight was associated with lower blood pressure at age 7 and have no effect on other cardio-metabolic traits.

Different levels of cardiometabolic indicators in multiple vs. singleton children.

BACKGROUND: We aimed to compare cardiometabolic indicators in singletons and multiples at age 7 and explore the birthweight mediation effect. METHODS: We studied 5431 singletons and 103 sets of multiples from Generation XXI birth cohort. Anthropometric measurements, body composition, and fasting blood samples were obtained. Age- and sex-specific z-scores were calculated (additionally height-specific for blood pressure). Adjusted regression coefficients and respective 95% confidence intervals [ß (95%CI)] were computed using path analysis. RESULTS: Multiples had lower weight [- 0.419 (- 0.616;-0.223)], height [- 0.404 (- 0.594;-0.213)], BMI [- 0.470 (- 0.705;-0.234)], fat mass index [- 0.359 (- 0.565;-0.152)], waist circumference [- 0.342 (- 0.537;-0.147)], and waist-to-height ratio [- 0.165 (- 0.326;-0.003)] z-scores. These results were explained by the indirect effect via birthweight, which was also negative and significant for all the aforementioned cardiometabolic indicators, while no direct effect was present. There were also significant indirect effects regarding fat-free mass index, glucose, insulin, and blood pressure, though the total effects were not significant, due to the balance between direct and indirect effects. The only significant direct effect was regarding diastolic blood pressure [- 0.165 (- 0.302;-0.028)]. CONCLUSIONS: At age 7, multiples presented better cardiometabolic indicators explained by lower weight at birth, except for the lower blood pressure which was independent of an effect via birthweight.

Prevalence and duration of breast milk feeding in very preterm infants: A 3-year follow-up study and a systematic literature review.

BACKGROUND: The World Health Organization recommends exclusive breast milk feeding until 6 months and continuing up to 2 years of age; little is known about whether very preterm infants are fed in accordance with these recommendations. We aimed to describe the prevalence and duration of breast milk feeding in very preterm children and to systematically review internationally published data. METHODS: We evaluated breast milk feeding initiation and duration in very preterm children born in 2 Portuguese regions (2011-2012) enrolled in the EPICE cohort and followed-up to the age of 3 (n = 466). We searched PubMed® from inception to January 2017 to identify original studies reporting the prevalence and/or duration of breast milk feeding in very preterm children. RESULTS: 91.0% of children received some breast milk feeding and 65.3% were exclusively breast fed with a median duration of 2 months for exclusive and 3 months for any breast milk; only 9.9% received exclusive breast milk for at least 6 months, 10.2% received any breast milk for 12 months or more, and 2.0% for up to 24 months. The literature review identified few studies on feeding after hospital discharge (n = 9); these also reported a low prevalence of exclusive breast milk feeding at 6 months (1.0% to 27.0%) and of any breast milk at 12 months (8.0% to 12.0%). CONCLUSIONS: The duration of breast milk feeding among Portuguese very preterm infants was shorter than recommended. However, this appears to be common globally. Research is needed to inform strategies to promote continued breast milk feeding.

Analysis of the substrate specificity of α-L-arabinofuranosidases by DNA sequencer-aided fluorophore-assisted carbohydrate electrophoresis.

