Conversion Of Lactate Research Articles

Efficient Conversion of Methyl Lactate to Methyl Acrylate over Cesium-Modified Al-Rich Beta Zeolites

Dehydration of methyl lactate (ML) obtained from biomass is a sustainable route for the production of methyl acrylate (MA). Herein, we demonstrate that cesium-exchanged Al-rich Beta zeolite (Cs-Beta) is highly efficient for the dehydration of ML to MA, exhibiting full ML conversion with 87.4% MA yield at 340 °C. In situ FT-IR spectra and kinetic studies provide evidence that the ML dehydration on the Cs-Beta zeolite follows a forward ion-exchange/dehydration/reverse ion-exchange reaction pathway. Model experiments, DFT calculations, TG-DSC analysis, and FT-IR spectra showed that coke formation competes with the dehydration of ML to MA. Kinetic studies and theoretical simulations demonstrate that cesium cations in the micropores of Beta zeolite have a strong steric hindrance, suppressing the formation of oligomers and coke. These results offer new insights for developing highly selective and stable catalysts for the dehydration of ML to MA in the future.

Biotic and abiotic insights into the storage of food waste and its effect on biohydrogen and methane production potential

The present study investigated the physicochemical and microbiological changes occurring during the storage of simulated restaurant food waste (FW) and how such changes affected its biohydrogen and biogas production potential. FW was stored for 72 h in a closed atmosphere under two different scenarios: i) without and ii) with inoculation of a mixed microbial culture harboring lactic acid bacteria (LAB). Both storage scenarios resulted in similar biotic and abiotic changes in FW. Particularly, FW was pre-acidified and pre-hydrolyzed to some extent during the storage, resulting in a feedstock enriched in LAB (≈ 95 % total relative abundance) and lactate (10.5–12.3 g/L, 87.0–90.5 % selectivity). Biochemical hydrogen potential tests revealed that the use of stored FW resulted in similar or even higher hydrogen production efficiencies compared to that of non-stored FW, achieving up to 60 NmL H2/g VS added and a maximum volumetric hydrogen production rate of 9.7 NL H2/L-d. Metabolically, the conversion of lactate into hydrogen was crucial regardless of the use of non-stored or stored FW, albeit the presence of fermentable carbohydrates in the substrate was also essential either to produce lactate or to co-produce extra hydrogen. On the contrary, biochemical methane potential tests showed that the biogas production potential of FW was not affected by storage, yielding on average 400 NmL CH4/g VS added and revealing that lactate oxidation to methane precursors represented an important step in FW biomethanization.

Spectrally selective bSSFP using off-resonant RF excitations permits high spatiotemporal resolution 3D metabolic imaging of hyperpolarized [1-13 C]Pyruvate-to-[1-13 C]lactate conversion.

To develop a high spatiotemporal resolution 3D dynamic pulse sequence for preclinical imaging of hyperpolarized [1-13 C]pyruvate-to-[1-13 C]lactate metabolism at 7T. A standard 3D balanced SSFP (bSSFP) sequence was modified to enable alternating-frequency excitations. RF pulses with 2.33 ms duration and 900 Hz FWHM were placed off-resonance of the target metabolites, [1-13 C]pyruvate (by approximately -245 Hz) and [1-13 C]lactate (by approximately 735 Hz), to selectively excite those resonances. Relatively broad bandwidth (compared to those metabolites' chemical shift offset) permits a short TR of 6.29 ms, enabling higher spatiotemporal resolution. Bloch equation simulations of the bSSFP response profile guided the sequence parameter selection to minimize spectral contamination between metabolites and preserve magnetization over time. Bloch equation simulations, phantom studies, and in vivo studies demonstrated that the two target resonances could be cleanly imaged without substantial bSSFP banding artifacts and with little spectral contamination between lactate and pyruvate and from pyruvate hydrate. High spatiotemporal resolution 3D images were acquired of in vivo pyruvate-lactate metabolism in healthy wild-type and endogenous pancreatic tumor-bearing mice, with 1.212 s acquisition time per single-metabolite image and (1.75 mm)3 isotropic voxels with full mouse abdomen 56 × 28 × 21 mm3 FOV and fully-sampled k-space. Kidney and tumor lactate/pyruvate ratios of two consecutive measurements in one animal, 1 h apart, were consistent. Spectrally selective bSSFP using off-resonant RF excitations can provide high spatio-temporal resolution 3D dynamic images of pyruvate-lactate metabolic conversion.

