Emerging Roles of tRNAs in RNA Virus Infections.

Viruses rely on the host cell translation machinery for efficient synthesis of their own proteins. Emerging evidence highlights different roles for host transfer RNAs (tRNAs) in the process of virus replication. For instance, different RNA viruses manipulate host tRNA pools to favor viral protein translation. Interestingly, specific host tRNAs are used as reverse transcription primers and are packaged into retroviral virions. Recent data also demonstrate the formation of tRNA-derived fragments (tRFs) upon infection to facilitate viral replication. Here, we comprehensively discuss how RNA viruses exploit distinct aspects of the host tRNA biology for their benefit. In light of the recent advances in the field, we propose that host tRNA-related pathways and mechanisms represent promising cellular targets for the development of novel antiviral strategies.

A comprehensive review on PFAS including survey results from the EFLM Member Societies.

OBJECTIVES: Per- and polyfluoroalkyl substances (PFASs) are a large class of synthetic chemicals widely used for their unique properties. Without PFAS, many medical device and in vitro diagnostic technologies would not be able to perform their intended purposes. Potential health risks associated with exposure to PFAS influence their use in IVD applications. This paper aims to assess the current situation concerning PFAS, including regulations and legislations for their use. It is important to know what happens to (PFAS) at the end of their lives in medical laboratories. METHODS: A survey was conducted in March 2023 to collect information on the potential emission and end-of-life of PFAS-containing medical technologies in the medical laboratories of the EFLM member societies. A series of questions were presented to the EFLM national societies and the results were documented. RESULTS: Eight respondents participated in the survey, representing EFLM member societies in seven different countries including hospital laboratories, university laboratories, and private laboratories. CONCLUSIONS: PFAS uses in MD and IVD are influenced by several factors, including evolving regulations, advances in technology, safety and efficacy of these substances. Advancements in analytical techniques may lead to more sensitive and precise methods for detecting and quantifying PFAS in biological samples, which can be essential for IVD applications related to biomarker analysis and disease diagnosis. Collaboration among regulatory agencies, industry, research institutions, hospitals, and laboratories on a global scale can aid in establishing harmonized guidelines and standards for the use of PFAS, ensuring consistency and safety within their applications.

Lack of country-wide systematic herpetology collections in Portugal jeopardizes future research and conservation.

Natural History Collections (NHCs) represent the world's largest repositories of long-term biodiversity datasets. Specimen collection and voucher deposition has been the backbone of NHCs since their inception, but recent decades have seen a drastic decline in rates of growth via active collecting. Amphibians and reptiles are amongst the most threatened zoological groups on the planet and are historically underrepresented in most worldwide NHCs. As part of an ongoing project to review the Portuguese zoological collections in the country's NHCs, herpetological data from its three major museums and smaller collections was gathered and used to examine the coverage and representation of the different taxa extant in Portugal. These collections are not taxonomically, geographically, or temporally complete. Approximately 90% of the Portuguese herpetological taxa are represented in the country's NHCs, and around half of the taxa are represented by less than 50 specimens. Geographically, the collections cover less than 30% of the country's territory and almost all of the occurring taxa have less than 10% of their known distribution represented in the collections. A discussion on the implications for science of such incomplete collections and a review of the current status of Portuguese NHCs is presented.

Cardiovascular risk factors during pregnancy impact the postpartum cardiac and vascular reverse remodeling.