Carbohydrate-active enzyme discovery is often not accompanied by experimental validation, demonstrating the need for techniques to analyze substrate specificities of carbohydrate-active enzymes in an efficient manner. DNA sequencer-aided fluorophore-assisted carbohydrate electrophoresis (DSA-FACE) is utmost appropriate for the analysis of glycoside hydrolases that have complex substrate specificities. DSA-FACE is demonstrated here to be a highly convenient method for the precise identification of the specificity of different α-L-arabinofuranosidases for (arabino)xylo-oligosaccharides ((A)XOS). The method was validated with two α-L-arabinofuranosidases (EC 3.2.1.55) with well-known specificity, specifically a GH62 α-L-arabinofuranosidase from Aspergillus nidulans (AnAbf62A-m2,3) and a GH43 α-L-arabinofuranosidase from Bifidobacterium adolescentis (BaAXH-d3). Subsequently, application of DSA-FACE revealed the AXOS specificity of two α-L-arabinofuranosidases with previously unknown AXOS specificities. PaAbf62A, a GH62 α-L-arabinofuranosidase from Podospora anserina strain S mat+, was shown to target the O-2 and the O-3 arabinofuranosyl monomers as side chain from mono-substituted ß-D-xylosyl residues, whereas a GH43 α-L-arabinofuranosidase from a metagenomic sample (AGphAbf43) only removes an arabinofuranosyl monomer from the smallest AXOS tested. DSA-FACE excels ionic chromatography in terms of detection limit for (A)XOS (picomolar sensitivity), hands-on and analysis time, and the analysis of the degree of polymerization and binding site of the arabinofuranosyl substituent.

Newborn weight change and childhood cardio-metabolic traits - a prospective cohort study.

BACKGROUND: Newborn weight change (NWC) in the first 4 days represents short-term adaptations to external environment. It may be a key developmental period for future cardio-metabolic health, but this has not been explored. We aimed to determine the associations of NWC with childhood cardio-metabolic traits. METHODS: As part of Generation XXI birth cohort, children were recruited in 2005/2006 at all public units providing obstetrical and neonatal care in Porto. Birthweight was abstracted from clinical records and postnatal anthropometry was obtained by trained examiners during hospital stay. NWC was calculated as ((minimum weight - birthweight)/birthweight) × 100. At age 4 and 7, children were measured and had a fasting blood sample collected. Fasting glucose, LDL-cholesterol, triglycerides, waist circumference, systolic and diastolic blood pressure were evaluated. This study included 312 children with detailed information on growth in very early life and subsequent cardio-metabolic measures. Path analysis was used to compute adjusted regression coefficients and 95% confidence intervals. RESULTS: NWC was not associated with any cardio-metabolic traits at ages 4 or 7. Strong associations were observed between each cardio-metabolic trait at 4 with the same trait at 7 years. The strongest associations were found for waist circumference [0.725 (0.657; 0.793)] and LDL-cholesterol [0.655 (0.575; 0.735)]. CONCLUSIONS: No evidence that NWC is related to childhood cardio-metabolic traits was found, suggesting that NWC should be faced in clinical practice as a short-term phenomenon, with no medium/long term consequences, at least in cardio-metabolic health. Our results show strong tracking correlations in cardio-metabolic traits during childhood.

Umbilical cord blood adipokines and newborn weight change.

PURPOSE: To ascertain the association between umbilical cord levels of adiponectin, leptin and resistin, and birth weight (BW) and newborn weight change (NWC) in the first 96 h of life. METHODS: 392 full-term singletons were recruited, in 2005/2006, at the five public units providing obstetrical and neonatal care in Porto. Information was collected by face-to-face interview and additionally from clinical records. Umbilical cord blood adipokines levels were determined and categorized using the 10th and 90th percentiles. Anthropometrics were obtained by trained examiners and NWC estimated as (weight - BW)/BW × 100, adjusted for newborn's age. Regression coefficients and 95% confidence intervals were calculated. RESULTS: Low leptin levels (≤5.6 ng/ml) were associated with lower BW (ß = -137.3 g 95%CI -268.6 g, -6.1 g) and high leptin levels (≥30.7 ng/ml) were associated with higher BW (ß = 276.3 g 95%CI 145.8 g, 406.8 g) and higher NWC (ß = 1.10% 95% CI 0.29%, 1.92%), comparing to newborns with normal leptin levels. Adiponectin and resistin were not associated with BW or NWC. CONCLUSION: High umbilical cord blood leptin levels predicted higher BW and lower weight loss in the immediate postnatal period.

Determinants of weight changes during the first 96 hours of life in full-term newborns.

