The central role of pyruvate metabolism on the epigenetic maturation and transcriptional profile of bovine oocytes.

In brief: Pyruvate metabolism is one of the main metabolic pathways during oocyte maturation. This study demonstrates that pyruvate metabolism also regulates the epigenetic and molecular maturation in bovine oocytes. Abstract: Pyruvate, the final product of glycolysis, undergoes conversion into acetyl-CoA within the mitochondria of oocytes, serving as a primary fuel source for the tricarboxylic acid (TCA) cycle. The citrate generated in the TCA cycle can be transported to the cytoplasm and converted back into acetyl-CoA. This acetyl-CoA can either fuel lipid synthesis or act as a substrate for histone acetylation. This study aimed to investigate how pyruvate metabolism influences lysine 9 histone 3 acetylation (H3K9ac) dynamics and RNA transcription in bovine oocytes during in vitro maturation (IVM). Bovine cumulus-oocyte complexes were cultured in vitro for 24 h, considering three experimental groups: Control (IVM medium only), DCA (IVM supplemented with sodium dichloroacetate, a stimulant of pyruvate oxidation into acetyl-CoA), or IA (IVM supplemented with sodium iodoacetate, a glycolysis inhibitor). The results revealed significant alterations in oocyte metabolism in both treatments, promoting the utilization of lipids as an energy source. These changes during IVM affected the dynamics of H3K9ac, subsequently influencing the oocyte's transcriptional activity. In the DCA and IA groups, a total of 148 and 356 differentially expressed genes were identified, respectively, compared to the control group. These findings suggest that modifications in pyruvate metabolism trigger the activation of metabolic pathways, particularly lipid metabolism, changing acetyl-CoA availability and H3K9ac levels, ultimately impacting the mRNA content of in vitro matured bovine oocytes.

Mitochondrial health index correlates with plasma circulating cell-free mitochondrial DNA in bipolar disorder.

Although mitochondrial dysfunction is known to play an essential role in the pathophysiology of bipolar disorder (BD), there is a glaring gap in our understanding of how mitochondrial dysfunction can modulate clinical phenotypes. An emerging paradigm suggests mitochondria play an important non-energetic role in adaptation to stress, impacting cellular resilience and acting as a source of systemic allostatic load. Known as mitochondrial allostatic load, this (phenomenon) occurs when mitochondria are unable to recalibrate and maintain cell homeostasis. This study aimed to evaluate the composite mitochondrial health index (MHI) in BD subjects and non-psychiatry controls. We will also explore whether lower MIH will be related to higher cell-free mtDNA (ccf-mtDNA) levels and poor clinical outcomes. In this study, 14 BD-I patients and 16 age- and sex-matched non-psychiatry controls were enrolled. Peripheral blood mononuclear cells (PBMCs) were used to measure the enzymatic activities of citrate synthase and complexes I, II, and IV and mtDNA copy number. Ccf-mtDNA was evaluated by qPCR in plasma. Mitochondrial quality control (MQC) proteins were evaluated by western blotting. After adjusting for confounding variables, such as age, sex, body mass index (BMI), and smoking status, patients with BD presented lower MHI compared to non-psychiatry controls, as well as higher ccf-mtDNA levels that negatively correlated with MHI. Because the MQC network is essential to maintain mitochondrial health, MHI and ccf-mtDNA were also examined in relation to several MQC-related proteins, such as Fis-1, Opa-1, and LC3. Our results showed that MHI correlated negatively with Fis-1 and positively with Opa-1 and LC3. Accordingly, ccf-mtDNA had a positive correlation with Fis-1 and a negative correlation with Opa-1 and LC3. Furthermore, we found a noteworthy inverse correlation between illness severity and MHI, with lower MHI and higher ccf-mtDNA levels in subjects with a longer illness duration, worse functional status, and higher depressive symptoms. Our findings indicate that mitochondrial allostatic load contributes to BD, suggesting mitochondria represent a potential biological intersection point that could contribute to impaired cellular resilience and increased vulnerability to stress and mood episodes. Ultimately, by linking mitochondrial dysfunction to disease progression and poor outcomes, we might be able to build a predictive marker that explains how mitochondrial function and its regulation contribute to BD development and that may eventually serve as a treatment guide for both old and new therapeutic targets.

