Concentrations Of Aspartate Research Articles

Biochemical composition and transcriptome analysis revealed nutrients consumption and physiological characteristics of Chinese sturgeon larvae during yolk-sac stage

During the yolk stage, the nutrients of larvae consumed are from the yolk sac, which greatly affect the survival and quality of fish. The aim of this study was to investigate the metabolic changes, physiological characteristics and the related molecular mechanism of Chinese sturgeon (Acipenser sinensis) larvae during yolk stage by biochemical analyses in combination with transcriptome analysis of larvae at three developmental stages (vertical, flat and benthic). The results showed that both total amino acid and protein contents showed a trend of first decreasing and then increasing. The lipid content was stable in the early stages, but decreased significantly in the benthic stage (P < 0.05). In terms of amino acid profile, the contents of leucine, isoleucine and lysine were relatively high in essential amino acids (EAAs), and the contents of glutamate, aspartate and serine were relatively high in non-essential amino acid (NEAA). As for fatty acid content, the most abundant fatty acids are C16:0 (palmitic acid), C18:1 (oleic acid), C18:2n-6 (linoleic acid) and C22:6n-3 (docosahexaenoic acid, DHA). The results of transcriptome analysis showed that 4936 genes were up-regulated and 3079 genes were down-regulated from vertical stage to flat stage, and 1703 genes were up-regulated and 837 genes were down-regulated from flat stage to benthic stage, respectively. According to KEGG enrichment analysis on the differentially expressed genes (DEGs), the up-regulated DEGs mGlu3, mGlu5, NMDAR, PepT1, TAT1, EAAT3, CYP51A1 and CYP7A1 were significantly enriched in pathways related to the nervous system, protein digestion and absorption, steroid synthesis, cholesterol metabolism and bile acid biosynthesis in early development of larvae. Down-regulated DEGs Drg1, NHP2, KRE33 and MPP10 were enriched in the ribosome biogenetic pathway. In late development, up-regulated DEGs, cGAS, IRF3, TRAF2, MHC-I, MHC-II and TAR2, were enriched in pathways associated with innate and adaptive immunity. Down-regulated DKK2 and FGFRs, ITGAs were enriched in Wnt signaling pathway and PI3K-AKT pathway, respectively. In summary, this study revealed that the larval fish consumed protein as the main nutrient from the vertical stage to the flat stage, while mainly consumed lipid from the flat stage to the benthic stage, and highlighted the pathways of DEGs involved in the nervous system and immune system, which would provide new ideal to the high quality larvae production in Chinese sturgeon, thus benefit species protection.

The Study of the Influence of Pyridine Oxime Derivative (GIZh-298) And Sodium Valproate on the Neurotransmitter Amino Acids Content in the Brain Structures of Mice in the Maximal Electroconvulsive Seizures Test

The effect of the antiepileptic compound GIZH-298 and the reference drug sodium valproate (VN) on the content of excitatory and inhibitory amino acids in the frontal cortex, hypothalamus, striatum, and hippocampus of the brain of mice after an electric shock-induced generalized tonic-clonic seizure (MES) was studied. It has been shown that the application of MES reduces the levels of aspartate, glycine, GABA and the ratio of “GABA/glutamate” in the hypothalamus. In the latter, a decrease in the concentration of glutamate, glycine and taurine was noted. VN (200 mg/kg/g) counteracts the effects of MES by interfering with the resulting decrease in GABA and the GABA/glutamate ratio in the hypothalamus. In the hypothalamus, striatum, and hippocampus, VN causes a decrease in the aspartate content both in intact control groups and in the group of mice that received MES. GIZH-298 (60 mg/kg/int. gastric) prevented the MES-induced decrease in the levels of GABA, glycine and taurine and the GABA/glutamate ratio in the hypothalamus.

Genome-Scale Modeling of CHO Cells Unravel the Critical Role of Asparagine in Cell Culture Feed Media.

Amino acids, including asparagine, aspartate, glutamine, and glutamate, play important roles in purine and pyrimidine biosynthesis as well as serve as anaplerotic sources fueling the tricarboxylic acid (TCA) cycle for mitochondrial energy generation. Despite extensive studies on glutamine and glutamate in CHO cell cultures, the roles of asparagine and aspartate, especially in feed media, remain underexplored. In this study, we utilized a CHO genome scale model to first deeply characterize the intracellular metabolic states of CHO cells cultured in different combinations of basal and feed media to understand the traits of asparagine/aspartate-dependent and glutamate-dependent feeds. Subsequently, we identified the critical role of asparagine and aspartate in the feed media as anaplerotic sources and conducted in silico simulations to ascertain their optimal ratios to improve cell culture performance. Finally, based on the model simulations, we reformulated the feed media by tailoring the concentrations of asparagine and aspartate. Our experimental data reveal a CHO cell preference for asparagine compared with aspartate, and thus maintaining an optimal ratio of these amino acids is a key factor for achieving optimal CHO cell culture performance in biopharmaceutical production.

