Comparison of α and ß-acid isomerization in hops and beer using HPLC, confocal microscopy, spectrofluorimetry and chemical analysis of metabolites and essential oils in flowers of different hop cultivars produced in Brazil.

We used confocal microscopy and spectrofluorescence to characterize the emission spectra in hop flowers, to follow the isomerization processes in different hop preparations, and beers, to compare with HPLC extracted samples. Flowers of different hop cultivars produced in three regions of Brazil, were quantitated by HPLC and GC-MS. The fluorescence spectra showed two characteristic emission bands evaluated from different preparations. The isomerization process leads to a gradual decrease in fluorescence intensity as the reaction progresses. This demonstrates the valuable use of confocal microscopy and fluorescence spectroscopy for analysis of the correlation between bitter acid indices with fluorescence intensity and lifetime microscopy. Such techniques can be used directly in the flowers allowing rapid monitoring of the brewing process. Twenty-nine substances were characterized in the essential oils and some cultivars presented quantities of bitter acids and essential oil levels close to those expected for plants after more than three years of cultivation.

T-cell immunity to SARS-CoV-2: what if the known best is not the optimal course for the long run? Adapting to evolving targets.

Humanity did surprisingly well so far, considering how unprepared it was to respond to the coronavirus disease 2019 (COVID-19) threat. By blending old and ingenious new technology in the context of the accumulated knowledge on other human coronaviruses, several vaccine candidates were produced and tested in clinical trials in record time. Today, five vaccines account for the bulk of the more than 13 billion doses administered worldwide. The ability to elicit biding and neutralizing antibodies most often against the spike protein is a major component of the protection conferred by immunization but alone it is not enough to limit virus transmission. Thus, the surge in numbers of infected individuals by newer variants of concern (VOCs) was not accompanied by a proportional increase in severe disease and death rate. This is likely due to antiviral T-cell responses, whose evasion is more difficult to achieve. The present review helps navigating the very large literature on T cell immunity induced by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection and vaccination. We examine the successes and shortcomings of the vaccinal protection in the light of the emergence of VOCs with breakthrough potential. SARS-CoV-2 and human beings will likely coexist for a long while: it will be necessary to update existing vaccines to improve T-cell responses and attain better protection against COVID-19.

Validity of self-reported oral conditions among Brazilian older women: Do socio-economic factors matter?

OBJECTIVE: To validate the level of agreement between self-report and clinical examination for oral conditions and evaluate the effect of sociodemographic conditions on the validity of self-report among women aged 60 and older. METHODS: A cross-sectional study was conducted in a social community center for seniors in Southern Brazil. Sociodemographic data (age, level of education, and income) were measured. Participants were interviewed and clinically examined for the number of teeth (DMF-T index) and the use of dental prostheses. The self-reported number of teeth in each arch and the use of dental prostheses were gathered through interviews. The level of agreement was estimated using the observed agreement, Kappa statistics, sensitivity/specificity (edentulism/prostheses) and Lin's concordance correlation coefficient, and related tests (number of teeth). The validity of the oral conditions was estimated according to sociodemographic information. RESULTS: Ninety-nine women participated in the study. High levels of agreement were observed for edentulism (97.8%; 95%CI 92.8;99.7; Kappa 0.947) and the use of dental prostheses (97.0%; 95%CI 91.3;99.4; Kappa 0.922). In both conditions, despite achieving similar concordance correlation coefficients (ranging from weak to moderate), the mean number of upper teeth was lower in clinical examination (7.1 ± 5.2) compared with self-reported (8.6 ± 3.6), while the opposite was observed for lower teeth (clinical examination: 9.1 ± 3.4; self-reported: 6.6 ± 5.3). Larger differences were found among women of low income and educational levels. CONCLUSIONS: Our findings suggest that the participants' socio-economic position might influence their self-reported number of teeth.

Combined exposure to alcohol and cannabis during development: Mechanisms and outcomes.