BACKGROUND: Newborn weight loss (NWL) in the first 3 days of life is around 6 percent of birthweight (BW). We aim to describe the determinants of an excessive and insufficient NWL in the first 96 hours of life. METHODS: A sample of 1,288 full-term singletons without congenital abnormality belonging to Generation XXI birth cohort was selected. Newborns were recruited in 2005-2006 at all public units providing obstetrical and neonatal care in Porto, Portugal. Information was collected by face-to-face interview and additionally abstracted from clinical records. Anthropometrics were obtained by trained examiners and newborn weight change (NWC) was estimated as (weight-BW)/BW × 100. We categorized NWL as excessive (below 10th percentile of the sample distribution of NWC: ≤-9.4% of BW), normal (between 10th and 90th percentiles: -9.3 to -4.2%) and insufficient (above 90th percentile: ≥ -4.1%). Adjusted odds ratios (OR) and 95 percent confidence intervals (CI) were calculated using multinomial regression models. RESULTS: Excessive NWL was positively associated with maternal age ≥40 years (OR = 3.32, 95%CI 1.19-9.25), maternal education (OR = 1.04, 95% CI 1.00-1.09), cesarean delivery (OR = 2.42, 95% CI 1.12-5.23), and phototherapy-treated jaundice (OR = 1.69, 95% CI 1.00-2.87). Insufficient NWL was positively associated with low BW (OR = 2.68, 95% CI 1.13-6.33), and formula/mixed feeding (OR = 1.74, 95% CI 1.13-2.66). CONCLUSION: Excessive NWL was positively associated with maternal age and education, cesarean delivery, and phototherapy-treated jaundice. Insufficient NWL reflected child's feeding. As breastfed newborns did not lose weight excessively, but newborns with formula/mixed feeding had insufficient NWL, our study supports that breastfeeding provides excellent nutrition during this period.

Expectations and basic knowledge in ultrasound screening among pregnant women: A questionnaire survey.

OBJECTIVE: This study aimed to evaluate the disparity of the expectations and basic knowledge of prenatal ultrasound (US) screening among pregnant women and make a comparison with the current scientific knowledge and national recommendations. We hypothesize that sociodemographic factors, including age, education, and professional occupation, may be associated with different levels of knowledge. METHODS: This was a cross-sectional study performed in 2021 of 336 women aged 18 to 46 years in a maternity facility in a tertiary hospital in Portugal. The main outcome measures were questionnaire data from questions divided into four categories (sociodemographic, expectations, knowledge, and final considerations/suggestions). The data were grouped according to the sociodemographic factors (age, educational level, and professional occupation) and analyzed and compared as a function of the social groups as well as overall tendencies. RESULTS: Our data confirmed a significant discrepancy between the expectations and general knowledge of pregnant women regarding prenatal US when compared with the current scientific knowledge. Importantly, we found that both depended greatly on sociodemographic factors, particularly educational level. This reiterated the importance of conducting effective dissemination actions of current scientific knowledge, focusing on the main objectives of US screening as well as the limitations of the existing technology. CONCLUSION: This study will help in defining strategies for future dissemination actions aiming to improve current practice and lead to a higher synchrony of expectations towards US between couples and practitioners.

Respiratory Syncytial Virus Hospital Admission Rates and Patients' Characteristics Before the Age of 2 Years in England, 2015-2019.

BACKGROUND: A granular understanding of respiratory syncytial virus (RSV) burden in England is needed to prepare for new RSV prevention strategies. We estimated the rates of RSV hospital admissions before the age of 2 years in England and described baseline characteristics. METHODS: A birth cohort of all infants born between March 1, 2015, and February 28, 2017 (n = 449,591) was established using Clinical Practice Research Datalink-Hospital Episode Statistics. Case cohorts included infants with admission for (1) RSV, (2) bronchiolitis, (3) any respiratory tract infection (RTI) <24 months and (4) RSV predicted by an algorithm <12 months. Baseline characteristics were described in the case and comparative cohorts (infants without corresponding admission). Cumulative incidence and admission rates were calculated. Multiple linear regression was used to estimate the proportion of RTI healthcare visits attributable to RSV. RESULTS: The RSV-coded/RSV-predicted case cohorts were composed of 4813/12,694 infants (cumulative incidence: 1.1%/2.8%). Case cohort infants were more likely to have low birth weight, comorbidities and to be born during RSV season than comparative cohort infants, yet >77% were term-healthy infants and >54% were born before the RSV season. During the first year of life, 11.6 RSV-coded and 34.4 RSV-predicted hospitalizations occurred per 1000 person-years. Overall, >25% of unspecified lower RTI admissions were estimated to be due to RSV. CONCLUSIONS: In England, 1 in 91 infants had an RSV-coded admission, likely underestimated by ~3-fold. Most infants were term-healthy infants born before the RSV season. To decrease the total burden of RSV at the population level, immunization programs need to protect all infants.