UHPLC-QQQ-MS/MS method for the simultaneous quantification of 18 amino acids in various meats.

Amino acids are an essential source of human protein, and their content and composition are the main factors determining food protein utilization rate. Determining amino acids is essential in the component analysis of food. Therefore, a groundbreaking technique was developed utilizing ultra-high performance liquid chromatography interfaced with a triple quadrupole mass spectrometer (UHPLC-QQQ-MS/MS) for concurrently quantifying 18 amino acids across various types of meat. According to the test results, it can be known that the average content of glutamate (2.03 × 104 ± 3.94 × 103 μg/g in pig feet) was the highest in all meat samples, and the content of aspartate (0.0945 ± 0.0950 μg/g in pork) was the lowest, which was not detected in some samples such as beef and lean meat. Orthogonal partial least-squares discrimination analysis (OPLS-DA) showed: (1) 13 amino acids (arginine, valine, serine, alanine, lysine, glycine, asparagine, methionine, proline, threonine, glutamate, phenylalanine, and leucine, VIP > 1) were used as characteristic amino acids between pork and pig feet; (2) serine, threonine, alanine, histidine, asparagine, and arginine (VIP > 1) were used as signature amino acids in different components of pork (lean meat, fat, and pigskin); (3) asparagine, glutamate, histidine, tyrosine, and valine (VIP > 1) were considered as signature amino acids in different types of meats (pork, mutton, beef, and chicken). This study provides a new UHPLC-QQQ-MS/MS method for the determination of amino acid content in meat and also provides data support for the comprehensive evaluation of the nutritional value of foods containing amino acids.

12339 Rotating Insulin Injection Sites Isn't Enough: A Challenging Case Of Insulin-Induced Lipoatrophic Panniculitis

Abstract Disclosure: J. Hand: None. S. Nemiroff: None. C. Zmijewski: None. C.J. Levy: None. Introduction: Insulin-induced localized lipoatrophy is a rare phenomenon that involves the loss of subcutaneous adipose tissue around sites of insulin injections, and it often presents as a deep, retracted scar on the skin. Its prevalence has dropped significantly given improvements in recombinant and purified insulin and its analogs, from 55% to more recently 1-2%. Lipoatrophy can cause erratic absorption of insulin, leading to suboptimal glycemic control and hypoglycemia. The pathophysiology is not completely understood but is thought to be immune-mediated. Treatment options are limited and often focus on rotating injection sites or changing the type of insulin, though some case reports alternatively describe using corticosteroid injections. Here, we report an unusual case where a patient developed a form of early, localized lipoatrophy called lipoatrophic panniculitis due to insulin. Clinical Case: A 28-year-old male with a 13-year history of type 1 diabetes presented for insulin optimization. The patient was using multiple daily injection (MDI) insulin therapy and later an insulin pump, with careful infusion site rotation. Seven years later, he experienced issues with his pump sites, developing nodules and bruises that became painful and caused labile glycemic control. He underwent dermatologic, rheumatologic, and allergic workup with two biopsies confirming insulin-induced lipoatrophic panniculitis. Immunosuppressive and varying insulin regimens were unsuccessful. Hydroxychloroquine and methotrexate showed limited improvement with intolerable gastrointestinal side effects. Trials of hydrocortisone in his insulin infusion or topical clobetasol with tacrolimus provided no benefit. Various MDI regimens with different concentrations of lispro, aspart, faster aspart, glargine, detemir, and degludec were tried; all elicited varying degrees of pain with suboptimal glycemic response. A supervised trial of inhaled insulin was associated with bronchospasms. An implantable insulin pump was sought through clinical trials but was not feasible due to eligibility. Ultimately, he was successfully referred for pancreas transplantation due to his labile and complicated diabetes, but the procedure was complicated by allograft thrombosis and failure. He is currently awaiting a second transplant. Clinical Lesson: There are few reports of patients with insulin-induced lipoatrophic panniculitis in the literature. It is an important differential to consider for a patient that is having unexpected glycemic excursions, and it is equally as challenging to select an adequate management regimen. Treatment is limited and a broad range of options need to be considered. When these patients fail mainstay immunosuppressive, corticosteroid, and insulin-based management, pancreas transplantation may be considered as an alternative feasible treatment. Presentation: 6/1/2024

Pathomechanism of infantile traumatic brain injury with a biphasic clinical course and late reduced diffusion evaluated by MR spectroscopy