Real-Time Pyruvate Chemical Conversion Monitoring Enabled by PHIP.

In recent years, parahydrogen-induced polarization side arm hydrogenation (PHIP-SAH) has been applied to hyperpolarize [1-13C]pyruvate and map its metabolic conversion to [1-13C]lactate in cancer cells. Developing on our recent MINERVA pulse sequence protocol, in which we have achieved 27% [1-13C]pyruvate carbon polarization, we demonstrate the hyperpolarization of [1,2-13C]pyruvate (∼7% polarization on each 13C spin) via PHIP-SAH. By altering a single parameter in the pulse sequence, MINERVA enables the signal enhancement of C1 and/or C2 in [1,2-13C]pyruvate with the opposite phase, which allows for the simultaneous monitoring of different chemical reactions with enhanced spectral contrast or for the same reaction via different carbon sites. We first demonstrate the ability to monitor the same enzymatic pyruvate to lactate conversion at 7T in an aqueous solution, in vitro, and in-cell (HeLa cells) via different carbon sites. In a second set of experiments, we use the C1 and C2 carbon positions as spectral probes for simultaneous chemical reactions: the production of acetate, carbon dioxide, bicarbonate, and carbonate by reacting [1,2-13C]pyruvate with H2O2 at a high temperature (55 °C). Importantly, we detect and characterize the intermediate 2-hydroperoxy-2-hydroxypropanoate in real time and at high temperature.

Symbiosis culture of probiotic Escherichia coli Nissle 1917 and Lactobacillus rhamnosus GG using lactate utilization protein YkgG

The abundance and biodiversity of intestinal bacteria play a critical role in immune system. Escherichia coli Nissle 1917 (EcN), a tractable probiotic cell has emerged as a ‘smart microbe’ that can provide therapeutic modalities at the sites of disease. Additionally, Lactobacillus rhamnosus GG (LGG) represents one of the key probiotics in clinical studies with the ability to enhance intestinal immunity in symbiosis with other cells. To fully exploit the advantages of EcN and LGG, we optimized the commensal culture medium for LGG and EcN, succeeded by the establishment of a novel fluorescence-based colony quantification. From the metabolite analysis, more lactate was consumed when LGG is co-cultured with EcN; thus, YkgG was further explored as it was foreseen to be the enzyme responsible for lactate conversion to pyruvate. The YkgG enzyme from EcN has Vmax of 64.5 mM min-1 and catalytic ability kcat/Km of 2055 mM-1 min-1, that is 6-fold compared to E. coli MG1655. Co-culture strategy of EcN with LGG in the mimic gastrointestinal tract increased the CFU by 50 % and 40 % within 5 mins and 10 mins, respectively. This practical, feasible, and promising symbiosis system aims to accelerate the application of probiotic in healthy food industries.

Multi-parametric hyperpolarized 13C/1H imaging reveals Warburg-related metabolic dysfunction and associated regional heterogeneity in high-grade human gliomas