Dynamics of transcription is affected by oxygen tension and developmental speed during in vitro production of bovine embryos.

This study examines the impact of oxygen tension and embryo kinetics on gene transcription dynamics in pathways crucial for embryonic preimplantation development, including lipid metabolism, carbohydrate transport and metabolism, mitochondrial function, stress response, apoptosis and transcription regulation. Bovine embryos were generated in vitro and allocated into two groups based on oxygen tension (20% or 5%) at 18 h post insemination (hpi). At 40 hpi, embryos were categorized into Fast (≥4 cells) or Slow (2 cells) groups, resulting in four experimental groups: FCL20, FCL5, SCL20 and SCL5. Embryo collection also occurred at 72 hpi (16-cell stage; groups FMO20, FMO5, SMO20 and SMO5) and at 168 hpi (expanded blastocyst (BL) stage; groups FBL20, FBL5, SBL20 and SBL5). Pools of three embryos per group were analysed in four replicates using inventoried TaqMan assays specific for Bos taurus, targeting 93 genes. Gene expression patterns were analysed using the K-means algorithm, revealing three main clusters: genes with low relative abundance at the cleavage (CL) and 16-cell morula (MO) stages but increased at the BL stage (cluster 1); genes with higher abundances at CL but decreasing at MO and BL (cluster 2); and genes with low levels at CL, higher levels at MO and decreased levels at BL (cluster 3). Within each cluster, genes related to epigenetic mechanisms, cell differentiation events and glucose metabolism were particularly influenced by differences in developmental kinetics and oxygen tension. Fast-developing embryos, particularly those cultured under low oxygen tension, exhibited transcript dynamics more closely resembling that reported in vivo-produced embryos.

Whole-genome DNA methylation analysis of the sperm in relation to bull fertility.

In brief: Bull fertility is an important economic trait, this study identified some DNA methylation biomarkers that are associated with bull fertility. Abstract: Subfertile bulls may cause huge economic losses in dairy production since their semen could be used to inseminate thousands of cows by artificial insemination. This study adopted whole-genome enzymatic methyl sequencing and aimed to identify candidate DNA methylation markers in bovine sperm that correlate with bull fertility. Twelve bulls were selected (high bull fertility = 6; low bull fertility = 6) based on the industry's internally used Bull Fertility Index. After sequencing, a total of 450 CpG had a DNA methylation difference higher than 20% (q < 0.01) had been screened. The 16 most significant differentially methylated regions (DMRs) were identified using a 10% methylation difference cut-off (q < 5.88 × 10-16). Interestingly, most of the differentially methylated cytosines (DMCs) and DMRs were distributed on the X and Y chromosomes, demonstrating that the sex chromosomes play essential roles in bull fertility. Additionally, the functional classification showed that the beta-defensin family, zinc finger protein family, and olfactory and taste receptors could be clustered. Moreover, the enriched G protein-coupled receptors such as neurotransmitter receptors, taste receptors, olfactory receptors, and ion channels indicated that the acrosome reaction and capacitation processes are pivotal for bull fertility. In conclusion, this study identified the sperm-derived bull fertility-associated DMRs and DMCs at the whole genome level, which could complement and integrate into the existing genetic evaluation methods, increasing our decisive capacity to select good bulls and explain bull fertility better in the future.

DOHaD: A Menagerie of Adaptations and Perspectives: The interplay between early embryo metabolism and mitoepigenetic programming of development.

In brief: This review discusses advances in the knowledge of epigenetic mechanisms regulating mitochondrial DNA and the relationship with reproductive biology. Abstract: Initially perceived simply as an ATP producer, mitochondria also participate in a wide range of other cellular functions. Mitochondrial communication with the nucleus, as well as signaling to other cellular compartments, is critical to cell homeostasis. Therefore, during early mammalian development, mitochondrial function is reported as a key element for survival. Any mitochondrial dysfunction may reflect in poor oocyte quality and may impair embryo development with possible long-lasting consequences to cell functions and the overall embryo phenotype. Growing evidence suggests that the availability of metabolic modulators can alter the landscape of epigenetic modifications in the nuclear genome providing an important layer for the regulation of nuclear-encoded gene expression. However, whether mitochondria could also be subjected to such similar epigenetic alterations and the mechanisms involved remain largely obscure and controversial. Mitochondrial epigenetics, also known as 'mitoepigenetics' is an intriguing regulatory mechanism in mitochondrial DNA (mtDNA)-encoded gene expression. In this review, we summarized the recent advances in mitoepigenetics, with a special focus on mtDNA methylation in reproductive biology and preimplantation development. A better comprehension of the regulatory role of mitoepigenetics will help the understanding of mitochondrial dysfunction and provide novel strategies for in vitro production systems and assisted reproduction technologies, as well as prevent metabolic related stress and diseases.

