UNASSIGNED: In dairy cattle, oxidative stress is a predominant problem associated with diseases and reproductive health issues. This study aimed to detect the variation in the antioxidant biomarkers by adding different concentrations of β-hydroxybutyric acid (BHBA) and sought to elucidate its effects on the gene expression levels of growth hormone (GH) and antioxidant biomarkers in bovine hepatocytes.
UNASSIGNED: Four antioxidant biomarkers, namely malondialdehyde (MDA), superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GSH Px) were evaluated using commercially available bovine ELISA kits. The expression levels of the bovine GH, its receptor (GHR), insulin-like growth factor (IGF), IGF-1, IGF-1 receptor, CAT, SOD, GSH-Px and β-actin (as a reference) genes in liver cell culture were determined by reverse transcriptase-PCR assay.
UNASSIGNED: With the increase of BHBA concentration and culture time, the activities of SOD, CAT, and GSH Px biomarkers in hepatocytes decreased. However, the content of MDA in hepatocytes increased gradually with the increase of hepatocyte culture time and BHBA concentration. The qPCR results revealed that after adding BHBA, gene expression levels of GSH-Px, SOD and IGF biomarkers in hepatocytes began to differ in the culture groups at 12 h, whereas the gene expression level of the CAT and GHR biomarkers in hepatocytes began to differ at 6 h.
UNASSIGNED: Quantitative PCR results showed that the BHBA significantly downregulated the expression levels of the GHR gene and CAT, GSH Px and SOD antioxidant biomarker genes.

BACKGROUND: Mitochondrial redox imbalance underlies the pathophysiology of type2 diabetes mellitus (T2DM), and is closely related to tissue damage and dysfunction. Studies have shown the beneficial effects of dietary strategies that elevate β-hydroxybutyrate (BHB) levels in alleviating T2DM. Nevertheless, the role of BHB has not been clearly elucidated.
METHODS: We performed a spectral study to visualize the preventive effects of BHB on blood and multiorgan mitochondrial redox imbalance in T2DM mice via using label-free resonance Raman spectroscopy (RRS), and further explored the impact of BHB therapy on the pathology of T2DM mice by histological and biochemical analyses.
RESULTS: Our data revealed that RRS-based mitochondrial redox states assay enabled clear and reliable identification of the improvement of mitochondrial redox imbalance by BHB, evidenced by the reduction of Raman peak intensity at 750 cm-1, 1128 cm-1 and 1585 cm-1 in blood, tissue as well as purified mitochondria of db/db mice and the increase of tissue mitochondrial succinic dehydrogenase (SDH) staining after BHB treatment. Exogenous supplementation of BHB was also found to attenuate T2DM pathology related to mitochondrial redox states, involving organ injury, blood glucose control, insulin resistance and systemic inflammation.
CONCLUSIONS: Our findings provide strong evidence for BHB as a potential therapeutic strategy targeting mitochondria for T2DM.

UNASSIGNED: The ketone body β-hydroxybutyric acid (BHB) plays critical roles in cellular proliferation and metabolic fuel utilization; however, its effects on the rumen microbiota remain unknown.
UNASSIGNED: Here, three doses of BHB (low, medium, and high) were supplemented to early-weaned goat kids.
UNASSIGNED: Compared with controls, the beneficial effects of BHB on growth and rumen development were observed in goats at 90 days of age (d). The low dose of dietary BHB increased the concentration of rumen acetate, propionate, and butyrate on d90. The sequencing results of the rumen microbiota revealed marked shifts in rumen microbial community structure after early-weaned goat kids consumed BHB for 2 months. The signature bacterial ASVs for each treatment were identified and were the main drivers contributing to microbial interactions in the rumen. The bacteria associated with rumen weight were also correlated with body weight. Some classified bacterial signatures, including Prevotella, Olsenella umbonate, and Roseburia faecis, were related to rumen volatile fatty acids and host development.
UNASSIGNED: Overall, dietary BHB altered rumen microbiota and environments in young goats, which contributed to rumen development and growth.