BackgroundInfantile traumatic brain injury (TBI) with a biphasic clinical course and late reduced diffusion (TBIRD) has recently been reported as a distinct type of TBI in infancy. However, the pathological and prognostic factors of TBIRD remain unknown. We aimed to compare patients with and without TBIRD and evaluate the pathomechanism of TBIRD using magnetic resonance spectroscopy (MRS). MethodsTen Japanese patients with TBI were admitted to our hospital and underwent MRS between September 2015 and September 2022 (age range, 3–15 months; median age, 8.5 months). TBIRD was diagnosed in six patients. MRS data were compared among patients with TBIRD, patients without TBIRD, and controls. Neurological prognosis was classified into grades 1 (normal) to 3 (severe). ResultsIn patients with TBIRD, MRS revealed an increase in the glutamine (Gln) level on days 3–29, which subsequently became close to normal. The degree of Gln elevation in the non-TBIRD group was smaller (117–158 % of controls) than that in the TBIRD group (210–337 %) within 14 days. MRS in the TBIRD group showed decreased N-acetyl aspartate (NAA) concentrations. The degree of NAA decrease was more prominent in grade 3 than in grades 1 and 2. NAA levels in the non-TBIRD group were almost normal. ConclusionsPatients with TBI and markedly elevated Gln levels on MRS may develop TBIRD. Neuro-excitotoxicity is a possible pathological mechanism of TBIRD. Decreased NAA levels may be useful for predicting the prognosis of patients with TBIRD.

Fecal microbiota transplantation alleviates mild-moderate COVID-19 associated diarrhoea and depression symptoms: A prospective study of a randomized, double-blind clinical trial.

Currently, the emergence of the endemic Coronavirus disease (COVID-19) situation still poses a serious threat to public health. However, it remains elusive about the role of fecal microbiota transplantation in treating COVID-19. We performed a randomized, double-blind, placebo-controlled clinical trial enrolling a cohort of 40 COVID-19 patients with mild-moderate symptoms. Our results showed that fecal microbiota transplantation provided an amelioration in diarrhoea (p = 0.026) of digestive system and depression (p = 0.006) of neuropsychiatric-related symptom in COVID-19 patients, respectively. Meanwhile, we found that the number of patients with diarrhoea decreased from 19 to 0 on day 7 after fecal microbiota transplantation treatment, and it was statistically changed compared to the placebo group (p = 0.047). Of note, the serum concentration of aspartate aminotransferase-to-alanine aminotransferase ratio (AST/ALT, fecal microbiota transplantation, pre vs. post: 0.966 vs. 0.817), a biomarker for predicting long COVID-19, was significantly reduced by fecal microbiota transplantation. In all, our study supports that fecal microbiota transplantation could be a novel therapeutic strategy for COVID-19 patients with diarrhoea and depressive symptoms, which is potentially valuable in ameliorating long COVID-19 symptoms.

Predicting performance in attention by measuring key metabolites in the PCC with 7T MRS

The posterior cingulate cortex (PCC) is a key hub of the default mode network and is known to play an important role in attention. Using ultra-high field 7 Tesla magnetic resonance spectroscopy (MRS) to quantify neurometabolite concentrations, this exploratory study investigated the effect of the concentrations of myo-inositol (Myo-Ins), glutamate (Glu), glutamine (Gln), aspartate or aspartic acid (Asp) and gamma-amino-butyric acid (GABA) in the PCC on attention in forty-six healthy participants. Each participant underwent an MRS scan and cognitive testing, consisting of a trail-making test (TMT A/B) and a test of attentional performance. After a multiple regression analysis and bootstrapping for correction, the findings show that Myo-Ins and Asp significantly influence (p < 0.05) attentional tasks. On one hand, Myo-Ins shows it can improve the completion times of both TMT A and TMT B. On the other hand, an increase in aspartate leads to more mistakes in Go/No-go tasks and shows a trend towards enhancing reaction time in Go/No-go tasks and stability of alertness without signal. No significant (p > 0.05) influence of Glu, Gln and GABA was observed.

NMR Fingerprinting of Conventional and Genetically Modified Soybean Plants with AtAREB1 Transcription Factors.