Association between dietary patterns and metabolic syndrome in a sample of Portuguese adults.

BACKGROUND: There is scarce evidence regarding the association between diet and metabolic syndrome (MetS) in Portuguese population. We aim to evaluate the association between a posteriori dietary patterns (DPs) and MetS and its features. METHODS: Using random digit dialing, a sample of 2167 adults was selected between 1999 and 2003, in Porto. During a face-to-face interview, a questionnaire was applied, anthropometric measures were taken, blood pressure measured and a fasting blood sample collected. Diet was assessed using a validated food frequency questionnaire, and four DPs were identified in each sex by multivariate finite mixture models. RESULTS: After adjustment for age and daily energy intake, comparing to the "healthy" DP, women with the "low fruit and vegetables" DP had a higher odds of high waist circumference (OR = 1.88 95% CI 1.17-3.01) and low HDL-cholesterol (OR = 1.78 95% IC 1.12-2.82) and women in the "red meat and alcohol" DP had higher odds of high waist circumference (OR = 1.45 95% CI 1.01-2.07) and of MetS (OR = 1.57 95% CI 1.07-2.29); men with the "fish" DP had a higher odds of high triglycerides (OR = 1.57 95% CI 1.05-2.35). After further adjustments (education, physical activity, smoking, alcohol drinking, BMI, and menopausal status) no significant associations remained. CONCLUSIONS: Four distinct DPs were identified in a community sample of Portuguese adults and there was no association with the prevalence of MetS.

Dental Pain in Cats: A Prospective 6-Month Study.

Dental pathology is among the most ubiquitous diseases in cats of all ages. Dental pain is yet to be fully understood in cats and therefore its presence is often missed. To better understand feline dental disease as a pain trigger during routine examination and whether disease severity correlates to the degree of pain, a 6-month prospective study in a cats' only veterinary hospital in Portugal was conducted. Sixty-four cats that randomly presented for different clinical procedures were evaluated. Dental and periodontal abnormalities (primary dental parameters, PDP), as well as clinical signs related to dental pain (secondary dental parameters, SDP), were assessed. All cats underwent an oral cavity examination, upon which, the Feline Acute Pain Scale from Colorado State University Veterinary Teaching Hospital (CPS), was used in order to assess pain. Six PDP (periodontal disease, gingival index, calculus index, tooth resorption, tooth fracture and missing teeth) and five SDP (mouth discomfort, halitosis, hypersalivation, difficulty in holding food and several attempts at prehension of food), were compared with CPS pain scores. All SDP were significantly associated to higher CPS pain scores (p < 0.05). The number of missing teeth was significantly associated to higher CPS pain scores (p < 0.0001). A trend was observed between higher CPS pain scores and tooth resorption (p = 0.08). This study concluded that cats with dental disease feel pain during clinical examination and the pain increases as the severity of the disease progresses.

Combinatorial assembly and optimisation of designer cellulosomes: a galactomannan case study.