Electrochemical detection of mephedrone using a graphene screen-printed electrode: a new sensitive and selective approach to identify synthetic cathinones in forensic analysis.

Mephedrone (MEP) is an illicit stimulant drug that belongs to the synthetic cathinone (SC) class, which has been widely used for recreational purposes and reported in forensic analysis. The preliminary identification of MEP and other SCs in seized samples is of great interest for forensic investigation and a fast and simple screening test for these drugs would be useful for on-site and in-house analyses. In this study, we present the electrochemical detection of MEP in forensic samples using, for the first time, independent redox processes of SCs on a graphene screen-printed electrode (SPE-GP). The proposed method for MEP detection on the SPE-GP was optimized in Britton-Robinson buffer solution (0.1 mol L-1) at pH 10.0 with adsorptive stripping differential pulse voltammetry (AdSDPV). The use of the SPE-GP with AdSDPV provides a wide linear range for MEP determination (2.6 to 112 µmol L-1) with a low limit of detection (LOD) (0.3 µmol L-1). The real surface area available for adsorption on the SPE-GP was estimated to be between 3.80 and 5.70 cm2, which provided high sensitivity for the proposed method. Furthermore, good stability of MEP electrochemical responses on the SPE-GP was obtained using the same or different electrodes (N = 3), with relative standard deviation (RSD) < 5.0% for both redox processes. Interference studies for a common adulterant (caffeine) and twelve other illicit drugs (phenethylamines, amphetamines, and other SCs) were performed with a highly selective response for MEP detection. Therefore, the SPE-GP with AdSDPV is demonstrated to be a selective and sensitive screening method to detect MEP and other SCs in forensic analysis, providing a fast and simple preliminary identification of these drugs in seized samples.

Microbiological analysis of oral biofilms in patients admitted to intensive care unit: Pilot study.

AIM: To identify Pseudomonas aeruginosa and Staphylococcus aureus in oral biofilms of intubated and non-intubated patients admitted to an Intensive Care Unit (ICU). METHODS: This was a cross-sectional study, with 30 biofilm sites sampled. S. aureus and P. aeruginosa were identified by conventional biochemical assays. Antimicrobial susceptibility was tested by disk-diffusion. RESULTS: Of 30 sites, 50% contained P. aeruginosa and 3.33% S. aureus. P. aeruginosa was detected in similar amounts in all 3 sample sites, with 5 colonized sites (50%). S. aureus colonized a single supragingival site (3.33%). There was resistance to multiple antimicrobial agents of P. aeruginosa in 7 sites (100%) and S. aureus in 1 (100%). CONCLUSIONS: This study revealed an important relationship between P. aeruginosa and S. aureus colonization at supragingival, subgingival and lingual sites and intubation, thus revealing antimicrobial resistant bacteria colonization of medical interest, which may contribute to the therapy choice directed to these microorganisms.

Assessment of River-Aquifer Interaction and Nitrogen Contamination in an Agricultural Zone Located in the Guarani Aquifer System Outcrop Area.

The excessive use of nitrogen fertilizers is responsible for an increase in nitrate concentrations in water bodies, which in the future could led to an irreversible contamination compromising the water resource quality. In this way, understand the water movement within a watershed and evaluate the impacts related to agricultural practices is relevant for water management, especially in an environmental fragile region, such as the outcrop area of the Guarani Aquifer. Water samples from a small watershed located at the Guarani Aquifer region in São Paulo state, representing surface water and groundwater discharge in riverbeds from two creeks, as well as groundwater (springs and wells) were collected for isotopic ratios (δ18O e δ2H) and nitrate determination. The results indicated that the river flow is mostly supplied by groundwater discharge, and despite the observed concentrations of nitrate in groundwater reaching the creeks, the current scenario indicate contamination in the surface water, above the regulatory levels. Therefore, the expansion in sugarcane production increases the possibility that the released nitrate reaches high levels in the future in this watershed.

