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Sultan Qaboos University Medical Journal Aug 2018This study aimed to establish lactate dehydrogenase (LDH) and β-glucuronidase as salivary biomarkers of periodontitis among smokers and non-smokers.
OBJECTIVES
This study aimed to establish lactate dehydrogenase (LDH) and β-glucuronidase as salivary biomarkers of periodontitis among smokers and non-smokers.
METHODS
This cross-sectional case-control study was conducted at the Aligarh Muslim University, Aligarh, India, between January and June 2017. A total of 200 participants were divided into four groups based on their periodontal and smoking statuses. Unstimulated mixed saliva samples were collected to estimate LDH and β-glucuronidase levels. In addition, total protein was estimated using Lowry's method.
RESULTS
There was a significant increase in enzyme activity in the periodontitis groups compared to the non-periodontitis groups ( <0.001). However, significantly lower enzyme activity was observed among smokers, irrespective of periodontal status ( <0.001). Nevertheless, a receiver operating characteristic curve analysis indicated the diagnostic potential of both enzymes to be fair-to-excellent.
CONCLUSION
Although smoking was found to significantly alter enzyme activity, LDH and β-glucuronidase were reliable salivary biomarkers of periodontitis among both smokers and non-smokers.
Topics: Adult; Area Under Curve; Biomarkers; Case-Control Studies; Cross-Sectional Studies; Female; Glucosylceramidase; Humans; India; L-Lactate Dehydrogenase; Male; Periodontal Index; Periodontitis; ROC Curve; Saliva; Smoking
PubMed: 30607272
DOI: 10.18295/squmj.2018.18.03.009 -
CNS Neuroscience & Therapeutics May 2024Despite the success of single-cell RNA sequencing in identifying cellular heterogeneity in ischemic stroke, clarifying the mechanisms underlying these associations of...
AIMS
Despite the success of single-cell RNA sequencing in identifying cellular heterogeneity in ischemic stroke, clarifying the mechanisms underlying these associations of differently expressed genes remains challenging. Several studies that integrate gene expression and gene expression quantitative trait loci (eQTLs) with genome wide-association study (GWAS) data to determine their causal role have been proposed.
METHODS
Here, we combined Mendelian randomization (MR) framework and single cell (sc) RNA sequencing to study how differently expressed genes (DEGs) mediating the effect of gene expression on ischemic stroke. The hub gene was further validated in the in vitro model.
RESULTS
We identified 2339 DEGs in 10 cell clusters. Among these DEGs, 58 genes were associated with the risk of ischemic stroke. After external validation with eQTL dataset, lactate dehydrogenase B (LDHB) is identified to be positively associated with ischemic stroke. The expression of LDHB has also been validated in sc RNA-seq with dominant expression in microglia and astrocytes, and melatonin is able to reduce the LDHB expression and activity in vitro ischemic models.
CONCLUSION
Our study identifies LDHB as a novel biomarker for ischemic stroke via combining the sc RNA-seq and MR analysis.
Topics: Animals; Humans; Genome-Wide Association Study; Ischemic Stroke; Isoenzymes; L-Lactate Dehydrogenase; Melatonin; Mendelian Randomization Analysis; Quantitative Trait Loci; Sequence Analysis, RNA; Single-Cell Analysis; Mice
PubMed: 38702940
DOI: 10.1111/cns.14741 -
Scientific Reports Sep 2023Biological properties of protein molecules depend on their interaction with other molecules, and enzymes are no exception. Enzyme activities are controlled by their...
Biological properties of protein molecules depend on their interaction with other molecules, and enzymes are no exception. Enzyme activities are controlled by their interaction with other molecules in living cells. Enzyme activation and their catalytic properties in the presence of different types of polymers have been studied in vitro, although these studies are restricted to only a few enzymes. In this study, we show that addition of poly-l-lysine (PLL) can increase the enzymatic activity of multiple oxidoreductases through formation of enzyme assemblies. Oxidoreductases with an overall negative charge, such as l-lactate oxidase, d-lactate dehydrogenase, pyruvate oxidase, and acetaldehyde dehydrogenase, each formed assemblies with the positively charged PLL via electrostatic interactions. The enzyme activities of these oxidoreductases in the enzyme assemblies were several-folds higher than those of the enzyme in their natural dispersed state. In the presence of PLL, the turnover number (k) improved for all enzymes, whereas the decrease in Michaelis constant (K) was enzyme dependent. This type of enzyme function regulation through the formation of assemblies via simple addition of polymers has potential for diverse applications, including various industrial and research purposes.