Pregnant women with cardiovascular risk (CVR) factors are highly prone to develop cardiovascular disease later in life. Thus, recent guidelines suggest extending the follow-up period to 1 yr after delivery. We aimed to evaluate cardiovascular remodeling during pregnancy and determine which CVR factors and potential biomarkers predict postpartum cardiac and vascular reverse remodeling (RR). Our study included a prospective cohort of 76 healthy and 54 obese and/or hypertensive and/or with gestational diabetes pregnant women who underwent transthoracic echocardiography, pulse-wave velocity (PWV), and blood collection at the 1st trimester (1T) and 3rd trimester (3T) of pregnancy as well as at the 1st/6th/12th mo after delivery. Generalized linear mixed-effects models was used to evaluate the extent of RR and its potential predictors. Pregnant women develop cardiac hypertrophy, as confirmed by a significant increase in left ventricular mass (LVM). Moreover, ventricular filling pressure (E/e') and atrial volume increased significantly during gestation. Significant regression of left ventricular (LV) volume, LVM, and filling pressures was observed as soon as 1 mo postpartum. The LV global longitudinal strain worsened slightly and recovered at 6 mo postpartum. PWV decreased significantly from 1T to 3T and normalized at 1 mo postpartum. We found that arterial hypertension, smoking habits, and obesity were independent predictors of increased LVM during pregnancy and postpartum. High C-reactive protein (CRP) and low ST2/IL33-receptor levels are potential circulatory biomarkers of worse LVM regression. Arterial hypertension, age, and gestational diabetes positively correlated with PWV. Altogether, our findings pinpoint arterial hypertension as a critical risk factor for worse RR and CRP, and ST2/IL33 receptors as potential biomarkers of postpartum hypertrophy reversal.NEW & NOTEWORTHY This study describes the impact of cardiovascular risk factors (CVR) in pregnancy-induced remodeling and postpartum reverse remodeling (up to 1 yr) by applying advanced statistic methods (multivariate generalized linear mixed-effects models) to a prospective cohort of pregnant women. Aiming to extrapolate to pathological conditions, this invaluable "human model" allowed us to demonstrate that arterial hypertension is a critical CVR for worse RR and that ST2/IL33-receptors and CRP are potential biomarkers of postpartum hypertrophy reversal.

Cumulus-oocyte complexes from sows show differences in lipid metabolism compared to cumulus-oocyte complexes from prepubertal gilts during in vitro maturation.

This study aimed to evaluate the effects of donor age on lipid metabolism during in vitro maturation (IVM) of pigs cumulus-oocyte complexes (COCs). We evaluated transcript levels of genes, the percentage of ooplasm occupied by lipid droplets (LD) and evaluated DNA methylation in COCs from sows and prepubertal gilts. Transcript levels of six genes (ACACA, ACSS2, FASN, FABP3, SLC27A4, PLIN2), which were analyzed in cumulus cells (CCs), increased after 44 h of IVM in the sow group. In the gilt group, only FASN expression increased, while NR3C1 expression decreased after IVM. The measurement of LD in oocytes showed an accumulation of lipids in sow oocytes during IVM, while gilt oocytes showed a decrease in LD. FABP3 and NR3C1 methylation patterns exhibited a demethylation pattern in CCs and oocytes from gilts and sows and showed statistical differences between groups. CCs from sows had a better capacity to change transcription levels of the major genes involved in lipid metabolism during IVM than CCs from gilts. This difference may be involved in accumulation of lipids, acquisition of competence, and maturation of enclosed oocytes. Our results contribute to a better understanding of mechanisms involved in lipid metabolism and acquisition of competence in porcine COCs.

Treatment patterns and outcomes for Hodgkin Lymphoma patients aged 60 and older: a report from the Brazilian Prospective Hodgkin Lymphoma Registry.

The treatment of older patients with Hodgkin lymphoma (HL) remains a challenge. We sought to identify the treatment patterns and outcomes in older HL patients included in the Brazilian HL registry (NCT02589548). A total of 136 patients with HIV-negative classic HL, aged ≥ 60 years, diagnosed between 2009 and 2018, were analyzed. The median age was 66 years old (60-90), 72% had advanced disease, 62% had a high IPS, and 49% had a nodular sclerosis subtype. Median follow-up was 64 months for alive patients. ABVD was the front-line treatment in 96% of patients. Twenty-one patients (15%) died during front-line treatment. The 5-year PFS and 5-year OS rates were 55% and 59%, respectively. The 5-year OS rates in localized and advanced disease were 81% and 51% (p=0.013). Lung toxicity developed in 11% of the patients treated with ABVD. Bleomycin was administered for > 2 cycles in 65% of patients. Compared with 2009-2014, there was a decrease in the use of bleomycin for > 2 cycles in 2015-2018 (88% × 45%, p<0.0001). The impact of socioeconomic status (SES) on outcomes was studied in patients treated with ABVD. After adjusting for potential confounders, lower SES remained independently associated with poorer survival (HR 2.22 [1.14-4.31] for OS and HR 2.84 [1.48-5.45] for PFS). Treatment outcomes were inferior to those observed in developed countries. These inferior outcomes were due to an excess of deaths during front-line treatment and the excessive use of bleomycin. SES was an independent factor for shorter survival.