Lipid availability during in vitro maturation alters oocyte lipid content and blastocyst development and metabolism.

Lipids play a crucial role in various biological functions, including membrane composition, energy storage, cell signalling, and metabolic and epigenetic processes. Abnormal lipid accumulation and metabolism during in vitro maturation (IVM) of oocytes have been linked to the use of fetal bovine serum (FBS), even though it provides several beneficial molecules, contributing to the oocyte competence. Delipidating agents have been used to mitigate these deleterious effects, but they can have adverse effects on embryonic development. In this study, we explored how lipids present in fetal bovine serum (FBS) can impact the composition of oocytes and their resulting blastocysts in vitro. For that, we used organic solvents to separate the polar and nonpolar (lipid enriched) phase of FBS. Oocytes were in vitro matured in the presence of 10% whole FBS (control), 10% FBS plus 10% nonpolar lipids (lipid enriched - OL) or 10% polar lipids only (partially delipidated - ODL). After 24 h, part of the matured oocytes was collected and those remaining in each group underwent in vitro fertilization (IVF) and culture (IVC) under the same conditions and expanded blastocysts were collected at day 7 (control, BL and BDL). Oocytes and embryos were analysed by Multiple Reaction Monitoring mass spectrometry (MRM-MS) to determine their lipid composition. Interestingly, principal component analysis (PCA) revealed a clear distinction in the lipid profile of oocytes and blastocysts from both treatments compared to the control group. Control oocytes and blastocysts had higher triacylglycerol and cholesterol ester enrichment while the OL, ODL, BL and BDL groups had higher amounts of free fatty acids (FFAs). The structural and signalling phospholipids also differed among groups. Our findings suggest that the lipid-enriched fraction of FBS can be manipulated for IVM to ensure proper maturation, resulting in oocytes and blastocysts with less accumulated intracellular lipids and an improved metabolic status.

Metabolic signature of spent culture media shows lipid metabolism as a determinant of pregnancy outcomes.

In the present study, we investigated the spent culture media of in vitro produced (IVP) bovine embryos which did (group Pregnant) or did not (group Non-pregnant) establish pregnancy after transfer. For that purpose, IVP embryos on D5 were transferred to individual droplets for the last 48 h of culture. Embryos at the blastocyst stage were then transferred to synchronized recipients, while respective culture media drops were collected and evaluated individually. The list of metabolites present in spent culture media was obtained by electrospray ionization mass spectrometry (ESI-MS) and analysed with Metaboanalyst® to characterize the metabolic profile of each group. The spectrometric analysis showed that pathways related to lipid metabolism, particularly fatty acids degradation via beta-oxidation, were more present in the Pregnant group whereas no significant pathway was identified in the group Non-pregnant. By using this method, we were able to identify a metabolic signature in culture media that allows for a better comprehension of preferential metabolic routes taken by the most viable embryos. These findings offer great insights into the biochemistry of embryo development and reveal a potential target for the development of better-quality IVP systems, as well as tools to identify bovine embryos with greater chances to establish and maintain pregnancy.

Early Intervention Involving Specific Task-Environment-Participation (STEP) Protocol for Infants at Risk: A Feasibility Study.

Aims: To verify the feasibility and preliminary effects of the STEP protocol, an intervention based on specific motor skills, environmental factors and participation, in infants at biological risk.Methods: Twenty-eight at-risk infants (STEP Protocol = 14; Standard Intervention = 14), aged 3-9 months and at risk for developmental delay. The following outcomes were assessed: motor skills (Alberta Infant Motor Scale-AIMS); frequency and involvement of participation (Young Children's Participation and Environment Measure-YC-PEM), and home environment opportunities (Affordances in the Home Environment for Motor Development-AHEMD-IS). For both groups, interventions were provided by parents. The intervention for group was based on the following principles: (1) standard intervention: stimulation of motor skills; (2) STEP: stimulation of motor skills, participation, mother-infant interaction, environmental enrichment. A mean comparison test was applied to verify difference between groups after the intervention.Results: The protocol showed good retention and recruitment rates. The STEP group had significantly higher outcomes after intervention on the AIMS (p = 0.014); frequency (p = 0.02) and engagement (p = 0.03) in participation, when compared to standard intervention.Conclusions: The results showed that the STEP protocol is feasible, and presents better results compared to the standard intervention, which reinforces the importance of promoting participation, specific motor skills and family involvement.