The continued increase in milk production during the last century has not been accompanied by an adequate dry matter intake (DMI) by cows, which therefore experience a negative energy balance (NEB). NEB is low and of minor importance at low milk yield (MY), such as for the nutrition of one calf, and under these circumstances is considered \"natural\". MY and low DMI around parturition are correlated and are the reason for the genetic correlation between increasing MY and increasing NEB up to 2000 MJ or more for 2-3 months postpartum in high-genetic-merit dairy cows. The extension and duration of NEB in high-producing cows cannot be judged as \"natural\" and are compensated by the mobilization of nutrients, particularly of fat. The released non-esterified fatty acids (NEFAs) overwhelm the metabolic capacity of the cow and lead to the ectopic deposition of NEFAs as triglycerides (TGs) in the liver. The subsequent lipidosis and the concomitant hampered liver functions cause subclinical and clinical ketosis, both of which are associated with \"production diseases\", including oxidative and endoplasmatic stress, inflammation and immunosuppression. These metabolic alterations are regulated by homeorhesis, with the priority of the physiological function of milk production. The prioritization of one function, namely, milk yield, possibly results in restrictions in other physiological (health) functions under conditions of limited resources (NEB). The hormonal framework for this metabolic environment is the high concentration of growth hormone (GH), the low concentration of insulin in connection with GH-dependent insulin resistance and the low concentration of IGF-1, the so-called GH-IGF-1 axis. The fine tuning of the GH-IGF-1 axis is uncoupled because the expression of the growth hormone receptor (GHR-1A) in the liver is reduced with increasing MY. The uncoupled GH-IGF-1 axis is a serious impairment for the GH-dependent stimulation of gluconeogenesis in the liver with continued increased lipolysis in fat tissue. It facilitates the pathogenesis of lipidosis with ketosis and, secondarily, \"production diseases\". Unfortunately, MY is still increasing at inadequate DMI with increasing NEB and elevated NEFA and beta-hydroxybutyric acid concentrations under conditions of low glucose, thereby adding health risks. The high incidences of diseases and of early culling and mortality in dairy cows are well documented and cause severe economic problems with a waste of resources and a challenge to the environment. Moreover, the growing public concerns about such production conditions in agriculture can no longer be ignored.

Heat stress has multiple potential effects on the brain, such as neuroinflammation, neurogenesis defects, and cognitive impairment. β-hydroxybutyric acid (BHBA) has been demonstrated to play neuroprotective roles in various models of neurological diseases. In the present study, we investigated the efficacy of BHBA in alleviating heat stress-induced impairments of adult hippocampal neurogenesis and cognitive function, as well as the underlying mechanisms. Mice were exposed to 43 ℃ for 15 min for 14 days after administration with saline, BHBA, or minocycline. Here, we showed for the first time that BHBA normalized memory ability in the heat stress-treated mice and attenuated heat stress-impaired hippocampal neurogenesis. Consistently, BHBA noticeably improved the synaptic plasticity in the heat stress-treated hippocampal neurons by inhibiting the decrease of synapse-associated proteins and the density of dendritic spines. Moreover, BHBA inhibited the expression of cleaved caspase-3 by suppressing endoplasmic reticulum (ER) stress, and increased the expression of brain-derived neurotrophic factor (BDNF) in the heat stress-treated hippocampus by activating the protein kinase B (Akt)/cAMP response element binding protein (CREB) and methyl-CpG binding protein 2 (MeCP2) pathways. These findings indicate that BHBA is a potential agent for improving cognitive functions in heat stress-treated mice. The action may be mediated by ER stress, and Akt-CREB-BDNF and MeCP2 pathways to improve adult hippocampal neurogenesis and synaptic plasticity.