Topics: L-Lactate Dehydrogenase; Catalysis; Industry; Lysine; Poly A; Polymers
PubMed: 37658129
DOI: 10.1038/s41598-023-41789-9 -
Parasitology Research Oct 2017Coenurus cerebralis, the larval stage (metacestode or coenurus) of Taenia multiceps, parasitizes sheep, goats, and other ruminants and causes coenurosis. In this study,...
Coenurus cerebralis, the larval stage (metacestode or coenurus) of Taenia multiceps, parasitizes sheep, goats, and other ruminants and causes coenurosis. In this study, we isolated and characterized complementary DNAs that encode lactate dehydrogenase A (Tm-LDHA) and B (Tm-LDHB) from the transcriptome of T. multiceps and expressed recombinant Tm-LDHB (rTm-LDHB) in Escherichia coli. Bioinformatic analysis showed that both Tm-LDH genes (LDHA and LDHB) contain a 996-bp open reading frame and encode a protein of 331 amino acids. After determination of the immunogenicity of the recombinant Tm-LDHB, an indirect enzyme-linked immunosorbent assay (ELISA) was developed for preliminary evaluation of the serodiagnostic potential of rTm-LDHB in goats. However, the rTm-LDHB-based indirect ELISA developed here exhibited specificity of only 71.42% (10/14) and sensitivity of 1:3200 in detection of goats infected with T. multiceps in the field. This study is the first to describe LDHA and LDHB of T. multiceps; meanwhile, our results indicate that rTm-LDHB is not a specific antigen candidate for immunodiagnosis of T. multiceps infection in goats.
Topics: Amino Acid Sequence; Animals; Cestode Infections; Cloning, Molecular; Computational Biology; Enzyme-Linked Immunosorbent Assay; Goat Diseases; Goats; L-Lactate Dehydrogenase; Larva; Models, Molecular; Molecular Conformation; Phylogeny; Sensitivity and Specificity; Sequence Alignment; Serologic Tests; Taenia
PubMed: 28766153
DOI: 10.1007/s00436-017-5568-x -
Physiological Reports Aug 2022It was hypothesized that the typical adult pattern of higher glycolytic capacity in skeletal muscle of males compared to females is not observed in children and that...
It was hypothesized that the typical adult pattern of higher glycolytic capacity in skeletal muscle of males compared to females is not observed in children and that fiber cross-sectional area (CSA) is a determinant of glycolytic capacity in children. Biopsies were performed in vastus lateralis in 9-12 years-old healthy boys and girls (N = 27). Fiber types were classified by myofibrillar ATPase staining and CSA was measured using planimetry. Citrate synthase (CS) and lactate dehydrogenase (LD) were analyzed using fluorometric and spectrophotometric methods. There was no significant difference between boys and girls in CS activity (0.45 ± 0.1 μkat g dry muscle in boys and 0.42 ± 0.1 in girls) or LD activity (24 ± 6 μkat g dry muscle in boys and 25 ± 7 in girls). CSA did not differ between boys and girls. CS was inversely related to type I CSA (r = -0.62, p < 0.001) and LD was directly related to type IIA (r = 0.63, p < 0.001) and type IIB CSA (r = 0.72, p < 0.001). CSA was a significant determinant of CS and LD, even after adjusting for sex and relative fiber type area in multiple regression analysis. This suggests that the typical adult pattern of higher muscle glycolytic capacity in males than in females, as estimated by LD activity, was not observed in children. Sex-specific patterns in glycolytic capacity thus appear to develop during the transition from childhood to adulthood. In addition, fiber CSA was a strong determinant of both muscle glycolytic and oxidative capacity in children, regardless of sex.
Topics: Adolescent; Adult; Biopsy; Child; Citrate (si)-Synthase; Female; Humans; L-Lactate Dehydrogenase; Male; Muscle Fibers, Skeletal; Muscle, Skeletal; Young Adult
PubMed: 35986491
DOI: 10.14814/phy2.15414 -
Frontiers in Cellular and Infection... 2022is a tick-borne apicomplexan hemoprotozoan responsible for bovine babesiosis. The current drugs used for bovine babesiosis treatment have several drawbacks, including...
