3-hydroxybutyrate Concentrations Research Articles

99 The Effects of β-caryophyllene on butyrate utilization and metabolism in Caco 2 cells

Abstract Butyrate is important for regulating energy status and cellular metabolism. Beta-caryophyllene (BCP) is a plant compound which may exert potentially beneficial effects on intestinal epithelial cells, such as barrier integrity and modulation of nutrient utilization. The goal of this preliminary study was to investigate the effects of BCP on butyrate utilization and metabolism in intestinal epithelial cells. Caco 2 cells were used as a model for the intestinal barrier. Cells were seeded in multi-well plates with hanging inserts (n = 4) and grown for 18 d. On d 19, 1 of 4 treatments was added to triplicate wells per plate. Treatments included vehicle control (VC), 40 µM BCP, 2 mM butyrate (BTR), and butyrate plus BCP (BB). Treatments were added to the apical side and incubated for 24 and 48 h. Transepithelial electrical resistance (TEER; ohms/cm2) readings and supernatant samples from both the apical and basolateral sides of the cell layer were taken at each time point, and butyrate and β- hydroxybutyrate (BHB) concentrations measured. Butyrate utilization was calculated as absolute nmol lost. Statistical analyses were performed using lm and emmeans packages of R, with treatment, time and side (of cell layer) as fixed effects. All wells started with the same statistical TEER values on h 0. After 24 and 48 h BCP showed no changes in TEER (P > 0.05). Compared with VC, the mean TEER for BTR and BB was greater at 24 h (579 vs 878 and 1,008 ± 35, respectively; P < 0.001). After 48 h, all groups returned to baseline, except BB which had a greater TEER (883 ± 35; P < 0.001) compared with other treatments. After 24 and 48 h BCP showed no changes for both sides of the cell layer (P > 0.05). Overall, absolute BHB concentrations increased from 24 to 48 h in all treatments (P < 0.05). Production of BHB in each of the VC and BC groups was less than 15 nmol at each time point. For BB and BTR groups, more nmol BHB was detected on the basolateral side (31.9 and 22.2 ± 1.3) vs apical (14.0 and 10.4 ± 1.3, respectively; P < 0.001) after 24 h. After 48 h, BHB was also greater on the basolateral side (58.2 and 45.2 ± 1.3 nmol for BB and BUT, respectively) vs apical side (30.2 and 24.0 ± 1.3 nmol; P < 0.001). When looking at BTR utilization, the BB treatment had greater butyrate disappearance than BTR in 24 h (254 vs 169 ± 12.6 nmol; P = 0.0005) and 48h (707 vs 573 ± 12.6 nmol; P < 0.0001). BCP may promote the metabolism of butyrate by intestinal epithelial cells. More research is needed to understand the mechanism of how BCP increases cellular butyrate utilization.

Gut Microbiota-Derived 3-Hydroxybutyrate Blocks GPR43-Mediated IL6 Signaling to Ameliorate Radiation Proctopathy.

Radiation proctopathy (RP) is a common complication of radiotherapy for pelvic malignancies with high incidence. RP accompanies by microbial dysbiosis. However, how the gut microbiota affects the disease remains unclear. Here, metabolomics reveals that the fecal and serous concentrations of microbiota-derived 3-hydroxybutyrate (3HB) are significantly reduced in RP mice and radiotherapeutic patients. Moreover, the concentration of 3HB is negatively associated with the expression of proinflammatory IL6 that is increased along with the severity of radiation damage. 3HB treatment significantly downregulates IL6 expression and alleviates IL6-mediated radiation damage. Irradiated cell-fecal microbiota co-culture experiments and in vivo assays show that such a radioprotection of 3HB is mediated by GPR43. Microbiome analysis reveals that radiation leads to a distinct bacterial community compared to untreated controls, in which Akkermansia muciniphila is significantly reduced in RP mice and radiotherapeutic patients and is associated with lower 3HB concentration. Gavage of A. muciniphila significantly increases 3HB concentration, downregulates GPR43 and IL6 expression, and ameliorates radiation damage. Collectively, these results demonstrate that the gut microbiota, including A. muciniphila, induce higher concentrations of 3HB to block GPR43-mediated IL6 signaling, thereby conferring radioprotection. The findings reveal a novel implication of the gut-immune axis in radiation pathophysiology, with potential therapeutic applications.

Role of GPR81 in regulating intramuscular triglyceride storage during aerobic exercise in rats.