BACKGROUND: Idiopathic epilepsy (IE) is a common, chronic brain dysfunction in dogs. Recently, the effect of feeding a diet enriched with medium-chain triglycerides (MCTs) on seizure frequency has been evaluated in several studies in dogs with IE. However, most dogs with IE in previous studies were treated with phenobarbital as the main antiseizure medication (ASM). In Japan, zonisamide (ZNS) is the most prescribed ASM for dogs with IE. The interaction between ZNS and various nutrients including MCTs and the potential effects on treatment efficacy resulting from combining these therapies have not been previously studied. A prospective, randomized, double-blinded, placebo-controlled, crossover dietary study was conducted. Dogs (n = 7) treated with ZNS were fed either a placebo diet (PL) or Purina ProPlan Veterinary Diet NeuroCare (NC) for 3 months, after which treatments were crossed over and continued for another 3 months. Seizure frequency (seizures/month; sz/m), blood tests including concentrations of ZNS and β-hydroxybutyric acid, and owner\'s visual analogue scale score were collected from all dogs for both treatment periods.
RESULTS: There was no significant difference in the seizure frequency between PL (2.95 ± 0.80 sz/m) and NC (1.90 ± 0.57 sz/m) during the 6 months of trial. Three of 7 dogs showed ≥ 50% seizure reduction, and 1 of those 3 dogs achieved seizure freedom in NC period. However, 2 of 7 dogs had no changes in epileptic seizure frequency, 2 of 7 dogs had a deterioration in seizure frequency in the NC period. Feeding the MCT diet concurrent with ZNS showed no apparent adverse effects and did not affect ZNS concentration.
CONCLUSIONS: This study indicated that the commercially available MCT-enriched diet (NC) can be safely used concurrently with ZNS for dogs with IE.

To investigate the association between circulating β-hydroxybutyric acid (βOHB) and diabetic kidney disease (DKD) risk in patients with type 2 diabetes (T2D).
A total of 1388 patients with T2D were recruited. Participants were divided into high and normal βOHB groups. Participants in the normal βOHB group were divided into four subgroups according to βOHB quartile (Q). The relationships of βOHB with DKD and DKD subtype were analysed using chi-square and binary logistic regression. Restricted cubic splines were used to explore the non-linear correlation between βOHB concentration and DKD risk in the total population.
A higher prevalence of DKD was detected in the high compared with the normal βOHB group (43.3% vs. 33.3%, P = .041). Participants in the Q4 group (βOHB, 0.12-0.30 mM) had the lowest prevalence of DKD (P = .001). In the binary logistic regression model, the multivariable-adjusted odds ratios (ORs) (95% confidence intervals [CIs]) for DKD risk were 2.30 (1.62-3.26) for Q1, 1.80 (1.23-2.62) for Q2 and 1.63 (1.10-2.41) for Q3 relative to Q4 (P < .001). Restricted cubic spline analyses suggested a J-shaped association of circulating βOHB concentration with DKD risk. DKD risk was lowest at a serum βOHB concentration of 0.183 mM (OR, 0.63; 95% CI, 0.52-0.77).
A J-shaped relationship between circulating ketone level and DKD risk in patients with T2D was determined. Circulating βOHB in the range of 0.12-0.30 mM was associated with a lower risk of DKD. Further studies are warranted to verify the causality and to elucidate the underlying mechanisms.

Metabolic and oxidative stress have been characterized as risk factors during the transition period from pregnancy to lactation. Although mutual relations between both types of stress have been suggested, they rarely have been studied concomitantly. For this, a total of 99 individual transition dairy cows (117 cases, 18 cows sampled during 2 consecutive lactations) were included in this experiment. Blood samples were taken at -7, 3, 6, 9, and 21 d relative to calving and concentrations of metabolic parameters (glucose, β-hydroxybutyric acid (BHBA), nonesterified fatty acids, insulin, insulin-like growth factor 1, and fructosamine) were determined. In the blood samples of d 21, biochemical profiles related to liver function and parameters related to oxidative status were determined. First, cases were allocated to 2 different BHBA groups (ketotic vs. nonketotic, N:n = 20:33) consisting of animals with an average postpartum BHBA concentration and at least 2 out of 4 postpartum sampling points exceeding 1.2 mmol/L or remaining below 0.8 mmol/L, respectively. Second, oxidative parameters [proportion of oxidized glutathione to total glutathione in red blood cells (%)], activity of glutathione peroxidase, and of superoxide dismutase, concentrations of malondialdehyde and oxygen radical absorbance capacity were used to perform a fuzzy C-means clustering. From this, 2 groups were obtained [i.e., lower antioxidant ability (LAA80%, n = 31) and higher antioxidant ability (HAA80%, n = 19)], with 80% referring to the cutoff value for cluster membership. Increased concentrations of malondialdehyde, decreased superoxide dismutase activity, and impaired oxygen radical absorbance capacity were observed in the ketotic group compared with the nonketotic group, and inversely, the LAA80% group showed increased concentrations of BHBA. In addition, the concentration of aspartate transaminase was higher in the LAA80% group compared with the HAA80% group. Both the ketotic and LAA80% groups showed lower dry matter intake. However, a lower milk yield was observed in the LAA80% group but not in the ketotic group. Only 1 out of 19 (5.3%) and 3 out of 31 (9.7%) cases from the HAA80% and LAA80% clusters belong to the ketotic and nonketotic group, respectively. These findings suggested that dairy cows vary in oxidative status at the beginning of the lactation, and fuzzy C-means clustering allows to classify observations with distinctive oxidative status. Dairy cows with higher antioxidant capacity in early lactation rarely develop ketosis.