Drought stress impacts soybean yields and physiological processes. However, the insertion of the activated form of the AtAREB1 gene in the soybean cultivar BR16, which is sensitive to water deficit, improved the drought response of the genetically modified plants. Thus, in this study, we used 1H NMR in solution and solid-state NMR to investigate the response of genetically modified soybean overexpressing AtAREB1 under water deficiency conditions. We achieved that drought-tolerant soybean yields high content of amino acids isoleucine, leucine, threonine, valine, proline, glutamate, aspartate, asparagine, tyrosine, and phenylalanine after 12 days of drought stress conditions, as compared to drought-sensitive soybean under the same conditions. Specific target compounds, including sugars, organic acids, and phenolic compounds, were identified as involved in controlling sensitive soybean during the vegetative stage. Solid-state NMR was used to study the impact of drought stress on starch and cellulose contents in different soybean genotypes. The findings provide insights into the metabolic adjustments of soybean overexpressing AREB transcription factors in adapting to dry climates. This study presents NMR techniques for investigating the metabolome of transgenic soybean plants in response to the water deficit. The approach allowed for the identification of physiological and morphological changes in drought-resistant and drought-tolerant soybean tissues. The findings indicate that drought stress significantly alters micro- and macromolecular metabolism in soybean plants. Differential responses were observed among roots and leaves as well as drought-tolerant and drought-sensitive cultivars, highlighting the complex interplay between overexpressed transcription factors and drought stress in soybean plants.

Decreased Cerebral Creatine and N-Acetyl Aspartate Concentrations after Severe COVID-19 Infection: A Magnetic Resonance Spectroscopy Study.

Background/Objectives: The aim of this study was to evaluate brain metabolism using MR spectroscopy (MRS) after recovery from Coronavirus disease (COVID-19) and to test the impact of disease severity on brain metabolites. Methods: We performed MRS on 81 individuals (45 males, 36 females, aged 40-60), who had normal MRI findings and had recovered from COVID-19, classifying them into mild (17), moderate (36), and severe (28) groups based on disease severity during the acute phase. The study employed two-dimensional spectroscopic imaging above the corpus callosum, focusing on choline (Cho), creatine (Cr), and N-acetylaspartate (NAA). We analyzed Cho/Cr and NAA/Cr ratios as well as absolute concentrations using water as an internal reference. Results: Results indicated that the Cho/Cr ratio was higher with increasing disease severity, while absolute Cho and NAA/Cr ratios showed no significant differences across the groups. Notably, absolute Cr and NAA levels were significantly lower in patients with severe disease. Conclusions: These findings suggest that the severity of COVID-19 during the acute phase is associated with significant changes in brain metabolism, marked by an increase in Cho/Cr ratios and a reduction in Cr and NAA levels, reflecting substantial metabolic alterations post-recovery.

P-158 Quantitative metabolomic profiling of follicular fluid identifies oocyte quality related biomarkers from infertile patients

Abstract Study question Is whether there is a metabolomic signature in the follicular fluid depending on oocyte quality and whether infertility may impact on this quality? Summary answer 46 metabolites were quantified via nuclear magnetic resonance spectroscopy in the follicular fluid, and each metabolite that was a biomarker of oocyte quality was pathology-specific. What is known already Literature confirmed that metabolomics had been valuable for exploring follicular fluid and the search for nutritive and non-nutritive biomarkers. There are currently no data in the literature concerning the homeostasis of this follicular fluid, and two options are possible: this fluid is a continuum of blood plasma, or it has its own homeostasis. Studies have highlighted follicular fluid qualitative and quantitative alterations in the metabolome due to one specific infertility pathology footprint and correlated in some instances to outcome measures of in vitro fertilization success but samples are pooled and there are no information concerning quality and competence oocyte assessment. Study design, size, duration 61 infertile patients who underwent in vitro fertilization (IVF) treatment with intracytoplasmic sperm injection (ICSI) from Femme Mère Enfant hospital (Bron, France) between 6th april 2022 and 12th may 2022 were included. Participants/materials, setting, methods 61 follicular fluids were collected individually and used for metabolomic profiling by nuclear magnetic resonance spectroscopy. A total of 46 metabolites were quantified. Metabolite concentrations in follicular fluid were compared with serum concentrations.Oocyte was considered mature if it has its first polar body and competent if it was able to develop into a blastocyst after culturing. In vitro fertilized oocytes were cultured and monitored with time-lapses incubation technology until day 5 or 6. Main results and the role of chance For 23 metabolites out of 46, follicular fluid exhibited lower concentrations than serum. Glucose and glutamine, which are among the most abundant metabolites, decreased by 54% (not significant) and 47% (P &amp;lt; 0.05) respectively. 7 metabolites exhibited higher concentrations in the follicular fluid than in the serum. An 85%-increase in lactate and a more-than-tenfold increase in pyruvate were noted (P &amp;lt; 0.05). The reverse ratios of lower substrates such as glucose and glutamine and higher waste products like lactate and pyruvate imply that the follicular fluid functions as a homeostatic compartment, being more influenced by exchange within follicular cells than by serum equilibration. The detected metabolic signatures of oocyte maturity and competence were all specific to the patient infertility-related pathology. The follicular fluid concentrations of aspartate, glucose, methionine and methylmalonate were significantly correlated to oocyte maturity in the control group (P &amp;lt; 0.05). For patients having endometriosis, asparagine concentration was significantly decreased by oocyte maturity (P &amp;lt; 0.05). For patients with diminished ovarian reserve, acetone concentration was significantly decreased by oocyte maturity (P &amp;lt; 0.05). The concentration of aspartate, that correlated with oocyte maturity, was also slightly decreased by oocyte competence, and for the patients with endometriosis the concentration of threonine was decreased by 30% by oocyte competence (P &amp;lt; 0.05). Limitations, reasons for caution Although 46 metabolites were well quantified, each metabolite that was a biomarker of oocyte maturity or competence was pathology specific. One major reason would be the high metabolite concentration variation observed between the different infertilities. This study indicates that the diagnosis of patients according to their infertility must be prior. Wider implications of the findings This study underlines the potential of follicular fluid metabolomics as a diagnostic avenue for oocyte-related becoming in order to enhance the selection of oocytes and improve outcomes in assisted reproductive and preservation fertility techniques. Trial registration number not applicable