Lactate, a metabolite of exercise, plays a crucial role in the body. In these studies, we aimed to investigate the role of G protein-coupled receptor 81 (GPR81), a specific receptor for lactate, in regulating lipid storage in the gastrocnemius muscle of rats. To achieve this, we measured the impact of sodium 3-hydroxybutyrate (3-OBA) concentration and time on the cAMP-PKA signaling pathway in the gastrocnemius muscles ofrats. Our investigation involved determining the effects of administering 3-OBA at a concentration of 3 mmol L-1 just 15 min before exercise. As expected, exercise led to a notable increase in intramuscular lactate concentration in rats. However, injecting 3-OBA prior to exercise yielded intriguing results. It not only further augmented the cAMP-PKA signaling pathway but also boosted the expression of lipolysis-related proteins such as hormone-sensitive lipase (HSL) and adipose triglyceride lipase (ATGL). Simultaneously, it decreased the expression of fat-synthesizing proteins, including acetyl CoA carboxylase (ACC) and fatty acid synthase (FAS), while increasing the protein expression of cytochrome c oxidase subunit Ⅳ(COX Ⅳ) and the activity of citrate synthetase (CS). Unfortunately, there was no significant change observed in intramuscular triglyceride (IMTG) content. In summary, our findings shed light on the role of lactate in partially regulating intramuscular triglycerides during exercise.

Effects of negative dietary cation-anion difference and calcidiol supplementation in transition diets fed to sows on piglet survival, piglet weight, and sow metabolism.

Diets that provide a negative dietary anion cation difference (DCAD) and supplement with a vitamin D metabolite 25-OH-D3 (calcidiol) may increase calcium availability at parturition, and enhance piglet survival and performance. This factorial study assessed the effects of DCAD, calcidiol (50 µg/kg), and parity (parity 1 or >1) and their interactions. Large White and Landrace sows (n = 328), parity 1 to 8 were randomly allocated in blocks to treatment diets from day 103 of gestation until day 3 postfarrow: 1) negative DCAD without calcidiol (negative DCAD + no CA), n = 84, 2) negative DCAD with calcidiol (negative DCAD + CA) n = 84, 3) positive DCAD without calcidiol (negative DCAD + no CA), n = 81, and 4) positive DCAD with calcidiol (positive DCAD + CA), n = 79. Negative DCAD diets were acidified with an anionic feed (2kg/t) and magnesium sulfate (2kg/t). All treatment diets contained cholecalciferol at 1,000 IU/kg. Dry sow diets contained 14.8% crude protein (CP), 5.4% crude fiber (CF), 0.8% Ca, and 83 mEq/kg DCAD. Treatment diets 1 and 2 contained 17.5% CP, 7.3% CF, 0.8% Ca, and -2 mEq/kg DCAD. Treatment diets 3 and 4 contained 17.4% CP, 7.4% CF, 0.8% Ca, and 68 mEq/kg DCAD. Before farrowing, all negative DCAD sows had lower urine pH than all sows fed a positive DCAD (5.66 ± 0.05 and 6.29 ± 0.05, respectively; P < 0.01); urinary pH was acidified for both DCAD treatments indicating metabolic acidification. The percentage of sows with stillborn piglets was not affected by DCAD, calcidiol, or parity alone but sows fed the negative DCAD + CA diet had a 28% reduction in odds of stillbirth compared to the negative DCAD + no CA diet and even lesser odds to the positive DCAD + CA diet. At day 1 after farrowing, blood gas, and mineral and metabolite concentrations were consistent with feeding a negative DCAD diet and that negative DCAD diets influence energy metabolism, as indicated by increased glucose, cholesterol, and osteocalcin concentrations and reduced nonesterified free fatty acids and 3-hydroxybutyrate concentrations. In the subsequent litter, total piglets born and born alive (14.7 ± 0.3 and 13.8 ± 0.3 piglets, respectively; P = 0.029) was greater for positive DCAD diets compared to negative DCAD diets; and there was an interaction between DCAD, calcidiol, and parity (P = 0.002). Feeding a negative DCAD diet influenced stillbirth, subsequent litter size, and metabolic responses at farrowing. More studies are needed to define optimal diets prefarrowing for sows.

Lipid metabolites index habitat quality for Canvasbacks on stopover areas during spring migration

Abstract Nutrients acquired by ducks on spring migratory stopover areas influence survival and subsequent reproduction. Accordingly, wetland loss and degradation on stopover areas can lead to reduced refueling efficiency and have demographic consequences. Lipid metabolite concentrations in blood provide a useful index of daily mass change in wild birds and can be used to make inferences about quality of stopover areas for refueling. We experimentally validated a model that uses lipid metabolites to predict daily mass change, and then we used those predictions to make inferences about foraging habitat quality for Canvasbacks (Aythya valisineria) at important stopover sites of the upper Mississippi and Illinois Rivers, USA. We measured plasma lipid metabolites and daily mass change of 60 wild Canvasbacks held in short-term captivity and subjected to feeding and fasting treatments. Respectively, triglyceride and β-hydroxybutyrate concentrations were positively and negatively related to mass change (R2 = 0.58). On average, Canvasbacks collected experimentally across our study area had positive index values indicating sufficient forage resources to allow energy acquisition. However, predicted daily mass increases were greater at more northerly pools of the Mississippi River (Pools 7–8 and 13) compared to more southerly locations (Pool 19 of the Mississippi River and the Illinois River Valley), which was likely due, in part, to more abundant submerged aquatic vegetation at more northerly pools. Our results affirm that lipid metabolites are useful predictors of daily mass change that in turn can provide inferences about habitat quality. Lower daily mass increases of Canvasbacks at Pool 19 are concerning because that area is considered a critically important spring stopover area for migratory diving ducks. Considering the challenges of managing large rivers for multiple uses, targeting restoration of wetlands isolated or isolatable from river systems could provide stopover areas with necessary forage for optimal mass gain.