Chemical strategies for sustainable medical laboratories.

Chemicals are essential components of our daily lives, for the well-being, high living standards and comfort of modern society. They are used in many sectors, including health. However, some chemicals have hazardous properties which can harm the environment and human health. Chemical pollution is significantly contributing to the current global problem of climate change and loss of biodiversity There is an increase in health problems that can be partially explained by the use of chemicals. Some man-made chemicals are found in the most remote places in the environment, but also in our bodies. Chemicals are everywhere. Chemicals strategy for sustainability towards a toxic-free environment will ensure better protection of human health and the environment from hazardous chemicals, boost innovation for safe and sustainable chemicals and enable the transition to chemicals that are safe and sustainable by design. It is a first step towards the Zero pollution ambition for a toxic-free environment announced in the European Green Deal. The strategy proposes comprehensive chemical legislations for the transformation of industry with the aim of attracting investment into safe and sustainable products and production methods. Clinical laboratories must choose safer, more sustainable alternatives to hazardous chemicals, address sustainability issues, and implement official guidelines on how to reduce their carbon footprint.

Digital transformation and sustainability in healthcare and clinical laboratories.

Healthcare, and in particular, clinical laboratories, are major contributors to carbon emissions and waste. Sustainability in healthcare has shifted from an environmental concern towards a holistic definition that includes balancing socio-ecological and socio-technical systems, including health services effectiveness and cost efficiency. Digital transformation can reduce waste and the cost of services by enhancing effectiveness while maintaining quality. Digital health interventions can provide personalized patient-centered care on a global scale and include decision support systems that have the potential to improve the performance and quality of healthcare. The right interfaces must be used so that the advantages of going digital are felt throughout the health system: a successful and sustainable implementation of digital innovation depends on its integration into a functional health ecosystem. Telehealth has the potential to reduce carbon emissions due to the reduced daily commute of health professionals, although research is limited. Recently, economic models have changed from the linear "take-make-dispose" to circular models based on recycling and upcycling that have the goal of keeping products, components, and materials at their highest utility and value. The previous linear models threaten human health and well-being and harm natural ecosystems.

Association between anti-Müllerian hormone levels and reproductive parameters in Wagyu cattle raised in Brazil.

The production of in vitro embryos has sped up the dissemination of superior genetic material. However, the variation among the cattle response to oocyte and embryo production is a challenging factor. This variation is even higher in the Wagyu cattle as the breed has a small effective population size. The identification of an effective marker related to reproductive efficiency would allow the selection of more responsive females to reproductive protocols. The objective of this study was to evaluate the blood levels of anti-Müllerian hormone and associate it with oocyte recovery and blastocyst rate of embryos produced in vitro in Wagyu cows, as well as observe the hormone circulating levels in males. Serum samples from 29 females with seven follicular aspirations and four bulls were used. AMH measurements were performed using the bovine AMH ELISA kit. A positive correlation was identified between oocyte production and blastocyst rate (r = 0.84, P = 9 × 10-9), and AMH levels with oocyte (r = 0.49, P = 0.006) and embryo (r = 0.39, P = 0.03) production. The mean levels of AMH were different between animals with low (11.06 ± 3.01) and high (20.75 ± 4.46) oocyte production (P = 0.01). Males showed high serological levels of AMH (3829 ± 2328 pg/ml) compared with other breeds. It is possible to use the serological measurement of AMH as a method to select Wagyu females with greater capacity for oocyte and embryo production. Further studies correlating AMH serological levels with Sertoli cell function in bulls are needed.

Successful lactation induction in non-pregnant gilts.

