Metabolic Status Research Articles

P-508 Management of incidental findings after preconception expanded carrier screening: identifying genetic predispositions to cancer and metabolic conditions in 8010 screened ART patients

Abstract Study question How do individuals and fertility clinics navigate incidental findings from Expanded Carrier Screening (ECS) in reproductive decision-making? Summary answer Incidental ECS findings prompted varied responses from patients and gamete donors, influencing diverse reproductive choices and underscoring the pivotal role of pre-test genetic counseling. What is known already ECS aims to assess reproductive risk in the general population for specific autosomal recessive and X-linked conditions. These manifest in infancy or early childhood and impact life expectancy or the quality of life. ECS results may guide interventions to avoid affected pregnancies, such as the implementation of preimplantation genetic testing of embryos (PGT-M). Additionally, ECS has the potential to uncover incidental findings with significant health implications, such as an elevated cancer risk or diagnosis. However, the lack of comprehensive reporting guidelines necessitates meticulous analysis to ensure effective clinical guidance for these individual findings. Study design, size, duration This retrospective descriptive study assessed the results of ECS tests performed in 5475 patients and 2535 oocyte and semen donors from four different fertility clinics between July 2017 and December 2023. ECS reports with incidental findings in the ATM, FH or LDLR genes were identified and further examined. We calculated individual and reproductive risks for affected individuals and recorded the subsequent clinical management pertaining to each case. Participants/materials, setting, methods Pre-test counselling was offered to both patients and gamete donors before consenting to ECS. Testing was outsourced to an external genetics laboratory using a panel of 309 genes analyzed through next generation sequencing and complementary molecular techniques. The laboratory reported pathogenic and likely pathogenic variants that were subsequently manually reviewed by an internal genetics committee. Personal and reproductive risks were assessed for each case and genetic counseling was provided in a subsequent meeting. Main results and the role of chance Genetic variants in ATM, FH, or LDLR genes were reported in 134 out of 8010 individuals undergoing ECS (1.67%). Among these, 108 variants were identified in patients: 53.7% (n = 58) in ATM, 19.4% in FH (n = 21), and 26.8% (n = 29) in LDLR. Out of the variants identified as incidental findings, 20/108 were actionable and eligible for PGT-M: 19 were associated with an increased risk for breast cancer and 1 for renal cancer. After being informed of the findings and their implications, 7/20 couples opted to revoke treatment as, according to internal policies, PGT-M was mandatory in such cases; 6/20 couples chose to undergo double gamete donation; 6/20 couples opted for PGT-M. Patients with pathogenic variants in LDLR linked to autosomal dominant inheritance for familial hypercholesterolemia (20/108, 18.5%), received follow-up guidelines and family genetic counseling. Pathogenic variants in ATM, FH, and LDLR were also identified in 26 gamete donors (24 oocyte donors and 2 sperm donors). Following internal review, 14 of these variants were considered incidental findings with an increased risk for breast cancer (13/14) or familial hypercholesterolemia (1/14), leading to the exclusion of these potential donors from the donation program. Limitations, reasons for caution The reproductive handling of incidental findings in our population relies on subjective decisions by the internal genetics committee, which may evolve with scientific advancements and recommendations from professional societies. Follow-up data on long-term reproductive outcomes are lacking. Wider implications of the findings This study reveals the diverse reproductive decisions made in response to incidental findings, underscoring the importance of pre-test genetic counseling. Our findings also highlight the subsequent impact on fertility programs, which may lead to the exclusion of gamete donors or patients choosing to revoke treatment. Trial registration number Not Applicable