BACKGROUND: Designer cellulosomes are self-assembled chimeric enzyme complexes that can be used to improve lignocellulosic biomass degradation. They are composed of a synthetic multimodular backbone protein, termed the scaffoldin, and a range of different chimeric docking enzymes that degrade polysaccharides. Over the years, several functional designer cellulosomes have been constructed. Since many parameters influence the efficiency of these multi-enzyme complexes, there is a need to optimise designer cellulosome architecture by testing combinatorial arrangements of docking enzyme and scaffoldin variants. However, the modular cloning procedures are tedious and cumbersome. RESULTS: VersaTile is a combinatorial DNA assembly method, allowing the rapid construction and thus comparison of a range of modular proteins. Here, we present the extension of the VersaTile platform to facilitate the construction of designer cellulosomes. We have constructed a tile repository, composed of dockerins, cohesins, linkers, tags and enzymatically active modules. The developed toolbox allows us to efficiently create and optimise designer cellulosomes at an unprecedented speed. As a proof of concept, a trivalent designer cellulosome able to degrade the specific hemicellulose substrate, galactomannan, was constructed and optimised. The main factors influencing cellulosome efficiency were found to be the selected dockerins and linkers and the docking enzyme ratio on the scaffoldin. The optimised designer cellulosome was able to hydrolyse the galactomannan polysaccharide and release mannose and galactose monomers. CONCLUSION: We have eliminated one of the main technical hurdles in the designer cellulosome field and anticipate the VersaTile platform to be a starting point in the development of more elaborate multi-enzyme complexes.

The fetal origins of disease: a prospective cohort study on the association of preeclampsia and childhood obesity.

Cardiovascular diseases are the main cause of mortality worldwide, and childhood excess weight/obesity are strong correlators of accumulated risk in later life. A relationship between maternal preeclampsia and offspring's childhood obesity is recognized, but most studies fail to control for strong confounders. Our goal is to analyze the association between preeclampsia and childhood excess weight/obesity, after accounting for important confounders. We recruited 5133 women with singleton pregnancies during admission for delivery. Sixty-seven pregnancies were complicated by preeclampsia. Maternal and children outcomes were assessed at 10 years of age. We analyzed the association between preeclampsia and childhood excess weight/obesity by fitting a linear regression model (using offspring body mass index (BMI) z-score at 10 years of age) and a logistic regression model (using excess weight/obesity status). We then controlled both models for known confounders, namely maternal prepregnancy BMI, parity, and smoking during pregnancy. At 10 years of age, offspring of preeclamptic mothers had a higher BMI z-score and were more likely classified as overweight/obese, but these differences were not statistically significant. After controlling for maternal prepregnancy BMI, parity, and smoking during pregnancy, there was a high magnitude change in the beta coefficient of preeclampsia in the linear (0.175; -0.014) and the logistic regression models (1.48; 1.23) suggesting that the association between preeclampsia and childhood excess weigh/obesity is significantly confounded by these variables. These confounders also showed a significant association with childhood obesity. This finding suggests that in utero exposure to preeclampsia seems to have less impact in childhood obesity than the previously described confounders.

Adiposity rebound and cardiometabolic health in childhood: results from the Generation XXI birth cohort.