A protocol to induce lactation was applied to non-pregnant gilts. In Experiment I, five gilts with oestrus synchronized through oral supplementation of 20 mg altrenogest for 18 days received: 10 mg oestradiol cypionate (EC) on the last day of oestrous expression (D0); 10 mg EC and 300 mg long-acting progesterone (P4) on D26; and two 0.53 mg doses of a prostaglandin F2α analogue (PGF) 12 h apart on D36. Blood was collected on D12, D19, D26 and D33. Milk secretion started in all gilts 24 h after PGF administration and lasted at least 8 days. Milk samples were collected from D37 to D45. The serum P4 concentration was lower on D12 than subsequently (p < .05), but the oestradiol concentration was unaltered (p > .05). The milk produced during the induced lactation was generally richer in protein and poorer in fat compared to the milk from the lactation of a reference sow. In Experiment II, the same protocol induced lactation in two gilts, which nursed fostered piglets for 22 days. Thus, lactation was induced in all treated gilts and the milk produced was capable to nurture fostered piglets.

Mean Platelet Volume Predicts Restenosis After Carotid Endarterectomy.

BACKGROUND: Carotid restenosis following carotid endarterectomy (CEA) has a cumulative risk at 5-years up to 32%, which may impact the well-being of patients following CEA. Haematological parameters in the standard complete blood cell count (CBC) are emerging as potential biomarkers, but their application in CEA is scarce. The primary aim of this study was to investigate haematological markers for restenosis following CEA. The secondary aim was to characterize clinical risk factors for restenosis. METHODS: From January 2012 to January 2019, 151 patients who underwent CEA under regional anaesthesia due to carotid stenosis were selected from a prospectively maintained cohort database. Patients were included if a preoperative CBC was available in the 2 weeks preceding CEA. Multivariable analysis was performed alongside propensity score matching (PSM) analysis, using the preoperative CEA parameters, to reduce confounding factors between categories. RESULTS: The study group comprised 28 patients who developed carotid restenosis. The remaining 123 patients without restenosis composed the control group. Mean age of the patients did not differ significantly between groups (70.25 ± 8.05 vs. 70.32 ± 9.61 YO, P = 0.973), neither did gender (male gender 89.3% vs. 78.9%, P = 0.206). Regarding haematological parameters, only MPV remained statistically significant within multivariable analysis (1.855, aOR [1.174-2.931], P = 0.008), a result supported by PSM analysis (2.072, aOR [1.036-4.147], P = 0.042). CONCLUSIONS: MPV was able to predict restenosis 2 years after CEA. Thus, MPV can be incorporated into score calculations to identify patients at greater risk of restenosis, who could benefit from specific monitoring during follow-up. While results are promising, more research is necessary to corroborate them.

NEMO-NDAS: A Panniculitis in the Young Representing an Autoinflammatory Disorder in Disguise.

ABSTRACT: A 15-month-old full-term boy of African descent with an asymptomatic sickle cell trait presented with episodes of transient erythematous subcutaneous nodules involving the entire body except the face, since 2 weeks of age. The skin lesions evolved to areas of lipoatrophy and hyperpigmentation. An initial skin biopsy, studied at a different department at 2 months, was initially misinterpreted as subcutaneous fat necrosis of the newborn, despite the lack of the typical radiated crystals and needle-shaped clefts characterizing that entity. At 4 months of age, he developed systemic inflammatory manifestations, including fever, a new rash, significant periorbital edema, and failure to thrive. An extensive workup showed leukocytosis, hypercalcemia, elevated inflammatory markers, hypertriglyceridemia, and transaminitis. A new skin biopsy of the eyelid was diagnosed as neutrophilic lobular panniculitis with necrotic adipocytes. An initial whole-exome sequencing did not identify any causative mutations, but a WES reanalysis focused on autoinflammatory disorders was requested based on additional clinicopathologic data and revealed a mosaic intronic mutation in IKBKG c. 671+3 G > C. This mutation encodes an mRNA missing exon 5 resulting in NF-kB essential modulator (NEMO) Δ-exon 5-autoinflammatory syndrome (NDAS). NEMO-NDAS is one of the systemic autoinflammatory diseases that may appear as an unexplained panniculitis in young children, who should be monitored for immunodeficiency and/or autoinflammatory diseases. The differential diagnosis of autoinflammatory disorders should be considered in such cases incorporating the use of the whole-genome/exome sequencing in the investigation. The inhibitor of kappa-B kinase regulatory subunit gamma (IKBKG) is located on chromosome Xq28 and encodes the NEMO, a critical molecule upstream of NF-kB activation.