CRISPR-Cas9: A Promising Therapeutic Approach for Diabetes Mellitus

Diabetes mellitus (DM) is a metabolic condition that raises blood glucose levels (hyperglycemia) without insulin or insulin receptor defects. Insulin is a peptide hormone the pancreatic beta cells produce and controls blood glucose levels. The defective immune system engulfs the beta cells of the pancreas, causing no insulin production or the insulin receptors to become faulty, resulting in insulin resistance. There are two main types of diabetes mellitus: type 1 diabetes mellitus and type 2 diabetes mellitus. In type 1 diabetes mellitus, the beta cells are engulfed by the defective immune system; hence the insulin production stops, and in type 2 diabetes mellitus, there is a defect in the insulin receptor that causes insulin resistance. Both types of diabetes cause uncontrolled blood sugar levels, which can lead to severe damage to vital body organs such as the heart, kidney, limbs, eyes, and nerves. The diseases of these organs are cardiovascular diseases, nephropathy, retinopathy, neuropathy, high blood pressure, and obesity. There are several methods to treat diabetes mellitus, such as islet cell transplant, tablets and medication, insulin pumps, weight loss surgery, insulin, diet and exercise, and emotional support. However, the medicines involved in traditionally treating diabetes mellitus are finerenone, tirzepatide, and, GLP-1 receptor, SGLT-2 inhibitors. These medicines cause severe damage to the body, such as cardiovascular death, transient ischemic attack, end-stage kidney diseases, and limb removal (amputation). Therefore, a new modern gene editing technology such as clustered regularly interspaced short palindromic repeats (CRISPR/Cas9) is involved in treating diabetes. In this technology, gene editing occurs in the human pluripotent stem cells (hiPSCs) taken from the patient’s body; these cells are converted into improved pancreatic beta cells. There are different methods involved, including cell-based therapies (such as stem cells and brown adipocytes), targeting specific genes associated with diabetes. This paper presents a critical review of the gene-editing tool CRISPR/Cas9 technology for the treatment of diabetes mellitus.

Bone microarchitecture and strength assessed by HR-pQCT in individuals with type 2 diabetes and prediabetes: the Maastricht study

Abstract Type 2 Diabetes (T2D) is a prevalent disease and has been associated with an increased fracture risk despite normal or even higher areal bone mineral density (BMD). The aim of this study was to estimate the association between glucose metabolism status (GMS) and measurements of glycemic control with high-resolution peripheral quantitative computed tomography (HR-pQCT) parameters of bone microarchitecture and strength. Participants of The Maastricht Study who underwent a HR-pQCT scan at the distal radius and tibia were included. GMS was determined by use of an oral glucose tolerance test and grouped into a normal glucose metabolism (NGM), prediabetes or T2D. Linear regression models were used, stratified by sex with multiple adjustments. This study incorporated cross-sectional data from 1569 (885 (56.4%) NGM, 251 (16.0%) prediabetes and 433 (27.6%) T2D) men, and 1606 (1159 (72.2%) NGM, 231 (14.4%) prediabetes and 216 (13.4%) T2D) women. The mean age was 60.1 ± 8.6 and 57.8 ± 9.0 years for men and women, respectively. After adjustment, T2D was associated with a higher total BMD measured by HR-pQCT and cortical thickness, and a smaller total and trabecular area in men and women compared to NGM. In women, T2D was additionally associated with a higher stiffness and failure load at the radius. Results were more pronounced at the distal radius than at the distal tibia. To conclude, these findings suggest that in this cohort of Maastricht Study participants, total and trabecular bone area are smaller, but bone microarchitecture, density and bone strength assessed by HR-pQCT are not impaired in individuals with T2D.

O-249 METAPHOR: METabolic imaging through AI-powered Phasor-based Hyperspectral analysis and Organelle recognition for the classification of human blastocysts