Plasma metabolomics reveals major changes in carbohydrate, lipid, and protein metabolism of abruptly weaned beef calves

1H NMR-based metabolomics was used to study the effect of abrupt weaning on the blood metabolome of beef calves. Twenty Angus calves (258 ± 5 kg BW; 5 to 6 months old) were randomly assigned to a non-weaned (NW) group that remained grazing with their dam or a weaned (W) group that underwent abrupt separation from their dam to a separate paddock on d 0 of the study. Body weight, behaviour, and blood samples for cortisol and metabolomics were measured at d 0, 1, 2, 7, and 14 of the study. On d 1 and 2, W calves spent less time grazing and ruminating, and more time vocalising and walking, had a greater concentration of cortisol, NEFA, 3-hydroxybutyrate, betaine, creatine, and phenylalanine, and lesser abundance of tyrosine (P < 0.05) compared to NW calves. Compared to NW calves at d 14, W calves had greater (P < 0.01) relative abundance of acetate, glucose, allantoin, creatinine, creatine, creatine phosphate, glutamate, 3-hydroxybutyrate, 3-hydroxyisobutyrate, and seven AA (alanine, glutamate, leucine, lysine, phenylalanine, threonine and valine) but lesser (P < 0.05) relative abundance of low density and very low-density lipids, and unsaturated lipids. Both PCA and OPLS-DA showed no clustering or discrimination between groups at d 0 and increasing divergence to d 14. Blood metabolomics is a useful tool to quantify the acute effects of stress in calves during the first 2 days after abrupt weaning, and longer-term changes in carbohydrate, lipid and protein metabolism due to nutritional changes from cessation of milk intake and greater reliance on forage intake.

Maintained imbalance of triglycerides, apolipoproteins, energy metabolites and cytokines in long-term COVID-19 syndrome patients.

Deep metabolomic, proteomic and immunologic phenotyping of patients suffering from an infection with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) have matched a wide diversity of clinical symptoms with potential biomarkers for coronavirus disease 2019 (COVID-19). Several studies have described the role of small as well as complex molecules such as metabolites, cytokines, chemokines and lipoproteins during infection and in recovered patients. In fact, after an acute SARS-CoV-2 viral infection almost 10-20% of patients experience persistent symptoms post 12 weeks of recovery defined as long-term COVID-19 syndrome (LTCS) or long post-acute COVID-19 syndrome (PACS). Emerging evidence revealed that a dysregulated immune system and persisting inflammation could be one of the key drivers of LTCS. However, how these biomolecules altogether govern pathophysiology is largely underexplored. Thus, a clear understanding of how these parameters within an integrated fashion could predict the disease course would help to stratify LTCS patients from acute COVID-19 or recovered patients. This could even allow to elucidation of a potential mechanistic role of these biomolecules during the disease course. This study comprised subjects with acute COVID-19 (n=7; longitudinal), LTCS (n=33), Recov (n=12), and no history of positive testing (n=73). 1H-NMR-based metabolomics with IVDr standard operating procedures verified and phenotyped all blood samples by quantifying 38 metabolites and 112 lipoprotein properties. Univariate and multivariate statistics identified NMR-based and cytokine changes. Here, we report on an integrated analysis of serum/plasma by NMR spectroscopy and flow cytometry-based cytokines/chemokines quantification in LTCS patients. We identified that in LTCS patients lactate and pyruvate were significantly different from either healthy controls (HC) or acute COVID-19 patients. Subsequently, correlation analysis in LTCS group only among cytokines and amino acids revealed that histidine and glutamine were uniquely attributed mainly with pro-inflammatory cytokines. Of note, triglycerides and several lipoproteins (apolipoproteins Apo-A1 and A2) in LTCS patients demonstrate COVID-19-like alterations compared with HC. Interestingly, LTCS and acute COVID-19 samples were distinguished mostly by their phenylalanine, 3-hydroxybutyrate (3-HB) and glucose concentrations, illustrating an imbalanced energy metabolism. Most of the cytokines and chemokines were present at low levels in LTCS patients compared with HC except for IL-18 chemokine, which tended to be higher in LTCS patients. The identification of these persisting plasma metabolites, lipoprotein and inflammation alterations will help to better stratify LTCS patients from other diseases and could help to predict ongoing severity of LTCS patients.

