Post-traumatic stress disorder and associated factors among adult war survivors in Northwest Ethiopia: Community-based, cross-sectional study.

Background: A person may endure or witness a traumatic incident, such as being exposed to war, and, as a result, develop post-traumatic stress disorder (PTSD). There is a lack of information about post-traumatic stress disorder in low and middle-income countries such as Ethiopia. However, armed conflict, abuse of human rights, and violence motivated by race are becoming more commonplace. This study aimed to assess the prevalence of PTSD and associated factors among war survivors in Nefas Meewcha Town, South Gondar Zone, Ethiopia, 2022. Methods: A community based cross-sectional study was carried out. 812 study participants were chosen using a multi-stage sampling process. A face-to-face interview used a post-traumatic stress disorder checklist (PCL-5) to evaluate PTSD. The association between PTSD and other demographic and psychosocial characteristics was investigated using bivariate and multivariable binary logistic regression analysis. A P-value of 0.05 was declared as statistical significance. Result: The prevalence of PTSD in this study was 40.8% with a 95% CI of 36.2 to 46.7. The likelihood of developing PTSD was significantly associated with the fallowing factors. A close family member killed or seriously injured (AOR = 4.53, 95% CI = 3.25-6.46), being female (AOR = 1.98, 95% CI = 1.3-3.0), moderate (AOR = 3.51, 95% CI = 2.52-4.68) and high perceived stress (AOR = 5.23, 95% CI = 3.47-8.26), depression symptoms (AOR = 4.92, 95% CI = 3.57-6.86), anxiety disorder symptoms (AOR = 5.24, 95% CI = 3.72-7.63), a chronic medical illness (AOR = 3.51, 95% CI = 2.52-5.41), physical assault (AOR = 2.12, 95% CI = 1.05-3.72) and being in a war fighting situation (AOR = 1.41, 95% CI = 1.21-3.14). Conclusion: This study reported that the prevalence of PTSD was high. Being female, having a previous history of chronic medical illness, depressive symptoms, anxiety symptoms, history of a family member or friend was injured or killed, poor social support, high perceived stress, physical assault, and being in a war fighting situation were statistically associated with PTSD. Hence, regular patient assessment by mental health organizations for those with a history of trauma and facilitation of ways to support such residents is highly recommended.

Hand-Washing at Critical Times and Associated Factors Among Mothers/Caregivers of Under-Five Year Children in Nefas Silk Lafto Sub-City, Addis Ababa, Ethiopia.

Background: Hand washing is the simplest, most affordable, and most effective means of limiting the spread of infections. Despite increasing efforts to improve hand washing at critical times (after defecation, after handling child/adult feces or cleaning child's bottom, after cleaning the environment, before preparing food, and before eating food), mothers/caregivers of under-five children fail to conduct it; but the reason appears unclear. Thus, this study sought to identify hand washing at critical times and associated factors among mothers/caregivers of under-five children in Nefas Silk Lafto Sub-City, Addis Ababa, Ethiopia. Methods: A facility-based cross-sectional study was conducted on April 1-15, 2019, and 312 mothers/caregivers participated. A pretested questionnaire was used to collect data from participants by interviewer-administered technique and the data were analyzed with the Statistical Package for Social Science version 20. The factors were determined by conducting logistic regression and the crude odds ratio (COR) and adjusted odds ratio (AOR) with their respective 95% confidence intervals. All statistical tests were conducted at a 5% level of significance. Results: The study revealed that 232 (74.4%; 95% CI [69.6%-79.2%]) mothers/caregivers washed their hands at critical times. The illiterate mothers/caregivers and mothers/caregivers who lacked tap water inside the home or the backyard had 66% (AOR = 0.34; 95%CI [0.17-0.69]) and 62% (AOR = 0.38; 95%CI [0.18-0.80]) reduced odds of washing hands at critical times, respectively. Mothers/caregivers from middle had (AOR = 4.56; 95%CI [1.84-11.33]), richer had (AOR = 5.61; 95%CI [2.11-15.30]), and the richest had (AOR = 6.14; 95%CI [2.24-16.72]) times increased likelihood of washing hands at critical times than the poorest. Conclusion: The majority of mothers/caregivers practiced hand washing at critical times, and improving maternal literacy, household economy, and availability of water sources in the backyard are needed to maintain and enhance the practice.