Abstract Study question Can we classify human blastocysts non-invasively using hyperspectral imaging? Summary answer HS imaging can significantly enhance our classification of human embryos and reveal numerous cellular features present in the embryo invisible to brightfield imaging. What is known already Non-invasive measurement of metabolism is a safe and powerful tool to reliably predict oocyte and embryo quality. We have previously shown that our METAPHOR technology, based on HS imaging coupled to artificial intelligence, was able to separate mouse oocyte categories according to maternal age with an average AUC of 96.2% and predict blastocyst formation (AUC 82.2%). Moreover, it was able to classify mouse embryos deprived from different nutrients with an average AUC of 93.7% showing clear superiority to grading provided by experienced embryologists (51%). Study design, size, duration A total of 74 human embryos donated for research at the blastocyst stage have been imaged to date. Embryos were warmed and imaged according to our established protocol and survival was statistically indistinguishable from control embryos. Participants/materials, setting, methods Embryos are imaged using multiphoton illumination near the infra-red. The HS detection covered the simultaneous measurement of the whole visible spectrum. Embryos are then allowed to implant in our proprietary 3D ex vivo platform. Implantation is monitored up until day 9 before immunostaining is performed to quantify presence and prevalence of lineage and implantation markers. Main results and the role of chance After imaging, embryos were placed on the 3D ex vivo implantation platform and let implant. Successful implantation (75.41%) was scored by observation of the characteristic deformation of collagen fibers within the matrix and staining for relevant cell lineages (OCT4, pMLC2). Interestingly, we observed that embryos with a higher trophectoderm (TE) grading tended to implant with a higher efficiency and penetrate deeper into the matrix than embryos with lower grading. HS images have unveiled a profound cellular diversity within embryos, elucidating distinctions between the Inner Cell Mass (ICM) and TE, as well as revealing different metabolic states defined by NADH dominance, FAD dominance, protoporphyrin levels, and apoptosis. HS raw images are processed using our HS-phasor approach which allows quick computing by converting the multidimensional HS data into comprehensive visual representation of bidimensional histograms and phasor plots. In summary, our analysis pipeline encompasses the algorithms to detect and segment the embryos, the computation of histogram and phasor-plot, and the classifier. AI allows us to classify embryos in distinct spectral spaces based on their metabolic features, paving the way to the establishment of a new grading system based on HS imaging. Limitations, reasons for caution The cohort size is still limited, and ongoing experiments will increase the dataset needed for more accurate prediction of embryo implantation. The implantation platform is an in vitro test that will require future clinical validation. Wider implications of the findings The safety and speed of METAPHOR warrants its incorporation at the IVF clinic, adding an additional layer of information to the current embryo selection methods. Moreover, the non-invasive measurement of the metabolic function opens new alleys of research such as the effect of media and culture supplements on embryo culture. Trial registration number Not applicable

Biochemical profile differences during the transition period based on different levels of non-esterified fatty acids at 7 weeks before parturition in Mediterranean Italian dairy buffaloes (Bubalus bubalis)

Metabolic adaptations to negative energy balance, as well as lipomobilization, influence inflammatory responses, immune function, and oxidative stress in animals. This study aimed to evaluate the biochemical profile of Mediterranean buffaloes with different levels of lipomobilization from the prepartum to the postpartum period. A total of 76 Mediterranean buffaloes were enrolled, and a weekly blood sample was taken from 7 weeks before to 6 weeks after calving. The concentration of non-esterified fatty acids (NEFAs) was determined in serum and was used to categorize buffaloes into three lipomobilization groups 7 weeks before calving: mild (NEFA-I; NEFA ≤ 0.29 mEq/L; n = 18), medium (NEFA-II; 0.29 < NEFA < 0.57 mEq/L; n = 20), and severe (NEFA-III; NEFA ≥ 0.57 mEq/L; n = 38). Two-way repeated measures ANOVA was used to assess changes within and between the groups and over time. Significant differences were found in the concentration levels of NEFA, β-hydroxybutyrate, glucose, cholesterol, protein profile, oxygen radicals, antioxidants, lysozyme, complement, and minerals. These results suggest that both medium and severe lipomobilization groups are associated with metabolic alterations. In conclusion, buffaloes with higher NEFA levels (>0.29 mEq/L; NEFA-II and NEFA-III) at 7 weeks before calving should be monitored more closely to reduce the risk of metabolic diseases. Furthermore, the medium (NEFA-II) and severe (NEFA-III) lipomobilization groups could be associated with differences in the animals’ ability to manage their metabolic status. Specifically, the severe mobilization group was most associated with a greater energy deficit during both the prepartum and postpartum periods without oxidative stress. On the contrary, the medium mobilization group was associated with a less severe energy deficit but was also associated with an inflammatory status and oxidative stress during the prepartum period. These distinctions highlight the need for tailored management strategies to address varying levels of metabolic stress in dairy buffaloes.

Spatial heterogeneity and functional zonation of living tissues and organs in situ.

In most organs, resources such as nutrients, oxygen, and physiologically active substances are unevenly supplied within the tissue spaces. Consequently, different tissue functions are exhibited in each space. This spatial heterogeneity of tissue environments arises depending on the spatial arrangement of nutrient vessels and functional vessels, leading to continuous changes in the metabolic states and functions of various cell types from regions proximal to these vessels to distant regions. This phenomenon is referred to as "zonation". Traditional analytical methods have made it difficult to investigate this zonation in detail. However, recent advancements in intravital imaging, spatial transcriptomics, and single-cell transcriptomics technologies have facilitated the discovery of "zones" in various organs and elucidated their physiological roles. Here, we outline the spatial differences in the immune system within each zone of organs. This information provides a deeper understanding of organs' immune systems.