Exposure to substances of abuse during pregnancy can have long-lasting effects on offspring. Alcohol is one of the most widely used substances of abuse that leads to the most severe consequences. Recent studies in the United States, Canada, and the United Kingdom showed that between 1% and 7% of all children exhibit signs and symptoms of fetal alcohol spectrum disorder (FASD). Despite preventive campaigns, the rate of children with FASD has not decreased during recent decades. Alcohol consumption often accompanies exposure to such drugs as tobacco, cocaine, opioids, and cannabis. These interactions can be synergistic and exacerbate the deleterious consequences of developmental alcohol exposure. The present review focuses on interactions between alcohol and cannabis exposure and the potential consequences of these interactions.

Efficacy of resveratrol in male urogenital tract dysfunctions: an evaluation of pre-clinical data.

Resveratrol is a polyphenol found naturally in fruits and plants. Recently, studies in humans and animal models have suggested beneficial properties of this polyphenol, such as improvements to metabolic and lipid profiles, along with antioxidant, anti-inflammatory and anti-proliferative effects. In the urogenital tract (UGT), resveratrol has also been tested clinically and experimentally as a therapeutic drug in several diseases; however, the translational efficacy of resveratrol, especially in UGT, is still a matter of debate. In the present review, we address the pre-clinical efficacy of resveratrol in UGT-related dysfunctions, focusing on lower urinary tract symptoms, non-cancerous prostatic disease (benign prostatic hyperplasia and prostatitis) and erectile dysfunction. In vitro studies indicate that resveratrol reduces inflammatory markers and oxidative stress, and improves endothelial function in UGT organs and cells isolated from humans and animals. Despite displaying low oral bioavailability, in vivo administration of resveratrol largely improves erectile dysfunction, benign prostatic hyperplasia, prostatitis and voiding impairments, as evidenced in different animal models. Resveratrol also acts as a microbiota modulator, which may explain some of its beneficial effects in vivo. In contrast to the large amount of pre-clinical data, there are insufficient clinical trials to establish resveratrol treatment efficacy in human UGT-related diseases. In summary, we provide an overview of the in vivo and in vitro efficacy of resveratrol in animal and human UGT dysfunctions, which may support future clinical trials.

Palliative Sedation at Home: A Scoping Review.

Palliative Sedation (PS) is an effective measure for the relief of refractory symptoms in end-of-life patients. This intervention can be performed at home, respecting the patients' and their families' decisions. A scoping review was performed to map the available evidence in the literature on the performance of PS at home. This review included 23 studies. Most were conducted in European countries with adult cancer patients. Patients, family members and healthcare providers participated in the decision making regarding the use of PS at home. PS was used primarily to manage refractory symptoms (pain, delirium, dyspnea, and others), and in 1 of the studies PS was mentioned as a possible intervention for shortening life. The most commonly used medication was midazolam and the average duration ranged from 4 h to 7 days. There are few reports on adverse events related to PS. This intervention seems to be a feasible possibility for the management of refractory symptoms in patients at the end of life, despite the fact that it can represent specific challenges for healthcare providers, patients and families. However, the literature is limited regarding PS in children and in people with diseases other than cancer, as well as on the evaluation of possible adverse effects related to this intervention. Furthermore, it is essential to have a broad ethical, clinical and legal debate on whether to consider the use of PS for the purpose of shortening life in specific cases.

Effects of pharmaceuticals compounds and calcium on granulation, microbiology, and performance of anaerobic granular sludge systems.