BACKGROUND: We aimed to evaluate the association of adiposity rebound (AR) timing on cardiometabolic health in childhood. METHODS: Participants were part of the Generation XXI birth cohort, enrolled in 2005/2006 in Porto. All measurements of the child's weight and height performed by health professionals as part of routine healthcare were collected. Individual body mass index (BMI) curves were fitted for 3372 children, using mixed-effects models with smooth spline functions for age and random effects. The AR was categorized into very early (<42 months), early (42-59 months), normal (60-83 months) and late (≥84 months). At age 10 years, cardiometabolic traits were assessed and age- and sex-specific z-scores were generated. Adjusted regression coefficients and 95% confidence intervals [ß (95% CI)] were computed. RESULTS: The mean age at AR was 61.9 months (standard deviations 15.7). Compared with children with normal AR, children with very early or early AR had higher z-scores for BMI [ß = 0.40 (95% CI: 0.28; 0.53); ß = 0.21 (95% CI: 0.12; 0.30)], waist circumference [ß = 0.33 (95% CI: 0.23; 0.43); ß = 0.18 (95% CI: 0.10; 0.25)], waist-height ratio [ß = 0.34 (95% CI: 0.24; 0.44); ß = 0.14 (95% CI: 0.07; 0.22)], fat mass index [ß = 0.24 (95% CI: 0.15; 0.33); ß = 0.14 (95% CI: 0.08; 0.21)], fat-free mass index [ß = 0.25 (95% CI: 0.14; 0.35); ß = 0.11 (95% CI: 0.03; 0.19)], systolic blood pressure [ß = 0.10 (95% CI: 0.01; 0.20); ß = 0.08 (95% CI: 0.01; 0.15)], insulin [ß = 0.16 (95% CI: 0.04; 0.29); ß = 0.10 (95% CI: 0.01; 0.19)], HOMA-IR [ß = 0.17 (95% CI: 0.04; 0.29); ß = 0.10 (95% CI: 0.03; 0.19)] and C-reactive protein [ß = 0.14 (95% CI: 0.02; 0.26); ß = 0.10 (95% CI: 0.01; 0.19)]. Children with very early AR also had worse levels of diastolic blood pressure [ß = 0.09 (95% CI: 0.02; 0.16)], triglycerides [ß = 0.21 (95% CI: 0.08; 0.34)] and high-density lipoprotein cholesterol [ß=-0.18 (95% CI: -0.31; -0.04)]. When analysed continuously, each additional month of age at the AR was associated with healthier cardiometabolic traits. CONCLUSION: The earlier the AR, the worse the cardiometabolic health in late childhood, which was consistently shown across a wide range of outcomes and in the categorical and continuous approach.

Does induction of labor at term increase the risk of cesarean section in advanced maternal age? A systematic review and meta-analysis.

BACKGROUND: Women of advanced maternal age, defined as ≥ 35 years at delivery, are at increased risk of multiple complications during pregnancy, with perinatal death being one of the most feared. For instance, the risk of stillbirth at term in this subgroup of women is higher than in younger women, and particularly high beyond 39 weeks of gestation. Induction of labor at 39-40 weeks might help prevent some cases of perinatal death, however, the fact that induction of labor has been historically associated with an increased risk of cesarean delivery and the knowledge that advanced maternal age is an independent risk factor for cesarean delivery are some of the major reasons why clinicians are reluctant to offer elective induction of labor in this particular group. OBJECTIVE: The aim of the study was to assess if induction of labor in advanced maternal age was associated with increased rates of cesarean delivery when compared to expectant management. MATERIAL AND METHODS: We performed an electronic search limited to published articles available between January 2000 and March 2020. Randomized clinical trials and retrospective studies with large cohorts comparing induction of labor with expectant management in singleton pregnancies at term, of women aged ≥ 35 years were included. The primary outcome was the rate of cesarean delivery in induction of labor versus expectant management, and secondary outcomes were the occurrence of assisted vaginal delivery and postpartum hemorrhage. RESULTS: Eight studies, including 81151 pregnancies (26,631 in the induction group and 54,520 expectantly managed), were included in the analysis. Six of the included studies were randomized clinical trials with the remaining two being observational and retrospective cohort studies. Induction of labor was not associated with a significant increased risk of cesarean delivery (OR 0.97, 95 % CI 0.86-1.1), assisted vaginal delivery (OR 1.12, 95 % CI 0.96-1.32) or postpartum hemorrhage (OR 1.11, 95 % CI 0.88-1.41). DISCUSSION: The belief that induction of labor is associated with an increased risk of cesarean delivery is based on the results of retrospective studies comparing induction with spontaneous labor at the same gestational age. However, at any point in a pregnancy, the comparison should be between induction of labor and expectant management, with the latter contributing to a pregnancy of greater gestation age and not always leading to spontaneous labor. When comparing induction to expectant management, our study shows no significant increase of cesarean section, assisted vaginal delivery or postpartum hemorrhage. Our study was not powered to assess neonatal outcomes, and additional research is needed to confirm whether induction of labor might have a positive effect in preventing stillbirth. CONCLUSION: Induction of labor at term in advanced maternal age has no significant impact on cesarean delivery rates, assisted vaginal delivery or postpartum hemorrhage, giving additional reassurance to obstetricians who would consider this intervention in this particular subgroup.