Measurement ofenergy availability in highly trained male endurance athletes and examination of its associations with bone health and endocrine function.

Despite the introduction of Relative Energy Deficiency in Sport (RED-s) in 2014, there is evidence to suggest that male endurance athletes still present with a high prevalence of low energy availability (LEA). Previous findings suggest that energy availability (EA) status is strongly correlated with impairments in endocrine function such as reduced leptin, triiodothyronine (T3), and insulin, and elevated bone loss. This study aimed to report the current EA status, endocrine function and bone health of highly trained Irish male endurance athletes. In this cross-sectional study, participants (n = 3 triathletes; n = 10 runners) completed a 7-day testing period during the competition season using lab-based measures, to ascertain EA status, hormone level and rates of bone metabolism. Serum blood samples were obtained to assess hormone levels and markers of bone metabolism. Mean EA was < 30kcal/kg lean body mass (LBM)/day in 76.9% of athletes. There was a strong association between LEA and low carbohydrate intake, and lower LBM. Mean levels of insulin, IGF-1 and leptin were significantly lower than their reference ranges. Elevated mean concentrations of β-CTX and a mean P1NP: β-CTX ratio < 100, indicated a state of bone resorption. The EA level, carbohydrate intake, hormone status and bone metabolism status of highly trained male endurance athletes are a concern. Based on the findings of this study, more frequent assessment of EA across a season is recommended to monitor the status of male endurance athletes, in conjunction with nutritional education specific to EA and the associated risks.

The inflammatory and metabolic status of patients with sudden-onset sensorineural hearing loss

IntroductionSudden sensorineural hearing loss (SSNHL) is a common emergency symptom in otolaryngology that requires immediate diagnosis and treatment. SSNHL has a multifactorial etiology, and its pathophysiologic mechanisms may be associated with inflammatory and metabolic changes that may affect the cochlear microenvironment or its nervous component, thus triggering the process or hindering hearing recovery. Therefore, the aim of this study was to assess metabolic and inflammatory changes to identify systemic parameters that could serve as prognostic factors for hearing recovery in patients with SSNHL.Materials and methodsThirty patients with a sudden hearing loss of at least 30 dB in three contiguous frequencies were enrolled in this study. Patients were followed up for 4 months and peripheral blood samples were collected at 7 days (V1), 30 days (V2) and 120 days (V3). Interleukins (IL)-1F7, IL-2, IL-4, IL-5, IL-6, IL-10, interferon γ (IFN-γ), tumor necrosis factor α (TNF-α) and adiponectin were quantified in serum. In addition, lipid and glycemic profiles as well as concentration of creatinine, uric acid, fructosamine, peroxide, total proteins and albumin were analyzed. Patients underwent weekly ear-specific hearing tests with standard pure tone thresholds for frequencies of 250–8,000 Hz, speech recognition threshold and word recognition score.ResultsPatients with SSNHL were divided into a group of patients who did not achieve hearing recovery (n = 14) and another group who achieved complete and significant recovery (n = 16). Most serologic parameters showed no significant changes or values indicating clinical changes. However, IFN-γ levels decreased by 36.3% between V1 and V2. The cytokine TNF-α showed a statistically significant decrease from V1 to V3 (from 22.91 to 10.34 pg./mL). Adiponectin showed a decrease from 553.7 ng/mL in V1 to 454.4 ng/mL in V3.DiscussionOur results show that serologic cytokine levels change in the acute phase of manifestation of SSNHL and establish a parallel between systemic changes and improvements in hearing, especially TNF-α, which showed differences in hearing recovery. The use of IFN-γ, TNF-α and adiponectin may elucidate the clinical improvement in these patients.

Inhibiting the NADase CD38 improves cytomegalovirus-specific CD8+ T cell functionality and metabolism.

Cytomegalovirus (CMV) is one of the most common and relevant opportunistic pathogens in immunocompromised individuals such as kidney transplant recipients (KTRs). The exact mechanisms underlying the disability of cytotoxic T cells to provide sufficient protection against CMV in immunosuppressed individuals have not been identified yet. Here, we performed in-depth metabolic profiling of CMV-specific CD8+ T cells in immunocompromised patients and show the development of metabolic dysregulation at the transcriptional, protein, and functional level of CMV-specific CD8+ T cells in KTRs with non-controlled CMV infection. These dysregulations comprise impaired glycolysis and increased mitochondrial stress, which is associated with an intensified expression of the nicotinamide adenine dinucleotide nucleotidase (NADase) CD38. Inhibiting NADase activity of CD38 reinvigorated the metabolism and improved cytokine production of CMV-specific CD8+ T cells. These findings were corroborated in a mouse model of CMV infection under conditions of immunosuppression. Thus, dysregulated metabolic states of CD8+ T cells could be targeted by inhibiting CD38 to reverse hypo-responsiveness in individuals who fail to control chronic viral infection.