Granular sludge is a promising biotechnology to treat sewage contaminated with pharmaceuticals due to its increased toxicity resistance. In this context, this study evaluated the potential of Ca2+ as a granulation precursor and how pharmaceutical compounds (loratadine, prednisone, fluconazole, fenofibrate, betamethasone, 17α-ethinyl estradiol, and ketoprofen) affect granulation. Continuous and intermittent dosages of Ca2+ in the presence and absence of pharmaceuticals were evaluated. The results showed that intermittent addition of Ca2+ reduces the time for anaerobic sludge granulation, and pharmaceuticals presence did not impair granulation. 10% of the granules presented mean diameters greater than 2.11 mm within 93 days with intermittent Ca2+ dosage in the pharmaceuticals' presence. In contrast, no granules higher than 2.0 mm were observed with no precursor addition. The pharmaceuticals' toxicity may have created a stress condition for the microbial community, contributing to more EPS production and a greater potential for granulation. It was also verified that pharmaceuticals' presence did not decrease organic matter, total alkalinity, and volatile fatty acids removals. The 16S rRNA gene analysis revealed taxa resistance to recalcitrant compounds when pharmaceuticals were added. Besides, the efficiency of a granular sludge bioreactor (EGSB) was evaluated for pharmaceuticals removal, and betamethasone, fenofibrate, and prednisone were effectively removed.

TP53 p.Arg337His geographic distribution correlates with adrenocortical tumor occurrence.

BACKGROUND: The p.Arg337His mutation of the TP53 is the most frequent germline missense variant associated with cancer described so far in this gene. It is mainly found in the South and Southeastern regions of Brazil, where it has been associated with a high incidence of pediatric adrenocortical (ACT) and choroid plexus tumors. The frequency and geographic distribution of this mutation is largely unknown, except for the Parana State, where a mean prevalence of 0.27% was reported. In the present study, we developed a high-throughput method for p.Arg337His genotyping, what allowed us to determine the frequency and geographic distribution of this mutation in a cohort from the most populous state in Brazil. METHODS: Consecutive samples from 31,612 newborns from São Paulo State were screened for p.Arg337His. The allelic discrimination was done by real-time polymerase chain reaction (PCR) and the presence of haplotype A3 in carriers was examined by using allele-specific oligonucleotide PCR, followed by nested-PCR to detect the SNP rs9894946. RESULTS: We found 67 (0.21%) samples positive for this mutation. The highest p.Arg337His frequencies were found in the cities close to the boundary between São Paulo and Minas Gerais State. No association could be found between p.Arg337His and gender, ethnicity, premature birth or twinning. Remarkably, a trend was found between the geographic distribution of p.Arg337His carriers and occurrence of ACT. CONCLUSION: We presented for the first time the p.Arg337His frequency among individuals unselected for any disease from a subset of the São Paulo State, the most populous in Brazil. The allele discrimination assay we presented here has proven to be a reliable and efficient method for high-throughput genotyping. ACT was found to be a good sentinel cancer to suppose p.Arg337His presence in our region.

Using online databases to produce comprehensive accounts of the vascular plants from the Brazilian protected areas: The Parque Nacional do Itatiaia as a case study.

BACKGROUND: Brazil is one of the most biodiverse countries in the world, with about 37,000 species of land plants. Part of this biodiversity is within protected areas. The development of online databases in the last years greatly improved the available biodiversity data. However, the existing databases do not provide information about the protected areas in which individual plant species occur. The lack of such information is a crucial gap for conservation actions. This study aimed to show how the information captured from online databases, cleaned by a protocol and verified by taxonomists allowed us to obtain a comprehensive list of the vascular plant species from the "Parque Nacional do Itatiaia", the first national park founded in Brazil. All existing records in the online database JABOT (15,100 vouchers) were downloaded, resulting in 11,783 vouchers identified at the species level. Overall, we documented 2,316 species belonging to 176 families and 837 genera of vascular plants in the "Parque Nacional do Itatiaia". Considering the whole vascular flora, 2,238 species are native and 78 are non-native. NEW INFORMATION: The "Parque Nacional do Itatiaia" houses 13% of the angiosperm and 37% of the fern species known from the Brazilian Atlantic Forest. Amongst these species, 82 have been cited as threatened, following IUCN categories (CR, EN or VU), seven are data deficient (DD) and 15 have been classified as a conservation priority, because they are only known from a single specimen collected before 1969.