A Clinical and Biochemical Comparative Study Of Diabetic Ketoacidosis (DKA) in Newly Diagnosed Vs Known Cases of Type 1 Diabetic Children

Objectives: We aimed to study the characterizing clinical and biochemical profiles of Diabetic Ketoacidosis (DKA) in children with newly diagnosed Type 1 Diabetes Mellitus (Type 1DM) compared to children with established diagnosis of Type 1DM presenting with DKA admitted to the pediatric intensive care unit of a large university hospital in the eastern region of Saudi Arabia. Methods: We retrospectively reviewed the medical records of 211 patients who were admitted to the pediatric intensive care unit with diabetic ketoacidosis between 2010 and 2019. The diagnosis of diabetic ketoacidosis was based on symptoms of polydipsia, polyurea, weight loss, vomiting, dehydration, abdominal pain, breathing problems, lethargy or coma, biochemical hyperglycemia (blood glucose level of >200 mg/dL), venous pH of <7.3, serum bicarbonate level of ≤15 mEq/L, and ketonemia (blood β -hydroxybutyrate concentration of ≥3 mM) or moderate or severe ketonuria (diagnosed as newly acquired type 1 diabetes). Results: The rate of newly diagnosed Type 1 DM with DKA was 41.7%, out of them who got severe and moderate diabetic ketoacidosis were 61.6% and 38.4%, respectively. We observed significantly increased heart and respiratory rates in patients newly diagnosed with diabetic ketoacidosis and in those with severe diabetic ketoacidosis (p<0.001) compared to known cases with Type 1DM presenting with DKA. We also identified significantly increased biochemical indices including HbA1c, random blood sugar, serum osmolality, blood urea nitrogen, creatinine, chloride, lactate, and anion gap in relation to severe diabetic ketoacidosis and newly diagnosed type 1 diabetes (p ≤ 0.05). Conclusions: We found that the clinical and biochemical profiles of patients with newly diagnosed Type 1 DM children were significantly affected compared to children who were known Type 1DM presenting with DKA.

Acidic ascites inhibits ovarian cancer cell proliferation and correlates with the metabolomic, lipidomic and inflammatory phenotype of human patients

BackgroundThe poor prognosis of ovarian cancer patients is strongly related to peritoneal metastasis with the production of malignant ascites. However, it remains largely unclear how ascites in the peritoneal cavity influences tumor metabolism and recurrence. This study is an explorative approach aimed at for a deeper molecular and physical–chemical characterization of malignant ascites and to investigate their effect on in vitro ovarian cancer cell proliferation.MethodsThis study included 10 malignant ascites specimens from patients undergoing ovarian cancer resection. Ascites samples were deeply phenotyped by 1H-NMR based metabolomics, blood-gas analyzer based gas flow analysis and flow cytomertry based a 13-plex cytokine panel. Characteristics of tumor cells were investigated in a 3D spheroid model by SEM and metabolic activity, adhesion, anti-apoptosis, migratory ability evaluated by MTT assay, adhesion assay, flowcytometry and scratch assay. The effect of different pH values was assessed by adding 10% malignant ascites to the test samples.Results The overall extracellular (peritoneal) environment was alkaline, with pH of ascites at stage II-III = 7.51 ± 0.16, and stage IV = 7.78 ± 0.16. Ovarian cancer spheroids grew rapidly in a slightly alkaline environment. Decreasing pH of the cell culture medium suppressed tumor features, metabolic activity, adhesion, anti-apoptosis, and migratory ability. However, 10% ascites could prevent tumor cells from being affected by acidic pH. Metabolomics analysis identified stage IV patients had significantly higher concentrations of alanine, isoleucine, phenylalanine, and glutamine than stage II-III patients, while stage II-III patients had significantly higher concentrations of 3-hydroxybutyrate. pH was positively correlated with acetate, and acetate positively correlated with lipid compounds. IL-8 was positively correlated with lipid metabolites and acetate. Glutathione and carnitine were negatively correlated with cytokines IL-6 and chemokines (IL-8 & MCP-1).ConclusionAlkaline malignant ascites facilitated ovarian cancer progression. Additionally, deep ascites phenotyping by metabolomics and cytokine investigations allows for a refined stratification of ovarian cancer patients. These findings contribute to the understanding of ascites pathology in ovarian cancer.

Diabetic Ketoacidosis and its Severity Predictors in Type 1 Diabetic Children; A 10-year Experience of A Teaching Hospital in Saudi Arabia.