The Increased aspartate levels in transgenic cotton (Gossypium hirsutum L.) lead to improved tolerance against whitefly (Bemisia tabaci, Gennadius).

The whitefly, a polyphagous insect pest feeding on nearly 1328 plant species, is a major threat to global cotton production and incurs up to 50% yield losses in cotton production in Pakistan. We investigated whether increased aspartate in phloem sap imparts whitefly toxicity and protects cotton plants from intense damage. The enzymatic step for aspartate production is carried through aspartate aminotransferase (AAT). In this study, we constitutively overexpressed the Oryza sativa cytoplasmic AAT (OsAAT2) under the CaMV35S promoter in Gossypium hirsutum cv. CIM-482. Real-time PCR analysis of the AAT transcripts revealed a 2.85- to 31.7-fold increase in mRNA levels between the different cotton lines. A substantial increase in the free-amino acid content of the major N-assimilation and transport amino acids (aspartate, glutamate, asparagine, and glutamine) was seen in the phloem sap of the transgenic cotton lines. The bioassay revealed that the two transgenic cotton lines with the highest free aspartate content in the phloem sap exhibited 97 and 94% mortality in the adult whitefly population and a 98 and 96% decline in subsequent nymph populations, respectively. There was also a significant change in the physiological behaviour of the transgenic cotton lines, with an increased net assimilation (A), gaseous exchange (Gs) and rate of transpiration (E). Improved morphological characteristics like plant height, total number of bolls and fiber yield were recorded in transgenic cotton lines. The AAT gene shows promise in mitigating whitefly infestations and enhancing the overall health and yield of cotton plants.

AspSnFR: A genetically encoded biosensor for real-time monitoring of aspartate in live cells

Aspartate is crucial for nucleotide synthesis, ammonia detoxification, and maintaining redox balance via the malate-aspartate-shuttle (MAS). To disentangle these multiple roles of aspartate metabolism, tools are required that measure aspartate concentrations in real time and in live cells. We introduce AspSnFR, a genetically encoded green fluorescent biosensor for intracellular aspartate, engineered through displaying and screening biosensor libraries on mammalian cells. In live cells, AspSnFR is able to precisely and quantitatively measure cytosolic aspartate concentrations and dissect its production from glutamine. Combining high-content imaging of AspSnFR with pharmacological perturbations exposes differences in metabolic vulnerabilities of aspartate levels based on nutrient availability. Further, AspSnFR facilitates tracking of aspartate export from mitochondria through SLC25A12, the MAS' key transporter. We show that SLC25A12 is a rapidly responding and direct route to couple Ca2+ signaling with mitochondrial aspartate export. This establishes SLC25A12 as a crucial link between cellular signaling, mitochondrial respiration, and metabolism.

Whole-Body Hypothermia vs Targeted Normothermia for Neonates With Mild Encephalopathy