Metabolism-epigenetic interactions on in vitro produced embryos.

Metabolism and epigenetics, which reciprocally regulate each other in different cell types, are fundamental aspects of cellular adaptation to the environment. Evidence in cancer and stem cells has shown that the metabolic status modifies the epigenome while epigenetic mechanisms regulate the expression of genes involved in metabolic processes, thereby altering the metabolome. This crosstalk occurs as many metabolites serve as substrates or cofactors of chromatin-modifying enzymes. If we consider the intense metabolic dynamic and the epigenetic remodelling of the embryo, the comprehension of these regulatory networks will be important not only for understanding early embryonic development, but also to determine in vitro culture conditions that support embryo development and may insert positive regulatory marks that may persist until adult life. In this review, we focus on how metabolism may affect epigenetic reprogramming of the early stages of development, in particular acetylation and methylation of histone and DNA. We also present other metabolic modifications in bovine embryos, such as lactylation, highlighting the promising epigenetic and metabolic targets to improve conditions for in vitro embryo development.

Tele-care intervention performed by parents involving specific task- environment- participation (STEP protocol) for infants at risk for developmental delay: protocol of randomized controlled clinical trial.

BACKGROUND: With the implementation of social distancing due to the Covid-19 pandemic, many at-risk infants are without therapy. An alternative mode of therapy in this situation is tele-care, a therapy in which assessments and interventions are carried out online, in the home environment. We describe a tele-care protocol involving parent delivered task and context specific movement training, participation and environmental adaptation for infants at risk for developmental delay. METHODS: Randomized controlled trial. Infants at risk, with 3 to 9 months corrected age, will be included, and randomized into two groups: control group (conventional guidelines) and experimental group (task, environment and participation in context-specific home program). Infants will be assessed for motor capacity (Infant Motor Profile and Alberta Infant Motor Scale); participation (Young Children's Participation and Environment Measure) and environment factors (Parent-Child Early Relational Assessment; Affordances in the Home Environment for Motor Development). The intervention period will be 10 weeks, and evaluations will be carried out before and after that period. All the assessment and intervention procedures will be carried out online, with instructions to parents for home therapy. The statistical analysis will be guided according to the distribution of the data, and a significance level of 5% will be adopted. All ethical approvals were obtained by the Ethics Committee of the University of São Carlos (Case number 31256620.5.0000.5504). The protocol will follow the SPIRIT statement. Findings will be disseminated in peer-reviewed publications and presented at national and international conferences. DISCUSSION: The results of this study will describe the effectiveness of a home intervention, focusing on specific activities, participation and environmental changes. These results will support the implementation of a remote protocol, with lower financial costs and focused on the particularities of the family. This type of care model can possibly help public policies to ensure equal access to evidence-based quality healthcare. TRIAL REGISTRATION: Brazilian Clinical Trials Registry: RBR8xrzjs , registered September 1, 2020.

Peptide-coating 2D and small chemical molecules prolong the passage of porcine spermatogonia stem cells.

Porcine spermatogonia stem cells (pSSCs) are the only type of somatic stem cell that can pass genetic information to the successive generations. Little is known about pSSCs vitality in vitro, and due to their increasing importance in stem cell research, here, we optimized a protocol to culture pSSCs and explored their potential fate in vitro. Utilizing a feeder-free culture system with a 2D peptide-coating and small chemical molecules (including CHIR99021, Repsox, vitamin C, folic acid, and CD lipid concentrate), we were able to prolong the culture time of pSSCs by at least three months compared with previous methods. Moreover, we found that pSSCs could proliferate and self-renew in the seminiferous tubules of infertile mice. However, they could not perform meiosis. Our study shows that this feeder-free culture system optimizes cell culture and may facilitate advanced research on SSC biology and genetic manipulation of germ cells.

Impact of the COVID-19 Pandemic on the Time of Utilitarian Walking and Walking as Exercise Among Brazilian Community-Dwelling Older Adults: The REMOBILIZE Study.