OBJECTIVE: Our objective was to determine the trend and precipitating factors of the severity of diabetic ketoacidosis (DKA) in the population admitted to the Pediatric Intensive Care Unit (PICU) in a large teaching hospital in the Eastern region of Saudi Arabia. METHODS: We conducted a retrospective, analytical study at King Fahad Hospital, Imam Abdulrahman Bin Faisal University, Alkhobar, Saudi Arabia. We retrieved the complete medical records of 2234 children who were admitted to the PICU during the 10-year period of 2010 through 2019. The children included those with polydipsia, polyurea, abdominal pain, vomiting, dehydration, and weight loss, as well as breathing disturbances due to acidosis and CNS issues such as lethargy or coma and elevated blood glucose level, > 200 mg/dL [> 11.1 mmol/L], venous pH 7.3, serum total CO2 15 mmol/L, and blood- hydroxybutyrate concentration 3 mmol/L or moderate or severe ketonuria. RESULTS: Out of 2234 PICU admissions, 211 (9.4%) were diagnosed with DKA. A persistent increase in the rate of DKA ended up at 14.1% in 2019 (p = .005). The incidence of DKA was 88/2234 (3.93%). The severity of DKA was as follows: 130 (61.6%) had severe and 81 (38.4%) had moderate DKA. Excessive sweet intake without adding insulin in 83 (39.3%) patients and unhealthy lifestyles (35.1%) were the best predictors of severe DKA (p = .001). CONCLUSION: Over a 10-year period, the DKA pattern was persistently rising and slightly falling, which ended up at the significantly highest rate of 14.1% in 2019. URTI, pneumonia, unhealthy lifestyle, and excess sweet intake were significant precipitating factors associated with severe DKA.

Menopause modulates the circulating metabolome: evidence from a prospective cohort study.

We studied the changes in the circulating metabolome and their relation to the menopausal hormonal shift in 17β-oestradiol and follicle-stimulating hormone levels among women transitioning from perimenopause to early postmenopause. We analysed longitudinal data from 218 Finnish women, 35 of whom started menopausal hormone therapy during the study. The menopausal transition was monitored with menstrual diaries and serum hormone measurements. The median follow-up was 14 months (interquartile range: 8-20). Serum metabolites were quantified with targeted nuclear magnetic resonance metabolomics. The model results were adjusted for age, follow-up duration, education, lifestyle, and multiple comparisons. Menopause was associated with 85 metabolite measures. The concentration of apoB (0.17 standard deviation [SD], 99.5% confidence interval [CI] 0.03-0.31), very-low-density lipoprotein triglycerides (0.25 SD, CI 0.05-0.45) and particles (0.21 SD, CI 0.05-0.36), low-density lipoprotein (LDL) cholesterol (0.17 SD, CI 0.01-0.34) and particles (0.17 SD, CI 0.03-0.31), high-density lipoprotein (HDL) triglycerides (0.24 SD, CI 0.02-0.46), glycerol (0.32 SD, CI 0.07-0.58) and leucine increased (0.25 SD, CI 0.02-0.49). Citrate (-0.36 SD, CI -0.57 to -0.14) and 3-hydroxybutyrate concentrations decreased (-0.46 SD, CI -0.75 to -0.17). Most metabolite changes were associated with the menopausal hormonal shift. This explained 11% and 9% of the LDL cholesterol and particle concentration increase, respectively. Menopausal hormone therapy was associated with increased medium-to-large HDL particle count and decreased small-to-medium LDL particle and glycine concentration. Menopause is associated with proatherogenic circulating metabolome alterations. Female sex hormones levels are connected to the alterations, highlighting their impact on women's cardiovascular health.

Metabolic Responses to Epinephrine by Periparturient Dairy Cows Fed Prepartum Diets Differing in Predicted Metabolizable Protein Supply

We determined the effects of prepartum dietary metabolizable protein (MP) supply on lipolytic and glucose response to epinephrine stimulus during the periparturient period. Twelve non-lactating cows in second or greater pregnancies were assigned to a low MP (LMP) diet formulated to provide ~900 g/day MP or a high MP (HMP) diet to provide ~1,100 g/day MP. Cows received prepartum diets from 28 days before expected parturition to day of parturition and then received a common lactation diet until 35 days postpartum. Blood was sampled weekly to day −7, daily to day 7, and weekly through day 35 relative to parturition for analysis of glucose, non-esterified fatty acids (NEFAs), 3-hydroxybutyrate (BHB), urea N, and total protein concentrations. Epinephrine challenges were conducted on day −10 before expected parturition and days 7 and 14 postpartum. Epinephrine [1.4 μg/kg body weight (BW)] was administered via jugular vein and blood was collected from the opposite jugular vein from −45 min to +120 min relative to epinephrine administration. Plasma was analyzed for glucose and NEFA concentrations. Prepartum dry matter intake (DMI) (14.7 and 12.6 ± 1.5 kg/day for LMP and HMP, respectively), postpartum DMI (21.4 and 19.4 ± 1.3 kg/day for LMP and HMP, respectively), and 4% fat-corrected milk (FCM) (37.0 and 36.8 ± 1.5 kg/day for LMP and HMP, respectively) did not differ significantly between diets. Cows fed HMP had higher plasma concentrations of urea N prepartum than cows fed LMP (14.0 vs. 7.9 ± 0.8 mg/dl). Cows fed HMP tended to have greater prepartum concentrations of total protein in plasma than those fed LMP (7.5 vs. 7.0 ± 0.2 g/dl). The area under the curve (AUC) for NEFA response to epinephrine did not differ between diets, but differed by day relative to parturition [8,284, 29,018, and 18,219 ± 2,302 min × (μeq/l) for days −10, 7, and 14, respectively]. Maximal response of NEFA concentration to epinephrine was greater for HMP than for LMP (744 vs. 438 ± 72 μeq/l). The glucose AUC did not differ between diets or among days. Lipolytic response, but not glucose response, to epinephrine was enhanced during the early postpartum period relative to the late dry period. However, those responses were not affected by prepartum MP supply.