Transcriptome analysis of acerola fruit ripening: insights into ascorbate, ethylene, respiration, and softening metabolisms.

KEY MESSAGE: The first transcriptome coupled to metabolite analyses reveals major trends during acerola fruit ripening and shed lights on ascorbate, ethylene signalling, cellular respiration, sugar accumulation, and softening key regulatory genes. Acerola is a fast growing and ripening fruit that exhibits high amounts of ascorbate. During ripening, the fruit experience high respiratory rates leading to ascorbate depletion and a quickly fragile and perishable state. Despite its growing economic importance, understanding of its developmental metabolism remains obscure due to the absence of genomic and transcriptomic data. We performed an acerola transcriptome sequencing that generated over 600 million reads, 40,830 contigs, and provided the annotation of 25,298 unique transcripts. Overall, this study revealed the main metabolic changes that occur in the acerola ripening. This transcriptional profile linked to metabolite measurements, allowed us to focus on ascorbate, ethylene, respiration, sugar, and firmness, the major metabolism indicators for acerola quality. Our results suggest a cooperative role of several genes involved in AsA biosynthesis (PMM, GMP1 and 3, GME1 and 2, GGP1 and 2), translocation (NAT3, 4, 6 and 6-like) and recycling (MDHAR2 and DHAR1) pathways for AsA accumulation in unripe fruits. Moreover, the association of metabolites with transcript profiles provided a comprehensive understanding of ethylene signalling, respiration, sugar accumulation and softening of acerola, shedding light on promising key regulatory genes. Overall, this study provides a foundation for further examination of the functional significance of these genes to improve fruit quality traits.

Oral health-related quality of life in older adults-Longitudinal study.

OBJECTIVE: The aim of the present study was to investigate oral health-related quality of life (OHRQoL) and determine associations with demographic, socio-economic and oral health variables at two evaluation times (2009 and 2015) among older adults assisted by Family Health Units in the Brazil. METHODS: A longitudinal study was conducted involving the evaluation (2009) and re-evaluation (2015) of 161 older adults (≥60 years of age). Demographic, socio-economic and general health variables were collected using a standardised questionnaire. Clinical oral health variables were also collected. OHRQoL was measured using the Oral Health Impact Profile-14 (OHIP-14). Multiple regression analysis was employed for the evaluation of OHRQoL considering the variables from the two evaluations. RESULTS: The severity of impact (mean OHIP-14) was 9.12 (SD: 10.81) in 2009 and 8.50 (10.52) in 2015. The greatest impact regarded the psychological discomfort domain at both evaluations (28.8% in 2009 and 22.9% in 2015). In the adjusted multiple linear regression, the mean OHIP-14 score increased 4.8 points (95% CI: 0.11-9.49) among individuals who experienced tooth loss between the two evaluations in comparison with those who maintained the same number of teeth. An increase in the OHIP-14 score occurred between 2009 and 2015 among 40.6% of the older adults. CONCLUSIONS: The findings of the present study reveal that the oral health status of older adults exerts an influence on OHRQoL, indicating the need for oral health policies directed at this portion of the population.

The role of atenolol in the modulation of the expression of genes encoding pro- (caspase-1) and anti- (Bcl2L1) apoptotic proteins in endothelial cells exposed to intestinal ischemia and reperfusion in rats