This study aimed to investigate behavioral changes related to utilitarian walking and walking as exercise among Brazilian older adults before and during the COVID-19 pandemic. Baseline data from the longitudinal REMOBILIZE study were used. The survey was online and comprised 1,482 adults aged 60 years and older. The outcome was the hours per week of walking time (for utilitarian purposes, as exercise, and total), estimated by the frequency and duration and analyzed by average and rate of decrease, comparing pre- and during the pandemic. Total walking time was used to estimate transition patterns between levels of walking intensity. A moderate reduction of 1.76 hr per week in the total walking time was observed, and 28.1% of the participants' walking transition patterns were from more to less physically active. Our findings demonstrate the need for gender-specific interventions and policies to increase the walking levels among older adults after the COVID-19 pandemic period.

Reconstructive dosimetry and radiation dose evaluation of workers and public due to a Brazilian radiological accident in industrial radiography.

Radiological accidents occur mainly in the practices recognized as high risk and which are classified by the International Atomic Energy Agency (IAEA) as Categories 1 and 2: radiotherapy, industrial irradiators and industrial radiography. In Brazil, five important cases in industrial gamma radiography occurred from 1985 to 2018, involving seven radiation workers and 19 members of the public. The accidents caused localized radiation lesions on the hands and fingers. One of these accidents is the focus of this paper. In this accident, a 3.28 TBq192Ir radioactive source was left unshielded for 9 h in a non-destructive testing (NDT) company parking lot, and many radiation workers, employees and public, including teachers of a primary school were exposed. The radioactive source was also directly handled by a security worker for about 1.5 min causing severe radiation injuries in the hand and fingers. This paper presents radiation dose estimates for all accidentally exposed individuals. Four scenarios were considered, and three internationally recognised and updated reconstructive dosimetry techniques were used, named, Brazilian visual Monte Carlo Dose Calculation (VMC), virtual environment for radiological and nuclear accidents simulation (AVSAR) and RADPRO Calculator®. The main radiation doses estimated by VMC were the absorbed dose of 34 Gy for the security worker's finger and his effective dose of 91 mSv; effective doses from 43 to 160 mSv for radiation workers and NDT employees; and effective doses of 9 mSv for teachers in the schoolyard.

Contrasting migratory journeys and changes in hippocampal astrocyte morphology in shorebirds.

Semipalmated sandpiper (Calidris pusilla) migration to the Southern Hemisphere includes a 5-day non-stop flight over the Atlantic Ocean, whereas semipalmated plover (Charadrius semipalmatus) migration, to the same area, is largely over land, with stopovers for feeding and rest. We compared the number and 3D morphology of hippocampal astrocytes of Ch. semipalmatus before and after autumnal migration with those of C. pusilla to test the hypothesis that the contrasting migratory flights of these species could differentially shape hippocampal astrocyte number and morphology. We captured individuals from both species in the Bay of Fundy (Canada) and in the coastal region of Bragança (Brazil) and processed their brains for selective GFAP immunolabeling of astrocytes. Hierarchical cluster analysis of astrocyte morphological features distinguished two families of morphological phenotypes, named type I and type II, which were differentially affected after migratory flights. Stereological counts of hippocampal astrocytes demonstrated that the number of astrocytes decreased significantly in C. pusilla, but did not change in Ch. semipalmatus. In addition, C. pusilla and Ch. semipalmatus hippocampal astrocyte morphological features were differentially affected after autumnal migration. We evaluated whether astrocyte morphometric variables were influenced by phylogenetic differences between C. pusilla and Ch. semipalmatus, using phylogenetically independent contrast approach, and phylogenetic trees generated by nuclear and mitochondrial markers. Our findings suggest that phylogenetic differences do not explain the results and that contrasting long-distance migratory flights shape plasticity of type I and type II astrocytes in different ways, which may imply distinct physiological roles for these cells.

Biochemical markers for pregnancy in the spent culture medium of in vitro produced bovine embryos†.