Time profiles of energy balance in dairy cows in association with metabolic status, inflammatory status, and disease

The early lactation period in dairy cows is characterized by complex interactions among energy balance (EB), disease, and alterations in metabolic and inflammatory status. The objective of this study was to cluster cows based on EB time profiles in early lactation and investigate the association between EB clusters and inflammatory status, metabolic status, oxidative stress, and disease. Holstein-Friesian dairy cows (n = 153) were selected and monitored for disease treatments during wk 1 to 6 in lactation. Weekly EB was calculated based on energy intake and energy requirements for maintenance and milk yield in wk 1 to 6 in lactation. Weekly plasma samples were analyzed for metabolic variables in wk 1 to 6, and inflammatory and oxidative stress variables in wk 1, 2, and 4 in lactation. Liver activity index (LAI) was computed from plasma albumin, cholesterol, and retino-binding protein concentration. First, cows were clustered based on time profiles of EB, resulting in 4 clusters (SP: stable positive; MN: mild negative; IN: intermediate negative; SN: severe negative). Cows in the SN cluster had higher plasma nonesterified fatty acids and β-hydroxybutyrate concentrations, compared with cows in the SP cluster, with the MN and IN cluster being intermediate. Cows in the SN cluster had a higher milk yield, lower dry matter intake in wk 1, lower insulin concentration compared with cows in the SP cluster, and lower glucose and IGF-1 concentration compared with cows in the SP and MN clusters. Energy balance clusters were not related with plasma haptoglobin, cholesterol, albumin, paraoxonase, and liver activity index (LAI). Second, cows were grouped based on health status [IHP: cows with treatment for inflammatory health problem (endometritis, fever, clinical mastitis, vaginal discharge or retained placenta); OHP: cows with no IHP but treatment for other health problem (milk fever, cystic ovaries, claw, and leg problems, rumen and intestine problems or other diseases); NHP: cows with no treatments, in the first 6 weeks after calving]. Energy balance was not different among health status groups. The IHP cows had lower nonesterified fatty acids and greater insulin concentration in plasma compared with OHP. The IHP cows had lower plasma albumin concentration, lower LAI and higher haptoglobin concentration compared with OHP and NHP. Overall, EB time profiles were associated with the metabolic status of dairy cows in early lactation, but were only limitedly related with markers of inflammation and oxidative stress status. Inflammatory and metabolic status were related to disease events in early lactation and caused prolonged effects on liver metabolism.

Differential reliance of CTD-nuclear envelope phosphatase 1 on its regulatory subunit in ER lipid synthesis and storage.

Lipin 1 is an ER enzyme that produces diacylglycerol, the lipid intermediate that feeds into the synthesis of glycerophospholipids for membrane expansion or triacylglycerol for storage into lipid droplets. CTD-Nuclear Envelope Phosphatase 1 (CTDNEP1) regulates lipin 1 to restrict ER membrane synthesis, but a role for CTDNEP1 in lipid storage in mammalian cells is not known. Furthermore, how NEP1R1, the regulatory subunit of CTDNEP1, contributes to these functions in mammalian cells is not fully understood. Here, we show that CTDNEP1 is reliant on NEP1R1 for its stability and function in limiting ER expansion. CTDNEP1 contains an amphipathic helix at its N-terminus that targets to the ER, nuclear envelope and lipid droplets. We identify key residues at the binding interface of CTDNEP1 and NEP1R1 and show that they facilitate complex formation in vivo and in vitro. We demonstrate that NEP1R1 binding to CTDNEP1 shields CTDNEP1 from proteasomal degradation to regulate lipin 1 and restrict ER size. Unexpectedly, NEP1R1 was not required for CTDNEP1's role in restricting lipid droplet biogenesis. Thus, the reliance of CTDNEP1 function on NEP1R1 depends on cellular demands for membrane production versus lipid storage. Together, our work provides a framework into understanding how the ER regulates lipid synthesis under different metabolic conditions.