Metabolomics: A Powerful Tool to Understand the Schizophrenia Biology.

ABSVTRACT: Schizophrenia, as any other psychiatric disorder, is a multifactorial and complex illness whose etiology is not completely established. Therefore, studies involving strategies that are able to describe the molecular alterations caused by the disease and, consequently, indicate the altered metabolic pathways are of increasing interest. Metabolomics is a very suitable approach that can be applied for this task, since it consists of the evaluation of the set of metabolites contained in a biological system undergoing a biological process, such as a disease or treatment. In metabolomics, state-of-the-art analytical techniques (mass spectrometry and nuclear magnetic resonance) are employed to identify and quantify the metabolites present in the studied biological samples, and chemometric and bioinformatic tools are applied to determine the specific metabolites and metabolic pathways that are relevant to the biological process under investigation. The aim of this chapter is to describe the basic principles of metabolomics, how this strategy can improve the understanding of the schizophrenia biology, and the findings obtained so far.

Impact of oral anticoagulation therapy on postoperative atrial fibrillation outcomes: a systematic review and meta-analysis.

BACKGROUND: Post-operative atrial fibrillation (POAF) is the most common complication after cardiac surgery. Recent studies had shown this phenomenon is no longer considered transitory and is associated with higher risk of thromboembolic events or death. The aim of this study was to systematically review and analyze previous studies comparing oral anticoagulation therapy with no anticoagulation, regarding these long-term outcomes. METHODS: PubMed/MEDLINE, EMBASE, Web of Science and Cochrane Database were systematically searched to identify the studies comparing the risk of stroke, or thromboembolic events or mortality of POAF patients who received anticoagulation compared with those who were not anticoagulated. Incidence of stroke, thromboembolic events and all-cause mortality were evaluated up to 10 years after surgery. Time-to-event outcomes were collected through hazard ratio (HR) along with their variance and the early endpoints using frequencies or odds ratio (OR). Random effect models were used to compute statistical combined measures and 95% confidence intervals (CI). Heterogeneity was evaluated through Q statistic-related measures of variance (Tau2, I2, Chi-squared test). RESULTS: Eight observational cohort studies were selected, including 15,335 patients (3492 on Oral Anticoagulants (OAC) vs 11,429 without OAC) that met the inclusion criteria for qualitative synthesis. Patients had a wide gender distribution (38.6-82.3%), each study with a mean age above 65 years (67.5-85). Vitamin K antagonists were commonly prescribed anticoagulants (74.3-100%). OAC was associated with a protective impact on all-cause mortality at a mean of 5.0 years of follow-up (HR is 0.85 [0.72-1.01]; p = 0.07; I2 = 48%). Thromboembolic events did not differ between the two treatment arms (HR 0.68 [0.40-1.15], p = 0.15). CONCLUSION: Current literature suggests a possibly protective impact of OAC therapy for all-cause mortality in patients with new-onset atrial fibrillation after cardiac surgery. However, it does not appear to impact thromboembolism rate.

Inflammatory Response Assessment in Patients with COVID-19 Under Extracorporeal Membrane Oxygenation Support.

BACKGROUND: Extracorporeal membrane oxygenation (ECMO), could be extremely helpful in the management of COVID-19 patients with refractory hypoxemic respiratory failure; however, to date, evidence on the true effecttiveness of ECMO in the COVID19 setting still hangs in the balance. METHODS: This was a prospective cohort study of 39 COVID-19 patients admitted to the intensive care unit (ICU) in an experienced ECMO center at a tertiary hospital during March/April 2020. Among the recruited participants, 10 (25.6%) required ECMO (ICU-ECMO group) and 29 (74.4%) did not have ECMO support (ICU group). Immunological parameters were assessed both at ICU admission and on a daily basis for 7 consecutive days. RESULTS: The absolute lymphocyte count increased significantly in the ICU-ECMO group compared to the ICU group in which it remained relatively stable: ß for the time variable was 127.1 [95% CI 68.9 - 185.3], p < 0.001 and for the interaction term -141.36 [-208.95 - -73.77], p < 0.001. On the other hand, globally, no significant differences were observed over time for the lymphocyte percentage, although it was higher in the ICU patients. Neutrophil counts were overall higher in the ICU-ECMO group (ß -4,275.38 [-6,845.21 - 1,705.55], p = 0.001). In regard to neutrophil percentage, a significant decrease over time was reported (ß -1.76 [-3.16 - -0.36], p = 0.014), namely in the ICU-ECMO group (ß for the interaction 2.09 [0.45 - 3.73], p = 0.013). CONCLUSIONS: Herein, we found ECMO support seems to provide a less aggressive immune response in COVID-19 patients with severe and refractory respiratory dysfunction.