The present study aimed to identify biomarkers to assess the quality of in vitro produced (IVP) bovine embryos in the culture media. IVP embryos on Day (D) 5 of development were transferred to individual drops, where they were maintained for the last 48 h of culture. Thereafter, the medium was collected and the embryos were transferred to the recipients. After pregnancy diagnosis, the media were grouped into the pregnant and nonpregnant groups. The metabolic profiles of the media were analyzed via electrospray ionization mass spectrometry, and the concentrations of pyruvate, lactate, and glutamate were assessed using fluorimetry. The spectrometric profile revealed that the media from embryos from the pregnant group presented a higher signal intensity compared to that of the nonpregnant group; the ions 156.13 Da [M + H]+, 444.33 Da [M + H]+, and 305.97 Da [M + H]+ were identified as biomarkers. Spent culture medium from expanded blastocysts (Bx) that established pregnancy had a greater concentration of pyruvate (p = 0.0174) and lesser concentration of lactate (p = 0.042) than spent culture medium from Bx that did not establish pregnancy. Moreover, pyruvate in the culture media of Bx can predict pregnancy with 90.9% sensitivity and 75% specificity. In conclusion, we identified markers in the culture media that helped in assessing the most viable IVP embryos with a greater potential to establish pregnancy.

Metabolism-driven post-translational modifications of H3K9 in early bovine embryos.

Metabolic and molecular profiles were reported as different for bovine embryos with distinct kinetics during the first cleavages. In this study, we used this same developmental model (fast vs slow) to determine if the relationship between metabolism and developmental kinetics affects the levels of acetylation or tri-methylation at histone H3 lysine 9 (H3K9ac and H3K9me3, respectively). Fast and slow developing embryos presented different levels of H3K9ac and H3K9me3 from the earliest stages of development (40 and 96 hpi) and up to the blastocyst stage. For H3K9me3, both groups of embryos presented a wave of demethylation and de novo methylation, although it was more pronounced in fast than slow embryos, resulting in blastocysts with higher levels of this mark. The H3K9ac reprogramming profile was distinct between kinetics groups. While slow embryos presented a wave of deacetylation, followed by an increase in this mark at the blastocyst stage, fast embryos reduced this mark throughout all the developmental stages studied. H3K9me3 differences corresponded to writer and eraser transcript levels, while H3K9ac patterns were explained by metabolism-related gene expression. To verify if metabolic differences could alter levels of H3K9ac, embryos were cultured with sodium-iodoacetate (IA) or dichloroacetate (DCA) to disrupt the glycolytic pathway or increase acetyl-CoA production, respectively. IA reduced H3K9ac while DCA increased H3K9ac in blastocysts. Concluding, H3K9me3 and H3K9ac patterns differ between embryos with different kinetics, the second one explained by metabolic pathways involved in acetyl-CoA production. So far, this is the first study demonstrating a relationship between metabolic differences and histone post-translational modifications in bovine embryos.

Anosognosia for memory in dementia with Lewy bodies compared with Alzheimer's disease.

OBJECTIVES: Anosognosia is the inability to recognize one's own symptoms. Although dementia with Lewy bodies (DLB) is the second most common degenerative dementia, there is little evidence of memory deficit awareness in this condition. The objectives of this research were to compare anosognosia between individuals with DLB and dementia due to Alzheimer's disease (AD) and to evaluate whether medial temporal atrophy, a marker of AD pathology, could help to explain different rates of anosognosia in DLB and dementia due to AD. METHODS/DESIGN: This is a cross-sectional study that took place at the Memory Clinic of D'Or Institute for Research and Education (IDOR). Twenty individuals with DLB and 20 with dementia due to AD were included in this study. We assessed anosognosia for memory using an index derived from subjective memory complaints (using the Memory Complaint Questionnaire) and from the performance in memory neuropsychological testing (Rey Auditory Verbal Learning Test). Thirty-one participants also underwent brain Magnetic Resonance Imaging to evaluate hippocampal atrophy with a visual scale (MTA-score [medial temporal atrophy score]). RESULTS: There was no significant difference between groups regarding age, years of education, sex or time of disease. Individuals with DLB had a higher index of anosognosia than dementia due to AD (2.92 and 1.87; p = 0.024), meaning worse awareness of memory deficits. MTA-score was slightly higher in dementia due to AD than in DLB, albeit without statistical significance. CONCLUSION: Our study was the first to demonstrate that anosognosia for memory is worse in DLB than in dementia due to AD. This finding supports the hypothesis that anosognosia in DLB is a heterogeneous phenomenon.