Intermittent fasting, calorie restriction, and a ketogenic diet improve mitochondrial function by reducing lipopolysaccharide signaling in monocytes during obesity: A randomized clinical trial

BackgroundMitochondrial dysfunction occurs in monocytes during obesity and contributes to a low-grade inflammatory state; therefore, maintaining good mitochondrial conditions is a key aspect of maintaining health. Dietary interventions are primary strategies for treating obesity, but little is known about their impact on monocyte bioenergetics. Thus, the aim of this study was to evaluate the effects of calorie restriction (CR), intermittent fasting (IF), a ketogenic diet (KD), and an ad libitum habitual diet (AL) on mitochondrial function in monocytes and its modulation by the gut microbiota. Methods and findingsA randomized controlled clinical trial was conducted in which individuals with obesity were assigned to one of the 4 groups for 1 month. Subsequently, the subjects received rifaximin and continued with the assigned diet for another month. The oxygen consumption rate (OCR) was evaluated in isolated monocytes, as was the gut microbiota composition in feces and anthropometric and biochemical parameters. Forty-four subjects completed the study, and those who underwent CR, IF and KD interventions had an increase in the maximal respiration OCR (p=0.025, n2p = 0.159 [0.05, 0.27] 95% confidence interval) in monocytes compared to that in the AL group. The improvement in mitochondrial function was associated with a decrease in monocyte dependence on glycolysis after the IF and KD interventions. Together, diet and rifaximin increased the gut microbiota diversity in the IF and KD groups (p=0.0001), enriched the abundance of Phascolarctobacterium faecium (p=0.019) in the CR group and Ruminococcus bromii (p=0.020) in the CR and KD groups, and reduced the abundance of lipopolysaccharide (LPS)-producing bacteria after CR, IF and KD interventions compared to the AL group at the end of the study according to ANCOVA with covariate adjustment. Spearman’s correlation between the variables measured highlighted LPS as a potential modulator of the observed effects. In line with this findings, serum LPS and intracellular signaling in monocytes decreased with the three interventions (CR, p=0.002; IF, p=0.001; and KD, p=0.001) compared to those in the AL group at the end of the study. ConclusionsWe conclude that these dietary interventions positively regulate mitochondrial bioenergetic health and improve the metabolic profile of monocytes in individuals with obesity via modulation of the gut microbiota. Moreover, the evaluation of mitochondrial function in monocytes could be used as an indicator of metabolic and inflammatory status, with potential applications in future clinical trials. Trial registrationThis trial was registered with ClinicalTrials.gov (NCT05200468)

Glycogen synthesis prevents metabolic imbalance and disruption of photosynthetic electron transport from photosystem II during transition to photomixotrophy in Synechocystis sp. PCC 6803.

Some cyanobacteria can grow photoautotrophically or photomixotrophically by using simultaneously CO2 and glucose. The switch between these trophic modes and the role of glycogen, their main carbon storage macromolecule, was investigated. We analysed the effect of glucose addition on the physiology, metabolic and photosynthetic state of Synechocystis sp. PCC 6803 and mutants lacking phosphoglucomutase and ADP-glucose pyrophosphorylase, with limitations in glycogen synthesis. Glycogen acted as a metabolic buffer: glucose addition increased growth and glycogen reserves in the wild-type (WT), but arrested growth in the glycogen synthesis mutants. Already 30 min after glucose addition, metabolites from the Calvin-Benson-Bassham cycle and the oxidative pentose phosphate shunt increased threefold more in the glycogen synthesis mutants than the WT. These alterations substantially affected the photosynthetic performance of the glycogen synthesis mutants, as O2 evolution and CO2 uptake were both impaired. We conclude that glycogen synthesis is essential during transitions to photomixotrophy to avoid metabolic imbalance that induces inhibition of electron transfer from PSII and subsequently accumulation of reactive oxygen species, loss of PSII core proteins, and cell death. Our study lays foundations for optimising photomixotrophy-based biotechnologies through understanding the coordination of the crosstalk between photosynthetic electron transport and metabolism.

Knowledge, attitudes and practices of sheep owners regarding abortion in Northern Tunisia.