A Prospective Cohort Study on the Periparturient Muscle Tissue Mobilisation in High Producing Dairy Cows.

Excessive periparturient fat mobilisation and its association with dairy cattle health and fertility is well documented; however, the role of muscle mobilisation has not been studied extensively. The objectives of this study were to (i) investigate the changes in the thickness of the longissimus dorsi muscle in high producing dairy cows during the periparturient period, (ii) identify factors associated with these changes, and (iii) describe their possible associations with cattle reproductive performance. Data were collected from a total of 500 lactations from 455 cows on three different UK farms. Muscle thickness (MT) (Longissimus dorsi) and back fat thickness (BFT) measurements were collected at three different time-points during the periparturient period using ultrasonography. Body condition score (BCS) was also assessed at the same time-points and blood samples were collected for the measurement of non-esterified fatty acids. Farm fertility records were used and genomically estimated breeding values were also available. Associations between variables were analysed with the use of multivariable linear and logistic regression models; Cox proportional hazard analysis was used for fertility outcomes. Muscle thickness decreased pre- to post-calving on all three farms, though they were notable between farm differences. Those animals with a lower BCS pre-calving had a higher MT loss; significant fat mobilisation occurred between the calving and early lactation period. Muscle thickness changes and fat mobilisation were not associated in this study. An increased time to first service was described for those animals that mobilised more muscle tissue. Our study advances the understanding of periparturient muscle tissue mobilisation in dairy cattle and highlights its potential associations with cattle fertility.

Fatty Hepatocytes Induce Skeletal Muscle Atrophy In Vitro: A New 3D Platform to Study the Protective Effect of Albumin in Non-Alcoholic Fatty Liver.

The liver neutralizes endogenous and exogenous toxins and metabolites, being metabolically interconnected with many organs. Numerous clinical and experimental studies show a strong association between Non-alcoholic fatty liver disease (NAFLD) and loss of skeletal muscle mass known as sarcopenia. Liver transplantation solves the hepatic-related insufficiencies, but it is unable to revert sarcopenia. Knowing the mechanism(s) by which different organs communicate with each other is crucial to improve the drug development that still relies on the two-dimensional models. However, those models fail to mimic the pathological features of the disease. Here, both liver and skeletal muscle cells were encapsulated in gelatin methacryloyl and carboxymethylcellulose to recreate the disease's phenotype in vitro. The 3D hepatocytes were challenged with non-esterified fatty acids (NEFAs) inducing features of Non-alcoholic fatty liver (NAFL) such as lipid accumulation, metabolic activity impairment and apoptosis. The 3D skeletal muscle tissues incubated with supernatant from fatty hepatocytes displayed loss of maturation and atrophy. This study demonstrates the connection between the liver and the skeletal muscle in NAFL, narrowing down the players for potential treatments. The tool herein presented was employed as a customizable 3D in vitro platform to assess the protective effect of albumin on both hepatocytes and myotubes.

NEFAs Influence the Inflammatory and Insulin Signaling Pathways Through TLR4 in Primary Calf Hepatocytes in vitro.

Transition dairy cows are often in a state of negative energy balance because of decreased dry matter intake and increased energy requirements, initiating lipid mobilization and leading to high serum ß-hydroxybutyrate (BHBA) and non-esterified fatty acid (NEFAs) levels, which can induce ketosis and fatty liver in dairy cows. Inflammation and insulin resistance are also common diseases in the perinatal period of dairy cows. What is the relationship between negative energy balance, insulin resistance and inflammation in dairy cows? To study the role of non-esterified fatty acids in the nuclear factor kappa beta (NF-κB) inflammatory and insulin signaling pathways through Toll-like receptor 4 (TLR4), we cultured primary calf hepatocytes and added different concentrations of NEFAs to assess the mRNA and protein levels of inflammatory and insulin signaling pathways. Our experiments indicated that NEFAs could activate the NF-κB inflammatory signaling pathway and influence insulin resistance through TLR4. However, an inhibitor of TLR4 alleviated the inhibitory effects of NEFAs on the insulin pathway. In conclusion, all of these results indicate that high-dose NEFAs (2.4 mM) can activate the TLR4/NF-κB inflammatory signaling pathway and reduce the sensitivity of the insulin pathway through the TLR4/PI3K/AKT metabolic axis.