Although whole-body hypothermia is widely used after mild neonatal hypoxic-ischemic encephalopathy (HIE), safety and efficacy have not been evaluated in randomized clinical trials (RCTs), to our knowledge. To examine the effect of 48 and 72 hours of whole-body hypothermia after mild HIE on cerebral magnetic resonance (MR) biomarkers. This open-label, 3-arm RCT was conducted between October 31, 2019, and April 28, 2023, with masked outcome analysis. Participants were neonates at 6 tertiary neonatal intensive care units in the UK and Italy born at or after 36 weeks' gestation with severe birth acidosis, requiring continued resuscitation, or with an Apgar score less than 6 at 10 minutes after birth and with evidence of mild HIE on modified Sarnat staging. Statistical analysis was per intention to treat. Random allocation to 1 of 3 groups (1:1:1) based on age: neonates younger than 6 hours were randomized to normothermia or 72-hour hypothermia (33.5 °C), and those 6 hours or older and already receiving whole-body hypothermia were randomized to rewarming after 48 or 72 hours of hypothermia. Thalamic N-acetyl aspartate (NAA) concentration (mmol/kg wet weight), assessed by cerebral MR imaging and thalamic spectroscopy between 4 and 7 days after birth using harmonized sequences. Of 225 eligible neonates, 101 were recruited (54 males [53.5%]); 48 (47.5%) were younger than 6 hours and 53 (52.5%) were 6 hours or older at randomization. Mean (SD) gestational age and birth weight were 39.5 (1.1) weeks and 3378 (380) grams in the normothermia group (n = 34), 38.7 (0.5) weeks and 3017 (338) grams in the 48-hour hypothermia group (n = 31), and 39.0 (1.1) weeks and 3293 (252) grams in the 72-hour hypothermia group (n = 36). More neonates in the 48-hour (14 of 31 [45.2%]) and 72-hour (13 of 36 [36.1%]) groups required intubation at birth than in the normothermic group (3 of 34 [8.8%]). Ninety-nine neonates (98.0%) had MR imaging data and 87 (86.1%), NAA data. Injury scores on conventional MR biomarkers were similar across groups. The mean (SD) NAA level in the normothermia group was 10.98 (0.92) mmol/kg wet weight vs 8.36 (1.23) mmol/kg wet weight (mean difference [MD], -2.62 [95% CI, -3.34 to -1.89] mmol/kg wet weight) in the 48-hour and 9.02 (1.79) mmol/kg wet weight (MD, -1.96 [95% CI, -2.66 to -1.26] mmol/kg wet weight) in the 72-hour hypothermia group. Seizures occurred beyond 6 hours after birth in 4 neonates: 1 (2.9%) in the normothermia group, 1 (3.2%) in the 48-hour hypothermia group, and 2 (5.6%) in the 72-hour hypothermia group. In this pilot RCT, whole-body hypothermia did not improve cerebral MR biomarkers after mild HIE, although neonates in the hypothermia groups were sicker at baseline. Safety and efficacy of whole-body hypothermia should be evaluated in RCTs. ClinicalTrials.gov Identifier: NCT03409770.

Effects of Bifidobacterium animalis subsp. lactis BB-12 and yogurt on mice during oral antibiotic administration

Probiotics have the potential to prevent disruptions to normal gastrointestinal function caused by oral antibiotic use. In this study, we examined the capacity of Bifidobacterium animalis subspecies lactis BB-12 (BB-12) and yogurt, separately and combined, to mitigate the effects of the antibiotic amoxicillin-clavulanate (AMC) on the gut microbiota and metabolomes of C57BL/6 J mice. Male and female mice were administered either BB-12, yogurt, BB-12 in yogurt, or saline for 10 days concurrent with the inclusion of AMC in the drinking water. Male mice exposed to AMC exhibited significant reductions (p<0.05) in body weight over the course of the study compared to sham (no AMC) controls whereas no such effects were observed for female mice. AMC administration resulted in rapid alterations to the intestinal microbiota in both sexes irrespective of BB-12 or yogurt treatment, including significant (p<0.05) losses in bacterial cell numbers and changes in microbial alpha-diversity and beta-diversity in the feces and cecal contents. The effects of AMC on the gut microbiota were observed within one day of administration and the bacterial contents continued to change over time, showing a succession marked by rapid reductions in Muribaculaceae and Lachnospiraceae and temporal increases in proportions of Acholeplasmataceae (day 1) and Streptococcaceae and Leuconostocaceae (day 5). By day 10 of AMC intake, high proportions of Gammaproteobacteria assigned as Erwiniaceae or Enterobacteriaceae (average of 63 %), were contained in the stools and were similarly enriched in the cecum. The cecal contents of mice given AMC harbored significantly reduced concentrations of (branched) short-chain fatty acids (SCFA), aspartate, and other compounds, whereas numerous metabolites, including formate, lactate, and several amino acids and amino acid derivatives were significantly enriched. Despite the extensive impact of AMC, starting at day 7 of the study, the body weights of male mice given yogurt or BB-12 (in saline) with AMC were similar to the healthy controls. BB-12 (in saline) and yogurt intake was associated with increased Streptococcaceae and both yogurt and BB-12 resulted in lower proportions of Erwiniaceae in the fecal and cecal contents. The cecal contents of mice fed BB-12 in yogurt contained levels of formate, glycine, and glutamine that were equivalent to the sham controls. These findings highlight the potential of BB-12 and yogurt to mitigate antibiotic-induced gut dysbiosis.