Abortion in ewes causes high economic losses and represents a threat for human health due to abortive zoonotic pathogens. The present study aimed to assess the knowledge, attitudes and practices (KAP) among sheep owners in the northern Tunisia regarding ewes' abortions. Between February 2021 and May 2022, a structured questionnaire containing both close and open-ended questions was applied to 120 sheep owners in northern Tunisia. The data collected were analysed by chi-square test using Epi info 6 software. The majority (75%) of participants reported a history of abortion in their sheep flocks. Sheep owners thought that the most frequent cause of abortion was physical factors, such as trauma, climate and stress (60%±5.5%; 48/80), followed by toxicity (15%±4%; 12/80), metabolic and nutritional conditions (12.5%±3.7%; 10/80), vaccination (5%±2.4%; 4/80) and infectious causes (7.5%±2.9%; 6/80) (p<0.001). The majority of animal owners reported that abortions occurred mainly during autumn (39.6%±5%; 38/96), followed by summer (27%±4.5%; 26/96), winter (23%±4.3%; 22/96) and spring (10.4%±3.1%; 10/96) (p<0.001). Approximately, half (45.8%±5%; 55/120) of interviewed farmers would not take any action if an abortion occurred. Half of the interviewed farmers (50.5%±5.1%; 48/95) did not apply any preventive measures when manipulating aborted ewes, and most of the sheep owners (77.3%±3.8%; 92/119) did not know that aborted ewes could transmit zoonotic pathogens. Our survey concluded that sheep owners in Northern Tunisia had poor knowledge and attitudes as well as applied limited actions concerning several health aspects related to abortion. Education programmes should be established in order to improve Tunisian sheep owners' KAP regarding abortion.

Integrated multi-omics reveals the relationship between growth performance, rumen microbes and metabolic status of Hu sheep with different residual feed intakes

Residual feed intake (RFI) is a metric that provides a more accurate measure of feed efficiency. The lower the RFI, the higher the feed efficiency. The changes in the host microbiome and metabolome contribute to the greater feed efficiency of low RFI (LRFI) animals. The aim of this study was to explore the differences in rumen microorganisms, rumen metabolites and plasma metabolites of Hu sheep with differing RFI through the microbiome and metabolome. A total of 80 Hu sheep were used. The experiment consisted of a 15-d pretrial period and a 128-d experimental period. The RFI in the experimental period was calculated for all sheep, and the sheep were screened into high RFI (HRFI, n = 8) and LRFI (n = 8) groups. The HRFI and LRFI sheep did not differ in their initial and final body weights, average daily gain and body measurements, but the dry matter intake of LRFI sheep was significantly decreased (28.4%, P < 0.001). The sheep with LRFI had higher digestibility of crude protein (P = 0.010) and ether extract (P = 0.010) compared to HRFI group. The concentrations of acetate (P = 0.036), propionate (P = 0.010), valerate (P = 0.027) and total volatile fatty acids (P = 0.048) in rumen of LRFI group were higher compared to HRFI group. The results of 16S rDNA sequencing indicated that the sheep with LRFI had higher proportions of Prevotella genus in rumen liquid (P = 0.031). The rumen metabolome and plasma metabolome results showed that the citrate cycle, pyruvate metabolism and alanine, aspartate and glutamate metabolism processes were more active for sheep in LRFI group, which provided more energy substrate such as malic acid, oxoglutaric acid and citric acid. In conclusion, sheep with LRFI can utilize feed more efficiently, and the more active energy metabolism pathway and the production of energy substances may account for the higher feed efficiency.

Exploration of bone metabolism status in the distal femur of mice at different growth stages

Exploration of bone metabolism status in the distal femur of mice at different growth stages

Differential regulation of mitochondrial uncoupling protein 2 in cancer cells

The persistent growth of cancer cells is underscored by complex metabolic reprogramming, with mitochondria playing a key role in the transition to aerobic glycolysis and representing new therapeutic targets. Mitochondrial uncoupling protein 2 (UCP2) has attracted interest because of its abundance in rapidly proliferating cells, including cancer cells, and its involvement in cellular metabolism. However, the specific contributions of UCP2 to cancer biology remain poorly defined. Our investigation of UCP2 expression in various human and mouse cancer cell lines aimed to elucidate its links to metabolic states, proliferation, and adaptation to environmental stresses such as hypoxia and nutrient deprivation. We observed significant variability in UCP2 expression across cancer types, with no direct correlation to their metabolic activity or proliferation rates. UCP2 abundance was also differentially affected by nutrient availability in different cancer cells, but UCP2 was generally downregulated under hypoxia. These findings challenge the notion that UCP2 is a marker of malignant potential and suggest its more complex involvement in the metabolic landscape of cancer.