is a tick-borne apicomplexan hemoprotozoan responsible for bovine babesiosis. The current drugs used for bovine babesiosis treatment have several drawbacks, including toxicity, the lack of effectiveness to clear the parasite, and potential to develop resistance. Identifying compounds that target essential and unique parasite metabolic pathways is a rational approach toward finding alternative drug treatments. Based on the genome sequence and transcriptomics analysis, it can be inferred that anaerobic glycolysis is the dominant adenosine triphosphate (ATP) supply for , and lactate dehydrogenase (LDH) is one of the essential enzymes in this pathway. Furthermore, the LDH sequence is distinct from its bovine homologue and thus a potential chemotherapeutic target that would result in decreasing the ATP supply to the parasite but not to the host. Gossypol is a known efficient specific inhibitor of LDH in the and the , among other related parasites, but no such data are currently available in the parasites. Hereby, we show that the LDH amino acid sequence is highly conserved among but not in spp. A predictive structural analysis of LDH showed the conservation of the key amino acids involved in the binding to gossypol compared to . Gossypol has a significant (P < 0.0001) inhibitory effect on the growth of , with IC of 43.97 mM after 72 h of treatment. The maximum IC (IC) was observed at 60 mM gossypol. However, a significant effect on the viability of cattle PBMC was observed when the cells were cultured with 60 mM (IC) gossypol compared with DMSO-exposed control cells. Interestingly, cultured at 3% oxygen expresses significantly higher levels of LDH and is more resistant to gossypol than the parasites maintained at ambient conditions containing ~20% oxygen. Altogether, the results suggest the potential of gossypol as an effective drug against infection, but the risk of host toxicity at therapeutic doses should be further evaluated in studies.
Topics: Adenosine Triphosphate; Animals; Babesia; Babesia bovis; Babesiosis; Cattle; Cattle Diseases; Gossypol; L-Lactate Dehydrogenase; Leukocytes, Mononuclear; Oxygen
PubMed: 35521220
DOI: 10.3389/fcimb.2022.870852 -
The Journal of Biological Chemistry Jul 2021Cellular pyruvate is an essential metabolite at the crossroads of glycolysis and oxidative phosphorylation, capable of supporting fermentative glycolysis by reduction to...
Cellular pyruvate is an essential metabolite at the crossroads of glycolysis and oxidative phosphorylation, capable of supporting fermentative glycolysis by reduction to lactate mediated by lactate dehydrogenase (LDH) among other functions. Several inherited diseases of mitochondrial metabolism impact extracellular (plasma) pyruvate concentrations, and [1-C]pyruvate infusion is used in isotope-labeled metabolic tracing studies, including hyperpolarized magnetic resonance spectroscopic imaging. However, how these extracellular pyruvate sources impact intracellular metabolism is not clear. Herein, we examined the effects of excess exogenous pyruvate on intracellular LDH activity, extracellular acidification rates (ECARs) as a measure of lactate production, and hyperpolarized [1-C]pyruvate-to-[1-C]lactate conversion rates across a panel of tumor and normal cells. Combined LDH activity and LDHB/LDHA expression analysis intimated various heterotetrameric isoforms comprising LDHA and LDHB in tumor cells, not only canonical LDHA. Millimolar concentrations of exogenous pyruvate induced substrate inhibition of LDH activity in both enzymatic assays ex vivo and in live cells, abrogated glycolytic ECAR, and inhibited hyperpolarized [1-C]pyruvate-to-[1-C]lactate conversion rates in cellulo. Of importance, the extent of exogenous pyruvate-induced inhibition of LDH and glycolytic ECAR in live cells was highly dependent on pyruvate influx, functionally mediated by monocarboxylate transporter-1 localized to the plasma membrane. These data provided evidence that highly concentrated bolus injections of pyruvate in vivo may transiently inhibit LDH activity in a tissue type- and monocarboxylate transporter-1-dependent manner. Maintaining plasma pyruvate at submillimolar concentrations could potentially minimize transient metabolic perturbations, improve pyruvate therapy, and enhance quantification of metabolic studies, including hyperpolarized [1-C]pyruvate magnetic resonance spectroscopic imaging and stable isotope tracer experiments.
Topics: Acids; Buffers; Carbon Isotopes; Cell Extracts; Cell Line, Tumor; Cell Membrane; Extracellular Space; Glycolysis; Humans; Inhibitory Concentration 50; Kinetics; L-Lactate Dehydrogenase; Lactic Acid; Monocarboxylic Acid Transporters; Pyruvic Acid; Substrate Specificity; Symporters
PubMed: 34022218
DOI: 10.1016/j.jbc.2021.100775 -
BMC Research Notes Dec 2018Rapid diagnostic tests have been of tremendous help in malaria control in endemic areas, helping in diagnosis and treatment of malaria cases. It is heavily relied upon... (Review)
Review
OBJECTIVE
Rapid diagnostic tests have been of tremendous help in malaria control in endemic areas, helping in diagnosis and treatment of malaria cases. It is heavily relied upon in many endemic areas where microscopy cannot be obtained. However, caution should be taken in the interpretation of its result in clinical setting due to its limitations and inherent weakness. This paper seeks to present the varying malaria RDT test results, the possible interpretations and explanation of these results common in endemic regions. Published works on malaria RDT studies were identified using the following search terms "malaria RDT in endemic areas", "Plasmodium falciparum and bacterial coinfection" "Plasmodium falciparum RDT test results in children in endemic areas" in Google Scholar and PubMed.