In recent times, advances in the field of metabolomics have shed greater light on the role of metabolic disturbances in neuropsychiatric conditions. The following review explores the role of ketone bodies and ketosis in both the diagnosis and treatment of three major psychiatric disorders: major depressive disorder, anxiety disorders, and schizophrenia. Distinction is made between the potential therapeutic effects of the ketogenic diet and exogenous ketone preparations, as exogenous ketones in particular offer a standardized, reproducible manner for inducing ketosis. Compelling associations between symptoms of mental distress and dysregulation in central nervous system ketone metabolism have been demonstrated in preclinical studies with putative neuroprotective effects of ketone bodies being elucidated, including effects on inflammasomes and the promotion of neurogenesis in the central nervous system. Despite emerging pre-clinical data, clinical research on ketone body effectiveness as a treatment option for psychiatric disorders remains lacking. This gap in understanding warrants further investigating, especially considering that safe and acceptable ways of inducing ketosis are readily available.

BACKGROUND: Cough variant asthma (CVA) is a chronic inflammatory disease characterized by cough as the main symptom. Suhuang antitussive capsule (Suhuang), one of traditional Chinese patent medicines, mainly treats CVA clinically. Previous studies have shown that Suhuang significantly improved CVA, post-infectious cough (PIC), sputum obstruction and airway remodeling. However, the effect of Suhuang on ovalbumin-induced (OVA-induced) metabolic abnormalities in CVA is unknown.
OBJECTIVE: This study aimed to identify potential metabolites associated with efficacy of Suhuang in the treatment of CVA, and determined how Suhuang regulates metabolites, and differential metabolites reduce inflammation and oxidative stress.
METHODS: Rats were given 1 mg OVA/100 mg aluminum hydroxide in the 1st and 7th days by intraperitoneal injection and challenged by atomizing inhalation of 1% OVA saline solution after two weeks to establish the CVA model. Rats were intragastrically (i.g.) administrated with Suhuang at 1.4 g/kg and β-hydroxybutyric acid (β-HB) were given with different concentrations (87.5 and 175 mg/kg/day) by intraperitoneal injection for 2 weeks. After 26 days, GC-MS-based metabolomic approach was applied to observe metabolic changes and search differential metabolites. The number of coughs, coughs latencies, enzyme-linked immunosorbent assay (ELISA), histological analysis and quantitative-polymerase chain reaction (Q-PCR) were used to investigate the effects of Suhuang. Then β-HB on CVA rats, NLRP3 inflammasome and GSK3β/AMPK/Nrf2 signalling pathway were detected by western blotting.
RESULTS: The results showed that Suhuang treatment significantly enhanced the serum level of β-HB. Interestingly, exposure to exogenous β-HB was also protective against OVA-induced CVA. β-HB significantly reduced the number of coughs and lengthened coughs latencies, improved lung injury, reduced the secretion of various cytokines, and directly inhibited the NLRP3 inflammasome. In addition, β-HB increased the nuclear accumulation of Nrf2 by activating the GSK3β/AMPK signaling axis, and then inactivating the NF-κB signaling pathway, effectively protecting OVA-induced CVA from oxidative stress and inflammation.
CONCLUSIONS: The results of this study shows that β-HB can reduce inflammation and oxidative stress, the increased production of β-HB in serum might be the crucial factor for Suhuang to exert its effect in the treatment of CVA.