A fluorogenic complementation tool kit for interrogating lipid droplet-organelle interaction.

Contact sites between lipid droplets and other organelles are essential for cellular lipid and energy homeostasis upon metabolic demands. Detection of these contact sites at the nanometer scale over time in living cells is challenging. We developed a tool kit for detecting contact sites based on fluorogen-activated bimolecular complementation at CONtact sites, FABCON, using a reversible, low-affinity split fluorescent protein, splitFAST. FABCON labels contact sites with minimal perturbation to organelle interaction. Via FABCON, we quantitatively demonstrated that endoplasmic reticulum (ER)- and mitochondria (mito)-lipid droplet contact sites are dynamic foci in distinct metabolic conditions, such as during lipid droplet biogenesis and consumption. An automated analysis pipeline further classified individual contact sites into distinct subgroups based on size, likely reflecting differential regulation and function. Moreover, FABCON is generalizable to visualize a repertoire of organelle contact sites including ER-mito. Altogether, FABCON reveals insights into the dynamic regulation of lipid droplet-organelle contact sites and generates new hypotheses for further mechanistical interrogation during metabolic regulation.

EFFECTIVENESS OF AUTOLOGOUS PLATELET-RICH PLASMA FOR HEALING DIABETIC FOOT ULCERS

Autologous platelet-rich plasma (APRP) is widely used to accelerate the healing of chronic wounds. However, the advisability of its use in patients with diabetic foot ulcers (DFU), which develop in various forms of the disease and are associated with complex metabolic conditions, is still being discussed. This study consisted in a comparative analysis of the effectiveness of local use of autologous platelet-rich plasma in the treatment of DFU in patients with neuropathic and neuroischemic forms of the disease. A total of 106 patients with type 2 diabetes mellitus undergoing hospital treatment were divided into a study group (n=48) and a traditional wound therapy group (n=58), similar in ankle-brachial index values ​​and ulcer size. Systemic therapy in both groups of patients included measures to regulate metabolism and nutrition, and infection control. Treatment results were assessed by the area of foot wounds and duration of healing. The duration of therapy and the mean platelet concentration in autologous plasma did not differ between groups. The wound healing time in patients with the neuropathic form (60.4±23.77 days) and the neuroischemic form of the disease (64.8±29.6 days) during treatment with APRP was significantly shorter than in the subgroups of the comparison group with traditional treatment of DFU (85.11±40.7 and 90.40±47.12 days respectively) (p&lt;0.01), but there was no significant difference between the subgroups receiving treatment with APRP. Although there was no significant difference in the average daily healing area among all subgroups (p&gt;0.05), the rate of wound healing in the subgroups with APRP was higher (14.85±10.66 and 15.33±14.91 mm2/day, respectively) than in the subgroups of the comparison group (8.39±6.02 and 8.79±7.13 mm2/day, respectively), which confirms the effectiveness of local treatment with APRP in combination with traditional systemic therapy.

Is Cancer Metabolism an Atavism?

The atavistic theory of cancer posits that cancer emerges and progresses through the reversion of cellular phenotypes to more ancestral types with genomic and epigenetic changes deactivating recently evolved genetic modules and activating ancient survival mechanisms. This theory aims at explaining the known cancer hallmarks and the paradox of cancer’s predictable progression despite the randomness of genetic mutations. Lineweaver and colleagues recently proposed the Serial Atavism Model (SAM), an enhanced version of the atavistic theory, which suggests that cancer progression involves multiple atavistic reversions where cells regress through evolutionary stages, losing recently evolved traits first and reactivating primitive ones later. The Warburg effect, where cancer cells upregulate glycolysis and lactate production in the presence of oxygen instead of using oxidative phosphorylation, is one of the key feature of the SAM. It is associated with the metabolism of ancient cells living on Earth before the oxygenation of the atmosphere. This review addresses the question of whether cancer metabolism can be considered as an atavistic reversion. By analyzing several known characteristics of cancer metabolism, we reach the conclusion that this version of the atavistic theory does not provide an adequate conceptual frame for cancer research. Cancer metabolism spans a whole spectrum of metabolic states which cannot be fully explained by a sequential reversion to an ancient state. Moreover, we interrogate the nature of cancer metabolism and discuss its characteristics within the framework of the SAM.