Effects of NEFAs during IVM on pig embryos from granulosa cell-cocultured oocytes.

Circulating levels of nonesterified fatty acids (NEFAs) are elevated in some females, which can impair oocyte maturation and embryo development, and may alter the phenotype of the progeny. However, the effects of NEFAs on human embryo development are not clear due to ethical limitations. Thus, we used pig as the model to investigate the impacts of NEFAs on oocyte and embryo due to their similar reproductive and metabolic physiologies to humans. In this study, porcine cumulus-oocyte complexes were in vitro maturated under a pathologically high concentration of NEFAs (468 µM palmitic acid, 194 µM stearic acid, and 534 µM oleic acid) with the presence of granulosa cell monolayer, in contrast to control without NEFAs. The mature oocytes were fertilized to produce embryos for further analysis of the transcriptome and DNA methylation patterns. The elevated level of NEFAs decreased the blastocyst rate and delayed the blastocyst development. Ingenuity pathway analysis showed that the most affected gene pathways were related mainly to cell activities, metabolism, and inflammation. These findings indicated that oocytes exposed to the exogenous high level of NEFAs during in vitro maturation resulted in altered gene expression and DNA methylation of early embryos, which have detrimental impacts on blastocyst quality.

Cocultured porcine granulosa cells respond to excess non-esterified fatty acids during in vitro maturation.

BACKGROUND: Non-esterified fatty acids (NEFAs) are one of the main lipid components of follicular fluid at concentrations that depend on circulating levels. Elevated levels of NEFAs impair oocyte quality, development potential, and may subsequently influence the metabolism and reproductive fitness of offspring. Granulosa cells (GCs) are the follicular cells that are closely communicating with the oocyte. However, the responses of GCs exposed to high levels of NEFAs when cocultured with cumulus-oocyte complexes (COCs), and how they attenuate the negative effects of NEFAs on oocytes, are unclear. RESULTS: To better understand this protective effect, monolayers of porcine GCs were cocultured with COCs during in vitro maturation (IVM) in the presence of elevated levels of NEFAs. Genomic expression analysis was conducted to explore the responses of the GCs to the elevated levels of NEFAs. After limma algorithm analysis, 1,013 genes were differentially expressed between GCs cultured with and without elevated NEFAs. Among them, 438 genes were upregulated and 575 were downregulated. The differentially expressed genes were enriched in pathways related to metabolism, inflammation, and epithelial-mesenchymal transition. CONCLUSIONS: The pathways and upstream regulators suggested that the cocultured GCs responded to the elevated NEFAs with (1) inhibition of the transition from granulosa to luteal cell, (2) interactions of metabolism change, anti-inflammation, mitochondrial function, and cell transition, (3) intercommunication with cocultured COCs of anti-inflammatory factors.

The effect of melatonin on glucose tolerance, insulin sensitivity and lipid profiles after a late evening meal in healthy young males.

The suppression of melatonin by light at night (LAN) has been associated with a disruption of SCN function and biological processes. This study aimed to explore the impact of melatonin on glucose and lipid metabolism before and after a late evening meal. Nine healthy male participants (26 ± 1.3 years, BMI 24.8 ± 0.8 kg/m2 (mean ± SD) were randomly categorised into a three-way cross-over design protocol: light (>500 lux) (LS), dark (<5 lux) + exogenous melatonin (DSC) and light (>500 lux) + exogenous melatonin (LSC). All participants were awake in a semi-recumbent position during each clinical session, which started at 18 00 h and ended at 06:00 h the following day. The meal times were individualised according to melatonin onset estimated from the participants' 48-h sequential urine collection. The administration of exogenous melatonin was conducted 90 min before the evening meal. Saliva and plasma samples were collected at specific time points to analyse the glucose, insulin, NEFAs, TAGs, cortisol and melatonin levels. Participants demonstrated a significant reduction in postprandial plasma glucose, insulin and TAGs levels in the presence of melatonin (LSC and DSC) compared to LS (p = .002, p = .02 and p = .007, respectively). Pre-prandial plasma NEFAs were significantly lower in LS than DSC and LSC as melatonin rose (p < .001). Exogenous melatonin administrated before an evening test meal improved glucose tolerance, insulin sensitivity and reduced postprandial TAGs. This study could have implications for shift workers who may have lower melatonin levels at night due to light suppression.