Multiscale modeling of the microbial production of polyhydroxyalkanoates using two carbon sources

Polyhydroxyalkanoates are a promising material for a broad range of plastic products. Due to the high production costs, the proportion of industrial produced polyhydroxyalkanoates is low compared to conventional plastics. One possibility to reduce the costs is to reduce the price of the substrates using organic carbon-rich wastes streams. Using mathematical modeling, the microbial production process can be optimized by adjusting process conditions like the oxygen supply or initial substrate ratios. This contribution outlines a multiscale model for the poly(3-hydroxybutyrate) production in Cupriavidus necator using the carbon-sources fructose and acetate. The model comprises a hybrid cybernetic model to describe the macroscopic dynamics and a polymerization model which describes the chain length dynamics. The multiscale model is used to analyze the effects of different initial carbon to ammonium ratios and dissolved oxygen levels on the poly(3-hydroxybutyrate) concentration and chain length distribution.

Ketogenic diet and chemotherapy combine to disrupt pancreatic cancer metabolism and growth

Ketogenic diet is a potential means of augmenting cancer therapy. Here, we explore ketone body metabolism and its interplay with chemotherapy in pancreatic cancer. Metabolism and therapeutic responses of murine pancreatic cancer were studied using KPC primary tumors and tumor chunk allografts. Mice on standard high-carbohydrate diet or ketogenic diet were treated with cytotoxic chemotherapy (nab-paclitaxel, gemcitabine, cisplatin). Metabolic activity was monitored with metabolomics and isotope tracing, including 2H- and 13C-tracers, liquid chromatography-mass spectrometry, and imaging mass spectrometry. Ketone bodies are unidirectionally oxidized to make NADH. This stands in contrast to the carbohydrate-derived carboxylic acids lactate and pyruvate, which rapidly interconvert, buffering NADH/NAD. In murine pancreatic tumors, ketogenic diet decreases glucose's concentration and tricarboxylic acid cycle contribution, enhances 3-hydroxybutyrate's concentration and tricarboxylic acid contribution, and modestly elevates NADH, but does not impact tumor growth. In contrast, the combination of ketogenic diet and cytotoxic chemotherapy substantially raises tumor NADH and synergistically suppresses tumor growth, tripling the survival benefits of chemotherapy alone. Chemotherapy and ketogenic diet also synergize in immune-deficient mice, although long-term growth suppression was only observed in mice with an intact immune system. Ketogenic diet sensitizes murine pancreatic cancer tumors to cytotoxic chemotherapy. Based on these data, we have initiated a randomized clinical trial of chemotherapy with standard versus ketogenic diet for patients with metastatic pancreatic cancer (NCT04631445).

Subclinical ketosis as a factor reduction of reproductive indicators of high productive cows

Purpose: The aim of the research is to study the effect of subclinical ketosis (SCK) on the effectiveness of artificial insemination of highly productive dairy cows in the postpartum period.Materials and methods. The studied animals were selected according to the principle of conditional analogs and were divided into 2 groups of 6 animals each. Group 1 - animals with a blood concentration of 3- hydroxybutyrate˂ 1.0 mmol / l, group 2 - animals with subclinical ketosis with a concentration of 3-hydroxybutyrate in the blood in the range of 1.0-1.4 mmol / l. Blood was collected from animals on the 5th, 15th and 33-35th days after calving. In the obtained blood serum samples, the content of total protein, albumin, urea, glucose, cholesterol, triglycerides, calcium, phosphorus, magnesium, creatinine, total bilirubin, the activity of transaminase enzymes and alkaline phosphatase was determined. The concentration of steroid hormones has been determined. Before artificial insemination, the animals were synchronized according to the pressinovsinh scheme.Results. Most of the biochemical blood parameters did not go beyond the reference values. By the time of hormonal stimulation of sexual heat in cows with SCK, a decrease in 3-hydroxybutyrate (˂ 1,0 mmol / l) and an increase in glucose (&gt; 3.0 mmol / l) were recorded. On the 15th day after calving, in the group with SCK, there was an increase in total bilirubin, cholesterol, triglycerides, and magnesium (p˂ 0,01). On the 15th day after calving, testosterone levels were 13% higher in the SCK group. By the time of synchronization, the level of progesterone in the group of animals with SCK was lower in comparison with the control group. In the group of animals with SCK, the lowest reproductive indices were recorded: the insemination index in the group with SSC was higher by 46%, the service period was 49 days longer, the interbody period was significantly longer by 68 days (p˂ 0,05).Conclusion. Subclinical ketosis, recorded on the 15th day after calving, helps to reduce the reproductive performance of high -yielding dairy cows. Normalization of the concentration of 3-hydroxybutyrate from subclinical to physiologically normal values, and subsequent hormonal synchronization do not reduce this negative effect of SCK.