BackgroundDynamic hyperpolarized (HP)-13C MRI has enabled real-time, non-invasive assessment of Warburg-related metabolic dysregulation in glioma using a [1-13C]pyruvate tracer that undergoes conversion to [1-13C]lactate and [13C]bicarbonate. Using a multi-parametric 1H/HP-13C imaging approach, we investigated dynamic and steady-state metabolism, together with physiological parameters, in high-grade gliomas to characterize active tumor. MethodsMulti-parametric 1H/HP-13C MRI data were acquired from fifteen patients with progressive/treatment-naïve glioblastoma [prog/TN GBM, IDH-wildtype (n = 11)], progressive astrocytoma, IDH-mutant, grade 4 (G4AIDH+, n = 2) and GBM manifesting treatment effects (n = 2). Voxel-wise regional analysis of the cohort with prog/TN GBM assessed imaging heterogeneity across contrast-enhancing/non-enhancing lesions (CEL/NEL) and normal-appearing white matter (NAWM) using a mixed effects model. To enable cross-nucleus parameter association, normalized perfusion, diffusion, and dynamic/steady-state (HP-13C/spectroscopic) metabolic data were collectively examined at the 13C resolution. Prog/TN GBM were similarly compared against progressive G4AIDH+ and treatment effects. ResultsRegional analysis of Prog/TN GBM metabolism revealed statistically significant heterogeneity in 1H choline-to-N-acetylaspartate index (CNI)max, [1-13C]lactate, modified [1-13C]lactate-to-[1-13C]pyruvate ratio (CELval > NELval > NAWMval); [1-13C]lactate-to-[13C]bicarbonate ratio (CELval > NELval/NAWMval); and 1H-lactate (CELval/NELval > NAWMundetected). Significant associations were found between normalized perfusion (cerebral blood volume, nCBV; peak height, nPH) and levels of [1-13C]pyruvate and [1-13C]lactate, as well as between CNImax and levels of [1-13C]pyruvate, [1-13C]lactate and modified ratio. GBM, by comparison to G4AIDH+, displayed lower perfusion %-recovery and modeled rate constants for [1-13C]pyruvate-to-[1-13C]lactate conversion (kPL), and higher 1H-lactate and [1-13C]pyruvate levels, while having higher nCBV, %-recovery, kPL, [1-13C]pyruvate-to-[1-13C]lactate and modified ratios relative to treatment effects. ConclusionsGBM consistently displayed aberrant, Warburg-related metabolism and regional heterogeneity detectable by novel HP-13C/1H imaging techniques.

Enzyme assembly on nanoparticle scaffolds enhances cofactor recycling and improves coupled reaction kinetics.

Enzyme activity can be many times enhanced in configurations where they are displayed on a nanoparticle (NP) and this same format sometimes even provides access to channeling phenomena within multienzyme cascades. Here, we demonstrate that such enhancement phenomena can be expanded to enzymatic cofactor recycling along with the coupled enzymatic processes that they are associated with. We begin by showing that the efficiency of glucose driven reduction of nicotinamide adenine dinucleotide (NAD+ → NADH) by glucose dehydrogenase (GDH) is enhanced ca. 5-fold when the enzyme is displayed on nanocrystalline semiconductor quantum dots (QDs) which are utilized as prototypical NP materials in our experimental assays. Coupling this enzymatic step with NADH-dependent lactate dehydrogenase (LDH) conversion of lactate to pyruvate also increases the latter's rate by a similar amount when both enzymes were jointly incorporated into self-assembled QD-based nanoclusters. Detailed agarose gel mobility assays and transmission electron microscopy imaging studies confirm that both tetrameric enzymes assemble to and crosslink the QDs into structured nanoclusters via their multiple-pendant terminal (His)6 sequences. Unexpectedly, control experiments utilizing blocking peptides to prevent enzyme-crosslinking of QDs resulted in even further enhancement of individual enzyme on-QD kinetic activity. This activity was also probed revealing that 200-fold excess peptide/QD addition enhanced individual GDH and LDH on-QD kcat a further 2- and 1.5×, respectively, above that seen just by QD display to a maximum of ∼10-fold GDH enhancement. The potential implications for how these enzyme kinetics-enhancing phenomena can be applied to single and multi-enzyme cascaded reactions in the context of cofactor recycling and cell-free synthetic biology are discussed.