Nanocarried antioxidants in freezing extenders for boar spermatozoa.

Post-thawing cryoinjuries in boar spermatozoa due to oxidative stress may be reduced by adding nanoencapsulated antioxidants to freezing extenders. This study evaluated post-thawing kinetics, structural and biochemical functions of boar spermatozoa frozen with extenders including resveratrol and vitamin E loaded into polymeric nanocapsules. Resveratrol was added at 0 (control), 5, 10, 20, 40 and 80 µg/ml, whereas Vitamin E was added at 0 (control), 50, 100, 200 and 400 µg/ml. Both antioxidants were tested in free and nanoencapsulated presentations. In contact with empty nanocapsules, some sperm kinetics parameters were impaired compared to the control (p < .05), whereas lipoperoxidation declined (p < .05). With inclusion of 40 µg/ml nanoencapsulated resveratrol, some sperm kinetics parameters were improved (p < .01), but sperm motility, structural and biochemical functions did not differ from the control (p > .05). No improvement in sperm quality occurred with inclusion of vitamin E, although sperm kinetics with 400 µg/ml nanoencapsulated vitamin E was reduced compared to the control (p < .01). Inclusion of 40 µg/ml nanoencapsulated resveratrol benefitted boar sperm kinetics after thawing, but no improvement resulted from inclusion of vitamin E.

Validity and reliability of a shorter version of the Geriatric Depression Scale in institutionalized older Portuguese adults.

OBJECTIVES: Depressive symptoms are common in older adults in institutional contexts; however, there is a lack of validated measures for these settings. Identifying depressive symptoms can help clinicians to manage them and to prevent or delay their complications. This study aimed to validate the Geriatric Depression Scale (GDS) in an institutionalized sample of older adults. METHOD: 493 institutionalized older people (73% women) aged 60 or over were evaluated through the GDS, the Mini International Neuropsychiatric Interview (MINI) (depression vs. no depression = 11% vs. 89%), the Geriatric Anxiety Inventory (GAI), the Positive Affect (PA) and Negative Affect (NA) Schedule, and the Satisfaction with Life Scale (SWLS). Test-retest reliability was assessed with 57 older adults. RESULTS: An 8-item version presented a Cronbach's alpha value of .87 with a single factor explaining its variance. The correlations (p < .01) attested the concurrent validity (GAI: r = .76; PA: r = -.22; AN: r = .62; SWLS: r = -.32). Test-retest reliability (6.51 months) was adequate (r = .52). ROC analysis (AUC = .82; sensitivity = 80%; specificity = 77%) and Youden index revealed a cutoff of 5/6 for the diagnosis of depression. CONCLUSION: Results support the validity and the screening capacity of a short version of GDS in institutional contexts. Short screening instruments for depressive symptoms may facilitate their identification, allowing for timely clinical interventions in institutional settings.

Adverse remodeling in atrial fibrillation following isolated aortic valve replacement surgery.

BACKGROUND: Atrial fibrillation (AF) is the most common arrhythmia with adverse clinical outcomes. Aortic valve replacement (AVR) is one of the most frequently performed cardiac surgeries, although there is scarce evidence on arrhythmic outcomes. We aimed to evaluate AF during the first year post- isolated aortic valve replacement surgery and its clinical, analytical, and echocardiographic predictors. METHODS: Severe aortic stenosis patients with no prior atrial fibrillation submitted to isolated aortic valve replacement surgery were included in our study, of which 316 remained in sinus rhythm and 24 developed AF. We performed logistic regression searching for AF predictors and a longitudinal comparison between pre and post-operative echocardiographic data. RESULTS: Postoperative AF (POAF), diabetes, and follow-up indexed Left Atrium Diameter (iLAD) were significantly higher in the group of patients developing AF. POAF and iLAD were independent AF predictors at follow-up. No differences between groups were found regarding baseline and follow-up echocardiographic data except for indexed Left Ventricle End-diastolic Diameter (LVED), which failed to decrease after surgery in the AF group. CONCLUSIONS: POAF and iLAD independently predicted AF at 1 year following isolated AVR surgery in aortic stenosis patients with no AF history. iLVED did not decrease significantly at follow-up in AF patients, possibly reflecting adverse ventricular remodeling.