Effect of per os administration of fucoxanthin alcoholic and oil extracts to Balb/c mice on biochemical parameters and serum cytokine level

Microalgae are a source of various biologically active substances, including carotenoids. One of the carotenoids of diatom microalgae is fucoxanthin, which has antioxidant, antimicrobial, antitumour and anti-inflammatory properties that can be used in the treatment of a wide range of human and animal pathologies. The aim of the study was to investigate the effect of alcohol and oil extract of fucoxanthin on biochemical parameters and levels of serum cytokines in mice during intragastric administration. Material and methods. Fucoxanthin extracts from the microalga Nanofrustulum shiloi were used. Mice of Balb/c strain were intragastrically administrated with 0.5 ml of oil (500 μg/ml) or alcoholic extract of fucoxanthin (187.5 μg/ml), mice in the comparison group were given 0.5 ml of olive oil or 45% ethyl alcohol once a day within 5 days. On the 8th day, blood serum was obtained and the level of albumin, triglycerides, cholesterol, low (LDL) and high density lipoproteins (HDL), urea, uric acid, creatinine, aspartate (AST) and alanine aminotransferases (ALT), lactate, stable nitric oxide (NO) metabolites (nitrites), cytokines (IL-1β, IL-6, IL-10, TNF-α, IFN-γ) was determined using ELISA with photometric detection. Results and discussion. In the comparison group, ethyl alcohol promoted an increase in serum triglyceride, cholesterol, LDL, HDL, ALT, AST, nitrite, IL-10, IFN-γ and TNF-α content compared to intact animals. Fucoxanthin alcoholic extract administration increased the level of LDL, HDL, uric acid, TNF-α and decreased albumin, cholesterol, uric acid, creatinine, ALT, AST, IL-6 content compared to control. The olive oil group showed a decrease in serum triglycerides, cholesterol, urea, uric acid, ALT and AST content but an increase in LDL, HDL, nitrite level compared to the control. Administration of fucoxanthin oil extract to animals decreased albumin, triglycerides, cholesterol, HDL, uric acid, ALT, AST, lactate, IL-6, IL-10 compared to control. Conclusions. Thus, it was discovered that fucoxanthin in Balb/c mice contributes to a decrease in hepatic synthetic function, as evidenced by the change in the concentration of albumin, triglycerides, uric acid), influences energy metabolism (creatinine, lactate), cell membrane stability (cholesterol, LDL, HDL), serum transaminase activity, pro- and anti-inflammatory cytokines content.

Challenges with hippocampal MR spectroscopy as a surrogate for pre-radiotherapy assessment of neurocognitive impairment in patients with brain metastasis.

Patients with multiple brain metastases (BM) benefit from hippocampal-avoiding whole brain radiotherapy (HA-WBRT), the challenging and less available form of WBRT. This study explores potential of pre-radiotherapy (pre-RT) hippocampal magnetic resonance spectroscopy (MRS) measuring hippocampal neuronal density as an imaging surrogate and predictive tool for assessing neurocognitive functions (NCF). 43 BM patients underwent pre-RT hippocampal MRS. N-acetyl aspartate (NAA) concentration, a marker for neuronal density (weighted by creatine (Cr) and choline (Cho) concentrations), and neurocognitive function (NCF) tests (HVLT and BVMT) performed by certified psychologists were evaluated. Clinical variables and NAA concentrations were correlated with pre-RT NCFs. HVLT and BVMT subtests showed pre-RT deterioration except for BVMT recognition. Significantly better NCFs were observed in women in HVLT subsets. Significantly higher NAA/Cr + Cho was measured in women (median 0.63 vs. 0.55; P=0.048) in the left hippocampus (no difference in the right hippocampus). In men, a positive correlation (0.51, P=0.018) between total brain volume and HVLT-TR, between left hippocampal NAA/Cr + Cho and HVLT-R (0.45, P=0.063), and between right hippocampal NAA/Cr + Cho and BVMT-recognition (0.49, P=0.054) was observed. In women, a borderline significant negative correlation was observed between left hippocampal NAA/Cr + Cho and BVMT-TR (-0.43, P=0.076) and between right NAA/Cr + Cho and HVLT-DR (-0.42, P=0.051). Borderline statistically significant correlations were observed with speculative interpretation underlying the challenges of hippocampal MRS as a surrogate for neurocognitive impairment. Further studies need to be done to ascertain the opportunities for imaging predictors of benefit from memory sparing radiotherapy.