Preparation of a Nanorod-like Mo-VOx Catalyst for Gas Phase Selective Oxidation of Methyl Lactate with Air

Selective oxidation of higher alcohols to carbonyls is one of the most important chemical transformations in research and industrial chemistry. Gas phase selective oxidation of alcohols with air as oxidant is a “green chemistry” synthesis path, although it is a great challenge to maintain high conversion with high selectivity. In this work, we synthesized nanorod-like Mo-VOx catalysts through regulation of the Mo/V ratio via a solid-state reaction method, and the group of Mo promoted VOx catalysts exhibited the combination of high catalytic activity and selectivity. The methyl pyruvate (MP) selectivity over 0.17Mo-VOx catalysts reached 79.6% at a relatively lower reaction temperature of 180 °C with 90.1% methyl lactate conversion. The characterization results revealed that the 0.17Mo-VOx catalysts showed a nanorod-like morphology with larger specific surface area and smaller crystalline size, generated active MoV2O8 crystalline phase, and could give rise to active V4+/V5+ redox couples as well as surface oxygen species. In terms of structure–activity relationship, when Mo was initially introduced into VOx, the MoV2O8 phase was gradually formed, and MP yield over yMo-VOx catalysts increased significantly together with V4+ proportion. However, excessive Mo would not form more active MoV2O8 phases, and the MP yield decreased.

Biochemical methane potential and active microbial communities during anaerobic digestion of biodegradable plastics at different inoculum-substrate ratios

The influence of the inoculum-substrate ratio (ISR) on the mesophilic and thermophilic biochemical methane potential test of two biodegradable plastics was evaluated. Poly(lactic acid) (PLA) and polyhydroxybutyrate (PHB) were selected for this study, the first for being recalcitrant to mesophilic anaerobic digestion (AD) and the second, by contrast, for being readily biodegradable. Several ISRs, calculated on the basis of volatile solids (VS), were tested: 1, 2, 2.85, 4, and 10 g(VS of inoculum).g(VS of substrate)−1. A high ISR was associated with an enhanced methane production rate (i.e., biodegradation kinetics). However, the ultimate methane production did not change, except when inhibition was observed. Indeed, applying the lowest ISR to readily biodegradable plastics such as PHB resulted in inhibition of methane production. Based on these experiments, in order to have reproducible degradation kinetics and optimal methane production, an ISR between 2.85 and 4 is recommended for biodegradable plastics. The active microbial communities were analyzed, and the active bacteria differed depending on the plastic digested (PLA versus PHB) and the temperature of the process (mesophilic versus thermophilic). Previously identified PHB degraders (Ilyobacter delafieldii and Enterobacter) were detected in PHB-fed reactors. Thermogutta and Tepidanaerobacter were detected during the thermophilic AD of PLA, and they are probably involved in PLA hydrolysis and lactate conversion, respectively.

Differential Metabolism of Glycerol Based on Oral versus Intravenous Administration in Humans.

Glycerol can be metabolized to glucose via gluconeogenesis or lactate via glycolysis. It is unknown if glycerol is metabolized similarly in the portal and systemic circulations in humans. Eight metabolically healthy overnight-fasted individuals received equimolar amounts of 13C3-glycerol orally and intravenously on two separate occasions with serial blood draws over four hours. Serum samples underwent liquid chromatography–mass spectrometry analysis. Oral 13C3-glycerol administration led to higher average serum glucose enrichment than intravenous administration (5.02 ± 1.43 versus 4.07 ± 0.79%, p = 0.009). In contrast, intravenous 13C3-glycerol administration yielded higher average serum lactate enrichment than oral administration (5.67 ± 0.80 versus 4.85 ± 1.30%, p = 0.032). Peak serum glucose enrichment was also higher with oral administration (9.37 ± 2.93 versus 7.12 ± 1.28%, p = 0.010). Glycerol metabolism across the portal and systemic circulations is not congruent. Orally administered labeled glycerol led to greater labeled glucose production, while intravenously administration yielded greater lactate production. These data support direct glycerol to lactate conversion in humans.

Is there any correlation between muscle fatigue and serum lactate dehydrogenase levels in prediabetic individuals?