Comprehensive clinically oriented workflow for nucleotide level resolution and interpretation in prenatal diagnosis of de novo apparently balanced chromosomal translocations in their genomic landscape.

We present a comprehensive clinically oriented workflow for large-insert genome sequencing (liGS)-based nucleotide level resolution and interpretation of de novo (dn) apparently balanced chromosomal abnormalities (BCA) in prenatal diagnosis (PND). Retrospective or concomitant with conventional PND and liGS, molecular and newly developed clinically inspired bioinformatic tools (TAD-GConTool and CNV-ConTool) are applied to analyze and assess the functional and phenotypic outcome of dn structural variants (dnSVs). Retrospective analysis of four phenotype-associated dnSVs identified during conventional PND precisely reveal the genomic elements disrupted by the translocation breakpoints. Identification of autosomal dominant disease due to the disruption of ANKS1B and WDR26 by t(12;17)(q23.1;q21.33)dn and t(1;3)(q24.11;p25.3)dn breakpoints, respectively, substantiated the proposed workflow. We then applied this workflow to two ongoing prenatal cases with apparently balanced dnBCAs: 46,XX,t(16;17)(q24;q21.3)dn referred for increased risk on combined first trimester screening and 46,XY,t(2;19)(p13;q13.1)dn referred due to a previous trisomy 21 pregnancy. Translocation breakpoints in the t(16;17) involve ANKRD11 and WNT3 and disruption of ANKRD11 resulted in KBG syndrome confirmed in postnatal follow-up. Breakpoints in the t(2;19) are within ATP6V1B1 and the 3' UTR of CEP89, and are not interpreted to cause disease. Genotype-phenotype correlation confirms the causative role of WDR26 in the Skraban-Deardorff and 1q41q42 microdeletion phenocopy syndromes, and that disruption of ANKS1B causes ANKS1B haploinsufficiency syndrome. In sum, we show that an liGS-based approach can be realized in PND care providing additional information concerning clinical outcomes of dnBCAs in patients with such rearrangements.

Atrial matrix remodeling in atrial fibrillation patients with aortic stenosis.

BACKGROUND: This study aimed to evaluate atrium extracellular matrix remodeling in atrial fibrillation (AF) patients with severe aortic stenosis, through histological fibrosis quantification and extracellular matrix gene expression analysis, as well as serum quantification of selected protein targets. METHODS: A posthoc analysis of a prospective study was performed in a cohort of aortic stenosis patients. Between 2014 and 2019, 56 patients with severe aortic stenosis submitted to aortic valve replacement surgery in a tertiary hospital were selected. RESULTS: Fibrosis was significantly increased in the AF group when compared to sinus rhythm (SR) patients (p = 0.024). Moreover, cardiomyocyte area was significantly higher in AF patients versus SR patients (p = 0.008). Conversely, collagen III gene expression was increased in AF patients (p = 0.038). TIMP1 was less expressed in the atria of AF patients. MMP16/TIMP4 ratio was significantly decreased in AF patients (p = 0.006). TIMP1 (p = 0.004) and TIMP2 (p = 0.012) were significantly increased in the serum of AF patients. Aortic valve maximum (p = 0.0159) and mean (p = 0.031) gradients demonstrated a negative association with serum TIMP1. CONCLUSIONS: Atrial fibrillation patients with severe aortic stenosis present increased atrial fibrosis and collagen type III synthesis, with extracellular matrix remodelling demonstrated by a decrease in the MMP16/TIMP4 ratio, along with an increased serum TIMP1 and TIMP2 proteins.