Supplementation of oleic acid, stearic acid, palmitic acid and ß-hydroxybutyrate increase H3K9me3 in endometrial epithelial cells of cattle cultured in vitro.

There is growing evidence that greater than homeostatic blood concentrations of nonesterified fatty acids (NEFAs) and ß-hydroxybutyrate (BHBA) have negative consequences on dairy cow's fertility, but effects on cell homeostasis in the reproductive system is not completely understood. In this study, lipids accumulation, reactive oxygen species (ROS) concentrations, abundance of gene transcripts, and immunofluorescence signal of H3K4me3 and H3K9me3 were evaluated in endometrial epithelial cells of cattle cultured with NEFAs (Oleic (OA), Stearic (SA) and Palmitic (PA) acids), BHBA, NEFAs + BHBA or each of the three NEFAs alone. The cellular lipids were in greater concentrations as a result of NEFAs + BHBA, NEFAs, SA or OA supplementation, but not by BHBA or PA. The ROS concentrations were greater when there were treatments with NEFAs + BHBA, NEFAs or BHBA. The relative mRNA abundance for genes involved in the regulation of apoptosis (XIAP), glucose transport (GLUT3), and DNA methylation (DNMT1) were greater when there were NEFAs + BHBA, but not NEFAs, BHBA, OA, SA or PA treatments. The immunofluorescence signal for H3K9me3 was greater when there were NEFAs + BHBA, NEFAs or PA, but not by BHBA, OA or SA treatments. These findings indicate that NEFAs and BHBA have an additive effect on endometrial cells of cattle by altering epigenetic markers and the expression of genes controlling important cellular pathways. Furthermore, there was cellular lipid accumulation and increased H3K9me3 in cultured bovine endometrial cells that was mainly induced by OA and PA treatments, respectively.

Curdlan intake changes gut microbial composition, short-chain fatty acid production, and bile acid transformation in mice.

Indigestible polysaccharides, such as dietary fibers, benefit the host by improving the intestinal environment. Short-chain fatty acids (SCFAs) produced by gut microbial fermentation from dietary fibers exert various physiological effects. The bacterial polysaccharide curdlan benefits the host intestinal environment, although its effect on energy metabolism and SCFA production remains unclear. Hence, this study aimed to elucidate the effect of curdlan intake on gut microbial profiles, SCFA production, and energy metabolism in a high-fat diet (HFD)-induced obese mouse model. Gut microbial composition of fecal samples from curdlan-supplemented HFD-fed mice indicated an elevated abundance of Bacteroidetes, whereas a reduced abundance of Firmicutes was noted at the phylum level compared with that in cellulose-supplemented HFD-fed mice. Moreover, curdlan supplementation resulted in an abundance of the family Bacteroidales S24-7 and Erysipelotrichaceae, and a reduction in Deferribacteres in the feces. Furthermore, curdlan supplementation elevated fecal SCFA levels, particularly butyrate. Although body weight and fat mass were not affected by curdlan supplementation in HFD-induced obese mice, HFD-induced hyperglycemia was significantly suppressed with an increase in plasma insulin and incretin GLP-1 levels. Curdlan supplementation elevated fecal bile acid and SCFA production, improved host metabolic functions by altering the gut microbial composition in mice.

Fatty acids and evolving roles of their proteins in neurological, cardiovascular disorders and cancers.