Aim: Lactate dehydrogenase (LDH) is an enzyme that acts as a catalyst in the conversion of lactate to pyruvate which is abundantly found in liver, kidney, heart and muscles. Previous studies have all shown a strong positive correlation between muscle fatigue and increased serum LDH levels with type 2 diabetes mellitus but no study has actually assessed the same for prediabetes. The basic objective of this study, thus, is to find out the correlation between muscle fatigue and serum LDH levels in prediabetic individuals. Methods: A cross-sectional study was conducted in adults between 24–60 years old who were classified as prediabetic individuals as per norms established by American Diabetes Association. A total of 100 prediabetic individuals were selected for the study. Fatigability was calculated as a function of work done by the pleximeter finger of the dominant hand using Mosso’s ergograph. The study was conducted at Rajarshi Dashrath Autonomous State Medical College, Ayodhya. Results: Out of 100 prediabetic participants, 50% were males with a mean age of 44.14 ± 10.91 years and remaining 50% were females with a mean age of 41.12 ± 11.5 years. Overall, the average work done by the participants was 2.9 ± 1.2 weight lifted•total distance moved (kg•m) with an average serum LDH level of 323.84 ± 26.82 unit/litre (U/L). Conclusions: This study aimed at assessing the correlation between muscle fatigue and serum LDH levels in prediabetic individuals so that further work can be initiated to improve the quality of life in prediabetics that maybe drastically hampered due to easy fatigability in prediabetic individuals.

Lentilactobacillus buchneri domination during the fermentation of Japanese traditional fermented fish (funazushi).

Funazushi is a Japanese traditional fermented fish made with boiled rice without the addition of microbial starter cultures. Isolates from various commercial funazushi products, as identified by 16S rDNA sequences, suggested that Lentilactobacillus buchneri strains are major lactic acid bacteria. Based on an analysis of the putative CRISPR (clustered regularly interspaced short palindromic repeat) region, the genetic diversity of L.buchneri strains was examined. The data suggested that the diversity of L.buchneri strains depended on the factories at which funazushi was produced. An analysis of samples during fermentation indicated that the transition of microbes occurred, and L.buchneri was the dominant species. To determine the factors associated with domination, bacteriocin production and environmental stress tolerance, including NaCl and organic acid (lactate and acetate) tolerance, were evaluated. L.buchneri isolates did not produce bacteriocin. Although the isolates did not exhibit NaCl tolerance, they displayed higher lactate tolerance than other lactic acid bacteria isolated during funazushi fermentation. Based on reports that L.buchneri can convert lactate to acetate, the previous and present results suggested that lactate tolerance and lactate conversion in L.buchneri could explain its domination in funazushi. Our study presented a model for the domination mechanisms of specific microbes in fermented foods by spontaneous fermentation.

Biorefining food waste through the anaerobic conversion of endogenous lactate into caproate: A fragile balance between microbial substrate utilization and product inhibition

New technologies development and renewable source exploitation are key tools to realize the European Green Deal and to boost the bio-based economy. In this context, fermentation of organic residues as food waste is an efficient method to obtain marketable products such as carboxylic acids widely applied in industrial production. Under favourable thermodynamic conditions, short chain fatty acids deriving from primary fermentation could be biologically converted into medium-chain fatty acids as caproate via chain elongation (CE) process, by using ethanol or lactate as electron donors.This study evaluates the effectivity of producing caproate from Food Waste extract rich in organics with in situ electron donor production. The test carried out at OLR 15 gCOD L−1d−1 showed high Volatile Fatty Acids (from acetic to caproic acid) yields (0.37 g g−1CODfed), with a maximum caproate concentration of 8 g L−1. The associated microbiome was composed by lactate-producing bacteria (Corynebacterium, Lactobacillus, and Olsenella) and by chain elongators (Clostridiaceae and Caproiciproducens). By stressing the system with OLR increase up to 20 gCOD L−1d−1, the CE process was inhibited by the high concentration of caproate (low occurrence of Clostridiaceae and Caproiciproducens). Nevertheless, after few days of stop-feeding regime imposed to the system, the microbiome restored its capability to proceed with lactate-based CE pathways. Different batch tests carried out with the inhibited biomass at increasing initial caproate concentration confirmed its impact on lactate utilization kinetics.