Abstract Purpose: To investigate the role of atenolol in the gene expression of caspase 1 (Casp1) and Bcl2L1 on vascular endothelium of rat intestine after ischemia and reperfusion (IR). Methods: Eighteen adult male Wistar rats were randomly divided into 3 groups (n=6): SG (Sham group): no clamping of the superior mesenteric artery; IRG: IR plus saline group: IRG+At: IR plus Atenolol group. Rats from IRG and IRG+At were subjected to 60 min of intestinal ischemia and 120 min of reperfusion. Atenolol (2mg/kg) or saline were injected in the femoral vein 5 min before ischemia, 5 min and 55 min after reperfusion. Thereafter, intestinal segments were appropriately removed and processed for Endothelial Cell Biology Rat RT2 Profiler PCR Array. Results: the anti-apoptotic Bcl2L1 gene expression was significantly down-regulated (-1.10) in the IRG and significantly up-regulated in the IRG+At (+14.15). Meanwhile, despite Casp1 gene expression was upregulated in both groups, it was significantly higher in the IRG (+35.06) than the IRG+At (+6.68). Conclusions: Atenolol presents antiapoptotic effects on rat intestine subjected to IR partly by the up-regulation of the anti-apoptotic Bcl2L1 gene expression. Moreover, atenolol can mitigate the pro-apoptotic and pro-inflammatory effects of Casp1 gene on rat intestine after IR.

The role of atenolol in the modulation of the expression of genes encoding pro- (caspase-1) and anti- (Bcl2L1) apoptotic proteins in endothelial cells exposed to intestinal ischemia and reperfusion in rats

Purpose:To investigate the role of atenolol in the gene expression of caspase 1 (Casp1) and Bcl2L1 on vascular endothelium of rat intestine after ischemia and reperfusion (IR).Methods:Eighteen adult male Wistar rats were randomly divided into 3 groups (n=6): SG (Sham group): no clamping of the superior mesenteric artery; IRG: IR plus saline group: IRG+At: IR plus Atenolol group. Rats from IRG and IRG+At were subjected to 60 min of intestinal ischemia and 120 min of reperfusion. Atenolol (2mg/kg) or saline were injected in the femoral vein 5 min before ischemia, 5 min and 55 min after reperfusion. Thereafter, intestinal segments were appropriately removed and processed for Endothelial Cell Biology Rat RT2 Profiler PCR Array.Results:the anti-apoptotic Bcl2L1 gene expression was significantly down-regulated (-1.10) in the IRG and significantly up-regulated in the IRG+At (+14.15). Meanwhile, despite Casp1 gene expression was upregulated in both groups, it was significantly higher in the IRG (+35.06) than the IRG+At (+6.68).Conclusions:Atenolol presents antiapoptotic effects on rat intestine subjected to IR partly by the up-regulation of the anti-apoptotic Bcl2L1 gene expression. Moreover, atenolol can mitigate the pro-apoptotic and pro-inflammatory effects of Casp1 gene on rat intestine after IR.(AU)

Herbarium collection of the Rio de Janeiro Botanical Garden (RB), Brazil.

BACKGROUND: This paper provides a quantitative and general description of the Rio de Janeiro Botanical Garden herbarium (RB) dataset. Created over a century ago, the RB currently comprises ca. 750,000 mounted specimens, with a strong representation of Brazilian flora, mainly from the Atlantic and Amazon forests. Nearly 100% of these specimens have been entered into the database and imaged and, at present, about 17% have been geo-referenced. This data paper is focused exclusively on RB's exsiccatae collection of land plants and algae, which is currently increasing by about twenty to thirty thousand specimens per year thanks to fieldwork, exchange and donations. Since 2005, many national and international projects have been implemented, improving the quality and accessibility of the collection. The most important facilitating factor in this process was the creation of the institutional system for plants collection and management, named JABOT. Since the RB is continuously growing, the dataset is updated weekly on SiBBr and GBIF portals. NEW INFORMATION: The most represented environments are the Atlantic and Amazon forests, a biodiversity hotspot and the world's largest rain forest, respectively. The dataset described in this article contains the data and metadata of plants and algae specimens in the RB collection and the link to access the respective images. Currently, the RB data is publicly available online at several biodiversity portals, such as our institutional database JABOT, the Reflora Virtual Herbarium, the SiBBr and the GBIF portal. However, a description of the RB dataset as a whole is not available in the literature.