The dysregulation of fat metabolism is involved in various disorders, including neurodegenerative, cardiovascular, and cancers. The uptake of long-chain fatty acids (LCFAs) with 14 or more carbons plays a pivotal role in cellular metabolic homeostasis. Therefore, the uptake and metabolism of LCFAs must constantly be in tune with the cellular, metabolic, and structural requirements of cells. Many metabolic diseases are thought to be driven by the abnormal flow of fatty acids either from the dietary origin and/or released from adipose stores. Cellular uptake and intracellular trafficking of fatty acids are facilitated ubiquitously with unique combinations of fatty acid transport proteins and cytoplasmic fatty acid-binding proteins in every tissue. Extensive data are emerging on the defective transporters and metabolism of LCFAs and their clinical implications. Uptake and metabolism of LCFAs are crucial for the brain's functional development and cardiovascular health and maintenance. In addition, data suggest fatty acid metabolic transporter can normalize activated inflammatory response by reprogramming lipid metabolism in cancers. Here we review the current understanding of how LCFAs and their proteins contribute to the pathophysiology of three crucial diseases and the mechanisms involved in the processes.

Alteration in the Lipid Profile and the Desaturases Activity in Patients With Severe Pneumonia by SARS-CoV-2.

The kidnapping of the lipid metabolism of the host's cells by severe acute respiratory syndrome (SARS-CoV-2) allows the virus to transform the cells into optimal machines for its assembly and replication. Here we evaluated changes in the fatty acid (FA) profile and the participation of the activity of the desaturases, in plasma of patients with severe pneumonia by SARS-CoV-2. We found that SARS-CoV-2 alters the FA metabolism in the cells of the host. Changes are characterized by variations in the desaturases that lead to a decrease in total fatty acid (TFA), phospholipids (PL) and non-esterified fatty acids (NEFAs). These alterations include a decrease in palmitic and stearic acids (p ≤ 0.009) which could be used for the formation of the viral membranes and for the reparation of the host's own membrane. There is also an increase in oleic acid (OA; p = 0.001) which could modulate the inflammatory process, the cytokine release, apoptosis, necrosis, oxidative stress (OS). An increase in linoleic acid (LA) in TFA (p = 0.03) and a decreased in PL (p = 0.001) was also present. They result from damage of the internal mitochondrial membrane. The arachidonic acid (AA) percentage was elevated (p = 0.02) in the TFA and this can be participated in the inflammatory process. EPA was decreased (p = 0.001) and this may decrease of pro-resolving mediators with increase in the inflammatory process. The total of NEFAs (p = 0.03), PL (p = 0.001), cholesterol, HDL and LDL were decreased, and triglycerides were increased in plasma of the COVID-19 patients. Therefore, SARS-CoV-2 alters the FA metabolism, the changes are characterized by alterations in the desaturases that lead to variations in the TFA, PL, and NEFAs profiles. These changes may favor the replication of the virus but, at the same time, they are part of the defense system provided by the host cell metabolism in its eagerness to repair damage caused by the virus to cell membranes.

Weight-loss-independent benefits of exercise on liver steatosis and stiffness in Japanese men with NAFLD.

BACKGROUND & AIMS: A weight-loss-independent beneficial effect of exercise on non-alcoholic fatty liver disease (NAFLD) management has been reported, but the underlying mechanism is unknown. To help determine this mechanism, the effects of exercise on individual tissues (liver, adipose tissue, and skeletal muscle) were retrospectively studied. METHODS: Data from Japanese obese men with NAFLD in a 3-month exercise regimen were analysed and compared with those in a 3-month dietary restriction program designed to achieve weight loss. The underlying mechanism was studied in a smaller subcohort. RESULTS: Independent of the effect of weight loss, the exercise regimen reduced liver steatosis by 9.5% and liver stiffness by 6.8% per 1% weight loss, and resulted in a 16.4% reduction in FibroScan-AST score. Improvements in these hepatic parameters were closely associated with anthropometric changes (reduction in adipose tissue and preservation of muscle mass), increases in muscle strength (+11.6%), reductions in inflammation and oxidative stress (ferritin: -22.3% and thiobarbituric acid: -12.3%), and changes in organokine concentrations (selenoprotein-P: -11.2%, follistatin: +17.1%, adiponectin: +8.9%, and myostatin: -21.6%) during the exercise regimen. Moreover, the expression of target genes of the transcription factor Nrf2, an oxidative stress sensor, was higher in monocytes, suggesting that Nrf2 is activated. Large amounts of high-intensity exercise were effective at further reducing liver steatosis and potentiating improvements in pathophysiological parameters (liver enzyme activities and organokine profiles). CONCLUSIONS: The weight-loss-independent benefits of exercise include anti-steatotic and anti-stiffness effects in the livers of patients with NAFLD. These benefits seem to be acquired through the modification of inter-organ crosstalk, which is characterised by improvements in organokine imbalance and reductions in inflammation and oxidative stress. LAY SUMMARY: We investigated the effects of exercise on non-alcoholic fatty liver disease (NAFLD) that were not related to weight loss. We found that exercise had considerable weight-loss-independent benefits for the liver through a number of mechanisms. This suggests that exercise is important for NAFLD patients, regardless of whether they lose weight.