Targeting lactate dehydrogenase B-dependent mitochondrial metabolism affects tumor initiating cells and inhibits tumorigenesis of non-small cell lung cancer by inducing mtDNA damage

Once considered a waste product of anaerobic cellular metabolism, lactate has been identified as a critical regulator of tumorigenesis, maintenance, and progression. The putative primary function of lactate dehydrogenase B (LDHB) is to catalyze the conversion of lactate to pyruvate; however, its role in regulating metabolism during tumorigenesis is largely unknown. To determine whether LDHB plays a pivotal role in tumorigenesis, we performed 2D and 3D in vitro experiments, utilized a conventional xenograft tumor model, and developed a novel genetically engineered mouse model (GEMM) of non-small cell lung cancer (NSCLC), in which we combined an LDHB deletion allele with an inducible model of lung adenocarcinoma driven by the concomitant loss of p53 (also known as Trp53) and expression of oncogenic KRAS (G12D) (KP). Here, we show that epithelial-like, tumor-initiating NSCLC cells feature oxidative phosphorylation (OXPHOS) phenotype that is regulated by LDHB-mediated lactate metabolism. We show that silencing of LDHB induces persistent mitochondrial DNA damage, decreases mitochondrial respiratory complex activity and OXPHOS, resulting in reduced levels of mitochondria-dependent metabolites, e.g., TCA intermediates, amino acids, and nucleotides. Inhibition of LDHB dramatically reduced the survival of tumor-initiating cells and sphere formation in vitro, which can be partially restored by nucleotide supplementation. In addition, LDHB silencing reduced tumor initiation and growth of xenograft tumors. Furthermore, we report for the first time that homozygous deletion of LDHB significantly reduced lung tumorigenesis upon the concomitant loss of Tp53 and expression of oncogenic KRAS without considerably affecting the animal’s health status, thereby identifying LDHB as a potential target for NSCLC therapy. In conclusion, our study shows for the first time that LDHB is essential for the maintenance of mitochondrial metabolism, especially nucleotide metabolism, demonstrating that LDHB is crucial for the survival and proliferation of NSCLC tumor-initiating cells and tumorigenesis.

Increasing of LDH Specific Activity and PEPCK Level Play a Role on Activation of Gluconeogenesis Pathway in Early Onset Pre-Eeclampsia Placenta.

Recent advancement on experiment concluded that etiology of pre-eclampsia (PE) could be explained by the "two-stage" theory. The theory of which explained that pre-eclampsia occurs due to abnormalities in spiral arteries development and release of inflammatory response. Failure of spiral arteries development, the lesion of damage could be due to ischemia-reperfusion or hypoxia-reoxygenation. Hypoxia in pre-eclamptic placenta leads to metabolic change to anaerobic in glycolysis. Lactate dehydrogenase (LDH) has important role in anaerobic glycolysis that catalyzes the conversion of lactate to pyruvate during hypoxia. On the other hand, phosphoenolpyruvate carboxy kinase (PEPCK) is merely an enzyme of gluconeogenesis. This research conduct to reveal that in early onset pre-eclampsia the placenta still hypoxic and undergoes gluconeogenesis even after delivery, through metabolic enzyme of LDH and PEPCK level. This cross-sectional study compared early onset PE (< 34 weeks) with normal term placenta. We measured LDH enzyme activity using colorimetric assay and PEPCK protein using ELISA method. Result show that placental LDH specific activity was increased significantly in PE with median 2.750 (0.030 - 5.680) U/mg compared to normal term placenta 0.255 (0.032 - 1.194) U/mg (Mann-Whitney, p< 0.001). PEPCK level was significantly increased in PE 8.998 (1.737-44.914) ng/mg compared to normal term placenta 1.552 (0.741-8.832) ng/mg (Mann-Whitney, p< 0.001). We conclude that anaerobic glycolysis and gluconeogenesis pathway are increased in early onset PE placenta as adaptation mechanism to hypoxic condition.

Mesenchymal stromal cells cultured in physiological conditions sustain citrate secretion with glutamate anaplerosis

Bone marrow mesenchymal stromal cells (MSCs) have immunomodulatory and regenerative potential. However, culture conditions govern their metabolic processes and therapeutic efficacy. Here we show that culturing donor-derived MSCs in Plasmax™, a physiological medium with the concentrations of nutrients found in human plasma, supports their proliferation and stemness, and prevents the nutritional stress induced by the conventional medium DMEM. The quantification of the exchange rates of metabolites between cells and medium, untargeted metabolomics, stable isotope tracing and transcriptomic analysis, performed at physiologically relevant oxygen concentrations (1%O2), reveal that MSCs rely on a high rate of glucose to lactate conversion, coupled with parallel anaplerotic fluxes from glutamine and glutamate to support citrate synthesis and secretion. These distinctive traits of MSCs shape the metabolic microenvironment of the bone marrow niche and can influence nutrient cross-talks under physiological and pathological conditions.