Development and Validation of a Reversed Phase HPLC Method for Determination of Anacardic Acids in Cashew (Anacardium occidentale) Nut Shell Liquid.

Cashew nut shell liquid (CNSL) contains phenolic lipids with aliphatic chains that are of commercial interest. In this work, a chromatographic method was developed to monitor and quantify anacardic acids (AnAc) in CNSL. Samples containing AnAc were analyzed on a high-performance liquid chromatograph coupled to a diode array detector, equipped with a reversed phase C18 (150 × 4.6 mm × 5 µm) column using acetonitrile and water as the mobile phase both acidified with acetic acid to pH 3.0 in an isocratic mode (80:20:1). The chromatographic method showed adequate selectivity, as it could clearly separate the different AnAc. To validate this method, AnAc triene was used as an external standard at seven different concentrations varying from 50 to 1,000 µg mL-1. The Student's t-test and F-test were applied to ensure high confidence for the obtained data from the analytical calibration curve. The results were satisfactory with respect to intra-day (relative standard deviation (RSD) = 0.60%) and inter-day (RSD = 0.67%) precision, linearity (y = 2,670.8x - 26,949, r2 > 0.9998), system suitability for retention time (RSD = 1.02%), area under the curve (RSD = 0.24%), selectivity and limits of detection (19.8 µg mg-1) and quantification (60.2 µg mg-1). The developed chromatographic method was applied for the analysis of different CNSL samples, and it was deemed suitable for the quantification of AnAc.

The role of atenolol in the modulation of the expression of genes encoding pro- (caspase-1) and anti- (Bcl2L1) apoptotic proteins in endothelial cells exposed to intestinal ischemia and reperfusion in rats.

PURPOSE: To investigate the role of atenolol in the gene expression of caspase 1 (Casp1) and Bcl2L1 on vascular endothelium of rat intestine after ischemia and reperfusion (IR). METHODS: Eighteen adult male Wistar rats were randomly divided into 3 groups (n=6): SG (Sham group): no clamping of the superior mesenteric artery; IRG: IR plus saline group: IRG+At: IR plus Atenolol group. Rats from IRG and IRG+At were subjected to 60 min of intestinal ischemia and 120 min of reperfusion. Atenolol (2mg/kg) or saline were injected in the femoral vein 5 min before ischemia, 5 min and 55 min after reperfusion. Thereafter, intestinal segments were appropriately removed and processed for Endothelial Cell Biology Rat RT2 Profiler PCR Array. RESULTS: the anti-apoptotic Bcl2L1 gene expression was significantly down-regulated (-1.10) in the IRG and significantly up-regulated in the IRG+At (+14.15). Meanwhile, despite Casp1 gene expression was upregulated in both groups, it was significantly higher in the IRG (+35.06) than the IRG+At (+6.68). CONCLUSIONS: Atenolol presents antiapoptotic effects on rat intestine subjected to IR partly by the up-regulation of the anti-apoptotic Bcl2L1 gene expression. Moreover, atenolol can mitigate the pro-apoptotic and pro-inflammatory effects of Casp1 gene on rat intestine after IR.

Evaluation of nutritional and chemical composition of yacon syrup using 1H NMR and UPLC-ESI-Q-TOF-MSE.

A complete characterization of yacon syrup was performed by analytical techniques, including NMR and UPLC-QTOF-MSE. The effect of the different stages of yacon syrup production on fructooligosaccharides (FOS) and chlorogenic acid (CGA) contents were also evaluated. As a result, in addition to higher levels of FOS and CGA, some mineral elements, such as K, Ca and P, and essential amino acids, such as tryptophan, valine, and threonine, were determined in yacon syrup. Twenty-five compounds were putatively identified, and the main compounds were phenolics derived from quinic and trans-cinnamic acids. Considering the different stages of yacon syrup production, the results indicate that the contents of FOS and CGA were maintained in the pulping, enzymatic maceration and microfiltration, leading to a concentration of these components in the last stage of processing (vacuum concentration). These results will be used to fortify this innovative and promising product in the area of functional foods.