RESULTS
The review results show that RDT positive results in febrile patients can either be true or false positive. True positive, representing either a possible single infection of Plasmodium or a co-infection of bacteria and P. falciparum. False RDT negative results can be seen in febrile patient with P. falciparum infection in prozone effect, Histidine rich protein 2 (HRP2) gene deletion and faulty RDT kits. Hence, a scale up of laboratory facilities especially expert microscopy and other diagnostic tools is imperative.
Topics: Adult; Antigens, Protozoan; Child; Diagnosis, Differential; Diagnostic Tests, Routine; False Negative Reactions; False Positive Reactions; Fever; Humans; L-Lactate Dehydrogenase; Malaria, Falciparum; Plasmodium falciparum; Protozoan Proteins; Sensitivity and Specificity
PubMed: 30509313
DOI: 10.1186/s13104-018-3967-4 -
Journal of Visualized Experiments : JoVE Jun 2018Mapping enzymatic activity in space and time is critical for understanding the molecular basis of cell behavior in normal tissue and disease. In situ metabolic activity...
Mapping enzymatic activity in space and time is critical for understanding the molecular basis of cell behavior in normal tissue and disease. In situ metabolic activity assays can provide information about the spatial distribution of metabolic activity within a tissue. We provide here a detailed protocol for monitoring the activity of the enzyme lactate dehydrogenase directly in tissue samples. Lactate dehydrogenase is an important determinant of whether consumed glucose will be converted to energy via aerobic or anaerobic glycolysis. A solution containing lactate and NAD is provided to a frozen tissue section. Cells with high lactate dehydrogenase activity will convert the provided lactate to pyruvate, while simultaneously converting provided nicotinamide adenine dinucleotide (NAD) to NADH and a proton, which can be detected based on the reduction of nitrotetrazolium blue to formazan, which is visualized as a blue precipitate. We describe a detailed protocol for monitoring lactate dehydrogenase activity in mouse skin. Applying this protocol, we found that lactate dehydrogenase activity is high in the quiescent hair follicle stem cells within the skin. Applying the protocol to cultured mouse embryonic stem cells revealed higher staining in cultured embryonic stem cells than mouse embryonic fibroblasts. Analysis of freshly isolated mouse aorta revealed staining in smooth muscle cells perpendicular to the aorta. The methodology provided can be used to spatially map the activity of enzymes that generate a proton in frozen or fresh tissue.
Topics: Animals; L-Lactate Dehydrogenase; Mice
PubMed: 29985359
DOI: 10.3791/57760 -
Oncogene Apr 2021Metabolic reprogramming is considered hallmarks of cancer. Aerobic glycolysis in tumors cells has been well-known for almost a century, but specific factors that...
Metabolic reprogramming is considered hallmarks of cancer. Aerobic glycolysis in tumors cells has been well-known for almost a century, but specific factors that regulate lactate generation and the effects of lactate in both cancer cells and stroma are not yet well understood. In the present study using breast cancer cell lines, human primary cultures of breast tumors, and immune deficient murine models, we demonstrate that the POU1F1 transcription factor is functionally and clinically related to both metabolic reprogramming in breast cancer cells and fibroblasts activation. Mechanistically, we demonstrate that POU1F1 transcriptionally regulates the lactate dehydrogenase A (LDHA) gene. LDHA catalyzes pyruvate into lactate instead of leading into the tricarboxylic acid cycle. Lactate increases breast cancer cell proliferation, migration, and invasion. In addition, it activates normal-associated fibroblasts (NAFs) into cancer-associated fibroblasts (CAFs). Conversely, LDHA knockdown in breast cancer cells that overexpress POU1F1 decreases tumor volume and [F]FDG uptake in tumor xenografts of mice. Clinically, POU1F1 and LDHA expression correlate with relapse- and metastasis-free survival. Our data indicate that POU1F1 induces a metabolic reprogramming through LDHA regulation in human breast tumor cells, modifying the phenotype of both cancer cells and fibroblasts to promote cancer progression.
Topics: Animals; Cellular Reprogramming; Disease Progression; Heterografts; Humans; L-Lactate Dehydrogenase; MCF-7 Cells; Mice; Mice, Inbred BALB C; Mice, Nude; Transcription Factor Pit-1; Transfection
PubMed: 33714987
DOI: 10.1038/s41388-021-01740-6