Trends in Changes of Automatic Milking System Biomarkers and Their Relations with Blood Biochemical Parameters in Fresh Dairy Cows.

The aim or this study was to determine the relationship between non-esterified fatty acids and biomarkers from an automatic milking system (AMS). Fresh dairy cows (n = 102) were selected and milked in Lely Astronaut® A3 milking robots. The rumination time (RT), body weight (BW), milk content and composition parameters, milk fat/protein ratio (F/P), and milk electrical conductivity were registered by the same milking robots. For examining non-esterified fatty acids (NEFAs), blood samples were acquired from cows in the dry period. According to the NEFA concentrations, all cows were divided into two groups: Group I, with <0.300 mEq/L (n = 66), and Group II, with ≥0.300 mEq/L (n = 36). Albumin (ALB), aspartate aminotransferase (AST), gamma-glutamyl transferase (GGT), and cortisol concentrations were also analyzed once a week up to 30 days in milking. The study revealed that the cows in Group I had higher concentrations of ALB, cortisol, and GGT, but the average concentration of AST was lower. In Group 1, the milk F/P was higher, but the milk yield was lower. We hypothesize that biomarkers from AMS could help in the early diagnosis of metabolic diseases after calving or to control negative energy balance before calving.

Nonesterified Fatty Acids and Kidney Function Decline in Older Adults: Findings From the Cardiovascular Health Study.

RATIONALE & OBJECTIVE: Circulating nonesterified fatty acids (NEFAs) make up a small portion of circulating lipids but are a metabolically important energy source. Excessive circulating NEFAs may contribute to lipotoxicity in many tissues, including the kidneys. We investigated the relationship between total circulating NEFA concentration and kidney outcomes in older, community-dwelling adults. STUDY DESIGN: Prospective cohort study. SETTING & PARTICIPANTS: 4,698 participants≥65 years of age in the Cardiovascular Health Study who underwent total fasting serum NEFA concentration measurements in 1992-1993. EXPOSURE: Fasting serum NEFA concentration at one time point. OUTCOME: Three primary outcomes: estimated glomerular filtration rate (eGFR) decline of≥30%, the composite of eGFR decline≥30% or kidney failure with replacement therapy, and change in eGFR. These outcomes were assessed over 4- and 13-year periods. ANALYTICAL APPROACH: Logistic regression for the dichotomous outcomes and mixed effects models for the continuous outcome, with sequential adjustment for baseline covariates. Inverse probability of attrition weighting was implemented to account for informative attrition during the follow-up periods. RESULTS: Serum NEFA concentrations were not independently associated with kidney outcomes. In unadjusted and partially adjusted analyses, the highest quartile of serum NEFA concentration (compared with lowest) was associated with a higher risk of≥30% eGFR decline at 4 years and faster rate of decline of eGFR. No associations were evident after adjustment for comorbidities, lipid levels, insulin sensitivity, medications, and vital signs: the odds ratio for the eGFR decline outcome was 1.33 (95% CI, 0.83-2.13), and the difference in eGFR slope in the highest versus lowest quartile of serum NEFA concentration was-0.15 (95% CI, -0.36 to 0.06) mL/min/1.73m2 per year. LIMITATIONS: Single NEFA measurements, no measurements of post-glucose load NEFA concentrations or individual NEFA species, no measurement of baseline urine albumin. CONCLUSIONS: A single fasting serum NEFA concentration was not independently associated with long-term adverse kidney outcomes in a cohort of older community-living adults.