Treatment of mandibular condyle fractures. A 20-year review.

BACKGROUND/AIM: The condylar fracture is among the most frequent facial fractures. Despite all the published studies, its treatment remains controversial. The aim of this retrospective study was to evaluate the epidemiology and complications of mandibular condyle fractures managed by surgical and conservative treatments, over a period of twenty years. MATERIAL AND METHODS: The files of 262 patients with 318 condyle fractures were included in this study. One hundred and seven patients had conservative management, and 155 had surgical management for the condylar fractures. The outcomes were assessed by reviewing the patient's clinical records. RESULTS: The prevalence was higher in males and most patients were Caucasians in the third decade of life. The mean age was 30.17 years old. The main causes were motorcycle accidents followed by bike accidents, and the mean follow-up time was 226.5 days. The number of complications found in the two modalities of treatment was low and similar. CONCLUSION: Both treatments achieved their goals. There was no statistical difference in the complication rate between the two groups.

Pretreatment of cashew apple bagasse using protic ionic liquids: Enhanced enzymatic hydrolysis.

To enhance the enzymatic digestibility of cashew apple bagasse (CAB) feedstock in order to produce sugar fermentation-derived bioproducts, the CAB was subjected to three different pretreatments with the ionic liquid 2-hydroxyl-ethylammonium acetate (2-HEAA) and characterized by FTIR, NMR and chemical methods. All conditions were able to delignify CAB, however the best lignin removal (95.8%) was achieved through the method performed with 8.7% w/w of CAB/2-HEAA ratio at 130°C for 24h. Although the cellulose crystallinity has been increased in CAB treated with the ionic liquid, but this fact did not influence its digestibility. Nevertheless, the pretreatment with 2-HEAA enhanced significantly the cellulose digestibility, increasing the glucose yield from 48 to 747.72mgglucose/gCAB. Furthermore, 2-HEAA pretreatment was efficient even with reused ionic liquid, obtaining high glucose concentration.

Lessons Learned From a Critical Analysis of the Fontan Operation Over Three Decades in a Single Institution.

BACKGROUND: The Fontan operation (FO) has evolved and many centers have demonstrated improved results relative to those from earlier eras. We report a single-institution experience over three decades, describing the outcomes and evaluating risk factors. METHODS: Successive patients undergoing primary FO were divided into era I (1984-1994), era II (1995-2004), and era III (2005-2014). Clinical and operative notes were reviewed for demographic, anatomic, and procedure details. End points included early and late mortality and a composite of death, heart transplantation (HTX), or Fontan takedown. RESULTS: A total of 420 patients underwent 18 atriopulmonary connections, 82 lateral tunnels (LT), and 320 extracardiac conduit (EC) Fontan procedures. Forty-six (11%) patients died; early and late mortality were 7.9% and 3.1%, respectively. Eight (1.9%) patients underwent HTX, 11 (2.6%) underwent Fontan conversion to EC, and 1 (0.2%) takedown of EC to bidirectional Glenn shunt. Prevalence of concomitant valve surgery ( P < .001) and pulmonary artery reconstruction ( P < .001) differed over the eras. Preoperative valve regurgitation was associated with likelihood of early mortality (odds ratio [OR] = 3.5, P = .002). Embolic events (OR = 1.9, P = .047), preoperative valve regurgitation (OR = 2.3, P = .029), diagnosis of unbalanced atrioventricular canal defect (OR = 1.14, P = .03), and concomitant valve replacement (OR = 6.9, P = .001) during the FO were associated with increased risk of the composite end point (death, HTX, or takedown). CONCLUSION: Technical modifications did not result in improved results across eras, due in part to more liberal indications for surgery in the recent years. Valve regurgitation, unbalanced atrioventricular canal, embolic events, or concomitant valve replacement were associated with FO failure.