Impact of tight glucose control on circulating 3-hydroxybutyrate in critically ill patients

BackgroundRecent evidence suggests a potentially protective effect of increasing ketone body availability via accepting low macronutrient intake early after onset of critical illness. The impact of blood glucose control with insulin on circulating ketones is unclear. Whereas lowering blood glucose may activate ketogenesis, high insulin concentrations may have the opposite effect. We hypothesized that the previously reported protective effects of tight glucose control in critically ill patients receiving early parenteral nutrition may have been mediated in part by activation of ketogenesis.MethodsThis is a secondary analysis of 3 randomized controlled trials on tight versus liberal blood glucose control in the intensive care unit, including 700 critically ill children and 2748 critically ill adults. All patients received early parenteral nutrition as part of the contemporary standard of care. Before studying a potential mediator role of circulating ketones in improving outcome, we performed a time course analysis to investigate whether tight glucose control significantly affected ketogenesis and to identify a day of maximal effect, if any. We quantified plasma/serum 3-hydroxybutyrate concentrations from intensive care unit admission until day 3 in 2 matched subsets of 100 critically ill children and 100 critically ill adults. Univariable differences between groups were investigated by Kruskal-Wallis test. Differences in 3-hydroxybutyrate concentrations between study days were investigated by Wilcoxon signed-rank test.ResultsIn critically ill children and adults receiving early parenteral nutrition, tight glucose control, as compared with liberal glucose control, lowered mean morning blood glucose on days 1–3 (P < 0.0001) via infusing insulin at a higher dose (P < 0.0001). Throughout the study period, caloric intake was not different between groups. In both children and adults, tight glucose control did not affect 3-hydroxybutyrate concentrations, which were suppressed on ICU days 1–3 and significantly lower than the ICU admission values for both groups (P < 0.0001).ConclusionTight versus liberal glucose control in the context of early parenteral nutrition did not affect 3-hydroxybutyrate concentrations in critically ill patients. Hence, the protective effects of tight glucose control in this context cannot be attributed to increased ketone body availability.

Metabolomics profiling predicts outcome of tocilizumab in rheumatoid arthritis: an exploratory study

To study metabolic signatures can be used to identify predictive biomarkers for a patient's therapeutic response. We hypothesized that the characterization of a patients' metabolic profile, utilizing one-dimensional nuclear magnetic resonance (1H-NMR), may predict a response to tocilizumab in patients with rheumatoid arthritis (RA). 40 active RA patients meeting the 2010 ACR/EULAR classification criteria initiating treatment with tocilizumab were recruited. Clinical outcomes were determined at baseline, and after six and twelve months of treatment. EULAR response criteria at 6 and 12months to categorize patients as responders and non-responders. Blood was collected at baseline and after six months of tocilizumab therapy. 1H-NMR was used to acquire a spectra of plasma samples. Chenomx NMR suite 8.5 was used for metabolite identification and quantification. SPSS v.27 and MetaboAnalyst 4.0 were used for statistical and pathway analysis. Isobutyrate, 3-hydroxybutyrate, lysine, phenylalanine, sn-glycero-3-phosphocholine, tryptophan and tyrosine were significantly elevated in responders at the baseline. OPLS-DA at baseline partially discriminated between RA responders and non-responders. A multivariate diagnostic model showed that concentrations of 3-hydroxybutyrate and phenylalanine improved the ability to specifically predict responders classifying 77.1% of the patients correctly. At 6months, levels of methylamine, sn-glycero-3-phosphocholine and tryptophan tended to still be low in non-responders. The relationship between plasma metabolic profiles and the clinical response to tocilizumab suggests that 1H-NMR may be a promising tool for RA therapy optimization. More studies are needed to determine if metabolic profiling can predict the response to biological therapies in RA patients.