Loss of Pkm2 Alters Myocardial Metabolism and Increases Oxidative Stress After Infarction

IntroductionPyruvate kinase (PKM1) directs pyruvate to the Krebs cycle for oxidative metabolism in the healthy heart. Our lab described a hypoxia‐mediated switch to the alternatively spliced isoform PKM2, enhancing pyruvate to lactate conversion. Recently, we have also found that Pkm2 knockout (KO) mice had profound depletion of basal glucose in the heart compared to control mice. Pkm2 has also been shown to reduce oxidative damage and promote cardiomyocyte cell proliferation after myocardial infarction (MI). We hypothesize that upregulation of PKM2 can alter metabolic pathways by promoting glycolysis, and that after injury, this can preserve ATP production, protecting the heart from the stresses of hypoxia and injury.MethodsGlobal Pkm2 KO mice were subjected to permanent ligation of the left anterior descending coronary artery to mimic an MI. RNA‐seq analysis of left ventricles from control (n=8) and Pkm2 KO mice (n=8) before and 3 days after sham or MI surgery was performed. Semi‐quantitative real‐time PCR was used to confirm changes in selected genes of interest.ResultsLoss of Pkm2 moderately altered gene expression at baseline (q&lt;0.05, FDR&lt;0.05). Notably, the mitochondrial gene COX3 was downregulated in Pkm2 KO hearts. 68 genes were differentially expressed in Pkm2 KO hearts after MI, not observed in control MI hearts. MI of Pkm2 KO hearts resulted in considerable reduction of transcripts of enzymes in the insulin signaling pathway, mitochondrial oxidative phosphorylation, mitochondrial uncoupling, fatty acid metabolism, and increase in transcripts encoding enzymes in the pentose phosphate pathway, response to oxidative stress, and apoptotic signaling. Semi‐quantitative PCR of selected genes involved in glucose metabolism confirmed RNA‐seq results.ConclusionsRNA‐seq analysis of Pkm2 KO hearts demonstrated that loss of Pkm2 altered gene expression of metabolic and mitochondrial enzymes. Pkm2 KO hearts also showed increased abundance of pro‐apoptotic markers which may be a result of increased oxidative stress.

Inter-species Metabolic Interactions in an In-vitro Minimal Human Gut Microbiome of Core Bacteria

Knowledge of the functional roles and interspecies interactions are crucial for improving our understanding of the human intestinal microbiome in health and disease. However, the complexity of the human intestinal microbiome and technical challenges in investigating it pose major challenges. In this proof-of-concept study, we rationally designed, assembled and experimentally tested a synthetic Diet-based Minimal Microbiome (Db-MM) consisting of ten core intestinal bacterial species that together are capable of efficiently converting dietary fibres into short chain fatty acids (SCFAs). Despite their genomic potential for metabolic competition, all ten bacteria coexisted during growth on a mixture of dietary fibres, including pectin, inulin, xylan, cellobiose and starch. By integrated analyses of metabolite production, community composition and metatranscriptomics-based gene expression data, we identified interspecies metabolic interactions leading to production of key SCFAs such as butyrate and propionate. While public goods, such as sugars liberated from colonic fibres, are harvested by non-degraders, some species thrive by cross-feeding on energetically challenging substrates, including the butyrogenic conversion of acetate and lactate. Using a reductionist approach in an in-vitro system combined with functional measurements, our study provides key insights into the complex interspecies metabolic interactions between core intestinal bacterial species.