Transcriptome and epigenome analysis of porcine embryos from non-esterified fatty acid-exposed oocytes.

Increasing evidence indicates that maternal malnutrition leads to decreased female fertility and dysregulated metabolic homeostasis in offspring. High levels of non-esterified fatty acids (NEFAs) in follicular fluid were reported to be involved in these maternal nutritional effects, but the mechanisms remain unclear. This study explored the mechanisms of action of abnormal NEFA levels during porcine oocyte in vitro maturation (IVM) on early embryo development (blastocysts) using phenotypic, transcriptomic, and epigenetic analysis. The oocytes were treated during IVM with, in addition to the 1% (v/v) porcine follicular fluid in the control group, a combination of 468 µmol/L palmitic acid, 194 µmol/L stearic acid, and 534 µmol/L oleic acid supplemented to North Carolina State University-23 (NCSU-23) maturation medium to achieve a high level of NEFAs. After IVM, oocytes were in vitro fertilized and then cultured in regular conditions for blastocysts. Expanded blastocysts were collected to complete transcriptomic and epigenetic analysis. Macroscopically, high level of NEFAs impaired embryo development by reducing the blastocyst rate. Analysis of the transcriptome revealed that pathways related to inflammation, apoptosis, metabolism, and oxidative stress were the most affected. Moreover, DNA methylation data demonstrated differentially methylated regions in genes related to cellular metabolism and inflammation processes. Therefore, our conclusion is that high level of NEFAs during IVM might affect porcine early embryo development by diminishing blastocyst rate and altering gene expression, especially at the metabolism and cell status levels, which could further decrease the embryo quality.

Intrafollicular injection of nonesterified fatty acids impaired dominant follicle growth in cattle.

Dairy cows frequently undergo a state of negative energy balance (NEB) after parturition and some have impaired ovarian functions that result in delayed resumption of estrous cyclicity and development of follicles without ovulation occurring. During the postpartum period, cows undergo body-fat store losses, hormonal changes, fat mobilization and increases in nonesterified fatty acid (NEFAs) concentrations in blood and follicular fluid. The effect of NEFAs on follicular development and function of follicular cells, however, is not fully understood. The aim of this study, therefore, was to study the effect of an intrafollicular injection of a mixture of oleic, stearic and palmitic NEFAs on dominant follicle development and function of granulosa cells in cows that were not in a NEB state. Follicular size was less at 24 and 48 h after administration of NEFAs compared to that of control follicles injected with vehicle only. At 24 h after intrafollicular injection, the relative mRNA transcript abundance for proteins involved in steroidogenesis (CYP19A1, 3BHSD, STAR, FSHR), metabolism (GLUT1, GLUT3, INSR, IRS1, IRS2, SLC27A1, PPARG), and cell proliferation and apoptosis (CCND2; XIAP) in granulosa cells, as well as estradiol concentrations in follicular fluid were similar in control and NEFA-treated follicles. In conclusion, the results of this study indicate increased intrafollicular concentrations of NEFAs in cows that are not in a NEB state has a detrimental effect on follicle development. We propose intrafollicular injection is a useful approach to further investigate the local effects of NEFAs on the function of follicular cells.

Non-esterified fatty acids in the ovary: friends or foes?