Effects of different sodium salts and nitrogen sources on the production of 3-hydroxybutyrate and polyhydroxybutyrate by Burkholderia cepacia

The effects of NaCl, Na2SO4, Na2HPO4, and Na3C6H5O7 on the production of 3-hydroxybutyrate, polyhydroxybutyrate, and by-products by Burkholderia cepacia. Proper addition of Na3C6H5O7 can significantly promote the production of 3-hydroxybutyric acid and polyhydroxybutyrate. The concentration, productivity, and yield of 3-hydroxybutyrate were increased by 48.2%, 55.6%, and 48.3% at 16 mM Na3C6H5O7. The increases of 80.1%, 47.1%, and 80.0% in the concentration, productivity, and yield of polyhydroxybutyrate were observed at 12 mM Na3C6H5O7. Na2SO4 and Na2HPO4 also have positive effects on the production capacity of 3-hydroxybutyrate and polyhydroxybutyrate within a certain range of concentration. NaCl is not conducive to the improvement of fermentation efficiency. Compared with a single nitrogen source, a mixed nitrogen source is more conducive to enhancing the production of 3-hydroxybutyrate and polyhydroxybutyrate.

Adenomyosis is associated with specific proton nuclear magnetic resonance (1H-NMR) serum metabolic profiles

To determine whether the adenomyosis phenotype affects the proton nuclear magnetic resonance (1H-NMR)-based serum metabolic profile of patients. Cohort study. University hospital-based research center. Seventy-seven patients who underwent laparoscopy for a benign gynecologic condition. Pelvic magnetic resonance imaging and collection of a venous peripheral blood sample were performed during the preoperative workup. The women were allocated to the adenomyosis group (n = 32), or the control group (n = 45). The adenomyosis group was further subdivided into two groups: diffuse adenomyosis of the inner myometrium (n = 14) and focal adenomyosis of the outer myometrium (n = 18). Other adenomyosis phenotypes were excluded. Metabolomic profiling based on 1H-NMR spectroscopy in combination with statistical approaches. The serum metabolic profiles of the patients with adenomyosis indicated lower concentrations of 3-hydroxybutyrate, glutamate, and serine compared with controls. Conversely, the concentrations of proline, choline, citrate, 2-hydroxybutyrate, and creatinine were higher in the adenomyosis group. The focal adenomyosis of the outer myometrium and the diffuse adenomyosis phenotypes also each exhibited a specific metabolic profile. Serum metabolic changes were detected in women with features of adenomyosis compared with their disease-free counterparts, and a number of specific metabolic pathways appear to be engaged according to the adenomyosis phenotype. The metabolites with altered levels are particularly involved in immune activation as well as cell proliferation and cell migration. Nevertheless, this study did find evidence of a correlation between metabolite levels and symptoms thought to be related to adenomyosis. Further studies are required to determine the clinical significance of these differences in metabolic profiles.

Effects of vaccination against brucellosis and clostridia on the intake, performance, feeding behavior, blood parameters, and immune responses of dairy heifers calves.

The aim of this study was to identify possible effects of different vaccination strategies (concomitantly or not) against brucellosis and clostridia on intake, performance, feeding behavior, blood parameters, and immune responses of dairy heifers calves. Fifty heifers calves were enrolled [38 Gyr (Zebu, Bos taurus indicus) and 12 5/8 Holstein × Gyr]. At 120 d of age, animals were randomly distributed among 3 groups: B (n = 18), vaccinated against brucellosis; C (n = 14), vaccinated against clostridia and CB (n = 18), vaccinated concomitantly for both. Rectal and thermographic temperatures were evaluated on days -1, 0, 1, 2, 3, 5, 7,10, 14, and 28 relatives to the vaccination day. Feed and water intake, body weight (BW), and feeding behavior were monitored daily by an electronic feeding system. Blood was sampled on days 0, 3, 7, 14, and 28, relative to the vaccination day for determination of glucose and β -hydroxybutyrate (BHBA) concentrations. Blood sampled on day 0 (prevaccination) and on days 28 and 42 were used to evaluate the immune response against Brucella abortus and clostridia. There was an increase in rectal temperature between the first and the third day postvaccination in the 3 groups. The thermography revealed an increase of local temperature for 7 d on groups B and CB. Group C had increased local temperature for a longer period, lasting for up to 14 d. Dry mater intake was reduced for groups B and CB, but no alteration was observed for group C. No alterations regarding initial BW, final BW, average daily weight gain, and feed efficiency were observed. No differences were observed for the 3 vaccination groups for blood parameters throughout the evaluation period. The concomitant vaccination against brucellosis and clostridia led to lower neutralizing antibody titers against epsilon toxin of Clostridium perfringens and botulinum toxin type C of C. botulinum (C > CB > B). When cellular proliferation assay and serological tests to B. abortus were evaluated, no differences were observed between groups B and CB. The present results indicate that the concomitant vaccination against brucellosis and clostridia has no relevant impact on the intake, performance, and feeding behavior of dairy calves. However, the concomitant vaccination of vaccines against these 2 pathogens impacts animal immunity against clostridial infections.