EVALUATION AND CORRELATION OF SERUM ELECTROLYTE PATTERN, MAGNESIUM, LDH AND VITAMIN D LEVELS IN PATIENTS OF HYPO THYROIDISM AT AT ERTIARY CARE CENTRE IN HARYANA

Introduction: Thyroid hormone is a central regulator of body hemodynamics, thermoregulation, and metabolic functions and in the disorders of the thyroid gland, signs and symptoms are often nonspecific. Both subclinical hypothyroidism and hyperthyroidism are associated with an increased risk of disease and alteration in biochemical and physiologic measures. In severe hypothyroidism and myxoedema, hyponatremia was a consequence of enhanced renal water retention mediated by vasopressin. Lactate dehydrogenase (LDH) is an enzyme found in nearly all living cells. It catalyzes the conversion of lactate to pyruvate and back, as it converts NAD+to NADH. However, only a few studies have investigated serum lactate dehydrogenase activity in patients with thyroid dysfunction. Vitamin D acts likea steroid hormone and is presently considered an immunomodulator affecting a wide range of functions. Aim and Objectives: To estimate the levels of serum electrolytes like sodium, potassium magnesium, LDH, and vitamin D in hypothyroid patients and to correlate with TSH. Materials and methods: The study population was serum specimens from 150 patients with hypothyroidism with ages ranging from 25-65 years. The samples were collected from the Thyroid laboratory, Department of Biochemistry, PGIMS, Rohtak. The different parameters of the study were estimated on the electrolyte analyzer, autoanalyzer, and radioimmunoassay. Results and conclusion: The mean values for Na, K, Mg, LDH, and vitamin D were found to be 128.13±5.09, 3.01±0.25, 1.87±0.32, 604.89±175.79, and 7.48±1.61, respectively. Our findings support that hypothyroid patients have subtle disturbances of electrolytes, LDH and vitamin D levels.

LDHB Deficiency Promotes Mitochondrial Dysfunction Mediated Oxidative Stress and Neurodegeneration in Adult Mouse Brain.

Age-related decline in mitochondrial function and oxidative stress plays a critical role in neurodegeneration. Lactate dehydrogenase-B (LDHB) is a glycolytic enzyme that catalyzes the conversion of lactate, an important brain energy substrate, into pyruvate. It has been reported that the LDHB pattern changes in the brain during ageing. Yet very little is known about the effect of LDHB deficiency on brain pathology. Here, we have used Ldhb knockout (Ldhb−/−) mice to test the hypothesis that LDHB deficiency plays an important role in oxidative stress-mediated neuroinflammation and neurodegeneration. LDHB knockout (Ldhb−/−) mice were generated by the ablation of the Ldhb gene using the Cre/loxP-recombination system in the C57BL/6 genetic background. The Ldhb−/− mice were treated with either osmotin (15 μg/g of the body; intraperitoneally) or vehicle twice a week for 5-weeks. After behavior assessments, the mice were sacrificed, and the cortical and hippocampal brain regions were analyzed through biochemical and morphological analysis. Ldhb−/− mice displayed enhanced reactive oxygen species (ROS) and lipid peroxidation (LPO) production, and they revealed depleted stores of cellular ATP, GSH:GSSG enzyme ratio, and downregulated expression of Nrf2 and HO-1 proteins, when compared to WT littermates. Importantly, the Ldhb−/− mice showed upregulated expression of apoptosis mediators (Bax, Cytochrome C, and caspase-3), and revealed impaired p-AMPK/SIRT1/PGC-1alpha signaling. Moreover, LDHB deficiency-induced gliosis increased the production of inflammatory mediators (TNF-α, Nf-ĸB, and NOS2), and revealed cognitive deficits. Treatment with osmotin, an adipoR1 natural agonist, significantly increased cellular ATP production by increasing mitochondrial function and attenuated oxidative stress, neuroinflammation, and neuronal apoptosis, probably, by upregulating p-AMPK/SIRT1/PGC-1alpha signaling in Ldhb−/− mice. In brief, LDHB deficiency may lead to brain oxidative stress-mediated progression of neurodegeneration via regulating p-AMPK/SIRT1/PGC-1alpha signaling, while osmotin could improve mitochondrial functions, abrogate oxidative stress and alleviate neuroinflammation and neurodegeneration in adult Ldhb−/− mice.