A majority of common metabolic diseases can result in excessive lipolysis, leading to elevated levels of non-esterified fatty acids (NEFAs) in the body fluids. In females, increased NEFA levels in the follicular fluid markedly alter the functions of intrafollicular cells such as granulosa cells (GCs) and oocytes. Therefore, elevated levels of NEFAs have been suggested to be a significant player of subfertility in females of both human and economically important animal species such as cattle, buffalo, sheep, pig, chicken, and dog. However, the effects imposed by saturated and unsaturated fatty acids (SFAs and UFAs) on ovarian follicles are controversial. The present review emphasizes that SFAs induce apoptosis in granulosa and cumulus cells of ovarian follicles in different species. They further could adversely affect oocyte maturation and developmental competence. Many types of UFAs affect steroidogenesis and proliferation processes and could be detrimental for follicular cells, especially when at elevated concentrations. Interestingly, monounsaturated fatty acids (MUFAs) appear to contribute to the etiology of the polycystic ovarian syndrome (PCOS) as they were found to induce the transcription and translation of the androgenic transcription factor SOX9 while downregulating its estrogenic counterpart FOXL2 in GCs. Overall, this review presents our revised understanding of the effects of different fatty acids on the female reproductive success, which may allow other researchers and clinicians to investigate the mechanisms for treating metabolic stress-induced female infertility.

Undernutrition-induced lipid metabolism disorder triggers oxidative stress in maternal and fetal livers using a model of pregnant sheep.

This study aimed to evaluate the oxidative status and antioxidant capacity in maternal and fetal livers upon undernutrition as well as the connection between oxidative stress and lipid metabolism disorder. Ten ewes, who were pregnant for 115 days, were restricted to a 30% level of ad libitum feed intake to develop an undernourished model, while another 10 pregnant ewes were fed normally as controls. Undernutrition induced severe lipid metabolism disorder and oxidative stress in blood, maternal liver, and fetal liver. RNA-sequencing data displayed that antioxidant capacity was changed and antioxidant genes were downregulated in maternal and fetal livers of the undernourished model. Non-esterified fatty acids (NEFAs) and beta-hydroxybutyrate (BHBA) levels showed a positive correlation with oxidative indices and negative correlation with the expression of antioxidant genes both in maternal and fetal livers. Primary hepatocytes experiments confirmed that both high levels of NEFAs and BHBA could elicit oxidative stress and decrease antioxidant capacity, and the peroxisome proliferator-activated receptor alpha (PPARA)/retinoid X receptor alpha (RXRA) signaling pathway played a vital role in enhancing antioxidant capacity and relieving oxidative stress. In conclusion, maternal undernutrition induced lipid metabolism disorder, which downregulated antioxidant genes, decreased antioxidant activity, and further triggered oxidative stress both in maternal and fetal livers. Activation of PPARA/RXRA signaling could enhance antioxidant capacity and mitigate oxidative stress. Our findings contribute to protecting the pregnant mother and her fetus from oxidative stress.

Choline and methionine regulate lipid metabolism via the AMPK signaling pathway in hepatocytes exposed to high concentrations of nonesterified fatty acids.

High concentrations of nonesterified fatty acids (NEFAs) and ß-hydroxybutyric acid (BHBA) induce lipid peroxidation, resulting in liver damage. Choline and methionine (Met) can promote energy balance and benefit liver health in transition dairy cows; however, the regulating mechanism remains unclear. In the present study, we established the hepatocyte damage model by 1.5 mM NEFAs or BHBA treatment, and examined lipid metabolism in hepatocytes. The results showed that 1.5 mM NEFAs and 1.5 mM BHBA significantly decreased the messenger RNA (mRNA) expression of AMP-activated protein kinase (AMPK)-α as well as its target genes carnitine palmitoyltransferase-1α (CPT-1α), acetyl-CoA carboxylase, fatty acid synthetase, and Apolipoprotein B100 (ApoB100). Choline and Met upregulated the phosphorylation level of AMPK-α, which was blocked by BML (an AMPK-α inhibitor). The mRNA expression level of peroxisome proliferator-activated receptor-α (PPAR-α), CPT-1α, and ApoB100 showed a similar trend. The expressions of liver X recptoer α (LXR-α) and sterol regulatory element-binding protein 1c (SREBP-1c) were decreased by choline and Met, while only the decrease of LXR-α was blocked by BML. These findings indicate that the high-level NEFAs and BHBA weaken the lipid metabolism by impairing the fatty acid oxidation, synthesis, and transport proteins. Choline and Met regulate PPAR-α and LXR-α transcriptional activity through AMPK-α phosphorylation and regulate SREBP-1c independently of AMPK-α to promote lipid oxidation and transport in NEFAs-treated hepatocytes.