Metabolism-dependent secondary effect of anti-MAPK cancer therapy on DNA repair.

Amino acid bioavailability impacts mRNA translation in a codon-dependent manner. Here, we report that the anti-cancer MAPK inhibitors (MAPKi) decrease the intracellular concentration of aspartate and glutamate in melanoma cells. This coincides with the accumulation of ribosomes on codons corresponding to these amino acids and triggers the translation-dependent degradation of mRNAs encoding aspartate- and glutamate-rich proteins, involved in DNA metabolism such as DNA replication and repair. Consequently, cells that survive MAPKi degrade aspartate and glutamate likely to generate energy, which simultaneously decreases their requirement for amino acids due to the downregulation of aspartate- and glutamate-rich proteins involved in cell proliferation. Concomitantly, the downregulation of aspartate- and glutamate-rich proteins involved in DNA repair increases DNA damage loads. Thus, DNA repair defects, and therefore mutations, are at least in part a secondary effect of the metabolic adaptation of cells exposed to MAPKi.

Neurometabolic activity in the thalamic and frontal cortex and cognitive functions in adolescents with generalized epileptic syndromes

The study aimed to analyse the mutual correlations between the metabolic activity of the brain evaluated by the magnetic resonance spectroscopy method (MRS) and intelligence and cognitive functions in adolescents with idiopathic generalised epilepsies (IGE). The focus of the study was placed on the activity of the thalamus and frontal cortex.Twenty newly diagnosed patients with juvenile absence epilepsy (JAE) and juvenile myoclonic epilepsy (JME) were included in the study. Adolescents underwent primary structural neuroimaging (1.5T) and advanced evaluation of the frontal cortex and thalamus by MRS. Concentrations of N-acetyl aspartate (NAA), creatine plus phosphocreatine (Cr) and choline (Cho) were measured and were expressed as ratios of NAA/Cr, NAA/Ch and NAA/Cr+Ch. Patients also completed a panel of IQ and cognitive tests (e.g., FePsy Battery – classification test, verbal fluency test, RAVLT memory test). The correlation between the level of metabolites and the results of intellectual and cognitive tests was examined.Strong correlations were found between IQ and some cognitive functions and metabolic ratios in the IGEs group. Overall the patients whose NAA/Cr, NAA/Ch, NAA/Cr+Ch ratios were lower, obtained worse results in IQ and in some of the neuropsychological tests.The metabolite activity in the thalamus and frontal cortex impacts the results of neuropsychological tests in the group of patients with IGE.

Effects of organic trace minerals chelated with oligosaccharides on growth performance, blood parameters, slaughter performance and meat quality in sheep.

The present study assessed the effects of oligosaccharide-chelated organic trace minerals (OTM) on the growth performance, digestive enzyme activity, blood parameters, slaughter performance, and meat quality indexes of mutton sheep. A total of 60 East Ujumuqin × small-tailed Han crossbred mutton sheep were assigned to two groups (10 duplicates per group) by body weight (26.12 ± 3.22 kg) according to a completely randomized design. Compared to the CON group, the results of the OTM group showed: (1) no significant changes in the initial body weight, final body weight, dry matter intake, average daily gain, and feed conversion ratio (p > 0.05); (2) the activities of trypsin, lipase, and amylase in the jejunum were significantly increased (p < 0.05); (3) serum total protein, albumin, and globulin of the blood were significantly increased (p < 0.05), and the growth factor interleukin IL-10 was significantly higher (p < 0.05), while IL-2, IL-6, and γ-interferon were significantly lower (p < 0.05). Immunoglobulins A, M, and G were significantly higher (p < 0.05); (4) the live weight before slaughter, carcass weights, dressing percentage, eye muscle areas, and GR values did not differ significantly (p > 0.05); (5) shear force of mutton was significantly lower (p < 0.05), while the pH45min, pH24h, drip loss, and cooking loss did not show a significant difference (p > 0.05). The content of crude protein was significantly higher (p < 0.05), while the ether extract content was significantly reduced (p < 0.05), but no significant difference was detected between moisture and ash content; (6) the total amino acids, essential amino acids, semi-essential amino acids, and umami amino acids were significantly increased (p < 0.05). Although umami amino acids were not significant, the total volume increased (p > 0.05). Among these, the essential amino acids, threonine, valine, leucine, lysine in essential amino acids and arginine were significantly increased (p < 0.05). Also, non-essential amino acids, glycine, serine, proline, tyrosine, cysteine, and aspartic acid, were significantly higher (p < 0.05). The content of alanine, aspartate, glutamic acid, phenylalanine, and tyrosine in umami amino acids was significantly higher (p < 0.05).